AGENC( FOR INTERNA7IOIAL DEVELOPMENT WASHINGTON DC 20523
DATE
MEMCRAINDUM
TO ATDPPCCDIEDI room 209 SA-18
FROM Al)Cr Victoria Or e
SUBJECT Trainrnitt~a of AIDSCI Progress Report(s)
Attache-d for periinert retent ionproper disposition is the following
AIDSCI Progre Report No
Attachment
P R O G R E S S REPORT NO 4
NATURAL INSECTICIDES FOR SPODOPTERA LITIJRA FABR
A RESEARCH PROJLCT
USA IDPSTC P4 OGRAM
GRANT NO 936-5512-gtO0-5032-00
SUBMITTED) BY
PICHE WIRIYACHITRA
PRINCE OF SONGKIA UNIVERS]
HAT YAI THAILAND
ROM r~irl
PROJECT PROFILE
Country Thailand
Grant No 936-5542-G-00-5032-00
Science and Technology CooperationProgram Program on
Project Title Natural Insecticides for Spooptera litura Fabr
Projct Leader Pichaet Wiriyachitra
Organization Prince of Songkla University Hat Yai
Co-invcstiqators Prachaval Sukumalanand
Total Froject Budget US $ 150000
Froject Dration 3 years
Poportinc Pirind I December 1986 to 31 May 1988
for This Period 133021179 BahtDudget Allocation
BACKGROUND
S itulta Pabr is a widespread insect pest It is polyphagous
and regarded as one of the most damaging pests in the world agriculture
Pandy and Srivastava (1967) reported that the insect attacks 112 species
of cultivated crops all over the world In Thailand at least 30
var-leties of plants are found to 1-e host plants of this insect
These include v-getables floral plants ornamental plants economic
field crops and weeds
The time required for egg larval and pupal development are
2-3 14 and 7-8 days respectively2 The adult longevity is 8-10 days
in the male and 10-12 days in the female The larva molts 5 times
It is at this stage that the insect is most damaging Since the early
larval instars are quite vunerable to external damage it has been
chcsen 3isthe target organism for this work
For self-defence it is not surprising that plants would contain
substances which are inst-ct antifeedants insect repellents and insectishy
cides Some naturally occurring antifeedants which have been published
3 4are glycosides of steroidal alkaloids demissine solacauline
5 6 tomatine leptines I and 11 ring A enone andor ring D aromatic
steroids the nicandrenones and other 7890 juglone (5-hydroxynapthoshy
1o 11 12quinone) the isoquinoline alkaloid isoboldine phenylpropanoids
germrcrane sesquiterpenes shiromodiol and shiromol 1314 efit-clerodane
16ndc 15and clerodane diterpenes clerodendrin caryoptin and others
mel iantriol 7 azadirachtin 18 xylomolin 9the hydroxylated steroe(i
ins 0 harrisonin21 polyqodii ugandensidial and warburganal 22
l 23 i n4 25~irfej 2rnd 24i rindid)id 2 and unedoside However
there is no report of any of these fr+ ostances being used effectively
in agriculture
- 2 -
A number of naturally occurring compounds have been found to
exhibit insecticidal activity These include alkaloids terpenoids
phenolics cinines chromenes prenylated guinones and sesqiiterpenes
esterified with phenolic acids 2 6 Commercially the substances which
have found their way to agricultuiral application are nicotine derris
27 pyrethrum and the newer product ryania Crude extracts of the
plant producing these r ubstances or dried and powdered plant tissue
have a long history of use i SE Asia as ii secticides However none
of there natural products is a universal insecticide Tobacco leaf
containing nicotine is subject to attack by several insects the root
of derris-containing plant is attacked by the larvae of a species of
Neetle the flower from which pyrethrins are extracted needs synthetic
invecticidros for protection f-om insects in the fields In addition
inrects can develop resistance to a certain chemicals after a long
exposure New and better insecticides will therefore have to be
constantly souriht
Since S UtU4( is one of the major pests in SE Asia the
6ircovery of a new and more effective insecticide with low mammalian
toxicity from local plants woul6 produce enormous benefit to the region
ar well as to ThMiand in general
ORJECTI VES
1 To isolate and identify natural insecticides and antifecdants
which have ow mammalian toxicity for S 4W1a
2 Extract or Fynthesizing those compounds on a practical
scale and design of industrial extraction procedure
3 Evaluation of their applicability in the field
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
P R O G R E S S REPORT NO 4
NATURAL INSECTICIDES FOR SPODOPTERA LITIJRA FABR
A RESEARCH PROJLCT
USA IDPSTC P4 OGRAM
GRANT NO 936-5512-gtO0-5032-00
SUBMITTED) BY
PICHE WIRIYACHITRA
PRINCE OF SONGKIA UNIVERS]
HAT YAI THAILAND
ROM r~irl
PROJECT PROFILE
Country Thailand
Grant No 936-5542-G-00-5032-00
Science and Technology CooperationProgram Program on
Project Title Natural Insecticides for Spooptera litura Fabr
Projct Leader Pichaet Wiriyachitra
Organization Prince of Songkla University Hat Yai
Co-invcstiqators Prachaval Sukumalanand
Total Froject Budget US $ 150000
Froject Dration 3 years
Poportinc Pirind I December 1986 to 31 May 1988
for This Period 133021179 BahtDudget Allocation
BACKGROUND
S itulta Pabr is a widespread insect pest It is polyphagous
and regarded as one of the most damaging pests in the world agriculture
Pandy and Srivastava (1967) reported that the insect attacks 112 species
of cultivated crops all over the world In Thailand at least 30
var-leties of plants are found to 1-e host plants of this insect
These include v-getables floral plants ornamental plants economic
field crops and weeds
The time required for egg larval and pupal development are
2-3 14 and 7-8 days respectively2 The adult longevity is 8-10 days
in the male and 10-12 days in the female The larva molts 5 times
It is at this stage that the insect is most damaging Since the early
larval instars are quite vunerable to external damage it has been
chcsen 3isthe target organism for this work
For self-defence it is not surprising that plants would contain
substances which are inst-ct antifeedants insect repellents and insectishy
cides Some naturally occurring antifeedants which have been published
3 4are glycosides of steroidal alkaloids demissine solacauline
5 6 tomatine leptines I and 11 ring A enone andor ring D aromatic
steroids the nicandrenones and other 7890 juglone (5-hydroxynapthoshy
1o 11 12quinone) the isoquinoline alkaloid isoboldine phenylpropanoids
germrcrane sesquiterpenes shiromodiol and shiromol 1314 efit-clerodane
16ndc 15and clerodane diterpenes clerodendrin caryoptin and others
mel iantriol 7 azadirachtin 18 xylomolin 9the hydroxylated steroe(i
ins 0 harrisonin21 polyqodii ugandensidial and warburganal 22
l 23 i n4 25~irfej 2rnd 24i rindid)id 2 and unedoside However
there is no report of any of these fr+ ostances being used effectively
in agriculture
- 2 -
A number of naturally occurring compounds have been found to
exhibit insecticidal activity These include alkaloids terpenoids
phenolics cinines chromenes prenylated guinones and sesqiiterpenes
esterified with phenolic acids 2 6 Commercially the substances which
have found their way to agricultuiral application are nicotine derris
27 pyrethrum and the newer product ryania Crude extracts of the
plant producing these r ubstances or dried and powdered plant tissue
have a long history of use i SE Asia as ii secticides However none
of there natural products is a universal insecticide Tobacco leaf
containing nicotine is subject to attack by several insects the root
of derris-containing plant is attacked by the larvae of a species of
Neetle the flower from which pyrethrins are extracted needs synthetic
invecticidros for protection f-om insects in the fields In addition
inrects can develop resistance to a certain chemicals after a long
exposure New and better insecticides will therefore have to be
constantly souriht
Since S UtU4( is one of the major pests in SE Asia the
6ircovery of a new and more effective insecticide with low mammalian
toxicity from local plants woul6 produce enormous benefit to the region
ar well as to ThMiand in general
ORJECTI VES
1 To isolate and identify natural insecticides and antifecdants
which have ow mammalian toxicity for S 4W1a
2 Extract or Fynthesizing those compounds on a practical
scale and design of industrial extraction procedure
3 Evaluation of their applicability in the field
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
PROJECT PROFILE
Country Thailand
Grant No 936-5542-G-00-5032-00
Science and Technology CooperationProgram Program on
Project Title Natural Insecticides for Spooptera litura Fabr
Projct Leader Pichaet Wiriyachitra
Organization Prince of Songkla University Hat Yai
Co-invcstiqators Prachaval Sukumalanand
Total Froject Budget US $ 150000
Froject Dration 3 years
Poportinc Pirind I December 1986 to 31 May 1988
for This Period 133021179 BahtDudget Allocation
BACKGROUND
S itulta Pabr is a widespread insect pest It is polyphagous
and regarded as one of the most damaging pests in the world agriculture
Pandy and Srivastava (1967) reported that the insect attacks 112 species
of cultivated crops all over the world In Thailand at least 30
var-leties of plants are found to 1-e host plants of this insect
These include v-getables floral plants ornamental plants economic
field crops and weeds
The time required for egg larval and pupal development are
2-3 14 and 7-8 days respectively2 The adult longevity is 8-10 days
in the male and 10-12 days in the female The larva molts 5 times
It is at this stage that the insect is most damaging Since the early
larval instars are quite vunerable to external damage it has been
chcsen 3isthe target organism for this work
For self-defence it is not surprising that plants would contain
substances which are inst-ct antifeedants insect repellents and insectishy
cides Some naturally occurring antifeedants which have been published
3 4are glycosides of steroidal alkaloids demissine solacauline
5 6 tomatine leptines I and 11 ring A enone andor ring D aromatic
steroids the nicandrenones and other 7890 juglone (5-hydroxynapthoshy
1o 11 12quinone) the isoquinoline alkaloid isoboldine phenylpropanoids
germrcrane sesquiterpenes shiromodiol and shiromol 1314 efit-clerodane
16ndc 15and clerodane diterpenes clerodendrin caryoptin and others
mel iantriol 7 azadirachtin 18 xylomolin 9the hydroxylated steroe(i
ins 0 harrisonin21 polyqodii ugandensidial and warburganal 22
l 23 i n4 25~irfej 2rnd 24i rindid)id 2 and unedoside However
there is no report of any of these fr+ ostances being used effectively
in agriculture
- 2 -
A number of naturally occurring compounds have been found to
exhibit insecticidal activity These include alkaloids terpenoids
phenolics cinines chromenes prenylated guinones and sesqiiterpenes
esterified with phenolic acids 2 6 Commercially the substances which
have found their way to agricultuiral application are nicotine derris
27 pyrethrum and the newer product ryania Crude extracts of the
plant producing these r ubstances or dried and powdered plant tissue
have a long history of use i SE Asia as ii secticides However none
of there natural products is a universal insecticide Tobacco leaf
containing nicotine is subject to attack by several insects the root
of derris-containing plant is attacked by the larvae of a species of
Neetle the flower from which pyrethrins are extracted needs synthetic
invecticidros for protection f-om insects in the fields In addition
inrects can develop resistance to a certain chemicals after a long
exposure New and better insecticides will therefore have to be
constantly souriht
Since S UtU4( is one of the major pests in SE Asia the
6ircovery of a new and more effective insecticide with low mammalian
toxicity from local plants woul6 produce enormous benefit to the region
ar well as to ThMiand in general
ORJECTI VES
1 To isolate and identify natural insecticides and antifecdants
which have ow mammalian toxicity for S 4W1a
2 Extract or Fynthesizing those compounds on a practical
scale and design of industrial extraction procedure
3 Evaluation of their applicability in the field
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
BACKGROUND
S itulta Pabr is a widespread insect pest It is polyphagous
and regarded as one of the most damaging pests in the world agriculture
Pandy and Srivastava (1967) reported that the insect attacks 112 species
of cultivated crops all over the world In Thailand at least 30
var-leties of plants are found to 1-e host plants of this insect
These include v-getables floral plants ornamental plants economic
field crops and weeds
The time required for egg larval and pupal development are
2-3 14 and 7-8 days respectively2 The adult longevity is 8-10 days
in the male and 10-12 days in the female The larva molts 5 times
It is at this stage that the insect is most damaging Since the early
larval instars are quite vunerable to external damage it has been
chcsen 3isthe target organism for this work
For self-defence it is not surprising that plants would contain
substances which are inst-ct antifeedants insect repellents and insectishy
cides Some naturally occurring antifeedants which have been published
3 4are glycosides of steroidal alkaloids demissine solacauline
5 6 tomatine leptines I and 11 ring A enone andor ring D aromatic
steroids the nicandrenones and other 7890 juglone (5-hydroxynapthoshy
1o 11 12quinone) the isoquinoline alkaloid isoboldine phenylpropanoids
germrcrane sesquiterpenes shiromodiol and shiromol 1314 efit-clerodane
16ndc 15and clerodane diterpenes clerodendrin caryoptin and others
mel iantriol 7 azadirachtin 18 xylomolin 9the hydroxylated steroe(i
ins 0 harrisonin21 polyqodii ugandensidial and warburganal 22
l 23 i n4 25~irfej 2rnd 24i rindid)id 2 and unedoside However
there is no report of any of these fr+ ostances being used effectively
in agriculture
- 2 -
A number of naturally occurring compounds have been found to
exhibit insecticidal activity These include alkaloids terpenoids
phenolics cinines chromenes prenylated guinones and sesqiiterpenes
esterified with phenolic acids 2 6 Commercially the substances which
have found their way to agricultuiral application are nicotine derris
27 pyrethrum and the newer product ryania Crude extracts of the
plant producing these r ubstances or dried and powdered plant tissue
have a long history of use i SE Asia as ii secticides However none
of there natural products is a universal insecticide Tobacco leaf
containing nicotine is subject to attack by several insects the root
of derris-containing plant is attacked by the larvae of a species of
Neetle the flower from which pyrethrins are extracted needs synthetic
invecticidros for protection f-om insects in the fields In addition
inrects can develop resistance to a certain chemicals after a long
exposure New and better insecticides will therefore have to be
constantly souriht
Since S UtU4( is one of the major pests in SE Asia the
6ircovery of a new and more effective insecticide with low mammalian
toxicity from local plants woul6 produce enormous benefit to the region
ar well as to ThMiand in general
ORJECTI VES
1 To isolate and identify natural insecticides and antifecdants
which have ow mammalian toxicity for S 4W1a
2 Extract or Fynthesizing those compounds on a practical
scale and design of industrial extraction procedure
3 Evaluation of their applicability in the field
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 2 -
A number of naturally occurring compounds have been found to
exhibit insecticidal activity These include alkaloids terpenoids
phenolics cinines chromenes prenylated guinones and sesqiiterpenes
esterified with phenolic acids 2 6 Commercially the substances which
have found their way to agricultuiral application are nicotine derris
27 pyrethrum and the newer product ryania Crude extracts of the
plant producing these r ubstances or dried and powdered plant tissue
have a long history of use i SE Asia as ii secticides However none
of there natural products is a universal insecticide Tobacco leaf
containing nicotine is subject to attack by several insects the root
of derris-containing plant is attacked by the larvae of a species of
Neetle the flower from which pyrethrins are extracted needs synthetic
invecticidros for protection f-om insects in the fields In addition
inrects can develop resistance to a certain chemicals after a long
exposure New and better insecticides will therefore have to be
constantly souriht
Since S UtU4( is one of the major pests in SE Asia the
6ircovery of a new and more effective insecticide with low mammalian
toxicity from local plants woul6 produce enormous benefit to the region
ar well as to ThMiand in general
ORJECTI VES
1 To isolate and identify natural insecticides and antifecdants
which have ow mammalian toxicity for S 4W1a
2 Extract or Fynthesizing those compounds on a practical
scale and design of industrial extraction procedure
3 Evaluation of their applicability in the field
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
MATERIALS AND METHODS
Plants investigated were collected in the southern region of
ThiIand S~imples were clascified taxonomically and vouchers were
deposited in tho herbariu in the DeparuiTit of Biology Prince of
Songk la Uni verit y
Ground plant materials were steam distilled or exhaustively
extracted with solvents The extractssteam ditiliates were screened
for inmsecticidal and antifeedant activity with S titula and the active
extracts reiolved into fractions Each fraction was screened for
activity aInd the active fraczin again separated into individual
constituerit uinq quick column and preparative layer chromatography
cnn silica geI
1 ich o the individual components was tested again for activity
and the LC value of thV active compound determined The structure of
the compound was elucidated using spectroscopic techniques
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
-4-
Ground
Plant Material
extracted with alcohol
-ive screncu for Crude Extract screened forgt -ive
lot investigated activity activity not investigated
screened for
activity
Activ2 Extrac
f ractiona tion
II Fraction IIIIraction~Xdctio
screened for activity
-ive -iye not investigated not investigated
LActive Fractiorl
iCompoundI rCompound II Coipound II
screened for activity
structure elucidated
-i____-N__ lActive compoundl
-ive structure elucidated
LC50 structure determined elucidated
syntheses comparison with degradation kno-wn a-ces
Diagram showing work plan
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 5 -
The leaf dipping (LD) method was used for the detection of
stomach poisor1ing and anti feeding activity the third instars of
Spodopvta etuaza 1lAvae were left to feed on the leaves of V-gna
Aadiata which hil ten immersed in the tested solution then airshy
dr i-d Ten larvaewre used in each of the four replications
(one as control ) The numbe-r of dead organisms was recorded at
the end of e-ch day The amount of the leaf consumption and the
larvae sine wi-re also estimatied
The topicFil application (TA) method was used for the detection
of dermal toxicity the tested rolution was dropped on the thorax
of the thi rd in5tar Iarvae The organisms were then left to feed
on untreateo leaveni of Vigna Aadiata in a plastic Tontainer Ten
larvae were und in each of the four replications (one as control)
The ncii-w-r of ci d organisms was recorded at the end of each day
Le1D coipl ed with TA method ws used for the detection of
the comlir cd effect of stomach poiconing and dermal toxicity The
tested 1ol tiin Wan drooped on trie thorax of the third instar larvae
The orqani w-wr( then left to feed on the leaves of Vigna adinta
which har] 1en imcm reId in the same tented solution then air-dried
Ten ]arvia were uicu in each o the four replications (one as control)
Thn numbr- of dead organisms was recorded at the end of each day
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
-6shy
RESULTSDISCUSSIONTABLES
Reinvestigation of the antifeeding activity of 4 Agaia
species eg A adokata A adarntica A a49entea A quamuto6a
revealed the significant acivities even at low concentration Attempt
was then rcade to fully explore the reaction of S L-tuta larvae to these
extractsfractions
A soy bean antificial diet was admixed with these crude
extractsfractions and used as feed for the larvae Outstanding
results were obtained frcm -xtractsfractions from A odottata
The feeding of larvae was significantly reduced the larval growth was
therefore slow They eventually died Table I 11 1 show these
resultf In detail
Ill 1
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
TABLE
MCRTALITY OF 5 LITUP F FED ON ARTIFICIAL DIET MIXED WITH EXTRAZT OF A SPECILS
ACCUNULATIVL MDRTAL1TY (ABBOTTS FORMULA)
PLANT SPECIES FPACTIOH CONC(PPM)
DAY 1 3 4 5 7 10
Aglaia d C H 10000 714 714 7 14 714 714 714 714 714 714 -
CH Cl 10000 667 1333 1667 10000 - - - - -
6000 337 6b7 3333 9667 10000 - -
6000 333 667 33 33 9667 10300 - -
4000 000 000 5333 10000 -- - -
2000 000 000 3333 7000 9333 10000 - -
1) QC(CH| 6000 000 000 000 000 0 00 000 000 000
2) QC(CHIICl) 10000 000 000 344 5714 92 l 9b43 10000 -
8000 000 000 1034 4285 7t57 10000 - -
6000 000 000 344 2459 7857 9643 10000 - -
4000 1000 1000 1723 2499 3929 5714 6785 8571 10000 -
2000 000 000 000 000 1425 3214 4285 75 uO 9260 9615
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
iABLi 1
MORTALITY OF LQ F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT UF 5 5FECIi
ACCuMULATIVE M RTALITY AbTT5 FORMULA) PLANT SPECIES FRACTON CCNC(Pk-H _
DAY 1 3 4 5 6 7 a 9 20
Aalaia cQrora 3 QCtC H C CI 31) 10000 000 000 000 OOu CO0 000 000 000 000 000
8000 000 000 000 000 000 OOu 00a 000 0O O
6000 334 334 334 334 334 334 334 334 334 334
4000 000 000 OOu 000 000 000 C00 000 000 000
2000 000 0Ou 000 000 010 000 000 000 000 000
41 QC(CHClI I0000 000 000 10000 - - - - - - -
8000 000 000 10000 - - - -
600C 000 333 9000 10000 - - -
4000 000 000 1334 10000 - - -
2000 000 000 3000 9311 10000 --
S QC HampOH) 10000 334 344 551 10000 - - -
8000 000 000 3000 9311 10000 - -
6000 000 000 6238 9310 10000 - -
2000 000 000 2758 8965 9643 10000 -
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
MORTALITY OF URA F FED ON ARTIFICIAL DIET MIXED WITH EATRACT Ok 6 SPECIES
ACCUMULATIVE I HCRTALITY (A-OTT- FORMULA)
PLANT SPECIESI FRACTICH CONC(PPM)
DAY 1 2 3 4 5 b 7 9 10 11
A______ oOH 10000 000 b67 2333 9333 10000 -
50-0 000 333 E66 7
66 67 89B5 9655 10(00
6000 00 000 6000 6b67 6620 9310 9b55 l00 00 - -
4000 000 000 333 333 1667 4462 6207 8276 8276 9655 0000
2000 000 000 333 333 344 689 2413 2658 3792 5555 -
EtOAc (insol) 10000 000 000 000 000 000 000 344 2758 3447 5172 -
000 000 000 000 030 000 OOG 344 20 EL 20E3 2758 -
6000 000 J00 000 000 000 000 3 44 344 10J4 IJ4 -
4000 000 0 00 000 0 00 000 00 0 00 0 0 000 000 -
2000 000 000 000 000 000 000 OCG 000 000 000 -
tOAc (601) 10000 000 034 4E27 a6 55 100 00 - - - - -
6000 000 689 5517 C000 - - -
6000 000 687 2758 965 10000 - -
4000 000 000 1034 9655 10000 - - -
2000 000 689 2069 6207 9365 10000 - - -
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
MORTALITY OF F FD ON AKTIFICIAL -4rLXFD WITH LtTPACT CF __ _pCI_
A ACCUNfULATIvE HCRTALI7Y (ABc-S pOprW LA)
Day 1 3 4 5 6 7 9 10 11 12 13
tCCl 30
600
400
200
000
000
000
000
000
000
000
000
000
500
000
000
500
500
000
000
3500
I000
000
000
6500
1100
500
000
9000
20c0
000
000
9500
3000
3000
500
10000
3530
4000
500
-
6000
4300
1500
-
6000
4000
1500
6500
4000
1500
7500
4000
1500
-HGOA Soo
So0
40
200
000
000
000
000
000
500
000
000
000
500
500
000
000
500
500
000
500
500
500
000
500
500
Scd
0clo
500
500
500
000
500
soo
500
000
500
500
500
000
500
500
500
000
500
500
500
000
10500
500
500
500
000
1500
500
500
o00
000
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
13 14 15 1 b 17 18 19 20 21 22 23 _4 25 26 27 28 9 30
3
3
3
75 00
40 00
1500
7500
-000
1000
8421
7894
1578
8947
8421
2631
9847
8947
3157
8947
9473
3157
8947
S473
8157
9473
9473
7b84
10000 -
9473 9473
421 4210
--
10000
5263 5263
-
5263 5263
-
- - -
3 500
5 00
C(00
1500
500
50000
000
1500
1000
500
000
3000
1000
500
000
4000
1000
500
000
400
1000
500
000
4000
1000
500
000
4000
000
500
000
5000
10 10
500
000
5500
1000
500
500
5500
1000
500
500
6000
1000
500
500
6500
1000
500
500
7000
1000
500
500
7000
1000
500
500
8000
1000
500
500
BO00
2500
500
500
8500
3000
500
500
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
TABLE
MORTALITY OF 2- LIMamp F FED ON ARTIFICIAL DrT 4 Pr 15WITH EXT ACT3F
PLANT SPECIES FRACTION
I
CONC (PPM)
Day 1
ACCUHULA2IL
3 4
HORTALITY
5
iAbbOT
b
5 FOPHULA)
7 9
holaiA andaani1 L C M 10000
8000
000
000
000
000
000
000
333
333
3 33
356
3j3
3 56
3 33
356
333
356
33
356
333
356
6000 000 000 000 000 000 000 000 0 U0 000 000
lCl
4000
2000
10000
000
000
000
000
000
000
000
000
333
000
000
1000
000
nO0
1071
000
( Go
1071
000
000
o 71
000
000
071
000
000
i01
000
0 o0
10 71
0000
6000
000
000
000
0uO
000
000
1667
0 0
16 67
000
1667
000
16b7
000
1b67
000
1 7
000
1667
000
4000 0O r
000 000 000 000 000 000 0 00 300 000
2000 000 000 0 00 000 000 000 O00 000 000 000
1ON10000
8000
000
000
000
000
333
000
1000
000
1334
000
1334
000
2000
000
2334
333
2334
333
2334
333
6000
4000
000
000
000
000
000
000
000
000
000
300
0 00
000
000
000
333
00G
333
0
333
2000 000 000 000 000 000 000 0 00 000 000 000
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
TMALhE i
tKDR-ALITY 0 S-L F FED ON ARTFICIAL DIET MIXED WITH Z TCT Ot AGIpound PECIES
ACCUV2-LTIVE 1-7RTALITY BABOf- ORMULA
PLA14T SPECIE5 FRACTICH o (PPM)
Djy I3 4 7 3 10
ampalsajZ C H 10000 000 0 00 000 000 0UO C00 000 010 000 000
8000 333 333 333 333 333 333 333 333 333 333
6000 000 000 000 000 000 000 000 0C 000 333
4000 000 000 000 000 000 000 000 00 0 00 000
2000 000 000 000 000 000 000 000 000 000 000
CHCl 10000 000 000 000 2000 6333 7000 6333 9333 10000 -
2000 000 00-0 000 333 4333 6333 n3 33 9667 10000 -
6000 000 000 D00 1667 2333 4-07 6000 667 332 966
4000 000 000 333 1000 2667 4333 1239000 i66b 10000
2000 000 000 333 1333 20CO 2000 b 00u 7333 d3 3^ 933
MaOH 10000 000 000 333 1000 2333 5333 7000 8333 86 67 9333
8000 333 333 667 1667 3667 5000 6000 -000 3 33 9333
6000 000 000 000 667 1000 3000 4667 70 00 8333 9667
4000 000 000 000 000 333 1000 2333 3 67 60 3u 3 13
2000 000 000 333 333 333 666 2000 4000 4b6 766
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
TABE I
MORTALITY OF 5_LIrUF F FED ON ARTIFICIAL DIET MIXED WITH EXTRACT OF A SPECES
ACCUMULATIVE- MORTALITY (ABBO--5 FOFLULAI
PLANT SPECIES I FRACTION CONC (PPr)
Day 1 2 3 4 5 6 7 8 9 10
S___ CH 10000 000 000 000 000 000 000 000 6b 7
667 67
8000 000 000 000 000 Cooo 000 000 0 00 0 0o 0 00
6000 000 000 000 000 000 000 000 0 00 000 000
4000 000 000 000 000 000 000 000 000 000 000
2000 000 000 000 000 000 000 000 000 000 000
CHC1 10000 000 000 000 000 000 000 000 000 000 000
8000 000 000 000 000 000 000 000 000 CO0 000
60uo 000 000 000 000 000 00 00 0 01) 000c 0 00
4000 000 000 000 000 000 000 u00 000 000 010
20v0 000 000 000 000 000 000 000 000 000 000
10000 000 000 000 000 000 000 OQO 3O 00c Oc0
8000 000 000 000 000 000 000 000 OCO 00 000
6000 000 000 000 000 000 000 00A 000 000 00
4000 000 000 000 000 000 000 000 000 0 Ou 000
2000 000 000 000 000 000 000 000 000 000 003
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
TABLE I
MORTALITY OF I U F FE ON ARTIFICIAL DIET MIXED WITH ExTRFCT OF n SPECIES
CONC ACCUMULATIVE MORTALITY (ABBOTS FIRIULA)
PLANT 5PECI2- FRACTION (PPM)
Day 1 2 3 4 5 0 7 8 9 10
pygnxuIm RZ2=L 1rC H 10000 000 000 000 000 000 000 000 666 666 1667
8000 000 000 000 000 000 000 000 666 1000 2000
6000 000 000 OO 000 0 00 000 333 667 1000 2667
CHCl 10000 000 000 000 000 000 000 000 333 333 333
8000 ocO 000 000 000 000 000 000 333 333 333
6000 000 000 000 000 000 000 000 333 1667 2333
- cryit l 10000 000 000 000 000 00m 000 333 667 667 1667
8000 000 00L 000 000 000 333 33 b67 667 l6b7
6000 000 000 000 000 000 030 000 333 333 1000
21non-crystAl 10000 000 000 000 000 000 000 1000 1000 1667 1667
8000 000 000 000 000 000 000 000 000 000 000
K MoOH
6000
10000
000
OO
000
000
OjU
000
000
000
000
000
000
000
000
1333
333
1333
1000
1333
1333
1333
80L0 000 000 000 000 0 G0 000 000 000 000 000
6000 000 000 000 333 333 667 667 667 667 667
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
MORTALITY OF _ITUF FED ON ARTIFICIAL VIET hIXED WITH LTACT oF YrXY SPECIES
PLA JT SPECIES FRACrION
P
COHC(PPI) ACCUMULATIVE MORTALIY
________________________________________
Day l 2 3 4 5
tABBOTS FOiKh JLAI
6 7 5 9 1D
iyxlu ap _ CH
CH Cl
10000
9000
6000
10000
333
000
000
000
333
000
000
000
33-
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
000
000
333
000
667
000
333
667
667
333
333
1000
1000
333
8000
6000
O00
000
000
000
000
000
000
000
000
000
000
000
OO
000
000
000
000
0 30
000
000
HMOH 10000
2000
000
000
000
333 333
000
333
000
333
000
667
0 00
667
0 00
b67
000
667
000
667
6000 000 000 000 000 000 C00 000 000 000 C00
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 7 -
Separation was carried out isolateto the active constituents
from A odota a using tI sche a hown in 11] 2 From the purified
fraction of high act ivi ty a bisan-id odorine(]) isolated
Its structure w evidence from its R spectra (I1 3) and the
terechemitry w i r~rf irmed 4)n y x-ray crystall ogr phy (111
11 2 - Diagram showing separation scheme for odorine
-- O
Ill 3 - 400 Hlz II -NXR spec trum of odortIne
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
I11 4 - Crystal itructure of odorine
Lpn rf- It inI th activity of this pure componert no
nf ic wt ant iieein ctivity a obs~ved Therefore this compound
cannot be the active inredient ard may only acts as a synerist in the
active fraction
then tnade to iloLate the minor contiLtuen s
in ti ictive frict ir I in I1 with exteniive chroratopraphy
(quici nlumn chrrn tctrr) sco letin (2) was isolated
further atit-WJ a
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
0 0
(2)
Prhi substance was confirmed as the active ingredient
since 1007 larval mortality resulted from an LD method using 160 ppm
solotion of thin ubstance
Experint was carried out tc pre-determine the lormulation
andplicat ion mothod for testing the antifeedin efficiency of the
crude extract in the f ield ten thi instar of S Ci tua were left
it itedcoan ech of 4 Vigta tadiaa plants in clay pots Ihe pots were
wrr nderl by water in clay trays to prevent the larvae to leave the
plant lo ts
rud rethanolic extract was admixed with bentonite in 13
by weight 3 parts of bentonite were added to I part of mcthanolic extract
in m hanol the solvent was then evaporated to dryness to give a brown
Pocwd er
An queous emulsion containing 005 Iriton and an equivalent
of 2 c rude methanolic extract wlt s made from the brown powder and sprayed
on tw of the Vina nadiata lat A a control the other two plants
We IV I d I h - - i1t on c rtaihi ton art c oncntratiou of Tiritn
and rtOr it A mrI-prt d i tince itt the I ve dlar pt between the two
(of rt wan h rv(ed W WIC d a b r -rtrnl1 lants were
cc i tt eato up while te treated pl1at reraiwred healthy
f F i ll1)
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 10 shy
111 5 Vigia kad ata 6 days after treatlrent With
crude methanolic extract (treated-coNtrol
tr ated-r-ntrol )
A smai 1 scale field test was then carried out with this
fcrrault ir 150 Vigna adiata p] ants were gron in cement pots
Ili ) After 30 day 300 third intar Iarvv of S -UJia were
l ft t r f It 1IeI i ri each pt (Ill 7) 1he plants in 4 pots
r ra ith If 1- 1d colution (Ill 8) A ii control another
p - r o raved vith n ITeuca (JUtiOn containini the same
cn ctitr a (f etvionit f l Jrilor A crr d diiference in the
v-- - I f din i wi (l 4r it (II 4 I11 )() at 24 hour
Vi a tI oa Ii-t ion of the arei of leaf dama fge and number counts of
r o riti ii a i tit t 4 and Z hrurs after spraying
qiirng tie ft l i++ini or t ul a shy
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 11 -
Calculation of area of leaf damage by S UIuka
leaf = 05) c7 of damage a(5 x + b (5 x 438)(5 x 018)
5x n
= a(25) 119) + c (09) x 100
a -- no of leaves which were d aped (r e than 507deg
b = no of leaves which were da ged ire than 25
c = no of leaves which were damiged maure than 10
(or no of leave coMtinued -pdud spots)with wei
d = no of leaven which wvr less th1r 1A damaged
(or no of lav wIth discontinued dtiraed spots)
a = no of leves which were not damaped
5 approx-- area of 4 cm 2 in which degree of damage was
est i ma ted
n = no of leaves randomly selected for checking in each
treatment
Similar test was also carried out with an aqueous extract
of 10 (ww) fresh leavo and h05 Ariton
The results of both tests were shown in 111 11 and 111 12
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 12 shy
11l 6 - Vigna adiala in cement iot 30 days old
S tua ere left to feed on Vigna kadcta lae
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 13 shy
Spraying With Lutet(I Uon
4shy
iii - Vi-a hadiata 24 hrs after spraying ith
Contro ol ion
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
10
- 14 shy
--J V ~gtwdiata 24 hr- afteir spraying with tested olution
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 15 -
Ill 1 - Comparison cI feedi ng a reai In
tested and control experiernt
i LE VEI I
0
HlOI4CI
I 12 FFeeding inhibition by tested solution
EDN -INIITO -MPR- OF LI
Q M3
RSH EYES-shy- ( 595
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 16 -
The offf icacy of the Ageaia 6p in crop protection was tested
in t h0 f ar( r plots
EIHld I t 1 111 13
e 10t 19H71 ecember
Place Farmer plots at Han Mae Tae Amphur Sun-Sai Chiang Mai
ro_p Cauliflowers 45 days after seedling transfer
Nubur of larva S MuPims eggs were attached to the cauliflowers
leavcs (Ill 14) After 7 days larvae hatched and grew to
third instar (Ill 15) at which Lime the number of larvae
was counted prior o spraying
Treatrents T l sprayed with control solution 005 Triton in aqueous
sol ut ion
12 sprayed with an tueous suspension containing 8aof
methanolic extract-hentonite (13) rixture and 005 Triton
T3 sprayed with 101 fresh leaf aqueous etract with 005
A i ton
14 sprayed with 10 dry leaf aqueous extract with 005
A1riton
Observation - count aumber rf larvae prior to spraying 12 24 and 48
hours after spraying
Pecsu I - um arized in Ill 16
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
17shy
11 13
Cauliflowers plots
used in Field Test I
Ill 14 ALtaching S Uulta eggs to cauliflowers leaves
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 18 shy
11 15 - Third instar larvae from the
attached eggs
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
II 16 - NUR OF INSECT AFER TREATMENT WITH AGLAIA SP
t
7-iV-
ur j$N
ETH
Z
A
H u4
A
CONTROL
A D S O R B E D O N
12
WITHl LEAF MLIIANOI EXTRACT
ADSORBED ON BENTONITE
N-ET
21 Hour
WITHI FRESHi LEAF
NETiIANO EXTRACT
X
48
WITII DRY LEAF
METHtANOL EXTRACT
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 20 -
Field Test 2
Dte 21 February 1988
P1ace Farrrer plot Leside road leading to Wat Sart Luang Chiang Mal
(rop PrBccor c i oI days aft r seedl ing tran sfer
2w _r ( relicatin - 3 rplications of 1 x 3 m plot
Ir 7et T1 prayed with control solution 005Triton in aqueous
solut Ion
T2sprayed with 101 dry leaf aqueous extract with 0057
Triton
T3 sprayed with 10 dry twig aqueous cxtract with 005
1riton
Results summarized in Ill 17 and Ill 18
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
Ill 17 - NUMBER OF SECT AFTER TREATIMENT W1H AGLAIA SP
I--
II
DAY
CONTROL WITH DRY LEAF WITH DRY TWIG
METHANOL EXTRACI METHANOL EXTRACT
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
ISI 8 - -LEAF DAAGE AFTER A 7LA i S
3-2i 4
-~~~ ~ ~~~ I-
14 t N
N)
CAA
CONT RO0L t-LTII DRY L AF wl -tiDRY T WG 11EITHANOL EXTRACT 1ITHANOL EXTRACT
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 23 -
Although some degree of efficacy of Agfaia 6p cxtract in
crop prottect on was observed accurate results could not be cheived
dup to the diminishing numbers of larvae both in tie treated and control
experiment
Another uxperiment was then carried out under the Ltter
ccmtr-)Id cond t ions
late 30 March 1988
plac Pon-harvest Centre Chiang Mal University
Sra-l Broccol (25 days after seedling transfer) in clay pot
urrrunded by 1 inch of water in clay tray to prevent the
l rva from escapirng
Nu7yor of I rva 0pot
Pepi arlana -replications of 10 pots
r -r_t II sprayed with control solution 005o Triton in
aqueous solution
2 sprayed with an aqueous emulsion of 2i leaf methanolic
etract and 00O Iriton
T3 prayed with an aqueous emulsion of 2 twig methanolic
extract and 0057 Iriton
Results sumn aried in Ill 19 20 21 22 23
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
24 -
Ill 19- Compariscn of leaves damage 4 days after
treatment with 27 leaf methanolic extract
iMd Con tro I
Ill 20 Corrp-ison of leaf dami ge 5 days
after treatment with 24 ]eiif
metharnaoIc extract anI conitrol
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 25 shy
1121 - CoPa r I Son of leaf darrage 5 (lays
after trmarment wi th 2 leaf
methanle] ic extrLct (T 2) 2 twig
netha-nclIc extract (T 3 ) and
control (
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
ATERATM ITH AGLAIA Sp
a a
CONTROL WITh LEAF METHANOL WITH TWIG METHANOL EXTRACT
EXTRACT
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
i 23 LEA DAA P G- F A - AGLAIA
Uii
-I-I
1 4
ct cl
CONTROL tjITH LEAF METHAINOL
EXTRACTEXTRACT
WITH TI IG M shy THAN O
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 28 shy
RRIEAwaO PLAN
It can be ccn Iuded from Ihe ( iermenLs on Agtaia sp
that thre exI nt a pa qihilIty o usin this plant as an antifeedant
for Srdopten Ulna in agriculture IPlans have been node for the
I 1l1itu i IL
1) Furdther asSess other irrnu latiorn of Agaa 6p in the
field
2) Compare the activity of scopoletin which is the active
antifeedant in thi plant with other analogues
Toward the end of this reporting period we had reassesed
the insecticidol activitv of the plants which had been pieviously
studied and recorded in our earlior report with various forul ations
Pr arable roult r obtained from the prolictinary studies
0 finl period of this project will then be also spent in evaluating
their Tplical ility ira the fields
IlFFpp[ cFE
I Sai Tikwattanit Mc Thesis Kasetsart University(1977)
2Vciha Patuoumotat lSc TIhILis aseLtL rt University
Si172)
5 Rtnolh I a I I h1 Ich~z br t~nt XIAIpp (195f1) 1I209
L an a I 1 I( I) _i 109C
7 thinbaton al LJ Ec-ktt JkrCtem Soc (1912) 94 825d
C i 1 2 a nd Ling JCS
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 29 shy
9 MJBHqluy 2Cruubju 1J lam andc LAWhj t iio JCSChumI
(012 (i12) 1
IU iil 12 ~cit I ~ J03 lnuut Ilivio ( Ijibt) 14 106 3
11 V--1 lni I1~~u Ji)141 L)ad4 -i
1 Al~Kuai NMzkhhi ACuizwk iiI sjuriai Ayr hiil Chem (073) 1y(09
3 K Wda Y Li ato aind FKmnaat Acjr Iiol ChUri 0) (9 34 941
14 EWaija Y Lnuotu and K151Maiio Ag~r ia l Chem (1970) 14 94b
IKatc andu Yjluzaka ta Tet rahedron Le tter (19 74)
17 i v i v a n a d9
(1175) 2 197S
19 KLO2) 1 Miu-a anO C 1iako2j hi 3 CLrCcd(97 6701
20 Iamp Lbo LY-0I KNaLkaishi and ACiiapya JCS
2 1 I ibo SNpjgti Y- Mi ura Nakanislii and AChay
22 Kul~ 1 -4I ( 1 Pet tLe I 1ijwicz and~t~nrih
J C L Cnc1 Coo ) 10 1
23 1 F aboampi F Ha an i sh i T I 1 soh and CillWKa k aW TKuuta
24 7 Iedil Hout- Plan~rt CCC 2 C t-dice to Io it A C S Sympos i Utn
Meriv Uc2 Ameiricajn ChenicoI a00201 gt4 WaiinqLyofl DC (1977)
1 73
PAeIini Plaiin~t JtIC1tacc to l(2tsi CS Stpi r
Series~ U inri can Chemical SOcact washington DC (1977) 374
0
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52
- 30 shy
26 Il Slurkil1 A Dictionary of the Economic Products of the
Hal ay enin-iula 2 VolMs Minitry of Agriculture and Cooprratives
huj ] Lupuira L-urpir (11)(6)
27 H Grair g t i]i I]ant Spc ie Prportedl y Possesing
I(rt-Control Propert ji-An IICUll )atabjaIe Vesource Systems
1nntjtitt irt-~e~ -tCrnter Hn)roluilu (1985)
23 P1 Tuntiwrwh tt ikurl -t al ht chremjftry (1981 )20 1164
29 T Imatayakul -t 11 A t Chrem (1979) 2 71
30 H HitnlI nll j Yrcwoto) Na tura llv Occurring Inecticides ed
7aconon H ann] Croby P( Marcel Dekker New York (1971) 62
31 W Itwers tatural Productf for Innovative Pest Management
ed Whitehead D L and flowers WS Pergqanon Press Oxford
(1903) 52