EXPERIMENTAL

Estelar

1. Survey of Gorakhpur Division for collection of essential oil yielding

plants (aromatic plants):

1.1 Geographical position of study area:

The sites of Gorakhpur Division taken in present investigation are

situated in Eastern part of Uttar Pradesh (Plate 1) between latitude of 27º05'

to 27º25' North and longitude of 83º20' to 84º10' East. The division

comprises Deoria, Gorakhpur, Kushinagar and Maharajganj districts; of

which Maharajganj and Gorakhpur districts have forests. Gorakhpur

Division is surrounded by Basti Division on the West & Mau on the East,

River Ghaghara on the South and Nepal Tarai on the North. The height

above sea level ranges from 107m in North-West to 93m in the South- East.

1.2 Topography:

The study area is like monotonous alluvial plane, the monotonous

being broken by presence of low watersheds, alluvial terraces and rivers.

Multinominal nalas, lakes, tanks and swamps are responsible for making the

local topography of the area. The land surface is apparently a level tract

sloping gently from North-West to South-East. Total absence of hills or

hillock is a remarkable feature of the area.

1.3 Soil:

The soil of this area is gangatic alluvial brought down by rivers

Ghaghara, Rapti, Rohin and Gandak from the Himalayas. Soil of the areas

ranges from low to medium in the organic matter with other soil nutrients.

Local topogaraphic and drainage variation have brought about significant

changes in the soil morphology, resulting in textural differences, grading

from sand through loam and silt to heavy clay. The major soil type includes

clayey, loam, sandy loam, silt laphicoam and usar or alkali soil.

39

Estelar

Estelar

1.4 Climate:

Gorakhpur Division includes Tarai region which is famous for its

monsoon type of climate with its characteristic three seasons such as rainy,

winter and summer. Besides three broad seasons, there are two periods viz.,

rainy winter and winter summer marking the transition from one season to

other. The rainfall varies considerably from year to year. The monsoon rains

commence during June and come to an end in September but may persist till

October. The minimum temperature goes down to 3ºC in the month of January

& maximum upto 43ºC in the month of June.

1.5 Vegetational composition:

Gorakhpur Division has dense forests cover close to the foothills of

Himalayas. All the forests of Gorakhpur Division viz., Achalgarh,

Anandnagar, Banki, Chowk, Doma, Kushmahawa, Kushmahi, Lehradevi,

Madanpur (out of Gorakhpur Division), Madhaulia, Nichlaul, Pakari,

Tehrighat and Tilkonia are rich in species composition of higher plants. These

forests are also inhabited by large number of wild animals such as Leopard,

Sambhar, Black langur, Monkey, Black buck, Cheetal, Wild dog, Jackal, Cat

etc. The vegetational flora constitutes herbs, shrubs, trees and climbers. Trees,

shrubs and climbers occur throughout the year and form permanent

vegetation, while herbaceous plants are seasonal which appearing during rainy

season and decrease during winter and finally become scarce in peak summer.

2. Methodology:

The present work is based on an intensive survey of aromatic plants

of Gorakhpur Division during 2008-2010. Excursions of different forests

(Achalgarh, Anandnagar, Banki, Chowk, Doma, Kushmahawa, Kushmahi,

Lehradevi, Madanpur (out of Gorakhpur Division), Madhaulia, Nichlaul,

40

Estelar

Pakari, Tehrighat and Tilkonia) and forest ranges (Bansgaon, Barahalganj,

Deoria, Gorakhpur, Khajani, Kushinagar, Partawal and Sahjanwa) of

Gorakhpur Division were made and aromatic plants were collected in their

flowering and fruiting conditions. The dominance of aromatic plants was

recorded on visual basis for presence and absence of species. The plant

specimens were assigned collection number. Location and other field

information (habit and habitat, flowering and fruiting, distribution pattern)

were recorded in field data book. During visits information regarding local

name, medicinal and traditional uses of plants were accessed through native

people and matched with earlier published literatures. The specimens were

pressed, dried and mounted on herbarium sheets with the help of adhesive,

following field and herbarium technique (Lawrence, 1951; Jain and Rao,

1977). Collected specimens were identified using flora (Duthie, 1903-1929;

Srivastava, 1976)

as well as by matching their specimens lodged in

departmental herbarium of Gorakhpur University and BSI (NRC) Dehradun.

The herbarium of collected plants was deposited at BSI, Dehradun. The

nomenclature of Taxa has been done upto date according to International

Code of Botanical Nomenclature (1988). The treatment of each species

includes correct name and citation of nomenclature, followed by description

of each species depicting diagnostic character in alphabetical order. The

photographs of wild plants are also provided (Plate No. 2-8). The essential oil

yielding plants (aromatic plants) collected during investigation are listed in

Table 5 which shows that species ordered alphabetically by family, place of

collection with collection number, intensity of occurrence, flowering/fruiting

and habit/habitat. Family wise distribution of plants is depicted in Fig. 1.

41

Estelar

Table 5: Essential oil yielding plants (Aromatic plants) of Gorakhpur Division

Plants name

(common name) Family Occurrence Place and collection number

Flowering &

fruiting Habit & habitat

1 2 3 4 5 6

Acorus calamus Linn.

(Bach) Araceae Uncommon

Doma forest, Kushmahi

forest/GKU4380 Apr.-Jul.

Rhizomatous herb / herb

undergrowth in sal forest

Adhatoda vasica Ness (Arusha)

Acanthaceae Common Tilkonia, Gorakhpur, Barahalganj/

GKU4334 Feb.-Apr. Shrub/along road side

Aegle marmelos (L.) Corr. (Bel)

Rutaceae ,, Partawal, Tilkonia/ GKU4313 Apr.-Aug. Tree/kitchen garden

Ageratum conyzoides Linn. (Ajagandha)

Asteraceae Abundant Gorakhpur, Tilkonia, Banki/

GKU4328 Nov.-May

Herb/along road side, waste

places

A. houstonianum Mill. (Ajagandha)

,, ,, Partawal, Deoria, Gorakhpur/

GKU4333 Aug.-May ‖

Anethum graveolens Linn. (Sowa)

Apiaceae Uncommon Bansgaon, Kushinagar/ GKU4367 Jan.-Mar. Herb/cultivated field

Anisomeles indica (L)Kuntz (Basiga)

Lamiaceae Common Gorakhpur, Tilkonia Barhalganj/

GKU4301 Nov.-Feb.

Herb or under shrubs/along

road side, waste land

Annona squamosa Linn. (Sitafal)

Annonaceae Uncommon Gorakhpur, Tilkonia, Partawal./

GKU4349 Apr.-Aug. Tree/ garden

Artemisia nilagirica Linn.

(Dauna) Asteraceae ,,

Nichlaul forest, Pakari forest,

Kushinagar /GKU4365 Dec.-Mar. Shrub/along road side

Ashphodelus tenuifolius Cav.

(Banpiyazi) Liliaceae

Occasionally

distributed Banki/ GKU4356 Dec.-June Herb/cultivated field

Azadirachta indica A. Juss

(Neem) Meliaceae Common

Gorakhpur, Khajani, Sahjanwa/

GKU4321 Mar.-Jul. Tree/along road side

Contd…..

42

Estelar

1 2 3 4 5 6

Blumea eriantha DC.

(Kukaraunha) Asteraceae Common

Achalgarh forest, Chowk forest,

Doma forest /GKU4306 Dec.-Apr.

Herb/dry waste places, along

road side

B. lacera (Burm.f.)DC

(Kukuraunha) ,, ,, Tilkonia, Partawal, Banki/ GKU4337 Feb.-May

Herb/dry waste places, along

road side

B. laciniata DC. (Kukaraunha) ,, ,, Kushmahi forest, Banki forest,

Lehradevi forest /GKU4363 Jan.-Apr. ,,

B. membranacea DC (Kukuraunha)

,, Abundant Bansgaon, Gorakhpur, Sahjanwa/

GKU4388 Jan.-May ,,

B. mollis (D.Don) Merr.

(Kukaraunha) ,, ,,

Nichlaul forest, Madhaulia forest,

Anandnagar/GKU4392 Feb.-May ,,

Caesulia axillaris Roxb.

(Bangra) ,, Uncommon

Banki forest, Kushmahi

forest/GKU4351 Sep.-Jan.

Herb/in the field of rice,

shady places Callicarpa macrophylla

(L.)Vahl.

(Priyangu) Verbenaceae ,,

Nichlaul forest, Kushmahawa forest,

Madanpur forest /GKU4338 Jul.-Jan.

Shrub/shrubby under growth

in sal forest

Callistemon

lanceolatus(R.Br)DC

(Bottlebrush) Myrtaceae ,, Gorakhpur, Kushinagar/GKU4373 Dec.-Mar. Tree/garden

Cannabis sativa Linn. (Bhang) Cannabinaceae Abundant Banki forest, Kushmahi forest,

Tilkonia forest /GKU4391 Jan.-Dec.

Herb/along road side,

railway track Chenopodium ambrosioides

Linn.

(Ban bhathuwa) Chenopodiaceae Uncommon

Kushmahi forest,

Kushinagar/GKU4346 Jun.-Sep.

Herb or under shrub/along

bank of canal

Chrysanthemum indicum (L)DC (Guldaudi)

Asteraceae ,, Barahalganj, Khajani, Gorakhpur/

GKU4319 Dec.-Mar. Herb/ garden

Citrus aurantifolia (Christm)

Swingle (Kaghzi nimbu)

Rutaceae Common Gorakhpur, Partawal/ GKU4315 Nov.-Jul. Small tree/ garden

Contd…

43

Estelar

1 2 3 4 5 6 Citrus aurantium Linn. (Khatta)

Rutaceae Common Sahjanwa, Tilkonia/ GKU4317 ‖ ‖

C. limon (L) Burm (Bara nimbu)

,, ,, Deoria, Barhalganj/ GKU4318 Nov.-Jul. Small tree/ garden

Clausena pentaphylla

(Roxb.)DC.

(Ratanjot) ,, Uncommon

Kushmahi forest, Pakari forest,

Nichlaul forest /GKU4354 Mar.-Jul.

Shrub/shrubby undergrowth

in sal forest

Cleome gynandra ( L.) Briq.

(Hulhul) Capparidaceae Common

Banki forest, Gorakhpur,

Kushmahawa forest/ GKU4350 Jul.-Oct.

Herb/ along bank of pond,

shady places Clerodendrum inerme

(L)Gaertn (Sangkupy)

Verbenaceae Uncommon Gorakhpur, Tilkonia, Banki/

GKU4374 Apr.-Nov. Shrub/along road side

C. viscosum (L) Vent. (Titbhant)

‖ ,, Sahjanwa, Bansgaon. Kushmahi/

GKU4330 Feb.-May

Shrub/shrubby undergrowth

in sal forest

Colebrookea oppositifolia Sm.

(Bantulsi, Pansra) Lamiaceae ,,

Tehrighat forest, Nichlaul forest,

Madanpur forest /GKU4336 Dec.-Mar.

Shrub/shrubby undergrowth

in sal forest

Cosmos sulphureus Cav. Asteraceae ,, Gorakhpur, Banki./ GKU4389 Dec.-Mar. Herb/ garden

Cotula anthemoides Linn. (Babuna)

,, ,, Tilkonia, Partawal /GKU4390 Nov..-Mar. Herb/ cultivated field, bank

of pond

Curcuma aromatica Salisb.

(Jangali-haldi) Zingiberaceae

Occasionally

present Nichlaul forest/GKU4316 Dec.-Mar.

Rhizomatous herb/ herb

under growth in sal forest

C. zedoaria Rosc. (Kachura) ,, Common Banki forest, Tilkonia forest,

Lehradevi forest/GKU4377 Jan.-June

Underground rhizomatous

herb/waste land

Cyperus brevifolius

(Rottb.)Hassk. (Mutha)

Cyperaceae ,, Kushmahi forest, Banki forest,

Gorakhpur/GKU4372 Aug.-Nov. Herb/grassland

Contd….

44

Estelar

1 2 3 4 5 6 Cyperus monocephalus

Endl.

(Musta) Cyperaceae Abundant

Pakari forest, Achalgarh forest,

Gorakhpur/GKU4329 Sep.-Nov. Herb/grassland

C. rotundus Linn.

(Motha) ,, Uncommon

Kushmahi forest, Banki

forest/GKU4370 Jul.-Oct. Herb/ in sal forest, grassland

C. triceps (Rottb.)Endl.

(Apavisha) ,, ,,

Kushmahi forest, Doma forest,

Gorakhpur/GKU4327 Sep.-Nov. Herb/ grassland

Erigeron bonariensis Linn.

(Bonaria) Asteraceae Common

Kushmahi forest, Kushinagar,

Achalgarh forest/GKU4305 Mar.-Aug. Herb/ grassland

E. canadensis Linn.

(Jarayupriya) ,, ,,

Banki forest, Kushinagar,

Gorakhpur/GKU4303 June-Sep. ,,

Eucalyptus citriodora Hook (Eucalyptus)

Myrtaceae Uncommon Tilkonia, Partawal/ GKU4361 Mar.-May Tree/along road side

Eugenia heyneana (L.)Wall.

(Kathjamun) ,, ,,

Tehrighat forest, Madhaulia forest,

Kushmahi forest /GKU4375 May-Aug.

Shrub/shrubby undergrowth

in sal forest

Eupatorium adenophorum

Spreng. (Bhenguar)

Asteraceae Occasionally

present Tehrighat forest,

Kushinagar/GKU4348 Feb.-May

Shrub/along bank of canal,

shady places

E. cannabinum Linn.

(Tangol-lati) ,, ,, Kushmahi forest/GKU4335 Jan.-Apr.

Herb/herb undergrowth in

sal forest

E. odoratum Linn.

(Ayapana) ,, Common

Chowk forest, Nichlaul forest,

Kushmahi forest /GKU4347 Dec.-Apr.

Shrub/shrubby undergrowth

in sal forest

Glycosmis pentaphylla

(Retz.) Corr; Hook.

(Karjeer) Rutaceae ,,

Nichlaul forest, Kushinagar,

Anandnagar /GKU4345 Aug.-Dec.

Shrub/shrubby undergrowth

in sal forest, along road side

Contd….

45

Estelar

1 2 3 4 5 6

Grangea maderaspatana

(L.) Poir.

(Jhinkimundi) Asteraceae Uncommon

Kushmahi forest,

Kushinagar/GKU4344 Dec.-May

Herb/ Along bank of pond,

shady places

Gynura crepidioides Benth.

(Jali) ,,

Occasionally

present Nichlaul forest/GKU4366 Nov.-Feb.

Herb/ shady places, along

road side

Hygrophila difformis Linn. (Sarpat)

Acanthaceae Uncommon Achalgarh forest, Doma forest,

Kushinagar/ GKU4309 Aug.-Mar.

Herb/ shady places, along

bank of rice field

H. pinnatifida Dalz. (Godadi)

,, Occasionally

present Kushmahi forest/GKU4343 Jan.-Mar. Herb/ along bank of canal

Hyptis suaveolens (L) Poit

(Wilayati tulsi) Lamiaceae Abundant Gorakhpur, Sahjanwa/ GKU4308 Nov.-Feb.

Shrub/shrubby undergrowth

in sal forest, along road side

Lantana camara Linn.

(Ghaneri) Verbenaceae ,,

Banki forest, Nichlaul forest,

Gorakhpur/GKU4376 Jan.-Dec.

Shrub/ along road side,

railway track

L. indica Roxb.

(Ghaneri) ,, ,,

Madhaulia forest, Doma forest,

Gorakhpur/GKU4355 Oct.-Dec. ,,

Lawsonia inermis Linn. (Mehndi)

Lythraceae Common Partawal, Khajani/ GKU4364 June-Nov. Shrub/ garden

Leonotis nepetaefolia R.Br

(Dhompo) Lamiaceae ,,

Kushmahi forest, Achalgarh forest,

Nichalul forest /GKU4314 Dec.-Mar.

Herb/along road side,

railway track

Leonurus sibiricus Linn.

(Guma) ,, Uncommon Gorakhpur, Sahjanwa /GKU4357 ‖

Herb/dry sandy places, along

road side

Leucas aspera Spreng.

(Goma) ,, Abundant

Tehrighat forest, Doma

forest/GKU4312 May-Jul. Herb/cultivated field

L. cephalotes Spreng.

(Goma) ,, Common

Chowk forest, Banki forest,

Kushmahawa forest /GKU4320 Jul.-Sep. Herb/moist sandy places

Contd…

46

Estelar

Contd….

47

1 2 3 4 5 6

Lippia alba Rich.

(Bhuiokra) Verbenaceae Common

Kushmahi forest, Pakari forest,

Gorakhpur/GKU4331 Jan.-Jul.

Shrub/shady places, along

road side

Melia azedarach Linn. (Bakain)

Meliaceae Uncommon Banki, Partawal, Tilkonia/ GKU4323 Feb-May Shrub/shady places, along

road side

Mentha arvensis Linn. (Pudina)

Lamiaceae Common Anandnagar, Gorakhpur,Tehrighat/

GKU4302 June-Aug. Herb/cultivated field

Murraya koenigii Spreng. (Kurry patta)

Rutaceae ,, Banki, Gorakhpur, Tilkonia/

GKU4339 Feb.-Apr.

Shrub, shrubby tree/in sal

forest, garden

M. paniculata (L)Jack (Kamini)

‖ Uncommon Tilkonia, Gorakhpur/ GKU4304 June-Aug. Shrubby tree/ garden, along

road side

Nepeta hindostana Linn.

(Bilaiyalotan) Lamiaceae Common

Madanpur forest, Kushmahi forest,

Gorakhpur/GKU4322 Jan.-Feb. Herb/damp places

Ocimum basilicum Linn. (Kali tulsi)

,, ,, Gorakhpur, Partawal, Tilkonia/

GKU4342 Aug.-Mar. Herb/shady places

Ocimum canum Sims.

(Bantulsi) ,, Abundant

Kushmahi forest, Gorakhpur, Chowk

forest /GKU4368 Dec.-Mar. Herb/waste places

O. gratissimum Linn. (Ramtulsi)

,, ,, Anandnagar, Barhalganj, Partawal/

GKU4352 Oct.-Feb. Herb/shady places

O. sanctum Linn. (Krishnatulsi)

,, ,, Gorakhpur, Khajani/ GKU4360 Aug.-Feb. ,,

Piper longum Linn. (Pipli)

Piperaceae Occasionally

distributed Gorakhpur, Banki, Madanpur/

GKU4324 Jul.-Aug.,

Dec.-Jan. Climber/shady places

Estelar

Contd…

48

1 2 3 4 5 6

Piper sylvaticum Roxb.

(Pahari-pipal) Piperaceae

Occasionally

distributed Madanpur/GKU4353

Jul.-Aug.

Dec.-Jan. Climber/ in babul forest,

along railway track

Pogostemon heyneanus

Benth.

(Pachouli) Lamiaceae Abundant

Kushmahi forest, Chowk forest,

Nichlaul forest /GKU4341 Jan.-Apr.

Shrub/shady places, along

road side

P. plectranthoides Desf.

(Pachouli) ,, ,,

Doma forest, Pakari forest,

Madhaulia forest/GKU4326

‖ ,,

Polyalthia longifolia

Sonner (Asoka) Annonaceae Common Tilkonia, Anandnagar/ GKU4358 Apr.-Jul. Tree/along road side

Polygonum glabrum Willd.

(Bihagni) Polygonaceae ,,

Kushmahi forest, Tilkonia forest,

Gorakhpur/GKU4379 Sep.-Apr. Herb/ shady moist places

Psidium guajava Linn. (Amrud)

Myrtaceae ,, Anandnagar, Barhalganj/ GKU4362 Dec.-Sep. Tree/ garden

Putranjiva roxburghii Wall. (Jiaputa)

Euphorbiaceae Uncommon Gorakhpur, Banki forest, Nichlaul

forest/ GKU4340 Nov.-Apr. Shrub, tree/along road side

Salvia plebeia R.Br. (Bhu-tulasi)

Lamiaceae ,, Achalgarh forest, Tehrighat forest,

Kushinagar/GKU 4307 Jan.-May

Herb/ along road side,

cultivated field

Saraca indica Linn. (Sitaasoke)

Caesalpiniaceae Common Khajani, Gorakhpur, Barhalganj/

GKU4384 Mar.-Apr. ‖

Siegesbeckia orientalis

Linn. (Katampam) Asteraceae

Occasionally

present Nichlaul forest/GKU4378 Oct.-Jan. Herb/ along road side

Syzygium cumini (L)Skeels (Jamun)

Myrtaceae Common Bansgaon, Banki, Sahjanwa/

GKU4332 May-Jul. Tree/along road side

Estelar

49

1 2 3 4 5 6

Tagetes erecta Linn. (Genda)

Asteraceae Common Anandnagar, Gorakhpur / GKU4369 Nov.-Apr. Herb/ garden

Vitex negundo Linn (Nirgundi)

Verbenaceae Uncommon Banki, Gorakhpur, Tilkonia/

GKU4310 May-Sep. Shrub/along road side

Xanthium strumarium Linn. (Lapetua)

Asteraceae Common Anandnagar, Banki, Barhalganj/

GKU4311 Sep.-June ‖

Zingiber officinale Rosc. (Adarak)

Zingiberaceae Uncommon Khajani, Kushinagar, Sahjanwa/

GKU4371 Sep.-Dec.

Underground herb/cultivated

field

Estelar

Fig. 1: Family wise distribution of aromatic plants

*Value represents number of aromatic plants

3. Observations:

Total 60 genera and 84 aromatic species were collected from different

forests and forest ranges of Gorakhpur Division (Table 5). The genera

of Blumea has largest number of species (5) followed by Cyperus and

Ocimum with 4 species. 9 genera were represented by two species; 2

genera represent 3 species while rests of the genera were represented by

only one species.

Out of 84 species, 41 species were herbs, 21 shrubs, 2 species under

shrubs, 4 species rhizomatous herbs, 6 species were shrubby trees and 8

species included trees while only two species such as Piper longum and

P. sylvaticum were climbers.

The collected aromatic plants belonged to 21 families (Fig. 1), out of

which Asteraceae (22) occupied topmost position followed by

Lamiaceae (16) and Rutaceae (8) while Araceae, Apiaceae,

50

3 2 1 1

22

1 1 1 1 4 1 16*

1 1 2

5 2 1

9

7 3 AcanthaceaeAnnonaceaeApiaceaeAraceaeAsteraceaeCaesalpiniaceaeCannabinaceaeCappiridaceaeChenopodiaceaeCyperaceaeEuphorbiaceaeLamiaceaeLiliaceaeLythraceaeMeliaceaeMyrtaceaePiperaceaePolygonaceaeRutaceaeVerbenaceaeZingiberaceae

Estelar

Caesalpiniaceae, Cannabinaceae, Chenopodiaceae, Capparidaceae,

Euphorbiaceae, Lythraceae, Liliaceae and Polygonaceae were

represented by only one species of each.

Out of total 84 aromatic species, 9 species belong to monocot family

(Table 6, Fig. 2) and 64 plants were wild nature.

Table 6: Aromatic plants distributed in Gorakhpur Division

Illustration Monocot Dicot

Total Number Per cent Number Per cent

Families 4 19.04 17 80.95 21

Genera 5 8.33 55 91.66 60

Species 9 10.71 75 89.28 84

Fig 2: Aromatic plants of Gorakhpur Division

Few species viz., Ageratum conyzoides, Anisomeles indica,

Cannabis sativa, Hyptis suaveolens, Lantana camara and Ocimum

51

0

10

20

30

40

50

60

70

80

Families Genera Species

4 5 9

17

55

75

Nu

mb

er

Monocot

DicotEstelar

canum were found to be abundantly distributed throughout the

surveyed places while the species like Clausena pentaphylla (Pakari,

Nichlaul and Kushmahi), Eupatorium adenophorum (Tehrighat,

Kushinagar), E. cannabinum, Hygrophila pinnatifida (Kushmahi),

Leonurus sibiricus (Gorakhpur, Sahjanwa), Piper longum (Banki,

Gorakhpur, Madanpur), P. sylvaticum (Madanpur), Curcuma

aromatica, Gynura crepidioides and Siegesbeckia orientalis

(Nichlaul) showed restricted distribution.

3.1 Distribution pattern:

The places of occurrence of each species were carefully noticed

during field survey. The places of occurrence of different species in the area

are given below:

a. Aquatic-Polygonum glabrum

b. Semi aquatic (Marshy/Swampy/Moist situation) – Acorus calamus,

Ageratum conyzoides, Caesulia axillaris, Eugenia heyneana, Eupatorium

adenophorum, Gynura crepidioides, Lippia alba, Nepeta hindostana,

Ocimum spp.

c. Dry places- Anisomeles indica, Azadirachta indica, Blumea eriantha, B.

lacera, B. membranacea, Chenopodium ambrosioides, Curcuma zedoaria,

Citrus spp., Hyptis suaveolens, Ocimum spp., Salvia plebeia, Vitex

negundo, Xanthium strumarium.

d. Riverian beds – Cotula anthemoides, Grangea maderaspatana,

Hygrophila pinnatifida.

e. In forests –Blumea eriantha, Callicarpa macrophylla, Colebrookea

oppositaefolia, Clerodendrum viscosum, Clausena pentaphylla, Curcuma

52

Estelar

aromatica, Eupatorium spp., Glycosmis pentaphylla, Gynura crepidioides,

Hyptis suaveolens, Murraya koenigii, M. paniculata, Piper longum, P.

sylvaticum, Pogostemon spp., Putranjiva roxburghii, Siegesbeckia

orientalis, Syzygium cumini.

f. Along road side, Railway track and Wasteland – Adhatoda vasica, Aegle

marmelos, Ageratum conyzoides, Anisomeles indica, Artemisia nilagirica,

Blumea spp., Cannabis sativa, Chenopodium ambrosioides, Cleome

gynandra, Clerodendrum inermae, C. viscosum, Cyperus spp., Eucalyptus

citriodora, Eugenia heyneana, Eupatorium odoratum, Glycosmis

pentaphylla, Hyptis suaveolens, Lantana spp., Lippia alba, Leonotis

nepetaefolia, Leonurus sibiricus, Melia azedarach, Nepeta hindostana,

Pogostemon spp., Polyalthia longifolia, Xanthium strumarium.

g. Cultivated field- Anethum graveolens, Blumea spp., Caesulia axillaris,

Hygrophila difformis, H. pinnatifida, Mentha arvensis, Leucas aspera,

Zingiber officinale.

h. Grassland/Garden- Aegle marmelos, Ageratum conyzoides, Callistemon

lanceolatus, Cannabis sativa, Clerodendrum inermae, Chrysanthemum

indicum, Citrus spp., Cosmos sulphureus, Cyperus spp., Erigeron spp.,

Lawsonia inermis, Mentha arvensis, Murraya koenigii, Ocimum spp.,

Polyalthia longifolia, Psidium guajava, Saraca indica, Syzygium cumini,

Tagetes erecta, Xanthium strumarium .

Thus, from above findings it is clear that few species have narrow

range of their distribution e.g. Polygonum glabrum an aquatic species, Cotula

anthemoides, Hygrophila pinnatifida and Grangea maderaspatana occur

along rivarian beds, Callicarpa macrophylla, Colebrookea oppositaefolia,

Clausena pentaphylla, Curcuma aromatica, Piper longum and P. sylvaticum

53

Estelar

occur in the forest only, have narrow range of their distribution. While

majority of species have wide range of distribution.

3.2 Phenological records:

The present data is based on survey and average observations of

three years (2008-2010) study on aromatic plants (essential oil yielding plants)

growing in different seasons at Gorakhpur Division. The flowering and

fruiting periods of individual plant species are reported earlier in Table 1 and

total number of species flowered and bore fruits in each month are given in

Fig. 3. The seasonal flowering and fruiting pattern are also depicted in Fig. 4.

Fig. 3: Flowering and fruiting in aromatic plants of Gorakhpur Division

54

0

10

20

30

40

50

60

70

80

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Num

ber

Months

No.of species in

flowering (Fl)

No. of species in

fruiting (Fr)

Total no. of species

in flowering and

fruiting(Fl + Fr)

Estelar

Fig. 4: Seasonal flower and fruit pattern of aromatic plants

As evident from Fig. 3 that most of the aromatic plant species were

found to be in flowering and fruiting during winter to summer transition

months (Dec- April). Maximum flowering occur during winter (October to

February), while minimum in summer peak. Majority of species bore fruits

during winter-summer season (26), while it was the minimum during summer-

monsoon (6) (Fig. 4). Besides 10 species were bore flowers and fruits

throughout the year.

3.3 Enumeration of aromatic plant species:

A total 84 aromatic plant species were collected from Gorakhpur

Division and are enumerated here alphabetically along with brief description

and medicinal uses. The family, habit/habitat, flowering & fruiting, place of

collection/collection number and occurrence are reported in Table 5.

Acorus calamus Linn. Sp. Pl. 324, 1753.

55

05

1015202530

6 9 8

13

25

13 10

14

6 9 8

13

26

10 Num

ber

No. of species

flowered

No. of species

fruit

Seasons

Estelar

A semi aquatic, rhizomatous perennial herb, rhizome creeping, much

branched, cylindrical, light brown, white and spongy within. Leaves bright

green, distichous, thickened in the middle, margin wavy. Flowers light brown,

densely packed in sessile cylindrical spadix. Fruits oblong, turbinate berries.

Dried powder of rhizome is given to children twice daily in fever

(Tomar, 2008).

Adhatoda vasica Nees in Wall. Pl. As. Rar. 3:103. 1832; FBI 4: 540; For Fl.

U.P. 3:522; FG: 240.1976. Justicia adhatoda Linn. Sp. Pl. 15. 1753; FUGP

2:76.

An evergreen bushy shrub. Leaves elliptic-lanceolate, acuminate.

Flowers white in dense, axillary peduncled spikes. Calyx and corolla 5 each,

corolla bilipped (2/3). Stamens 2. Fruits capsule, clavate, pubescent.

Decoction of leaves and bark is given in dose of 20ml, twice a day

for 2-3 days in cough and cold (Bera et al., 2008).

Aegle marmelos Corr. in TLS 5: 223,1800; FBI 1: 516; FUGP 1: 134; BBO 2:

164; For. Fl. U.P. 2: 115; FG: 66. Crataeva marmelos Linn. Sp. Pl. (ed. 1):

444, 1753.

A medium- sized deciduous tree with sharp, straight axillary spines.

Leaves alternate, 3- foliolate; terminal leaflet largest, ovate, lanceolate or

rhomboid, acuminate, long stalked. Flowers greenish white, in short lateral

panicles. Calyx and corolla 4-5 each. Stamens numerous. Fruits large,

globose, aromatic, yellow when ripe, seeds many, embedded in clear and

shining mucilage.

The roasted unripe fruits pulp is given in bowel complaints and ripe

fruits are taken as cooling drinks, leaf paste is taken in jaundice with sugar

(Singh & Mall, 2007).

56

Estelar

Ageratum conyzoides Linn. Sp. Pl. 1:839, 1753; FBI 3: 243; FUGP 1: 405;

For. Fl. U.P. 3: 392; FG: 171, 1976.

An erect, hairy, annual herb. Leaves opposite, ovate, crenate, acute.

Flower heads white blue or purplish, homogamous, discoid in dense or lax

terminal corymbs. Calyx modified into pappus. Corolla 5, tubular. Stamens 5,

syngenesious. Fruits cypsela, black with scaly pappus.

Decoction of leaves is applied externally on the affected part of the

skin disease, leaf juice taken in menstrual disorders (Natrajan et al., 1999).

A. houstonianum Mill. Gard. Dict. Ed 8. N. 2.

An erect, hairy, annual herb. Leaves opposite, ovate, crenate, acute.

Flower heads white, homogamous, discoid in dense or lax terminal corymbs.

Calyx modified into pappus. Corolla 5, tubular. Stamens 5, syngenesious.

Fruits cypsela, black with scaly pappus.

Decoction of leaves is used in skin disease (Ambasta, 2006).

Anethum graveolens Linn. Sp. Pl. 263. 1753; FG: 154. 1976. Peucedanum

graveolens (L.) Hiern, Fl. Trop. Afri. 3:19. 1871; FBI 2: 709; FUGP 1: 363.

An erect, glabrous, branched annual herb. Leaves 2-3 pinnate;

leaflets lanceolate to ovate, toothed or entire. Flowers yellow, in umbels.

Calyx and corolla, 5 each. Stamens 5. Fruits winged dorsally compressed

ellipsoid.

Seeds used as carminative and stomachic (Bhattacharjee, 2005).

Anisomeles indica L. Kuntze, Rev. Gen. Pl. 2:512. 1891; Mukerjee in RBSI

14: 152. 1940; Anisomeles ovata R.Br. in W. Ait. Hort. Kewed. 2:364. 1811;

FBI 4: 672; FUGP 2:109. - Epimeredi indicus (Linn.) Rothm. In Fedde

Repert. 53. 12. 1944.

A suffruticose, pubescent, aromatic annual herb or under shrub with

57

Estelar

acutely quadrangular stems. Leaves ovate, acute, crenate-serrate, base rounded

or subcordate petiolated densely hairy. Inflorescence verticillaster, flowers

bluish purple, in axillary whorls, forming dense interrupted spikes. Calyx 5,

ovoid or tubular. Corolla 5, united, 2-lipped. Stamens 4, didynamous. Fruits

nutlet black in colour.

The leaf juice is used twice a day for dysentery (Singh & Mall,

2007).

Annona squamosa Linn. Sp. Pl. 537. 1753; FBI 1: 78; FUGP 1:23; BBO 2:9;

For. Fl. U.P. 2:13; FG: 32.1976.

A shrub or small tree. Leaves oblong-lanceolate, dark green, obtuse

or subacuminate, glabrous. Flowers greenish-yellow, solitary or fascicled

terminal. Calyx 3. Corolla 3-6. Stamens many. Fruits fleshy syncarps,

globose, green, yellow with many black seeds.

Thalamus is used in antifertility. Seeds are useful in abortion and

menstrual. Paste of leaves is taken in jaundice (Tomar, 2008).

Artemisia nilagirica (Clarke) Pamp. in Nuov. Giorh. Bot Ital n. s. 33. 452

(1926).

An aromatic, perennial shrub, often gregarious, pubescent or villous

throughout. Leaves ovate, bipinnate to tripinnate, deeply pinnatisect,

pubescent above, white tomentose beneath. Heads homogamous, flowers

yellowish white, sessile in sub-erect or horizontal panicled racemes. Calyx

modified into pappus. Corrola 5, tubular. Anther 5, syngenesious. Fruits

achene.

Used in chronic fever, swelling and inflammations of liver also

employed as tonic and stimulant (Ambasta, 2006).

Ashphodelus tenuifolius Cav. in An. Cienc. Nat. 3: 46, t. 27, f.2.1801; FBI 6:

58

Estelar

332; FUGP 2 :333; FG: 318. 1976.

An erect scapigerous annual herb. Leaves terete. Scapes numerous,

branching above. Flowers white or slightly pinkish, in lax racemes. Perianth

6(3+3), petaloid. Stamens 6. Fruits globose capsule, loculicidal.

Seeds used as diuretic (Ambasta, 2006).

Azadirachta indica A. Juss Mem. Mus. Hist. Nat. Paris. 19:220. T.2.f.5. 1830;

BBO 2:176; For. Fl.U.P.2:124; FG: 70.1976. - Melia azadirachta Linn. Sp. Pl.

385. 1753; FBI 1: 544; FUGP 1: 150.

A large deciduous tree. Leaves unipinnate, crowded near the end of

branches; leaflets sub-opposite, serrate, acuminate, dark green above and paler

beneath. Flowers white, in axillary panicles. Calyx and corolla 4-5 each, calyx

of obtuse or rounded lobes; petals spathulates-oblong. Stamens 10. Fruits

drupes, ovoid-oblong, smooth.

Leaves extract orally administered for blood purification & removal

of intestinal worms. Leaf paste is applied on skin diseases, in small pox and in

pyorrhea (Ghazanfar & AL-Sabahi, 1993).

Blumea eriantha DC. in weight Contrib. Bot. Ind. 15, 1834; FBI, 3: 266,

1881.

An erect, puberulous, aromatic herb. Leaves simple, petiolate,

alternate, obovate or elliptic, apiculate, margin irregularly toothed,

membranous, pubescent on both surfaces. Heads heterogamous, yellow

clustered into axillary and terminal cymes. Calyx modified into pappus.

Corolla 5. Stamens 5, syngenesious. Fruits cypsela, dark brown.

Half cup of leaf infusions taken twice a day for diuretic (Chopra et

al., 2006).

B. lacera (Burm.f.) DC in Weight Contrib. 14, 1834; FBI 3: 263; FUGP 1:

59

Estelar

414; Randeria in Blumea 10: 237, 1960. Conyza lacera Burm. F. Fl. Ind.:

180. T. 59. FG: 1. 1968.

An erect aromatic annual herb. Leaves ovate–oblong or elliptic,

obtuse, entire or coarsely dentate, glandular and densely hairy on both

surfaces, often incised or lyrate. Heads, heterogamous, yellow in short

axillary cymes and terminal lax panicles. Calyx modified into pappus. Corolla

5. Stamens 5, syngenesious. Fruits cypsela, grey oblong with white pappus.

Leaf juice is applied on bruises of toe, cuts and wounds

(Bhattacharjee, 2005).

B. laciniata (Roxb.) DC. Prodr. 5: 436, 1834; FBI 3: 264; FUGP 1: 415; FG:

172, 1976. - Conyza laciniata Roxb. Fl. Ind. 3: 427, 1832.

An erect, aromatic, pubescent, annual herb. Leaves simple,

alternate, variable, usually large, runcinate-lyrate below and sub-pinnatifid,

spinulose above, hairy on both surfaces. Heads heterogamous, yellow in dense

to lax panicles. Calyx modified into pappus. Corolla 5. Stamens 5,

syngenesious. Fruits cypsela, flattened.

Plants used against mouth disease of cattle. The essential oils used

as antifungal and antibacterial agent (Asolkar et al., 2005).

B. membranacea DC. Prodr. 5: 440, 1836; FBI 3: 265; FUGP 1: 415 FG:

172, 1976.

A slender spreading annual herb. Leaves thinly membranous ovate

or lyrate, acute or obtuse, serrate, pubescent on both surfaces. Heads many

small, heterogamous, yellow, in glandular hairy terminal and axillary panicles.

Fruits cypsela with white pappus.

The essential oil obtained from the plants produced a marked and

long lasting fall in blood pressure in anaesthetized dogs. The oil exerted a

60

Estelar

direct depressant action on frog’s heart and spasmolytic effect on rabbit’s

ileum. Leaves used as antibacterial agent (Vanila et al., 2008).

B. mollis DC. Merr. In Philip. Journ. Sci (Bot.) 5: 395, 1910, FG: 172, 1976.

Erigeron molle. D. Don. Prodr. Fl. Nepal, 172, 1825 Blumea wightiana DC.

In Wt. Contrib. 14, 1834; FBI 3: 261; FUGP 1: 413.

An erect, annual, aromatic leafy herb. Leaves simple, alternate,

petiolate, elliptic-lanceolate or obovate, irregularly toothed or serrate,

glandular on both surface. Heads heterogamous, purplish in terminal or

axillary spiciform dense cymes or panicles. Calyx modified into pappus.

Corolla 5, tubular. Stamens 5, syngenesious. Fruits cypsela, linear.

The decoction of whole plant used to treat diarrhea (Asolkar et al.,

2005).

Caesulia axillaris Roxb. Pl. Cor. 1: 64, t. 93, 1795; FBI 3: 291; FUGP 1: 422.

An erect or sub-erect glabrous marshy annual herb. Leaves

alternate, sessile, lanceolate, acuminate, tapering to the auricled base. Heads

globose, homogamous, flowers purplish or white, connate together in

compound head. Calyx modified into pappus, axillary sessile. Corolla 5.

Anther 5, syngenesious. Fruits cypsela, dark brown.

A mixture of this plant and khesari in water is given to cattle in

stomach disease; leaf is used in treatment of goiter, plant used in baldness and

diarrhoea (Asolkar et al., 2005).

Callicarpa macrophylla (L.) Vahl. Symb. Bot. 3: 13.1794; FBI 4:568; FUGP

2: 86; For. Fl. U. P. 3: 533; FG: 253. 1976.

An erect shrub. Leaves elliptic or ovate-lanceolate, crenate to

serrate, acuminate, upper surface wrinkled, lower cottony. Flowers pink in

dense axillary, globose, cymes. Calyx 5, companulate. Corolla 5, tubular.

61

Estelar

Stamens 4. Fruits drupe, white.

Roots yield an essential oil used in stomach disorder, leaves

warmed and applied to rheumatic joints (Ambasta, 2006).

Callistemon lanceolatus (R.Br) DC. Sm. Sweet, Fl. Austr. Sub. T. 29. 1827;

FG: 133.

A large evergreen shrub or small trees with twisted trunk and

drooping branches. Leaves alternate, linear-lanceolate, acute and rigid.

Flowers crimson, in terminal drooping spikes. Calyx and corolla 4-5 each,

sepals deciduous. Stamens numerous. Fruits capsule, woody with truncate

apex.

The extract showed strong elastase inhibition and radical

scavenging activities, used for the development of skin antiwrinkle ingredients

in cosmetic formulation (Kim et al., 2009).

Cannabis sativa Linn. Sp. Pl. 1027. 1753; FBI 5: 487; FUGP 2: 200; FG:

301-302. 1976.

A scarsely branched, aromatic herb. Leaves upper 1-3 and lower 5-

11 partite, lobes lanceolate, serrate. Flowers small, greenish white, dioecious.

The male flowers in axillary panicled cymes while female flowers in axillary

racemes. Fruits compressed, crustaceous nut.

A poultice of leaves is applied externally around the anus for one

month to cure piles (Tomar, 2008).

Chenopodium ambrosioides Linn. Sp. Pl. 219. 1753; FBI 5:4; FG: 278. 1976;

Raizada Suppl. FUGP 336. 1976.

An erect, much branched, aromatic herb or under shrub. Leaves

oblong–lanceolate or oblong ovate, obtuse or acute, sinulate-dentate. Flowers

green, minute in axillary and terminal simple or paniculated leafy spikes.

62

Estelar

Perianth 5-lobed enclosing the fruits. Stamens 5. Fruits utricle membranous

brown.

Used as an anthelmintic against many forms of intestinal parasite

(Mishra et al., 2008a).

Chrysanthemum indicum (L)DC. Sp. Pl. 889; Thunb FL. Jap. 320.

An erect bushy, aromatic annual herb. Leaves aromatic, foliage,

deeply cut and lobed, oblong-broadly ovate, pinnatifid. Heads pink or white,

heterogamous, solitary terminal. Stamens 5, syngenesious. Fruits cypsela

oblong, dorsally compressed.

Used as stomachic and aperients (Ambasta, 2006).

Citrus aurantifolia (Christm) Swingle in Journ. Wash. Acad. Sci. 3:465.

1912; Webber & Batchelor, Citrus Ind. 1: 401; FG: 67. 1976- Limon

aurantifolia Christm. in Linn.

A thorny shrub or small tree. Leaves alternate, elliptic-oblong or

ovate, serrate, obtuse or pointed, petiole winged. Flowers white, in solitary

axillary. Calyx and corolla 4-5 each. Stamens numerous. Fruits small, globose

or ovoid.

Fruits juice contains citric acids and is useful in stomach disorder

and gastric trouble. (Bhattacharjee, 2005)

C. aurantium Linn. Sp pl. 783, 1753.

A small shrub or tree. Leaves large, ovate-oblong, emerginate,

usually downy beneath, petiole broadly winged. Flowers large, white in

solitary axillary. Calyx and corolla 4-5 each. Stamens numerous. Fruits

globose and pear shaped.

Used in the preparation of confections and other drinks. It is

cultivated for its nutritious fruits which is a cardio tonic and refrigerant

63

Estelar

(Ambasta, 2006).

C. limon (L). Burm. f. Fl. Ind. 173, 1768; Webber & Batchclor. Citrus Ind. 1:

398; FG: 67. 1976. C. medica Linn. var. limon Linn. Sp. Pl. 2. 782. 1753. FBI

1: 514; FUGP 1: 131; For. Fl. U.P.2:110

A small, thorny shrub or tree. Leaves pale green, oblong, elliptic-

ovate, pointed or obtuse, crenate, petiole winged. Flowers white, tinged with

pink in cymes or solitary. Calyx and corolla 4-5 each. Stamens numerous.

Fruits medium sized, ovoid or oblong with a terminal nipple, yellowish green

or green.

The fruits are rich in acidic juice which is used medicinally in

rheumatism, dysentery and diarrhoea (Ghazanfar & AL-Sabahi, 1993).

Clausena pentaphylla (Roxb.) DC. Prodr. 1: 538, 1824; FBI 1: 503; FUGP 1:

129; For. Fl. U.P. 2: 109.

A small aromatic deciduous shrub. Leaves imparipinnate, tri-hepta

foliolate; leaflets alternate or sub-opposite, ovate-elliptic, oblong, lanceolate,

margins crenulate, glabrous, gland punctate. Flowers yellowish or greenish

white in terminal, downy panicles. Calyx and corolla 4-5 each. Stamens 10.

Fruits berry ovoid, orange.

Bark is applied to fresh wounds in powdered form for quick

healing (Ali and Dixit, 1989).

Cleome gynandra Linn. Sp. Pl. (ed.1): 671. 1753; FG: 42.1976.-

Gynandropsis gynandra (L.) Briq. In Ann. Cons. Jard. Bot. Geneve

17:382.1914; Santapau in R BSI, 16 (1):9. 1960.

An erect, spreading glandular-pubescent annual herb. Leaves

digitately pentafoliolate, obovate, acute, entire or serrulate. Flowers white,

sub-corymbose, solitary in the axis of leafy bracts, raceme. Calyx 4. Corolla 5.

64

Estelar

Stamens 4-many. Gynandrophore present. Fruits capsule, oblong, dark

brown.

Bruised leaves used in headache, rheumatism and other local pains.

Seeds rubefacient and anthelmintic (Chopra et al., 2006).

Clerodendrum inerme (L) Gaertn Fruct. 1:271. T. 57 f. 1.1788; Clarke in

Hook. F. FBI 4: 589; Bor & Raizada, 146. F. 92. 1954; Raizada Suppl. FUGP.

212. 1976.-Volkoameria inermis L. Sp. Pl. 6:889. 1753.

A straggling or subscandent shrub. Leaves ovate, elliptic, oblong,

rounded or retuse. Flowers white, in axillary cymes. Calyx 5, companulate, 5

toothed, corolla 5, tube slender, subtended at the base by 2 lipped. Stamens 4,

didynamous. Fruits drupe, pyriform subtended at the base by accrescent calyx.

Leaves used as febrifuge, poultice of leaves used to resolve buboes

(Bedi et al., 2008).

C. viscosum (L) Vent. Jard. Malm. t. 25. 1803; For. Fl. U.P. 545; FG: 254.

1976. C. infortunatum auct. (non. L. 1753); FBI 4: 594; FUGP 2: 93; Bor &

Raizada, 150. f. 95. 1954.

An erect shrub with quadrangular branches. Leaves ovate usually

cordate, sparingaly hairy on both sides. Flowers white, tinged with pink in

peduncled cymes and forming a large terminal panicles. Calyx 5 toothed,

companulate, corolla 5. Stamens 5. Fruits drupe, black when ripe.

Leaf paste rubbed over scalps, stops falling hairs, relieve headache

and promotes hair growth. Roots used in gastric disorder, as well as pains and

bodyaches (Bhattacharjee, 2005).

Colebrookea oppositifolia Sm. Exot. Bot. 2: III, t. 115.1805; FBI 4: 642;

FUGP 2: 107, For. Fl. U.P.3: 558; FG: 262. 1976.

An erect, aromatic, spreading, white tomentose shrub with 65

Estelar

grooved, sub-quadrangular branches. Leaves elliptic, oblong, crenate-serrate,

acute hairy. Flowers white, small in clustered spikes. Calyx deeply 5-lobed,

acrescent. Corolla small, 4 lobes. Stamens 4. Fruits nutlet black.

Leaves used in cuts, wounds and burns. Roots used in hysteria and

epilepsy (Ambasta, 2006).

Cosmos sulphureus. Cavan. Icon. Descr. Pl. 1: 56, 1791; Sherff. In Field

Mus Nat. Hist. Bot. Ser 7:401, 1932.

An erect branched annual aromatic herb, with grooved and glabrous

stem. Leaves opposite, 2-3 pinnatipartiate; segments lanceolate, obtuse, entire,

glabrous. Heads golden yellow-white, heterogamous, solitary or combined

into corymbs. Calyx modified into pappus. Corolla 5, tubular. Stamens 5,

syngenesious. Fruits cypsela, fusiform.

Leaves and aerial part used in jaundice, intermittent fever,

splenomegaly. Tonic, hepatic, hepatoprotective (Botsaris, 2007).

Cotula anthemoides Linn. Sp. Pl.: 891, 1753; FBI 3: 316; FUGP 1: 434; FG:

175, 1976.

A small prostrate or diffused glabrous aromatic herb. Leaves

alternate 1-2 pinnatifid, segments with lanceolate lobes. Heads yellow,

heterogamous, solitary terminal on filiform peduncles. Corolla 5, yellow,

united. Stamens 5, syngenesious. Fruits cypsela. Pappus absent.

The whole plant heated with mustard oil and decoction is applied

externally on affected parts 2-3 times a day in rheumatism (Ambasta, 2006).

Curcuma aromatica Roxb. Salisb. Parad. Lond t 96.

A rhizomatous herb with a thick tuberous aromatic root stocks.

Leaves linear, lanceolate, an apparent stem is formed by rolled up leaf sheath.

Inflorescence raceme, flowers red in colour. Calyx 3, united. Corolla 3,

66

Estelar

unequal, tubular. Stamen 1. Fruits loculicidal cypsela.

Rhizome powder used in fever, contusions and sprains (Ambasta,

2006).

C. zedoaria Rosc. Scitam t. 109. Roxb. in As. Res. xi (1810) 332. Ct etiam

165.

A rhizomatous underground herb. An apparent stem is formed by

rolled up leaf sheath. Leaves alternate spirally arranged, linear. Inflorescence

spikes arise from the sheaths. Flowers yellow. Calyx 3, united. Corolla 3,

unequal, tubular. Fruits loculicidal cypsela.

Used as stimulant tonic, stomachic and relieve from joints pains

(Chopra et al., 2006).

Cyperus brevifolius (Rottb.) Hassk. Cal. Hort. Bogor. 24.1844; Singh &

Dixit in Bull. Bot. Soc. Bengal 23 (2): 199. 1969; FG: 335. 1976.

An erect, slender perennial sedge with horizontal rhizome. Leaves

linear, erect, spikes aggregated into terminal solitary globose green heads.

Spikelets lanceolate or ovate-lanceolate, 1- flowered. Perianth absent

represented by bristles or scales or hairs. Stamens 3. Fruits nut, yellowish

brown.

Readily eaten by cattle. Leaves used in diarrhoea (Asolkar et al.,

2005).

C. monocephalus Endl. F. Muell Fragm. 8. 271. Roxb. Hort. Beng (5): Fl.

Ind. i 188. Singh & Dixit in Bull. Bot. Soc. Beng. 23 (2): 200. 1969 & Ind.

For. 98 (2): 122. 1972.

An erect, glabrous sedge with slender, elongated creeping rhizomes.

Leaves linear, acuminate. Spikes solitary, sub-globose, white, spikelets 1-

flowered. Rachilla deciduous. Stamens 3. Fruits nut, oblong.

67

Estelar

Decoction of aromatic rhizomes used as diuretic, demulcent and

tonic, also given in fever and diabetes (Ambasta et al., 2006).

C. rotundus Linn. Cent. Pl. 2:6. 1756; Singh & Dixit in Bull. Bot. Soc. Beng.

23(2): 201. 1969 and Ind. For. 98(2): 123. 1972; FG: 241. 1976.

A glabrous, annual sedge with numerous tufted, fibrous roots.

Leaves arising near base of stem, usually shorter than stems, linear, tapering in

acute apex. Inflorescence umbellate with at least one sessile head like spikes

of spikelets. Heads globose or oblong, spikelets oblong, brownish, 10-12

flowered. Stamen 1. Fruits narrowly obovoid.

Decoction of whole plants useful in diarrhoea, plants diuretic,

astringent (Chopra et al., 2006).

C. triceps (Rottb.) Endl. Cat. Hort. Ac. Vindob.1:94.1842; Singh & Dixit in

Ind. For., 98 (2): 124. 1972; FG: 342. 1976.

An erect, small, glabrous, annual sedge with small rhizome. Leaves

narrow, acuminate, spikes 3-5, ovoid or oblong, white aggregated into a

compact head. Rachilla deciduous, Perianth absent represented by bristles,

scales or hairs. Spikelets, 1-flowered. Fruits nut oblong, ellipsoid and pale

brown.

Decoction of plants is given in fever, the root oil used for

stimulating liver and to relieve pruritus (Choudhury et al., 2010).

Erigeron bonariensis Linn. Sp. Pl.: 863, 1753; Hara in Fl. E. Himalaya: 136,

1971; Raizada, Suppl. FUGP (ed.1): 112, 1976 E. linifolius Willd. Sp. Pl. 3:

1955, 1804; FBI 3: 254.

An erect, hairy, deep rooted annual herb. Leaves simple, alternate,

upper sessile, obovate-oblong, coarsely toothed. Heads purplish, in corymbose

panicles, heterogamous. Calyx modified into pappus. Corolla 5, tubular.

68

Estelar

Stamens 5, syngenesious. Fruits cypsela, oblong, hairy.

Plants used as stimulating diuretic in febrile condition (Kala, 2005).

E. canadensis Linn. Sp. Pl.:863, 1753; FBI 3: 254; FUGP 1: 408.

An erect, slender, much branched, pubescent hairy annual herb.

Leaves simple, alternate, upper sessile, narrowly linear or linear–lanceolate,

entire, acute, base cuneate. Heads yellowish–white, heterogamous in

elongated branched panicles. Calyx modified into white hairy pappus.

Corolla 5, tubular. Anther 5, syngenesious. Fruits cypsela, dirty white.

Decoction of whole plant given for diarrhoea and dysentery (Chopra

et al., 2006).

Eucalyptus citriodora Hook. in Mitch. Journ. Trop. Austral. 235.

A tall tree. Leaves large, lanceolate. Flowers in lateral or supra

axillary umbels. Calyx and corolla 4-5 each, united, corolla fuses with calyx to

form cup called operculum. Stamens numerous. Fruits capsulated.

Oil used in perfumery, wood used in preparation of furniture

(Bhattacharjee, 2005).

Eugenia heyneana (L.) Wall. Duthie in Hook. Fl. Br. Ind. 2: 500. 1879; FBI

2:500

A small to medium sized tree. Leaves narrow, lanceolate,

acuminate, glabrous. Flowers greenish white in panicles. Calyx and corolla 4-

5 each. Stamens numerous. Fruits berry, crowned by cup like calyx-limb,

brown in colour.

Root chewed for relief from tooth ache, used as vermicide, flowers

used in inflammation (Mishra et al., 2008a).

Eupatorium adenophorum Spreng. Syst. iii. 420, Hort. Berol. ex Kunth, Ind.

Sem. Hort. Berol. 13, 1847.

69

Estelar

An erect perennial herb. Leaves simple, opposite, oblong, ovate,

sessile or petiolate, crenate or coarsely serrate. Heads homogamous, white,

discoid, arranged in dense panicles or corymbs. Calyx modified into pappus.

Corolla 5, tubular. Anther 5, syngenesious. Fruits cypsela, brown with

glandular tubercles.

The extract of plant is used as diuretic, crushed leaves applied to

cuts and wounds, and also to treat bites of poisonous insects and caterpillars

(Chopra et al., 2006).

E. cannabinum Linn. auct. Non L., FBI 3:247, 1881. E. heterophyllum DC

Prodr. 5: 180, 1836.

An erect, suffrutescent, perennial herb. Leaves simple, opposite,

petiolate, lanceolate, acute or acuminate, coarsely serrate, attenuate at base.

Heads purplish, homogamous in terminal or axillary corymbs. Calyx modified

into hairy pappus. Corolla 5, tubular. Anther 5, syngenesious. Fruits cypsela,

cylindrical, black.

Diuretic, herb employed as purgative (Ambasta, 2006).

E. odoratum Linn. Syst. ed. 10: 1205, 1759; FBI 3: 244, 1881.

An erect perennial shrub, stem pubescent. Leaves simple, petiolate,

opposite, decussate, ovate, dentate, acute or acuminate, pubescent on both

surfaces. Heads purplish, homogamous, in branched corymbose cymes. Calyx

modified into hairy pappus. Corolla 5, tubular. Anther 5, syngenesious. Fruits

cypsela, fusiform black.

Decoction of leaves haemostatic, aqueous extract of shoots cardiac

stimulants (Awasthi, 1991).

Glycosmis pentaphylla Corr. DC. Prodr. 1: 538, 1824; FBI 1: 499; FUGP 1:

136; For. Fl.U.P.2:106.

70

Estelar

An evergreen shrub. Leaves tri-pentafoliolate; leaflets ovate-

lanceolate, acute or acuminate, entire, dark green, glabrous. Flowers small,

white in terminal or axillary pubescent panicles. Calyx & corolla 4-5 each.

Stamens 10. Fruits berry, pink or blue.

Brush of stem is used for clean the teeth in pyorrhea. Juice is

applied for fever and liver complaints, leaves considered good antidote for

skin trouble & stomach ache (Natrajan et al., 1999)

Grangea maderaspatana (L.) Poir in Lamk. Encycl. Suppl.2:825, 1811;

FBI3:243; FUGP 1: 407; FG: 178, 1976. Artemisia maderaspatana L. Sp. Pl.

(ed. 1):849, 1753.

A prostrate or sub-erect, villous, annual aromatic herb, forming

circular patches at the ground. Leaves alternate, sessile, pinnatifid or lobulate,

coarsely toothed, pubescent on both surfaces. Heads yellow, heterogamous,

globose, solitary or rarely in pairs on leaf opposed peduncles. Calyx modified

into pappus. Corolla 5, tubular. Anther 5, syngenesious. Fruits cypsela, pale

brown.

Half cup of leaf infusion taken 2 times a day in obstructed

menstruation, infusion of leaves also considered as stomachic, deobstruent and

antispasmodic (Choudhury et al., 2010).

Gynura crepidioides Benth. in Hook. Niger Fl. 438. Afr. Trop. Id

211243OVIF2.

An erect, annual herb. Leaves simple, longe, alternate, ramale and cauline,

lanceolate or ovate, obovate-lanceolate, unequally, tripinnatifid, dentate or

entire, acute. Heads pinkish disciform, homogamous in terminal panicled

corymbs. Calyx modified into pappus. Corolla 5, tubular. Anther 5,

syngenesious. Fruits cypsela, brown.

71

Estelar

Leaves used in digestion, lotion of leaf employed as mild stomachic

(Kala, 2005).

Hygrophila difformis Linn. Shreemadhavan et Bennet in Bull. Bot. Surv.

Ind. 10 (2):222. 1968; Clark in Hook. f. FBI 4: 405. 1884.- Synnema

triflorum (Roxb.)O. Kuntze. Rov. Gen. Pl. 2: 500. 1891; Dixit & Singh in Ind.

For. 94(10): 769. 1968; FG: 250. 1976.

An erect or decumbent, pubescent aromatic, annual herb with

quadrangular stems swollen at the nodes and rooting below. Leaves petiolate,

ovate, crenate-serrate on both sides. Flowers bluish white or purplish, in

axillary whorls. Calyx tubular, 5-fid. Corolla 2-lipped. Stamens 4,

didynamous. Fruits capsule, narrow, sessile.

Leaves edible, used for poulticing wounds and in tooth ache,

antioxidant activity (Debasish et al., 2010).

H. pinnatifida Dalz. Shreemadhavan in Bull. BSI 10 (2):222. 1968; O.

Kuntze Gen. Pl. 2: 500. 1891; Synnema pinnatifidum (Benth.exC.B. Clarke)

O. Kuntze Rev. FG: 250. 1976.- Cardanthera pinnatifida Benth. in Gen. Pl.

2: 1074. 1875; Clarke in HK.f. FBI 4: 405.

An aromatic herb with tetragonous stems glandular, pubescent,

swollen at the nodes. Leaves opposite, oblong-lanceolate, sub-obtuse or acute,

deeply pinnatifid. Flowers purplish, solitary or in lax spikes. Calyx 5. Corolla

bilipped. Stamens didynamous. Fruits capsule, sessile.

Decoction of whole plant taken in diarrhoea (Chopra et al., 2006).

Hyptis suaveolens (L) Poit. in Ann. Mus. Paris 7: 472, t. 29,f.2. 1836; FBI 4:

630; Raizada Suppl. FUGP: 220. 1976.- Ballota suaveolens Linn. Syst. (ed.

10): 1100. 1759.

A suffruticose branched, aromatic, annual herb or under shrub.

72

Estelar

Leaves ovate, sub-orbicular, acute, mucronate, base obtuse-subcordate, hairy

above. Flowers blue small, on axillary peduncles or in bracteates axillary

cymes. Calyx 5, densely glandular, hairy, 10 ribbed. Corolla 5, united,

differentiated into tube and limb. Stamens 4. Fruits nutlet, ovoid black.

Root paste is applied to relieve pain in scorpion stings. Paste of

leaves is applied in skin infection (Ambasta, 2006).

Lantana camara Linn. Sp. Pl. 627. 1753; Clarke in Hook. f.l.C.562; FUGP 2:

84; Meeuse, Blumea 5:69. 1942.-L. aculeata L.Sp. Pl. 627. 1753.- L. camara

var. aculeate (Linn) Moldenke. Torreya 34:9.1934.

A straggling or scandent, aromatic shrub with recurved prickles.

Leaves ovate or ovate-oblong with cordate or sub-cordate base, scabrid on

both side, crenate-serrate. Flowers orange yellow or pink in axillary, spicate

heads. Calyx small, membranous 4-5 toothed. Corolla – tube cylindrical, 4-5.

Stamens 4, didynamous. Fruits drupe, black.

Used for itch, an antiseptic for wounds, decoction given in tetanus

and malaria (Mishra et al., 2008a).

L. indica Linn. Roxb. Fl. Ind. 3: 89. 1832; FBI 4: 562; FUGP 2: 83; For. Fl.

U.P. 3: 531; FG: 256. 1976.

A hairy scandent shrub, branches-straggling, armed with scattered,

prickles. Leaves ovate-oblong, crenate-serrate, acute or sub-obtuse. Flowers

white, purple or yellow in axillary head or spike. Calyx 4-5 toothed, small.

Corolla 4-5, tube cylindrical. Stamens 4. Fruits drupe purple when ripe.

Leaves used as a cure for snake-bite (Asolkar et al., 2005).

Lawsonia inermis Linn. Sp. Pl. 349. 1753; For. Fl. U.P.2: 349; FG: 138.

1976- L. alba Lamk. Encyl. 3: 106. 1789; FBI 2: 573; FUGP 1: 323.

A bushy shrub or small tree, with thorny branchlets. Leaves sub-

73

Estelar

sessile, elliptic or narrow, rhomboid. Flowers cream coloured, in corymbose

branched, terminal panicles. Calyx deciduous, 4 lobed. Corolla 4. Stamens 8.

Fruits coriaceous capsule with persistent calyx and style.

The paste of leaves is used by local inhabitants to dye nails, palms

and soles and to relieve burning of palms and soles. Barks are given in

jaundice and enlarge spleen. (Tomar, 2008).

Leonotis nepetaefolia R.Br. Prodr. : 504.1810; FBI 4: 691; FUGP 2: 116;

Mukejee l.c. 185; For. Fl. U.P.3:562; FG: 263. 1976. - Phlomis nepetaefolia

L. Sp. Pl. 586. 1753.

An erect annual herb with grooved, puberulous, quadrangular stem.

Leaves ovate, coarsely crenate-serrate, acute. Flowers orange-scarlet in dense

globose axillary whorls. Calyx with 8-9 rigid teeth, teeth sharply spinescent.

Corolla bilipped. Stamens 4. Fruits nutlet black, linear to oblong.

Inflorescence put in 50gm ghee, boiled to viscous paste and orally

administered two spoon full a day for cough. Plants boiled in mustered oil and

applied over waist to relieve pain, flowers and seeds used in cuts, wounds and

burns (Chopra et al., 2006).

Leonurus sibiricus Linn. Sp. Pl. 584. 1753; FBI 4: 678; FUGP 2: 110; For.

Fl. U.P. 563; FG: 263. 1976.

An erect, glabrous or pubescent, annual herb. Stems bluntly

quadrangular. Leaves palmately pinnatifid, linear, lanceolate. Flowers bluish-

red in axillary whorls. Calyx turbinate, 5- toothed. Corolla 5, bilabiate.

Stamens 4, didynamous. Fruits nutlet, black.

Dried leaves and flowering tops diuretic, prescribed in hysteria and

heart palpitation (Chopra et al., 2006).

Leucas aspera Spreng. Syst. 2: 743.1825; FBI 4: 690; FUGP 2: 113; FG 263,

74

Estelar

1976.-Phlomis aspera Willd. In Link. Enum. Pl. Hort. Berol. 2: 621. 1822.

An erect or diffused branched annual herb with hispid, quadrangular

stems. Leaves elliptic-oblong, linear, lanceolate, entire or crenate, acute.

Flowers white in terminal and axillary whorls. Calyx 5, striate, 6-10 toothed.

Corolla bilabiate (2/3). Stamens 4, didynamous. Fruits nutlet, brown.

Juice of leaves applied externally in chronic skin eruptions and

painful swellings, anti-inflammatory (Natarajan et al., 1999).

L. cephalotes (Wild) Link. Spreng. Syst. 2: 743. 1825; FBI 4: 689; FUGP 2:

114; FG: 264. 1976-Phlomis cephalotes Roth., Nov. Sp. Pl.: 262. 1821.

An erect, hairy, annual herb. Stem obtusely quadrangular. Leaves

ovate, linear-lanceolate, crenate-serrate, sub-acute, and pubescent. Flowers

white in large dense terminal whorls. Calyx striate, 6-10 toothed. Corolla 2/3.

Stamens 4, didynamous. Fruits nutlet, brown.

Juice of leaf mixed with honey is taken in cough, the aqueous

extract of leaves in constipation. Flowers and leaves chewed in tooth-ache and

gum disorder (Ambasta, 2006).

Lippia alba Mill. Raizada Suppl. FUGP. 214. 1976.-Lippia geminate H.B.K.

Nov.Gen.et Sp.2:266:1818;NEBr.ex Britt. & Wilson in Sc. Surv. Porto Rice

and Virgin Islands 6:141. 1925.

A straglling aromatic shrub. Leaves ovate, crenate-serrate, base

obtuse, decurrent, rugose. Flowers pink, in dense cylindrical spikes. Calyx

lobed or toothed. Corolla 5, united. Stamens 4. Fruits small, dry. Seed

pyrenes.

Considered as stomachic (Ambasta, 2006).

Melia azedarach Linn. Sp.Pl. 384. 1753; FBI 1:544; FUGP 1: 141 FG: 70.

1976.

75

Estelar

A small deciduous tree. Bark greyish brown, smooth. Leaves

bipinnate; leaflets serrate, ovate-lanceolate and acuminate. Flowers blue or

purple in lax panicles of cymes. Calyx and corolla 4-5 each. Stamens 10.

Fruits drupe, yellow, globose, 5-seeded.

Fruits are used in leprosy and bark as anthelmintic (Bhattacharjee,

2005).

Mentha arvensis Linn. Sp. Pl. 577, 1753.

A small herb, stem short, branched, creeping rhizomes. The leaves

are narrow below stalked, oblong, lanceolate, toothed. Flowers white, in

terminal spikes. Calyx 5, united. Corolla 5, bilipped. Stamens 4, didynamous.

Fruits nutlet.

Stimulant and carminative, infusion of leaves used as digestive and

in rheumatism (Bedi et al., 2008).

Murraya koenigii Spreng. Syst. 2: 315, 1826; Santapau in Fl. Khandala (ed.

3): 83, 1967; FBI 1: 503; FUGP 1: 129; For. Fl. U.P.2:108; FG: 69. Bergera

koenigii Linn. Mant. 1: 565, 1767; Roxb. Pl. Cor.2 :t 112; wight, I cont. 13.

A strongly scented, pubescent deciduous shrub or small tree. Leaves

imparipinnate; leaflets ovate, lanceolate, acuminate or retuse, oblique at base,

olive green. Flowers white in terminal and axillary corymbose panicles. Calyx

and corolla 4-5 each. Stamens 10, obdiplostemonous. Fruits berry, pale

yellow, black when ripe.

Stomachic and carminative, leaves used in diarrhoea and dysentery

& for checking vomiting (Bera et al., 2008).

M. paniculata (L) Jack in Mal. Misc. 1(5): 31.1820; FG: 69.1976.-Chaleas

paniculata Linn. Mant. 1: 68. 1967.- M. exotica Linn. Mant. 2:563. 1771

(Murraea); FBI 1: 502; FUGP 1: 128.

76

Estelar

An evergreen shrub or small tree. Leaves large, rhomboid or elliptic

lanceolate, base oblique, entire or obscurely crenulate. Flowers united, white,

fragrant in terminal and axillary corymbs. Calyx and corolla 4-5 each.

Stamens 10, obdiplostemonous. Fruits berry, red or orange, when ripe.

Powdered leaves applied to cut, root & bark used in cough and

hysteria (Bhattacharjee, 2005).

Nepeta hindostana (Roth) Haines. BBO 4: 744. 1922; FG: 265.1976.-

Glechoma hindostana Roth, Nov. Sp. Pl.: 258. 1821. - Nepeta runderalis

Buch.- Ham. Ex. Benth. in Wall. Pl. As. Rar. 1: 64. 1829; FBI 4: 661; FUGP 2:

117.

An erect or ascending herb with quadrangular, grooved, pubescent

branches. Leaves ovate or sub-orbiculate, crenate, obtuse, base-cordate or

truncate. Flowers bluish purple, in long peduncled axillary cymes. Calyx

tubular, 5 toothed. Corolla bilipped. Stamens 4, didynamous. Fruits nutlet,

light brown with white spot.

The whole plant used as a cardiac tonic, decoction used as gargle in

cough (Chopra et al., 2006).

Ocimum basilicum Linn. Sp. Pl. 597. 1753; FBI 4: 608; FUGP 2: 235; For. Fl.

U.P. 3:555; FG: 266. 1976.

An erect, spreading, aromatic herb. Stem purplish/violets,

quadrangular covered with glandular hairs. Leaves ovate or ovate to

lanceolate, entire or toothed, acute, gland dotted, hairy. Flowers white, in

whorls on simple or branched racemes. Calyx 2 lipped, ovoid or companulate.

Corolla bilabiate (4/1), 4 in posterior lip and 1 in anterior lip. Stamens 4,

didynamous. Fruits nutlet, not smooth, without black margin.

Juices of leaves used for ringworm, seeds are used in habitual

77

Estelar

constipation, diarrhoea (Ghazanfar & AL-Sabahi, 1993).

O. canum Sims. Botmag. Sl:rt. 2452. 1823; FBI 4:607; FUGP 2:99; Ocimum

americanum Linn. in Juslenius. Cent. I. Pl. 15. 1755; Keng, Gard. Bull.

Straits Settlem. 24: 126. 1969.

An erect, much branched, pubescent, annual herb. Leaves elliptic-

lanceolate, entire or serrate, acute at both end, glabrous. Flowers white or

purple in close whorls of terminal racemes. Calyx 5, bilipped, companulate.

Corolla also bilabiate. Stamens 4. Fruits nutlet, ellipsoid, black.

Decoction of leaves used in epilepsy and hysteria (Asolkar et al.,

2005).

O. gratissimum Linn. Sp. Pl. 1197, Sum Fl. vit 191.

A much branched perennial woody shrub. Leaves ovate cuneate,

acute, serrate and glabrous. Flowers white to pale yellow in simple or panicled

racemes. Calyx 5, companulate, 2-lipped (1/4). Corolla bilabiate (4/1).

Stamens 4. Fruits nutlet.

Plant used as mosquito repellent, in aromatic baths of fumigations

for rheumatism and paralysis (Natrajan et al., 1999)

O. sanctum Linn. Mant. 1:85. 1767; FBI 4:609; FUGP 2:98; For. Fl.U.P.

3:553; FG: 267. 1976.

An erect much branched annual woody herb or under shrub. Stems

and branches usually purplish or violets, covered with spreading hairs. Leaves

ovate or elliptic oblong, crenate–serrate, acute or obtuse, glabrous or

pubescent. Flowers pinkish or purplish, small, in whorls of simple or panicled

racemes. Calyx 5, companulate, 2 lipped (1/4). Corolla bilabiate (4/1).

Stamens 4, didynamous. Fruits nutlet, smooth, with black margin.

Leaf is given in cough, decoction of roots given as diaphoretic in

78

Estelar

malarial fever (Tomar, 2008).

Piper longum Linn. Sp. Pl. 29.1753; FBI 5: 83; For. Fl.U. P. 3:576; Raizada

Suppl. FUGP 242. 1976.

A slender, creeping, undershrub. Leaves alternate, shortly

acuminate, ovate or ovate- oblong, glabrous. Flowers greenish, dioecious in

axillary and terminal spikes. Stamens 2-4. Fruits berry minute crowed in

cylindrical spikes, red when ripe.

Sun dried unripe fruits are used as stimulant, carminative and tonic

(Ali and Dixit, 1989).

P. sylvaticum Roxb. Hort Beng 4 (1814) romen; Fl. Ind. ed. Carey & Ball, I

158 (1820); Fl. Ind. ed. Carey; 156 (1832).

A slender, creeping undershrub. Leaves alternate, ovate or cordate,

caudate, shortly acuminate. Flowers spicate, greenish, spikes dioecious.

Stamens 2-4, arranged in axillary and terminal. Fruit berry crowded in

cylindrical spikes, red when ripe.

Fruits carminative, used in food preparations (Chopra et al., 2006).

Pogostemon heyneanus Benth. in Wall Pl As. Rar. i 31.

A large aromatic herb. Leaves lanceolate, serrate-acute or

acuminate, base obtuse. Flowers white tinged with pink in pubescent

cylindrical spikes. Calyx 5- toothed. Corolla bilipped 4-lobed, bracts

foliaceous. Stamens 4. Fruits nutlet, reddish brown.

Decoction of leaves given in cough and asthma (Mishra et al., 2008a).

P. plectranthoides Desf. in Ann. Mus. Paris 2:155.1803;FBI 4:632;FUGP 2:

105; For Fl.U.P. P. benghalensis (Burm. f.) Kuntze, Rev. Gen. Pl. 529-1891;

Raizada in Ind. For. 92 (5):321. 1966; FG: 268. 1976.

A large aromatic herb. Leaves ovate, serrate, acute or acuminate,

79

Estelar

base sub-obtuse, pubescent beneath. Flowers white tinged with pink, in

pubescent cylindrical spikes. Calyx 5. Corolla 2-lipped. Stamens 4,

didynamous. Fruits nutlet brown.

Decoction of leaves used as stimulant and styptic (Ambasta, 2006).

Polyalthia longifolia Sonner. Thw. Enum. 398. 1864; FBI 1: 62; FUGP 1:25;

FG: 33. 1976. Uraria longifolia Sonn. Voy. Indes. 2:233, t 131. 1782.

An evergreen tree with straight trunk and triangular crown. Leaves

glossy green, lanceolate, glabrous, undulate. Flowers greenish, yellow,

numerous in umbellate cymes. Calyx 3. Corolla 6, biseriate. Stamens

cuneate. Fruits ovoid, purplish black when ripe.

Fruit eaten in times of scarcity. Bark febrifuge. (Ambasta, 2006)

Polygonum glabrum Willd. Sp. Pl. 2: 447. 1799; FBI 5: 34; FUGP 2: 153;

FG: 282. 1976.

An erect or decumbent herb. Leaves lanceolate or linear, shining,

gland dotted, ochreate stipules, tubular closely sheathing the stem. Flowers

pink or white, in terminal panicles. Bracts tubular, glabrous. Perianth 4-5,

coloured. Stamens 6-9. Fruits nutlets dark brown, shining.

Infusion of leaves given in colic and as a febrifuge. Root stocks

used for piles, jaundice debility and consumption (Chopra et al., 2006).

Psidium guajava (Linn.) Sp.Pl. 470. 1753; FBI 2: 468; Benthall. Trees Calc.

244; FG: 134. 1976.- P. pyriferum Linn. Sp. Pl. 672. 1762.

A large shrub or small tree. Leaves opposite, elliptic to oblong,

acute or rounded at apex, aromatic pellucid dotted. Flowers white, 1-3 on

slender axillary peduncles. Calyx and corolla 4-5 each. Stamens many. Fruits

fleshy berry, ovoid or pyriform, seeds many.

Leaves used as an astringent for bowel trouble, decoction of bark

80

Estelar

given in diarrhoea (Chhabra and Mahunnah, 1994).

Putranjiva roxburghii Wall. Tent. Fl. Nap. 61.

A dioecious evergreen tree or shrub with pendent branches. Leaves

elliptic-oblong, lanceolate, acute, margins wavy or serrulate. Flowers yellow,

dioecious. Male flowers in dense axillary clusters. Female flowers solitary, 1-

3 in axil. Fruits drupe, yellow when ripe, pointed at both end.

Wood used for house building, leaves and stones given in decoction

for cold fever and rheumatism (Ambasta, 2006).

Saraca indica (L) Mant. 1: 98. 1767; FBI 2:271; Benthall, trees calc. 178;

FUGP 1:273; FG: 123. 1976.-Jonesia asoca Roxb. in As. Res. 4: 355. 1795

An evergreen tree. Leaves paripinnate, dark green; leaflets 4-6

pairs, oblong or oblong-lanceolate, intrapetiolar stipules. Flowers orange or

orange yellow, in dense axillary corymbs. Calyx petaloid tubular. Corolla

absent. Stamens 10. Pod linear oblong, compressed, black.

Astringents used in uterine infection (Bera et al., 2008).

Salvia plebeia R.Br. R. Br. Prodr.: 501.1810; FBI 4: 655; FUGP 2: 118; FG:

268. 1976.

An erect pubescent annual herb with obtusely grooved quadrangular

stems. Leaves ovate to lanceolate, crenate, obtuse base, acuminate, glabrous or

hairy. Flowers white to bluish-white in spicate racemes. Calyx tubular or

companulate, bilipped. Corolla 2-lipped. Stamens 2. Fruits small nutlet, ovoid,

brown.

Seeds used in diarrhoea, plant diuretic and anthelmintic (Ambasta,

2006).

Siegesbeckia orientalis Linn. Sp. Pl. 900. 1753; FBI 3: 304, 1881.

An erect, branched annual herb. Leaves simple, petiolate, opposite,

81

Estelar

ovate with cuneate base, acute, at the apex irregularly serrate-dentate,

pubescent on both surfaces. Heads yellowish, heterogamous in lax panicles.

Receptacle convex with elliptic-ovate, glandular-hairy palaeaceous bracts.

Corolla 5, tubular. Stamens 5, syngenesious. Fruits cypsela, dark brown.

Pappus absent.

Said to be possesses healing properties in gangrenous ulcers and

sores. Also diaphoretic and cardiotonic (Chopra et al., 2006).

Syzygium cumini (L.) Skeels in U.S. Dept. Agric. Bur. Pl. Ind. Bull. 248: 2.

1912; FG: 134-135. 1976; Myrtus cumini Linn. Sp. Pl. 471. 1753. - Eugenia

jambolina Lamk. Encycl. 3: 198. 1789; FBI 2: 499; FUGP: 342.- Syzygium

jambolanum (Lamk.) DC. Prodr. 3: 259. 1828.

A medium to large-size tree. Leaves oblong or elliptic oblong, acute

or acuminate, coriaceous shining. Flowers greenish white, in lateral panicles.

Calyx and corolla 4-5 each, calyx companulate, limbed at 1st 4 lobed and later

truncate. Stamens many. Fruits berry, ovoid or oblong, dark purple, one

seeded.

The powder of stem bark mixed with curd and taken for treatment

of blood dysentery (Singh & Mall, 2007).

Tagetes erecta Linn. Sp. Pl. 887. 1753. T. major Gaertn. Fruct. 2: 437, 1791.

An erect, annual, stout, aromatic herb. Leaves pinnatifid, opposite

below, alternate above, long, imparipinnate; leaflets lanceolate, acute to

acuminate, margin sharply serrate to sub-entire. Heads yellow, heterogamous,

solitary terminal. Calyx modified into pappus. Corolla 5, united. Stamens 5,

syngenesious. Fruits cypsela, angular black.

Infusions used in rheumatism, cold and bronchitis. Leaves used in

muscular pains (Tomar, 2008).

82

Estelar

Vitex negundo Linn. Sp.Pl.638. 1753; FBI 4: 583; FUGP2:90; For. Fl. U.P. 3:

541; FG: 259. 1976.

A large deciduous shrub or small tree with white tomentose,

quadrangular branches. Leaves digitately 3-5 foliolate; leaflets petiolate,

linear, lanceolate, acuminate. Flowers bluish or purplish white in opposite

lateral cymes. Calyx 5, companulate. Corolla 2 lipped. Stamens 4,

didynamous. Fruits drupe black when ripe.

Used as insect repellent, leaves in hot water applied on the body to

relieve pain and swelling, also used in dermatological problems ( Bera et al.,

2008).

Xanthium strumarium Linn. Sp. Pl. 987, 1753; FBI 3: 303, 1881.

A scabrous, erect annual herb or under shrub. Leaves alternate,

petiolated, triangular, cordate, irregularly toothed, scabrous on both surfaces.

Flowers unisexual in terminal and axillary racemes, monoecious with male

heads in the upper axils and female heads in lower axils. In male heads corolla

tubular and stamens 5, syngenesious. In female heads corolla absent. Fruits

cypsela, ovoid, thick, covered with hooked prickles. Pappus absent.

Fruits used as tonic and are given in small pox (Ali and Dixit,

1989).

Zingiber officinale Rosc. in Trans Linn. Soc. 8 (1897) 348.

A slender perennial rhizomatous herb. Leaves linear, sessile,

glabrous, arrange in two ranks of each stem. The inflorescence grows on a

separate stem from the leaf stem and forms a dense spike, flowers yellowish

green ensheathed in a few scarious, glabrous bracts. Calyx 3, united. Corolla 3

tubular. Stamen 1. Fruits loculicidal capsule.

The raw ginger is carminative and stimulant. It is useful in dropsy,

83

Estelar

Estelar

Estelar

Estelar

Estelar

Estelar

Estelar

Estelar

asthama, cough and inflammations (Ghazanfar & AL- Sabahi, 1993).

4. Extraction of essential oils from collected plants:

The volatile constituents in the form of essential oil from each collected

aromatic plant were extracted separately through hydrodistillation by Clevenger’s

apparatus (Clevenger, 1928) at 90±20C for 4h. Each essential oil was dried over

anhydrous sodium sulfate and was stored in clean sealed glass vials separately at

4ºC for further use.

5. Procurement of bacterial culture and its maintenance:

The test bacteria viz., Erwinia herbicola (MTCC 3609) and

Pseudomonas putida (MTCC 1190) taken in present study were obtained from

MTCC (Microbial Type Culture Collection), IMTECH (Institute of Microbial

Technology), Chandigarh. The purity and identification of these bacteria were

verified by culturing them on selective as well as differential media (Kado and

Heskett, 1970).

Erwinia herbicola (Lohnis) Dye includes an aerobic, gram –ve, motile,

straight rod shape organism having petritrichous flagella. The colonies of Erw.

herbicola are yellow non pigmented form. It was identified on the basis of their

distinctive morphological growth on certain selective and differential media. Its

colony appeared yellow to brown on MM2CuSO4 growth medium, whitish yellow

on Luria Bertani medium (LB) and orange-yellow without pits on Czapek’s Dox

agar (CDA) medium (Bereswill et al., 1998)

Pseudomonas putida (Kris Hamilton) includes gram-ve, aerobic, single

cell, straight or curved rods, motile having polar monotrichous flagella. Culture of

Ps. putida produced diffusible fluorescent pigments, particularly on iron deficient

media. It was identified on the basis of fluorescent pigment produced on King’s B

medium and on Nutrient agar it has circular, convex and yellow

colonies (King et al., 1954; Colyer and Mount, 1984).

The axenic culture of both test bacteria were maintained initially on

84

Estelar

Growth Medium No. 3 (Ps. putida) and LB agar ( Erw. herbicola) slants in general

and recommended MTCC growth medium separately for further experimentation

and incubated in dark places at 4ºC. Routine sub culturing of both strains was

performed in recommended MTCC growth medium. Bacterial inoculum for

experimentation was prepared from working slant of each bacterium. An inoculating

loop was touched to four or five isolates of bacterial colony growing on working

agar slant and then inoculated in fresh tube of peptone broth. A control set was also

kept which included the respective broth without inoculum. The culture broths were

incubated aerobically at 35 ± 2ºC for about 32-72h and growth was confirmed by

comparing its growth in terms of turbidity with that of control set. The

suspension was standardized and adjusted by diluting it to match the turbidity of the

0.5 McFarland Standards (Appendix I) as suggested and recommended by Lorian

(1996) and Andrews (2001a). The inoculation of plates was done within 20min of

standardization thus, ensuring the maintenance of inoculum throughout the

experiments.

For seeding the bacterial inoculum on agar plate spread plate method was

adopted. In this method 0.1ml inoculum (Standardized using 0.5 McFarland

Standard) was spread on the solidified and dried surface of medium with the help of

sterile L-shape glass spreader. The plates were allowed to dry for 10-15min before

applying the disc. From time to time both bacterial strains were evaluated for their

purity by their specific characteristic

growth on various differential and selective media.

6. Isolation of deteriorating fungi and insects from stored samples of pigeon pea

seeds:

Samples of stored seeds of pigeon pea were collected in pre-sterilized

polyethylene bags from different places of four districts viz., Pakri, Shahpur,

Tarkulawa (Deoria), Chaurichaura, Gayatrinagar, Mohanpur (Gorakhpur), Barwa,

Fazilnagar, Mahasoon (Kushinagar) and Parsauna, Mithaura, Shikarpur

(Mahrajganj). 85

Estelar

The samples were brought to the laboratory and kept separately after labeling the

name of places along with districts for analysis of associated fungi and insects.

6.1 Mycoflora analysis:

The mycoflora of sterilized and unsterilized stored seeds of pigeon pea

was analyzed by Standard blotter paper (Tempe, 1953) and Agar plate (Muskett,

1948) methods.

7 unsterilized seeds of Cajanus cajan of each sample were kept

equidistantly in each pre-sterilized Petri plate containing solidified agar medium and

moistened blotter paper separately. The prepared assay plates were incubated at 28 ±

2ºC in triplicate for 7 days. The appearance of fungal colonies was observed after 7th

day.

For analysis of fungi from sterilized seeds, 7 seeds of each sample were

surface sterilized with 0.1% mercuric chloride solution upto 5min. The seeds were

then washed thoroughly with sterilized distilled water and were placed on solidified

agar medium as well as moistened blotter paper separately. The Petri plates were

incubated and observed similarly.

The fungal colonies appeared on seeds were isolated, purified and

identified by comparing their morphological and cultural characteristics with

the help fungal keys (Raper and Thom, 1949; Raper and Fennel, 1965; Booth, 1971

and Ellis, 1976). The single spore culture of each purified fungal species was

maintained on slants for 6-8 months at 4ºC. The per cent incidence of individual

fungal species isolated by agar as well as blotter paper methods from sterilized and

unsterilized pigeon pea seeds was calculated following the formula of Gowda and

Sullia (1987).

Number of seeds on which the fungus was

encountered in each sample

Per cent Incidence (PI) = x 100

Total number of seeds tested in each sample

Results are depicted in Fig 5a and 5b.

86

Estelar

Fig. 5a: Per cent incidence of fungi of stored pigeon pea seeds on agar plate

Fig. 5b: Per cent incidence of fungi of stored pigeon pea seeds on blotter

paper

87

11

100 100

80

6

83

14 23

71 66

34

54 46

63 57

0

20

40

60

80

100

120

Per

cen

t in

ciden

ce

Isolated fungal species

6

91 100

54

11 9

57

3

31

57 51

29

40

17

43 49

0

20

40

60

80

100

120

Per

cen

t in

ciden

ce

Isolated fungal species

Estelar

As evident from Fig. 5a & 5b that total seventeen fungal species

belonging to eight genera were isolated from pigeon pea seeds (sterilized &

unsterilized) by agar plate and blotter paper methods. The fungal species

identified as Aspergillus flavus Link, A. niger van Tieghem, A. ochraceus

Wilhelm, A. parasiticus Speare, A. terreus Thom, Cladosporium sp.,

Curvularia lunata (Wakker) Boedijin, Fusarium nivale (Fries) Cesati, F.

oxysporum von Schlechtendal, Penicillium sp., P. chrysogenum Thom, P.

italicum Wehmer, Mucor sp. and Rhizopus arrhizus Fischer were isolated by

both agar plate as well as blotter paper methods. Aspergillus flavipes Thom &

Charch and A. restrictus G.Smith were obtained by blotter paper method only

while Alternaria alternata (Fr.)Keissler was only isolated by agar plate

method. Out of 17 fungal species isolated, Aspergillus flavus, A. niger, A.

ochraceus and A. terreus showed higher per cent incidence on pigeon pea

seeds.

6.2 Study of collected pigeon pea samples for associated insects:

The collected seeds samples were observed separately with hand

lens for associated insects. Samples were heavily infested (Fig. 6) with

numerous small chocolate coloured beetles of about 0.30cm long with serrated

antennae, i.e. Callosobruchus chinensis L. and 1-8inch long, reddish brown

slightly elongated beetles with two black spots near middle, i.e. C. maculatus

F. (identified with the help of literatures, Drees and Jackman, 1999; Beck and

Blumer, 2007). The insects were authenticated at Entomology Lab,

Department of Zoology, DDU Gorakhpur University, Gorakhpur.

88

Estelar

Estelar

Fig. 6: Per cent infestation of pigeon pea seeds due to pulse bruchids

Gorakhpur and Mahrajganj samples exhibited higher per cent

infestation (Fig. 6). Insects were reared separately on insecticide free fresh

pigeon pea seeds at 28 ± 2ºC for further experimentation.

6.3 Selection of test organisms:

During mycoflora analysis Aspergillus flavus, A. niger, A.

ochraceus and A. terreus were found to be dominant. Largely the samples

showed presence of Callosobruchus chinensis and C. maculatus hence, these

fungi and insects were selected as test organisms.

7. Antibacterial activity of essential oils against test bacteria:

The antibacterial activity of essential oils against Erwinia herbicola

and Pseudomonas putida were evaluated following Andrews (2001a). In this

method 10ml agar medium were poured in 80mm glass Petri plates. The plates

were allowed to solidified and then seeded aseptically by spreading 0.1ml of

inoculum prepared in water (equivalent to 107-10

8 cfu/ml) and allowed to set

for 10-15min. 5µl of individual essential oil was soaked in 6mm diam filter

89

12

45

21

48

15

50

17

47

0

10

20

30

40

50

60

Deoria Gorakhpur Kushinagar Maharajganj

Per

cen

t in

fest

atio

n

Districts visited

Callosobruchus

chinensis

C. maculatus

Estelar

paper disc and such two presoaked filter paper discs were placed in each Petri

plate 30mm apart using sterile forceps and the gently pressed down onto agar

surface. Wilbrink agar (WA) medium was used for Erw. herbicola (Koike,

1965) while Nutrient agar (NA) for Ps. putida. Two additional control sets

+ve and –ve were also run parallel so as to check growth of bacteria and

possible contamination. The positive control plates were inoculated with test

bacteria while negative control plates were without bacteria. Each experiment

was maintained in triplicate and assays were performed twice. Result in terms

of zone of inhibition in mm (an apparent inhibition zone around 6mm diam

disc) was recorded after 32-72h of incubation at 35 ± 2ºC. The antibacterial

activities of essential oils are reported in Table 7.

Table 7: Antibacterial potency of essential oils against test bacteria

Plant species (essential oils)* Part used Zone of inhibition

$

Erwinia herbicola Pseudomonas putida

1 2 3 4

Acorus calamus Rhizome 13.97±2.12 15.34±1.23

Adhatoda vasica Leaf 10.28±0.98 1.92±0.43

Aegle marmelos ,, 14.33±3.78 10.92±1.94

Ageratum conyzoides Whole Part 5.58±1.12 4.50±2.5

A. houstonianum ,, 6.86±0.94 5.04±1.00

Anethum graveolens Shoot 8.42±1.52 10.42±0.53

Anisomeles indica Leaf 24.25±2.17 23.25±0.14

Annona squamosa ,, No inhibition 6.25±1.32

Artemisia nilagirica Shoot 13.58±0.94 14.88±3.0

Ashphodelus tenuifolius Whole part 2.98±0.67 5.67±1.02

Azadirachta indica Leaf 6.83±0.52 5.08±1.66

Blumea eriantha ,, 14.83±1.15 8.92±0.87

B. lacera ,, 15.67±2.23 16.42±4.90

B. laciniata Whole Part 7.98±1.20 5.67±0.94

B. membranacea ,, 8.58±0.94 No inhibition

B. mollis ,, No inhibition 7.58±0.62

Caesulia axillaris Shoot 10.17±2.70 10.58±0.87

Callicarpa macrophylla Leaf 10.36±4.29 16.13±5.68

Callistemon lanceolatus ,, 7.17±0.62 18.9±2.69

Cannabis sativa Shoot 12.0±1.23 10.5±3.76

Chenopodium ambrosioides ,, # #

Chrysanthemum indicum Leaf No inhibition 8.58±0.87

90 Contd……

Estelar

1 2 3 4

Citrus aurantifolia Leaf # 12.08±0.88

C. aurantium ,, # #

C. limon ,, 7.33±2.12 10.88±3.55

Clausena pentaphylla ,, # #

Cleome gynandra ,, No inhibition 7.13±1.23

Clerodendrum inermae ,, 16.0±2.46 17.13±194

C. viscosum ,, 17.58±0.72 14.92±1.37

Colebrookea oppositaefolia ,, 13.0±0.5 9.83±1.62

Cosmos sulphureus ,, 9.30±2.21 4.35±0.21

Cotula anthemoides ,, 3.89±0.56 9.0±2.3

Curcuma aromatica ,, 7.17±0.87 9.83±1.18

C. zedoaria Rhizome 25.83±0.28 25.17±1.87

Cyperus brevifolius Whole Part No inhibition 12.50±2.89

C. monocephalus ,, 6.33±2.70 6.58±1.90

C. rotundus ,, 3.09±0.94 6.08±1.29

C. triceps ,, 3.42±0.94 8.63±0.53

Erigeron bonariensis Shoot 1.42±0.23 0.92±2.47

E. canadensis ,, 1.50±0.70 8.17±0.88

Eucalyptus citriodora Leaf 7.33±0.88 5.17±0.59

Eugenia heyneana ,, 15.92±1.08 11.75±0.35

Eupatorium adenophorum ,, # #

E. cannabinum Whole part 2.83±0.14 5.67±1.33

E. odoratum Shoot 1.75±0.88 9.75±1.76

Glycosmis pentaphylla Leaf 7.25±2.0 3.42±1.25

Grangea maderaspatana Shoot 2.08±1.76 8.25±0.66

Gynura crepidioides ,, 2.75±0.88 9.0±0.00

Hygrophila difformis ,, 14.25±1.28 10.75±3.69

H. pinnatifida ,, 3.17±2.67 No inhibition

Hyptis suaveolens Leaf # #

Lantana camara ,, 9.08±1.28 7.91±3.12

L. indica ,, 11.76±0.34 9.03±2.3

Lawsonia inermis ,, 3.09±1.02 2.12±0.34

Leonotis nepetaefolia ,, 7.83±0.62 No inhibition

Leonurus sibiricus ,, 2.33±1.80 No inhibition

Leucas aspera Shoot 5.75±0.13 3.83±0.62

L. cephalotes ,, 11.33±3.13 13.17±2.23

Lippia alba Leaf # #

Melia azedarach ,, 4.92±2.15 No inhibition

Mentha arvensis ,, # #

Murraya koenigii ,, 7.5±0.75 0.17±0.53

M. paniculata ,, 2.92±0.03 4.75±0.86

Nepeta hindostana Shoot 14.58±3.95 4.58±0.53

Ocimum basilicum ,, 9.92±2.83 24.63±3.35

O. canum ,, 6.67±0.80 7.25±3.17

91 Contd…

Estelar

1 2 3 4

Ocimum gratissimum Shoot 2.92±0.28 6.63±0.38

O. sanctum ,, # #

Piper longum Leaf 10.17±1.61 13.08±1.18

P. sylvaticum ,, 7.08±0.62 11.75±1.52

Pogostemon heyneanus ,, 3.42±1.59 3.17±1.01

P. plectranthoides ,, 3.75±0.61 7.33±1.42

Polyalthia longifolia ,, 7.58±0.17 No inhibition

Polygonum glabrum Shoot 4.23±1.02 1.89±0.45

Psidium guajava Leaf 11.67±1.44 7.08±0.80

Putranjiva roxburghii ,, 6.0±1.8 8.33±0.91

Salvia plebeia Shoot 2.58±0.28 5.25±2.03

Saraca indica Leaf 5.67±1.0 7.58±1.12

Siegesbeckia orientalis Shoot No inhibition No inhibition

Syzygium cumini Leaf 3.33±1.66 6.17±0.25

Tagetes erecta ,, 7.75±0.17 16.42±2.78

Vitex negundo Shoot # #

Xanthium strumarium Leaf 8.42±0.62 No inhibition

Zingiber officinale Rhizome 5.08±2.98 14.25±3.04

* Oil concentration 5µl, # oil exhibited full plate inhibition of bacteria,

$Values given are mean of three replicates ± Standard Deviation

As evident from Table 7 that 9 oils such as Chenopodium

ambrosioides, Clausena pentaphylla, Citrus aurantium, Eupatorium

adenophorum, Hyptis suaveolens, Lippia alba, Mentha arvensis, Ocimum

sanctum and Vitex negundo exhibited full plate inhibition of both test bacteria.

No inhibition zone was incurred by Annona squamosa, Blumea mollis,

Chrysanthemum indicum, Cleome gynandra, Cyperus brevifolius and

Siegesbeckia orientalis essential oils against Erw. herbicola and Blumea

membranacea, Hygrophila pinnatifida, Leonotis nepetaefolia, Leonurus

sibiricus, Melia azedarach, Polyalthia longifolia, Siegesbeckia orientalis,

Xanthium strumarium oils were ineffective against Ps. putida. Additionally,

essential oils of Acorus calamus, Aegle marmelos, Anisomeles indica,

Artemisia nilagirica, B. eriantha, B. lacera, Clerodendrum inermae, C.

viscosum, Curcuma zedoaria, Eugenia heyneana and Hygrophila difformis

92

Estelar

were moderately effective with zone of inhibition ranging from 11.33-

25.83mm against Erw. herbicola and 10.75-25.17mm against Ps. putida while

rest of the oils were least effective against test bacteria.

8. Evaluation of essential oils against test fungi:

The toxicity of essential oils against Aspergillus flavus, A. niger, A.

ochraceus and A. terreus was tested separately by Inverted Petri plate

technique of Bocher (1938) with slight modification (Tripathi and Kumar,

2007). Czapek’s Dox Agar plates were prepared using 80mm diam glass Petri

plates (average volume of 60 ± 2ml). Each plate contained 10ml medium.

Mycelial disc (6mm diam) of test organism was cut from a one week old

culture and placed upside down on the centre of the agar plate. The inoculated

plates were inverted. Requisite amount of each essential oil (500ppm) was

impregnated onto filter paper disc (pre-sterilized disc of 10mm diam of

Whatman paper No. 44) separately. The impregnated filter paper discs were

aseptically transferred to lower lid of the inoculated inverted Petri plate. In

control set sterilized distilled water was used in place of oil. Both treated and

control plates were incubated at 28 ± 2ºC for 7 days in an Incubator. Each

experiment was revised and maintained in triplicate. The diameter of hyphae

(mm) from centre to inside of plates was measured after 7 days. The

fungitoxicity was recorded in terms of per cent mycelial inhibition and

calculated following the formula of Tripathi and Kumar (2007).

DC - DT

Per cent inhibition of mycelial growth = x 100

DC

Where DC = Average increase in dimension of fungal colony in control set

DT =Average increase in dimension of fungal colony in treatment set

The results are reported in Table 8.

93

Estelar

Table 8: Evaluation of essential oils for their toxicity against test fungi

94 Contd…

Plant species (essential oils)*

Per cent mycelial inhibition$

Aspergillus

flavus A. niger A. ochraceus A. terreus

1 2 3 4 5

Acorus calamus 30.56±2.10 32.24±1.23 17.34±1.12 45.0±3.0

Adhatoda vasica 96.86±1 94.52±8.6 98.39±2.02 90.78±4.53

Aegle marmelos 16.6±5.68 45.0±3.0 20.78±2.47 17.35±4.26

Ageratum conyzoides 53.3±3.90 75.6±0.76 74.11±0.76 65.28±3.5

A. houstonianum 61.6±2.45 72.6±1.10 4.66±2.08 37.20±1.80

Anethum graveolens 42.6±3.21 21.4±1.15 22.2±2.01 64.31±2.14

Anisomeles indica 18.15±0.42 17.32±0.91 65.64±1.44 57.69±1.44

Annona squamosa 67.14±1.53 19.33±3.5 31.22±1.27 22.13±2.5

Artemisia nilagirica 67.28±3.75 58.51±3.18 77.92±2.29 71.37±5.53

Ashphodelus tenuifolius 45.31±1 40.0±1.15 53.3±3.90 61.04±1.2

Azadirachta indica 28.34±5.0 67.57±6.21 41.21±1.03 19.84±0.05

Blumea eriantha 13.21±1.52 18.57±2.36 57.17±4.93 33.94±4.44

B. lacera 17.23±1.0 61.21±2.0 30.56±2.10 13.19±1.10

B. laciniata 17.43±0.68 22.04±2.10 32.45±5.04 23.21±0.92

B. membranacea 87.85±6.23 17.34±1.12 64.85±1.44 72.48±2.88

B. mollis 34.21±0.50 13.39±0.70 16.66±4.76 9.92±0.57

Caesulia axillaris 66.1±1.21 50±3.39 58.82±2.21 49.32±1.60

Callicarpa macrophylla 4.66±0.09 61.3±3.08 13.21±7.45 18.34±2.21

Callistemon lanceolatus 40±2.21 32.4±1.23 19.95±1.15 19.72±5.01

Cannabis sativa 45.0±0.08 30.0±2.0 40.0±2.0 45.0±1.23

Chenopodium ambrosioides 100±0.00 100±0.00 100±0.00 100±0.00

Chrysanthemum indicum 38.78±2.51 22.61±5.35 55.44±1.15 31.38±3.2

Citrus aurantifolia 100±0.00 100±0.00 100±0.00 100±0.00

C. aurantium 100±0.00 100±0.00 100±0.00 100±0.00

C. limon 100±0.00 100±0.00 100±0.00 100±0.00

Clausena pentaphylla 100±0.00 100±0.00 100±0.00 100±0.00

Cleome gynandra 66.61±2.9 69.04±3.0 60.68±2.80 62.23±2.0

Clerodendrum inermae 23.21±0.15 34.0±1.94 39.04±1.15 10.56±1.93

C. viscosum 44.59±1.16 13.89±3.95 14.49±2.16 48.13±1.32

Colebrookea oppositaefolia 56.57±4.35 34.32±2.30 54.66±1.25 20.63±2.29

Cosmos sulphureus 20.78±2.47 53.3±3.90 32.21±0.65 0.00±0.09

Cotula anthemoides 30.56±4.31 41.23±1.15 53.3±3.90 40.31±4.5

Curcuma aromatica 74.32±2.46 48.13±2.59 74.11±0.57 55.36±0.76

C. zedoaria 15.55±1.75 14.32±0.7 69.29±3.61 59.63±3.27

Cyperus brevifolius 54.56±1.21 45.53±3.09 32.08±1.15 23.31±2.47

C. monocephalus 52.0±1.73 68.0±1.60 54.0±0.28 30.0±3.50

C. rotundus 34.0±0.03 64.76±3.38 9.57±0.32 24.32±2.0

C. triceps 65.46±2.12 12.89±2.0 21.09±0.00 34.64±2.14

Erigeron bonariensis 7.59±1.73 3.21±2.17 68.02±4.36 37.1±0.86

E. canadensis 66.66±1.23 4.87±3.0 40.54±3.20 9.54±1.53

Estelar

* Oil concentration 500ppm, $Values given are mean of three replicates ± Standard Deviation

95

1 2 3 4 5

Eucalyptus citriodora 30.60±4.5 23.8±1.21 81.41±4.64 78.60±2.36

Eugenia heyneana 31.84±4.27 30.01±3.4 58.22±0.76 40.38±2.02

Eupatorium adenophorum 11.23±3.0 19.21±1.60 12.32±1.0 44.95±0.5

E. cannabinum 11.21±3.27 21.84±1.80 5.55±2.29 24.48±3.78

E. odoratum 55.5±2.13 58.50±3.98 7.46±0.8 37.69±4.07

Glycosmis pentaphylla 68.14±3.12 39.48±1.75 69.56±3.28 61.12±1.0

Grangea maderaspatana 41.03±2.0 30.67±1.0 9.68±0.5 20.81±2.0

Gynura crepidioides 40.65±1.0 67.43±4.65 56.43±2.21 34.21±1.20

Hygrophila difformis 84.34±2.24 89.42±3.0 91.72±2.15 92.68±3.0

H. pinnatifida 32.76±2.35 23.7±0.00 17.78±1.21 21.05±1.1

Hyptis suaveolens 48.5±3.23 38.4±5 24±1.04 44.34±0.58

Lantana camara 42.8±1.21 64.1±0.5 53.96±5.22 51.79±7.09

L. indica 58.4±2.30 57.1±0.5 61.21±0.22 8.34±2.0

Lawsonia inermis 40.69±1.5 19.2±3.21 20.31±1.5 17.34±1.15

Leonotis nepetaefolia 18.31±2.02 8.54±0.28 31.32±1.29 55.24±4.0

Leonurus sibiricus 13.85±2.78 9.56±2.59 38.23±1.46 13.79±0.5

Leucas aspera 38.89±1.0 32.14±0.5 55.45±6.36 64.40±1.44

L. cephalotes 4.41±3.0 38.12±4.0 4.31±0.51 2.31±1.0

Lippia alba 57.8±2.52 40.5±3.0 53.2±0.76 54.60±3.25

Melia azedarach 18.75±1.67 38.31±1.26 56.50±0.87 76.92±1.60

Mentha arvensis 100±0.00 100±0.00 100±0.00 100±0.00

Murraya koenigii 0.00±0.09 9.4±2.0 62.42±1.26 35.74±1.0

M. paniculata 39.4±2.23 42.4±1.11 39.39±1.80 29.87±1.04

Nepeta hindostana 11.23±4.25 21.84±1.89 15.77±2.08 54.56±3.78

Ocimum basilicum 90±2.0 93.13±1.2 98.13±2.30 91.32±1.23

O. canum 59.1±3.0 64.5±5.0 21.6±0.29 85.68±3.21

O. gratissimum 18.14±2.21 68.29±3.0 23.44±1.0 15.75±3.5

O. sanctum 100±0.00 100±0.00 100±0.00 100±0.00

Piper longum 3.14±1.12 17.17±2.41 66.67±2.0 49.36±1.44

P. sylvaticum 38.14±4.04 13.53±3.21 20.83±1.44 24.35±0.29

Pogostemon heyneanus 13.87±4.33 38.21±1.5 54.51±4.19 55.56±2.88

P. plectranthoides 33.33±3.25 31.23±1.75 57.14±2.30 61.54±5.19

Polyalthia longifolia 79.86±3.25 54.38±2.30 76.86±2.0 46.54±3.05

Polygonum glabrum 59.53±3.43 78.65±4.0 54.45±2.0 40.0±1.05

Psidium guajava 60.86±2 52.42±2.36 59.09±1.89 68.06±0.76

Putranjiva roxburghii 48.48±1.75 38.75±2.18 15.71±2.64 48.23±0.58

Salvia plebeia 54.54±2.21 55.45±1.73 66.66±5.0 33.38±1.44

Saraca indica 67.93±3.32 57.89±1.26 54.94±2.29 71.26±5.01

Siegesbeckia orientalis 42.31±0.34 40.54±1.98 23.43±2.23 34.21±0.93

Syzygium cumini 10±1.12 9.5±0.09 61.39±2.51 58.56±2.56 Tagetes erecta 21.49±1.15 20.78±2.47 22.2±2.21 13.23±0.15

Vitex negundo 8.54±1.32 11.23±1.26 11.18±2.29 13.22±0.29

Xanthium strumarium 74.74±1.60 57.12±1.44 49.50±2.36 54.84±4.82

Zingiber officinale 45.43±2.36 59.61±2.18 79.76±2.64 63.92±4.04

Estelar

As evident from Table 8, the essential oils of Chenopodium

ambrosioides, Citrus aurantifolia, C. aurantium, C. limon, Clausena

pentaphylla, Mentha arvensis and Ocimum sanctum exhibited absolute

toxicity inhibiting the mycelial growth of all the test fungi completely. The

oils from Adhatoda vasica, Hygrophila difformis and Ocimum basilicum

showed moderate level of fungitoxicity, while other essential oils exhibited

lower level of toxicity.

9. Repellent activity of essential oils against test insects-

The repellent activity of essential oils against Callosobruchus

chinensis and C. maculatus was assessed using Olfactometer following

Tripathi and Kumar (2007). 5µl of oil from each plant was soaked in cotton

swabs and introduced in one arm of Y tube of Olfactometer separately. Water

soaked cotton swab was placed in other arm as control set. 20 individuals of

each test insects were obtained from reared culture, and introduced separately

into basal arm of Y tube Olfactometer in 4 batches at interval of 10min each

to avoid mutual interference. Each experiment was revised and carried out in

triplicate. The repellent activity was recorded in terms of per cent repellency

by following formula.

NC - NT

Per cent repellency = x 100

NC

Where NC= number of individual in control arm of Olfactometer

NT= number of individual in treatment arm of Olfactometer

The results are reported in Table 9.

96

Estelar

Table 9: Repellent activity of essential oils against test insects

Plant species (essential oils)*

Per cent repellency$

Callosobruchus chinensis C. maculatus

1 2 3

Acorus calamus 57±1.27 10.7±1.0

Adhatoda vasica 100±0.00 100±0.00

Aegle marmelos 52.17±1.52 45.00±1.52

Ageratum conyzoides 65.38±1.0 66.67±1.52

A. houstonianum 50.00±1.52 24.33±3.05

Anethum graveolens 54.55±1.15 47.83±2.0

Anisomeles indica 83.33±0.58 78.10±0.58

Annona squamosa 40.00±2.0 34.78±2.64

Artemisia nilagirica 43.48±1.53 64.00±1.0

Ashphodelus tenuifolius 34.0±2.0 9.0±0.8

Azadirachta indica 60.86±1.0 71.42±1.0

Blumea eriantha 57.14±1.0 36.36±1.52

B. lacera 26.08±2.08 26.00±1.0

B. laciniata 26.66±1.53 45.0±1.52

B. membranacea 10.7±1.0 64.00±1.0

B. mollis 71.42±1.0 59.09±1.0

Caesulia axillaris 50.00±1.0 54.00±1.05

Callicarpa macrophylla 58.33±1.52 47.83±1.0

Callistemon lanceolatus 69.56±1.15 59.38±2.0

Cannabis sativa 40.00±2.0 26.04±1.15

Chenopodium ambrosioides 100±0.00 100±0.00

Chrysanthemum indicum 50.00±2.0 64.00±1.0

Citrus aurantifolia 61.90±1.52 64.74±1.73

C. aurantium 26.66±1.53 60.87±1.0

C. limon 78.94±0.577 65.38±1.0

Clausena pentaphylla 100±0.00 100±0.00

Cleome gynandra 73.91±1.0 43.33±1.52

Clerodendrum viscosum 65.22±0.57 57.17±1.52

Clerodendrum inermae 4.0±0.20 20.0±2.0

Colebrookea oppositaefolia 59.09±1.0 54.54±0.58

Cosmos sulphureus 52.17±0.58 35.32±0.5

Cotula anthemoides 44.44±1.52 40.0±4.5

Curcuma aromatica 45.45±1.73 55.00±1.0

C. zedoaria 57.89±1.15 57.82±1.0

Cyperus brevifolius 47.37±0.58 59.09±1.0

C. monocephalus 61.53±0.57 61.90±1.15

C. rotundus 30.04±2.02 32.9±2.32

C. triceps 21.29±1.52 20.5±2.0

Erigeron bonariensis 30.00±1.73 63.64±1.15

E. canadensis 42.85±1.73 62.50±1.0

Eucalyptus citriodora 47.62±1.52 68.18±1.52

97 Contd…

Estelar

1 2 3

Eugenia heyneana 56.25±1.52 63.64±0.58

Eupatorium adenophorum 34.78±1.53 65.00±0.58

E. cannabinum 52.17±0.58 31.81±3.60

E. odoratum 44.44±1.52 56.55±3.05

Glycosmis pentaphylla 45.00±2.08 62.50±1.0

Grangea maderaspatana 48.96±2.19 23.04±1.2

Gynura crepidioides 34.59±1.0 5.8±1.0

Hygrophila difformis 79.16±1.15 75.0±1.0

H. pinnatifida 13.83±0.2 21.7±2.3

Hyptis suaveolens 60.87±1.0 42.10±1.52

Lantana camara 56.52±1.15 60.00±1.52

L. indica 63.63±1.52 36.36±0.58

Lawsonia inermis 7.0±0.5 30.0±2.0

Leonotis nepetaefolia 71.43±1.15 57.14±0.58

Leonurus sibiricus 52.00±2.0 53.84±2.0

Leucas aspera 50.00±1.52 57.14±2.0

L. cephalotes 75.00±1.0 45.83±1.52

Lippia alba 60.87±1.0 55.00±1.0

Melia azedarach 31.03±1.0 59.09±1.0

Mentha arvensis 100±0.00 100±0.00

Murraya koenigii 36.04±1.0 3.0±0.5

M. paniculata 31.57±0.57 30.00±1.52

Nepeta hindostana 70.00±1.0 35.00±2.08

Ocimum basilicum 83.33±0.57 80.95±0.58

O. canum 83.63±2.51 78.27±1.52

O. gratissimum 59.09±1.0 52.17±1.0

O. sanctum 100±0.00 100±0.00

Piper longum 47.82±1.0 52.17±1.15

P. sylvaticum 33.33±1.0 65.22±0.58

Pogostemon heyneanus 47.36±1.15 52.17±2.08

P. plectranthoides 38.88±1.15 59.09±1.0

Polyalthia longifolia 75.00±1.0 29.16±2.88

Polygonum glabrum 13.4±1.52 40.0±2.0

Psidium guajava 45.45±1.0 50.00±1.52

Putranjiva roxburghii 56.52±0.58 40.00±1.0

Salvia plebeia 86.67±0.58 89.09±1.0

Saraca indica 58.33±2.0 57.89±1.15

Siegesbeckia orientalis 9.5±0.4 45.73±2.51

Syzygium cumini 37.50±3.60 53.33±2.51

Tagetes erecta 31.57±0.57 20.0±2.0

Vitex negundo 100±0.00 100±0.00

Xanthium strumarium 80.00±1.52 31.81±1.0

Zingiber officinale 70.83±1.52 52.38±0.58

* Oil concentration 5µl, $

Values given are mean of three replicates ± Standard Deviation

98

Estelar

The oils of Adhatoda vasica, Chenopodium, ambrosioides,

Clausena pentaphylla, Mentha arvensis, Ocimum sanctum and Vitex negundo

showed 100% repellency against both test insects (Table 9). Anisomeles

indica, Hygrophila difformis, Ocimum basilicum, O. canum and Salvia plebeia

oils showed more than 75% insect repellency. Rest of the essential oils

exhibited lower level of repellent activity.

As such, 9 essential oils exhibited potentiality against bacteria, 7

oils against fungi and 6 oils against insects. Out of them, only Clausena

pentaphylla was selected for detailed in vitro and in vivo investigation due to

high yield, perennial, wild nature and unexplored capabilities as botanical

toxicants.

10. Antibacterial properties of Clausena pentaphylla oil:

10.1 Determination of minimum inhibitory concentration (MIC):

Minimum inhibitory concentration of Clausena oil was determined

following Agar dilution method of Andrews (2001b). A range of double

dilution of oil (0.5-16.0µl/ml) was prepared in different Petri plates. Similarly

plates containing recommended sets range of antibiotics (Agromycin &

Streptomycin) were also run parallel. Now 10ml of sterilized medium was

poured aseptically in each Petri plates containing oil and antibiotics

separately. Tween 80 was added to the final concentration of 0.5% v/v prior to

pouring in order to enhance the solubility of oil in the medium (Christoph et

al., 2001). The pour plates were thoroughly agitated in order to mix the

oil/antibiotics with the medium on a level surface. These plates were allowed

to solidify and were dried so that no drops of moisture remain on agar surface.

Now three circles (2cm diam) were marked on reverse side of each plate and

circle was inoculated by 5µl (approximately 104 cfu/spot) of overnight

99

Estelar

bacterial culture (adjusted to 0.5 McFarland Standard) using an inoculating

loop (double wound, 2mm diam, Himedia). Two control sets viz., Nutrient

Agar/Wilbrink Agar (antimicrobial drug free, with Tween 80); inoculated with

test bacteria as +ve control and un-inoculated same as -ve control were

prepared simultaneously. The inoculum spots were allowed to be absorbed

and dried at room temperature before inverting the plates for 32-72h of

incubation at 35 ± 2ºC. Each experiment was revised and carried out in

triplicate.

The concentration of essential oil/antibiotics that completely inhibit

the visible growth of test bacteria was taken as MIC value. Results are

reported in Table 10.

Table 10: Minimum inhibitory concentration (MIC) of Clausena oil and

antibiotics against test bacteria

Concentrations of

oil and antibiotics

mixed in medium*

Bacterial growth on agar plate

Erw. herbicola Ps. putida

Oil Agromycin Streptomycin Oil Agromycin Streptomycin

0.06 + + + + + +

0.12 + + + + + +

0.25 + + + - + +

0.50 - + + - + +

1.0 - + - - + -

2.0 - - - - - -

4.0 - - - - - -

8.0 - - - - - -

16.0 - - - - - -

*Values in µl/ml for oil & µg/ml for antibiotics; + Growth present; - Growth absent

MIC value of Clausena oil was 0.5µl/ml for Erw. herbicola and

0.25µl/ml for Ps. putida (Table 10), and was lower than Agromycin and

Streptomycin antibiotics used in present study.

10.2 Determination of minimum bactericidal concentration (MBC):

The MBC of oil and antibiotics was determined following Mishra et

100

Estelar

al. (2008b). Fresh NA/WA medium (without oil and antibiotics) was prepared

and aseptically poured into 80mm Petri plates separately. The plates were

allowed to be solidified and dried. The Petri dishes of earlier MIC experiments

showing negative growth were used for inoculum. The inoculum of test

bacteria were picked from various poisoned plates of MIC experiments

showing no growth and streaked on the freshly prepared and solidified

NA/WA plates. The inoculum of test bacteria was picked from the point of

original inoculation (identified by pre marked circle on the back of the plates

of earlier MIC experiments) using sterile inoculating loop each time. Control

sets were maintained by streaking the test bacteria on fresh NA/WA plates.

The streaks of inoculum free sterilized loop after and before experiments were

kept as negative control and used for monitoring the contaminations. The

experiments were performed in triplicate. All the plates were incubated at 35 ±

2ºC for 32-72h. The lowest concentration of antimicrobial agents from which

the bacteria do not recover on the fresh medium was taken as MBC. Results

are reported in Table 11.

Table 11: Minimum bactericidal concentration (MBC) of Clausena oil and

antibiotics against test bacteria

Concentrations of

oil and antibiotics

mixed in medium*

Bacterial growth on agar plate

Erw. herbicola Ps. putida

Oil Agromycin Streptomycin Oil Agromycin Streptomycin

0.06 NA NA NA NA NA NA

0.12 NA NA NA NA NA NA

0.25 NA NA NA + NA NA

0.50 + NA NA + NA NA

1.0 + NA + + NA +

2.0 + + + - + +

4.0 - + + - + +

8.0 - + - - + -

16.0 - - - - + -

*Values in µl/ml for oil & µg/ml for antibiotics; + Growth present; - Growth absent, NA-

not applicable

101

Estelar

Clausena oil was cidal in nature with MBC value of 4µl/ml for Erw.

herbicola and 2µl/ml for Ps. putida (Table 11).

10.3 Effect of physical factors on bacterial toxicity of Clausena oil:

The effect of different physical factors viz., storage, temperature,

autoclaving on bacterial toxicity of oil was studied following Tripathi and

Kumar (2007) while effect of pH was carried out by method of Singh (2009).

10.3.1 Effect of storage-

Toxicity of one year stored oil sample was evaluated against test

bacteria separately by agar dilution method at regular interval of three months

at its MIC. The appearance of bacterial growth was recorded at interval of 32-

72h of incubation at 35 ± 2ºC. Results are reported in Table 12.

10.3.2 Effect of temperature-

Three lots of Clausena oil, (each of 1ml) were taken in airtight glass

vials of 5ml and kept at different temperatures viz., 40, 60, 80, 100 and 120 ºC

in an electric oven for 10, 15, 20, 25 and 30min respectively. The vials were

allowed to cool at room temperature. The oil of each tube was tested for its

antibacterial activity at MIC by agar dilution method. The appearance of

bacterial growth was recorded at interval of 32-72h of incubation at 35 ± 2ºC.

Results are reported in Table 12.

10.3.3 Effect of autoclaving-

1ml Clausena oil was kept in a screw capped glass vial (5ml) and

was subjected to autoclaving (15lb/inch2 pressure at 121ºC) for 20min. The

tube was then allowed to cool at room temperature for 1h. Autoclaved oil was

tested for its antibacterial activity at MIC by usual agar dilution method. The

appearance of bacterial growth was recorded at interval of 32-72h at

incubation of 35 ± 2ºC. Results are reported in Table 12.

102

Estelar

10.3.4 Effect of pH-

pH can change the antibacterial properties of essential oil due to

differential proton concentration in the medium. Requisite amount of oil was

mixed in polyethylene glycol (PEG) and pH was adjusted at 5, 6, 7 and 8 by

addition of either 1N HCl (acidic pH) or 1N NaOH (alkaline pH) solutions.

HCl/NaOH solution was gradually added drop by drop with constant

monitoring with a pH meter. The toxicity of pH adjusted oil was assessed at

its MIC value against test bacteria by usual agar dilution method. The

appearance of bacterial growth was recorded at interval of 32-72h of

incubation at 35 ± 2ºC. Results are reported in Table 12.

All the experiments were done in triplicate and revised. During each

experiment non poisoned plates were kept as control set.

Table 12: Effect of physical factors on antibacterial toxicity of Clausena oil

Physical factors

Appearance of growth present (+)/absent (-) on

agar plate

Erw. herbicola Ps. putida

Effect of storage period (in months)

3 - -

6 - -

9 - -

12 - -

Effect of temperature (in 0C)

40 - -

60 - -

80 - -

100 - -

120 - -

Effect of autoclaving (15 lb/inch2

pressure for 15 min) - -

Effect of pH

5 - -

6 - -

7 - -

8 + +

103

Estelar

Table 12 reveals effect of storage period, temperature, autoclaving

and pH on antibacterial toxicity of Clausena oil. The oil retained its toxicity

against both test bacteria such as Erw. herbicola and Ps. putida after

exposure upto 120ºC indicating its thermostable nature. Autoclaving had no

adverse effect on the potency of oil. The oil retained its potency at pH 5, 6

and 7 against both test bacteria but at pH 8 the toxicity declined. Besides the

toxicity of oil persisted upto 12 months of storage (Table 12).

11. Fungitoxic properties of Clausena oil:

Various fungitoxic properties viz., minimum inhibitory

concentration (MIC), minimum fungicidal concentration (MFC), effect of

increased inoculum and effect of physical factors on toxicity of Clausena oil

were studied as following:

11.1 Minimum inhibitory concentration (MIC):

The minimum amount of oil needed to emit vapours sufficient

enough to inhibit complete mycelial growth of test fungi was determined

following the Inverted Petri plate method of Bocher (1938). In control set, in

place of oil sterilized water was used. Each experiment was revised and

contained three replicates. The incubation temperature was at 28 ± 2ºC. The

per cent inhibition of mycelial growth was recorded after 7 days and results

are depicted in Fig 7.

11.2 Minimum fungicidal concentration (MFC):

For determination of nature (Static/cidal) of the oil, the technique

prescribed by Garber and Houston (1959) was followed. The oil treated discs

of test fungi showing complete inhibition of mycelial growth were washed

with sterilized water and placed again on fresh solidified CDA medium to

observe revival of mycelial growth. Each experiment was revised and

104

Estelar

maintained in triplicate. Revival of fungal growth was recorded by visible

growth of discs. MFC was the lowest concentration of essential oil from

which fungus can not revive. Results are depicted in Fig. 7.

Fig. 7: Minimum inhibitory and minimum fungicidal concentration of Clausena

oil against Aspergillus spp.

MIC of Clausena oil was found to be 100ppm for all test fungi,

while MFC value for A. flavus, A. niger, A. ochraceus and A. terreus was 3200,

3100, 2700 and 3300ppm respectively (Fig. 7).

11.3 Effect of increased inoculum of test fungi on potency of oil:

The effect of increased inoculum density of test fungi on the

potency of the Clausena oil at its MIC (100ppm) was studied by increasing

the number of inoculum discs in arithmetical progression of 2 upto maximum

numbers of 10 discs and increasing diameter of discs in arithmetical

105

0

500

1000

1500

2000

2500

3000

3500

MIC MFC MIC MFC MIC MFC MIC MFC

100

3200

100

3100

100

2700

100

3300

Val

ues

in

pp

m

Test fungi

C. pentaphylla

Aspergillus flavus A. niger A. ochraceus A. terreus

Estelar

progression of 5mm upto a maximum of 25mm diameter. Each experiment

replicated three times and was revised. The incubation temperature was at 28

± 2ºC. The observations were recorded after 7 days for presence (+) and

absence (-) of mycelial growth of test fungi. The results are reported in Table

13.

Table 13: Effect of increased inoculum on toxicity of Clausena oil

Effect of increased inoculum

Appearance of mycelial growth present (+)/absent

(-) on agar plate

Aspergillus

flavus A. niger

A.

ochraceus A. terreus

Number of inoculum disc of 5mm

2 - - - +

4 - - - +

6 - - - +

8 - - - +

10 - - - +

Diameter of inoculum disc in mm

10 - - - +

15 - - - +

20 - - - +

25 - - - +

Clausena oil (100ppm) could sustain inoculum load upto ten

fungal discs (of three test fungi) except A. terreus. Oil at 100ppm was

inhibited growth of single mycelial disc of 25mm diam of three test fungi

except A. terreus (Table 13).

11.4 Fungitoxic spectrum of Clausena oil:

The fungitoxic spectrum of Clausena oil at its MIC (100ppm) and

at higher MIC (200, 300ppm) was observed by Inverted Petri plate technique

against other isolated fungi of pigeon pea seeds viz., Alternaria alternata,

Aspergillus flavipes, A. parasiticus, A. restrictus, Cladosporium sp.,

Curvularia lunata, Fusarium nivale, F. oxysporum, Penicillium sp., P.

chrysogenum, P. italicum, Mucor sp. and Rhizopus arrhizus. Each

106

Estelar

experiment was carried out in triplicate and revised. The incubation

temperature was at 28 ± 2ºC for 7 days. The diameter of hyphae (mm) of

each fungal species from centre to inside of plates was measured after 7

days. The results are reported in terms of per cent mycelial inhibition in

Table 14.

Table 14: Fungitoxic spectrum of Clausena oil

Fungal species

Per cent mycelial inhibition at various

concentration (ppm) of oil

100 200 300

Alternaria alternata 100 100 100

A. flavipes 85.80 100 100

A. parasiticus 96.72 100 100

A. restrictus 81.76 100 100

Cladosporium sp. 100 100 100

Curvularia lunata 79.04 100 100

Fusarium nivale 80.48 100 100

F. oxysporum 79.09 100 100

Mucor sp. 74.04 100 100

Penicillium sp. 100 100 100

P. chrysogenum 78.16 100 100

P. italicum 71.15 100 100

Rhizopus arrhizus 100 100 100

The Clausena oil showed 100% per cent mycelial inhibition

against Alternaria alternata, Cladosporium sp., Penicillium sp. and

Rhizopus arrhizus while less than 100% against other fungi. Oil at 200ppm

was able to check the growth of all fungi, thus proved their broad range of

activity (Table 14).

11.5 Effect of physical factors on fungal toxicity of Clausena oil:

The effect of different physical factors viz., storage, temperature

and autoclaving on fungal toxicity of the oil was studied following the method

of Tripathi and Kumar (2007) while effect of pH was done following Shahi et

al. (1999). 107

Estelar

11.5.1 Effect of storage-

Experiments were carried out to ascertain the duration for which the

oil can be stored without losing its fungitoxicity. The oil was stored upto 12

months and was tested separately at regular interval of 3 months at its MIC

(100ppm) by usual Inverted Petri plate technique. The fungitoxicity was

calculated in terms of per cent mycelial inhibition. The results are reported

in Table 15.

11.5.2 Effect of temperature-

The fungitoxicity of oil exposed at desired temperature (40, 60, 80,

100 & 1200C) in an electric oven for 10, 15, 20, 25 and 30min respectively

was tested at its MIC (100ppm) against test fungi by usual Inverted Petri plate

technique. The results were recorded in terms of per cent mycelial inhibition.

The results are reported in Table 15.

11.5.3 Effect of autoclaving-

The toxicity of Clausena oil (autoclaved at 15 lb/inch2 pressure for

15min) was tested against the test fungi by usual Inverted Petri plate

technique. Result in terms of per cent mycelial inhibition was reported in

Table 15.

11.5.4 Effect of pH-

Requisite amount of oil was mixed in Polyethylene glycol and pH

was adjusted at 5, 6, 7 and 8 (using NaOH and HCl). The toxicity of pH

adjusted oil was tested at its MIC (100ppm) against the test fungi by usual

Inverted Petri plate technique. The result in terms of per cent mycelial

inhibition was reported in Table 15.

All the experiments were replicated three times and revised. During

each experiment a control set without oil was maintained. The incubation

108

Estelar

temperature was at 28 ± 2ºC for 7 days.

Table 15: Effect of physical factors on fungal toxicity of Clausena oil

Physical factors

Per cent mycelial inhibition

Aspergillus

flavus A. niger A. ochraceus A. terreus

Effect of storage period (in months)

3 100 100 100 100

6 100 100 100 100

9 100 100 100 100

12 100 100 100 100

Effect of temperature (in 0C)

40 100 100 100 100

60 100 100 100 100

80 100 100 100 100

100 100 100 100 100

120 100 100 100 100

Effect of autoclaving (15 lb/inch2

pressure for 15min) 100 100 100 100

Effect of pH

5 100 100 100 100

6 100 100 100 100

7 100 91.48 89.80 100

8 100 100 100 88.23

The oil remained effective after 12 months of storage showing long

shelf life. The oil was thermostable after temperature exposure upto 1200C.

Autoclaving had no adverse effect on the toxicity of the oil against all test

fungi. The oil retained its potency at pH 5 and 6, however, the toxicity for A.

niger, A. ochraceus and A. terreus was reduced at pH 7 and 8 respectively

(Table 15).

12. Pesticidal properties of Clausena oil:

12.1 Minimum repellency dose of Clausena oil:

Minimum repellency dose of oil against Callosobruchus chinensis

and C. maculatus was determined following Tripathi and Kumar (2007).

Different amounts of oil (10, 5 and 2.5µl) were soaked in sterilized cotton

109

Estelar

swabs and placed in experimental arm of Y tube Olfactometer separately. The

other arm of the Olfactometer which has cotton swab without oil was served

as control arm. 20 adults of each test insect were introduced in the base arm of

the Olfactometer in five batches at an interval of five min. Repellency was

observed after 30min. The number of insects in the base arm of the

Olfactometer was considered as non-reacting insects and excluded from the

data recorded. Experiments were performed in triplicate and revised. The

minimum repellency dose of oil is reported in Table 16.

Table 16: Minimum repellency dose of Clausena oil

Test insects Amount of oil

in µl

No. of insect in

base arm

No. of insect in

control arm

No. of insect in

experimental arm

Callosobruchus

chinensis

10 12 (60%) 08 (40%) 0 (0%)

5 07 (35%) 13 (65%) 0 (0%)

2.5 09 (45%) 08 (40%) 03 (15%)

C. maculatus

10 13 (65%) 07 (35%) 0 (0%)

5 14 (70%) 06 (30%) 0 (0%)

2.5 11 (55%) 08 (40%) 01 (5%)

No insect was found in experimental arm at 5µl oil dose indicating

that Clausena oil exhibited minimum repellency dose for both the test insects

(Table 16).

12.2 Ovipositional activity of Clausena oil:

This experiment was conducted using the modified method of

Huang et al. (2000b). 30 fresh seeds of Cajanus cajan were placed in Petri

plate (80mm diam) and filter papers soaked with different doses (5, 10 and

20µl) of Clausena oil were pasted separately on inner surface of upper lid

cover of Petri plate for a duration of 24 and 48h exposure. After exposure

these seeds were transferred to fresh Petri plate and five pairs of test insects

110

Estelar

(male + female) were released on pigeon pea seeds in each Petri plate

separately for egg laying and progeny emergence. In parallel a control set was

also maintained by introducing test insects on fresh non fumigated pigeon pea

seeds. Experiment was done separately for both insects and incubation

temperature was at 28 ± 2ºC. The observations were made after 10 days for

egg laying (oviposition) and after 21 days for progeny emergence. Each

experiment was revised and maintained in triplicate. The mean number of egg

laid and progeny emerged are depicted in Fig. 8.

Fig. 8: Ovipositional activity of Clausena oil against test insects

5µl dose and 48h exposure of Clausena oil significantly inhibited

the oviposition and progeny emergence of both insect species. At 5µl dose and

24h exposure although eggs were laid by C. maculatus but progenies

emergence failed completely (Fig. 8).

111

0

2

4

6

8

10

A B A BA B A B

1

0.3 0 0

1.33

0 0 0

0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

10

5

8

3

10

4

8

2

Mea

n n

o. of

egg l

aid (

A)

and p

rogen

y

emer

ged

(B

)

Test insects

5µl

10µl

20µl

Ethanol

Concentration

of oil

Estelar

12.3 Ovicidal activity of Clausena oil:

Ovicidal activity of Clausena oil was studied following Huang et al.

(2000b). During experiment newly emerged adults of both test insects were

sorted out from reared culture jars and released separately on fresh pigeon pea

seeds for egg laying. 20 eggs on infested seeds were taken in each Petri plate.

Further the different doses of Clausena oil (5, 10 and 20µl) were soaked onto

filter paper discs separately and pasted on the inner surface of the cover of

Petri plates. The eggs were exposed for 24 and 48h. After stipulated exposure

the treated eggs (alongwith seeds) were transferred separately onto clean pre-

sterilized Petri plate to observe adult emergence. For each concentration three

replicates were run along with a control set, having eggs treated with blank

filter paper. Experiment was done separately for both insects and incubation

temperature was at 28 ± 2ºC. Number of adult emerged from treated and

control set was recorded after three week. The results were based on mean

values are depicted in Fig. 9.

Fig. 9: Ovicidal activity of Clausena oil against eggs of test insects

112

0

2

4

6

8

10

12

14

24 h 48 h 24 h 48 h

3

0

4 3.66

0.66 0 0.33 0 0 0 0 0

14 13

12 13

Dev

elopm

ent

of

adult

fro

m f

um

igat

ed

eggs

5µl

10µl

20µl

Control

Callosobruchus chinensis C. maculatus

Test insects

Estelar

No insects (both species) appeared in the treatment set at 10µl dose of oil after

48h post treatment period indicating ovicidal nature of Clausena oil (Fig. 9).

12.4 Insecticidal activity of Clausena oil:

The insecticidal activity of Clausena oil was evaluated against C.

chinensis and C. maculatus following Kumar et al. (2008). The different

amounts of oil (5, 10 and 20µl) were soaked in filter paper discs (10mm diam)

separately and pasted on inner surface of the cover of the pre-sterilized Petri

plate containing 5g of pigeon pea seeds alongwith a group of ten insects.

Similarly a control set was also maintained without oil. In order to get cidal

nature of the oil, the insect revival was observed after transferring them onto

fresh Petri plate. Experimental set up for both insects was done separately.

Experiments were carried out in triplicate and revised. The incubation

temperature was at 28 ± 2ºC. The per cent mortality of insects was recorded at

intervals of 24 and 48h of exposure. Results are depicted in Fig. 10.

Fig. 10: Insecticidal activity of Clausena oil against test insects

113

5µl

10µl

20µl

75

80

85

90

95

100

24h48h

24h48h

86.66

100

93.33

100

100 100 100 100

100 100 100 100

Per

cen

t m

ort

alit

y o

f in

sect

s

Test insects

Estelar

Clausena oil exhibited insecticidal activity against both the test

insects with 100% mortality at 5µl dose of 48h exposure indicating

insecticidal nature of Clausena oil (Fig. 10).

12.5 Effect of physical factors on repellent activity of Clausena oil:

Effect of physical factors viz., storage, temperature and autoclaving

on repellent activity of oil was studied following Tripathi & Kumar (2007)

while effect of pH was done following Shahi et al. (1999).

12.5.1 Effect of storage-

The repellency of 12 months stored Clausena oil was tested against

both test insects at regular interval of three months using Olfactometer. The

results are reported in terms of per cent repellency in Table 17.

12.5.2 Effect of temperature-

The repellent activity of oil treated at desired temperature (40, 60,

80, 100 & 1200C) for 10, 15, 20, 25 and 30min respectively in an electric oven

was tested against both insects using Olfactometer. Results are reported in

terms of per cent repellency in Table 17.

12.5.3 Effect of pH and autoclaving-

The repellent activity of pH adjusted and autoclaved oil (mentioned

earlier) against both test insects was assessed separately. Results are reported

in Table 17.

All the experiments were carried out in triplicate and revised.

114

Estelar

Table 17: Effect of physical factors on insect repellent activity of Clausena

oil

The repellency of Clausena oil remained unaltered upto 12

months, thermostable upto 1200C and had no adverse effect on autoclaving.

The oil showed 100 per cent repellent activity at pH 5, 6 and 7 against both

the insects (Table 17).

13. Gas Chromatography-Mass Spectrometry analysis of Clausena oil:

GC-MS analysis of Clausena oil was performed by using Perkin

Elmer Clarus 500 GC, coupled with RTX-5 (60m X 0.32mm, ID X

film thickness 0.25μm) capillary column. The carrier gas was Helium (1

ml/min). Injector temperature was 210ºC and oven temperature was

programmed 60-210ºC at the rate of 3ºC/min, finally held isothermally for

115

Physical factors Per cent repellency

Callosobruchus chinensis C. maculatus

Effect of storage period (in months)

3 100 100

6 100 100

9 100 100

12 100 100

Effect of temperature (in 0C)

40 100 100

60 100 100

80 100 100

100 100 100

120 100 100

Effect of autoclaving (15 lb/inch2

pressure for 15min) 100 100

Effect of pH

5 100 100

6 100 100

7 100 100

8 100 96

Estelar

15min. The ionization of the sample components was performed in EI mode

of 70 eV.

The constituents of the oil were identified by calculation of their

retention indices under temperature programmed condition for n-alkanes (C6-

C24) comparison of their mass spectra with those of computer library search

(NIST/PFLEGER/WILEY) and confirmed by comparison of their retention

indices either with those of authentic compounds or with data published in the

literature (Adams, 2009). Results are reported in Table 18.

Table 18: Chemical composition of Clausena pentaphylla essential oil

S.No. Components KI % in oil

1. α-thujene 924 0.1

2. α-pinene 932 0.3

3. Sabinene 969 24.7

4. β-pinene 972 0.2

5. β-myrcene 974 1.2

6. α -phellandrene 1002 0.1

7. α-terpinene 1014 0.9

8. p-cymene 1020 0.1

9. Limonene 1024 7.8

10. β-ocimene 1032 2.0

11. -terpinene 1054 0.5

12. Trans-sabinene hydrate 1098 0.1

13. α-terpinolene 1186 13.8

14. 4-terpineol 1188 0.8

15. Safrole 1285 6.7

16. β-elemene 1389 0.1

17. Methyl eugenol 1403 38.1

Total identified 97.5

Monoterpene hydrocarbons 51.7

Oxygenated monoterpenes 45.7

Sesquiterpenes hydrocarbons 0.1

KI-Kovat’s Index

GC-MS analysis of the oil revealed presence of 125 major and minor

compounds (Appendix II), of which seventeen compounds representing

116

Estelar

around 97.5 % of the oil sample were identified (Table 18). Table shows

relative per cent content of components alongwith their Kovat’s index. The oil

was characterized by presence of eleven monoterpene hydrocarbons (57.7%),

five oxygenated monoterpenes (45.7%) and one sesquiterpene hydrocarbon

(0.1%). The major constituents were methyl eugenol (38.1%), sabinene

(24.7%), α-terpinolene (13.8%), limonene (7.8%) and safrole (6.7%).

14. In vivo comparative efficacies of Clausena oil and synthetic fumigants

in preservation of pigeon pea seeds during storage:

Experiment was conducted following the method of Shukla (2009).

To find out practical applicability of Clausena oil as fumigant for protection

of pigeon pea seeds from fungal and insect infestation, freshly harvested

pigeon pea seeds of local variety (Ramarhara) were collected from village of

Gorakhpur district in polyethylene bags. Different amounts of Clausena oil

soaked in cotton swabs so as to procure concentration of 400 and 800ppm and

introduced separately in pre-sterilized gunny bags and glass containers with

500g pigeon pea seeds for fumigation. Likewise two other sets were prepared

by treating pigeon pea seeds with synthetic fumigants (aluminium phosphide

and ethylene dibromide) for comparison purpose. The non fumigated seeds

were kept as control set. Each experiment was maintained in triplicate. The

containers were made airtight and stored in a seed storage cabinet in

laboratory at room temperature and 75 ± 5% humidity for six months.

Efficacy of oil and synthetic fumigants on pigeon pea infestation was assessed

by weight loss, seed damage, pest analysis, protein content and phytotoxicity

test.

14.1 Per cent weight loss and per cent seed damage:

The weight loss of stored seeds was calculated by formula of Parkin (1956).

117

Estelar

W-w

Per cent weight loss = X 100

W

Where W= weight of seeds before incubation

w = weight of seeds after six months of incubation

Seed damaged/injured was evaluated by weighing seeds with feeding

injuries and emergence hole on the surface of the seeds occurred due to pest

attack during six months of storage. Results are reported in Table 19.

Table 19: Per cent weight loss & per cent seed damage of stored pigeon pea

seeds treated with Clausena oil and synthetic fumigants after six

months of storage

Treatment Concentration

in ppm

Per cent weight loss Per cent seed damage

Gunny bag Glass

container Gunny bag

Glass

container

Clausena oil 400 0 0 0 0

800 0 0 0 0

Aluminium

phosphide

400 20 10 26 21.6

800 10 5 16.6 18

Ethylene

dibromide

400 5 2 10 8

800 2 0 5 0

Control 30 25 40 36.8

Clausena oil proved its seeds preserving capacity in control of pest

infestation of pigeon pea seeds upto six months of storage. The weight loss

and seed damage of control sets due to pest attack were 30 & 40% when

stored in gunny bags and 25 & 36.8% when stored in glass containers

respectively. Ethylene dibromide was more effective than aluminium

phosphide (Table 19).

14.2 Fungi and insects associated with pigeon pea seeds after storage:

Pigeon pea seeds (treatment and control) were observed after

storage for associated fungi and insects. Presence and absence of insects on

118

Estelar

seeds were observed by hand lens while fungal analysis was made by

Standard blotter paper as well as agar plate methods. The results are reported

in Tables 20-22.

Table 20: Mycoflora of pigeon pea seeds treated with Clausena oil and

synthetic fumigants after six months of storage by Agar plate technique

Fungal species

Treatment in ppm

Clausena oil Aluminium

phosphide

Ethylene

dibromide Control

400 800 400 800 400 800

A B A B A B A B A B A B A B

Aspergillus flavus - - - - + + + - + + + + + +

A. niger - - - - + + - - - + + + + +

A. ochraceus - - - - + + - + - - - - + +

A. terreus - - - - + + - - - - - - + +

Cladosporium sp. - - - - + - + + + - - - + +

Fusarium nivale - - - - + + - - + - - - + +

F. oxysporum + - - - + + - + - + - - + +

Mucor sp. - - - - + - + + - - - - + +

Penicillium italicum - - - - + - + + - + - - + +

Rhizopus arrhizus - - - - - + - + - - - - + +

Storage system: A- Gunny bag; B-Glass container; + presence of fungi; - absence of fungi

Table 21: Mycoflora of pigeon pea seeds treated with Clausena oil and

synthetic fumigants after six months of storage by Standard

blotter paper technique

Fungal species

Treatment in ppm

Clausena oil Aluminium

phosphide

Ethylene

dibromide Control

400 800 400 800 400 800

A B A B A B A B A B A B A B

Alternaria alternata - - - - + + - - + - - - + +

Aspergillus flavus - - - - + + + + + + - + + +

A. niger - - - - + + + - - - - - + +

A. ochraceus - - - - + + - - + - - + + +

A. parasiticus - - - - + + + - + - + - + +

A. terreus - - - - + + - - + - + - + +

Cladosporium sp. - - - - + + + + + - - - + +

Fusarium oxysporum + - - - - + - + - + + - + +

Penicillium sp. - - - - + + + - - + - - + +

Rhizopus arrhizus - - - - + + + + - + - - + +

Storage system: A- Gunny bag; B- Glass container; + presence of fungi; - absence of fungi 119

Estelar

Table 22: Insects analysis of pigeon pea seeds treated with Clausena oil and

synthetic fumigants after six months of storage

Insect species

Treatment in ppm

Control Clausena oil Aluminium

phosphide Ethylene dibromide

400 800 400 800 400 800

A B A B A B A B A B A B A B

Callosobruchus

chinensis - - - - - - - - - - - - - -

C. maculatus - - - - - - - - - - - - + -

Storage system: A- Gunny bag; B- Glass container; + presence; - absence

i. Mycoflora analysis-

Fumigant free pigeon pea samples (control set) stored in gunny bags

and glass containers showed appearance of 13 fungal species. The seeds

fumigated with 800ppm concentration of Clausena oil were free from fungal

infestation. The oil at 400ppm concentration was unable to control appearance

of F. oxysporum in gunny bags. Amongst synthetic fumigants, 800ppm

concentration of ethylene dibromide was more effective than aluminium

phosphide inhibited the appearance of all fungal species except A. flavus, A.

niger and A. ochraceus stored in glass containers while 400ppm concentration

of both commercial fumigants was ineffective leads to proliferation of several

fungal species (Tables 20 & 21).

ii. Insect pests-

In control sets only C. maculatus was found to be associated with

pigeon pea samples stored in gunny bags while no species of insect pest

could be isolated from glass containers. The treated seeds (oil/commercial

pesticides) were free from insects attack (Table 22).

14.3 Estimation of protein content-

Protein content of fumigated and non fumigated stored pigeon pea

120

Estelar

seeds was estimated by method of Lowry et al. (1951). Results are reported

in Table 23.

Table 23: Protein estimation of pigeon pea seeds treated with Clausena oil

and synthetic fumigants after six months of storage

Six months

stored

pigeon pea

seeds

Treatment in ppm

Control Clausena oil Aluminium

phosphide Ethylene dibromide

400 800 400 800 400 800

A B A B A B A B A B A B A B

Protein (%) 19.8 20 21.5 21.7 18.5 19 18.5 18 19 19.5 18 20 17 16.8

Protein content of freshly harvested pigeon pea seeds: 22.09%

Storage system: A- Gunny bag; B- Glass container.

There is no significant change in protein content of stored pigeon

pea seeds fumigated with Clausena oil in comparison to fresh seeds, however,

there was reduction in protein content of pigeon pea seeds in control set

(Table 23).

14.4 Phytotoxicity test:

To see the toxic effect of Clausena oil, stored pigeon pea seeds upto

six months (treated as well as control) and fresh healthy seeds were taken in

Petri plate for seed germination and seedling growth experiments.

i. Effect on seed germination-

40 fumigated and non fumigated seeds (control-1) of pigeon pea

from each set were selected randomly and aseptically placed on pre-sterilized

Petri plates separately containing three layers of moistened blotter paper for

germination. In control set (control-2) 40 fresh seeds were placed on blotter

paper in similar manner for comparison purpose. The Petri plates of all the

sets were incubated at 28 ± 2ºC temperature in dark chamber. Blotter papers

were moistened after interval of 3 days. The experiments were repeated twice

121

Estelar

and each contained three replicates. The observations were recorded on 3rd

, 5th

and 7th

day. The per cent germination was calculated and results are reported

in Table 24.

Table 24: Germination percentage of pigeon pea seeds treated with Clausena

oil and synthetic fumigants after six months of storage

Treatment Dose

(ppm)

Seed germination(%) at days$

3 5 7

Gunny

bag

Glass

container

Gunny

bag

Glass

container

Gunny

bag

Glass

container

Clausena oil 400 38±2.34 52±5.26 48±2.35 65±4.79 60±1.92 74±6.50

800 59±5.46 51±5.26 71±4.72 63±3.53 84±4.21 74±3.54

Aluminium

phosphide

400 20±0.89 20±0.00 24±1.58 28±1.0 28±1.78 36±1.94

800 26±1.22 22±1.31 32±1.81 35± 2.12 32±1.58 40±2.91

Ethylene

dibromide

400 31±2.12 30±2.21 36±2.04 42±0.83 50±1.67 44±1.67

800 30±1.03 21±0.23 34±1.41 34±0.89 44±1.67 37±1.67

Control-1 30±3.46 28±1.21 34±2.0 36±1.98 40±2.65 45±3.21

Control-2 50±2.21 69±0.98 82±1.21

$ Values given are mean of three replicates ± Standard Deviation

Control-1: Six months’ stored seeds (non fumigated)

Control-2: Fresh seeds

The Clausena oil at 800ppm did not inhibit the germination of

pigeon pea seeds as compared to germination of fresh healthy seeds (control-

2). Oil treated seeds showed 60-84% seed germination, aluminium phosphide

32-40%, ethylene dibromide 37-50% when stored in gunny bags and glass

containers, however, seeds of control sets (control-1) exhibited 30-45%. Thus,

the oil had no significant ill effect on seed germination in comparison to

control-2 (Table 24).

ii. Effect on seedling growth-

The length of radicle and plumule of the germinated seeds of

control (1 & 2) and treatment sets were measured regularly upto 5th

, 7th

and 9th

day and data are reported in Tables 25 & 26.

122

Estelar

Table 25: Effect of Clausena oil and synthetic fumigants on seedling growth

of pigeon pea seeds stored in gunny bags for six months

Treatment Dose

(ppm)

Length of radicle and plumule (cm)$

5 7 9

Radicle Plumule Radicle Plumule Radicle Plumule

Clausena

oil

400 2.94±0.36 8.78±2.46 3.72±0.30 9.40±2.72 4.38±0.49 9.9±2.65

800 4.78±0.85 9.10±0.89 5.0±0.07 9.42±1.27 5.78±1.23 10.3±1.78

Aluminium

phosphide

400 1.02±1.15 4.02±0.82 1.22±0.87 4.8±0.27 3.2±1.5 5.0±0.85

800 1.38±1.13 0.9±0.24 1.98±1.41 2.2±1.18 2.5±0.20 5.0±0.85

Ethylene

dibromide

400 1.8±0.94 4.9±0.32 2.5±1.11 5.2±0.85 3.0±1.03 6±1.09

800 1.9±0.89 3.2±1.5 3.0±0.92 4.0±4.3 4.3±1.03 5.6±1.2

Control-1 1.22±0.99 2.98±0.74 3.84±0.35 4.72±0.97 4.02±0.32 5.10±0.89

Control-2 4.67±0.15 9.5±1.23 5.2±1.0 10.2±0.9 6.0±0.34 11.0±1.3

$Values given are mean of three replicates ± Standard Deviation

Control-1: Six month’s stored seeds (non fumigated)

Control-2: Fresh seeds

Table 26: Effect of Clausena oil and synthetic fumigants on seedling growth of

pigeon pea seeds stored in glass containers for six months

$Values given are mean of three replicates ± Standard Deviation

Control-1: Six months’ stored seeds (non fumigated)

Control-2: Fresh seeds

123

Treatment Dose

(ppm)

Length of radicle and plumule (cm)$

5 7 9

Radicle Plumule Radicle Plumule Radicle Plumule

Clausena

oil

400 3.22±0.45 8.16±0.54 3.94±0.21 8.62±0.54 4.75±0.23 10.16±0.74

800 5.48±1.23 6.30±1.42 5.58±1.49 6.8±1.78 6.48±1.68 7.12±1.73

Aluminium

phosphide

400 1.0±0.83 3.2±0.50 1.4±0.03 4.6±0.57 1.8±1.23 5.0±0.78

800 1.2±1.3 2.0±0.32 1.8±1.51 3.0±1.0 1.8±1.51 4.0±2.23

Ethylene

dibromide

400 2.5±0.83 4±0.32 3.0±1.34 4.8±1.3 4.0±1.64 5.2±1.26

800 2.8±1.3 5±2.12 3.5±1.51 6.0±1.0 5.0±1.64 6.7±0.52

Control-1 3.12±1.78 5.14±1.07 4.08±1.33 6.16±1.47 4.35±1.27 6.42±1.27

Control-2 4.67±0.15 9.5±1.23 5.2±1.0 10.2±0.9 6.0±0.34 11.0±1.3

Estelar

Data indicates that oil did not show any adverse effect on the length

of radicle and plumule of seeds stored in gunny bag and glass container as

compared to synthetic fumigants and control set-1 (Tables 25 & 26).

124

Estelar

Estelar