Degree Research Plan and Proposal- Tran Ngoc Tuan(1) (1)

8/15/2019 Degree Research Plan and Proposal- Tran Ngoc Tuan(1) (1)

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HUAZHONG AGRICULTURAL UNIVERSITY

DEGREE RESEARCH PLAN AND PROPOSAL

NAME OF THE STUDENT: TRAN NGOC TUAN

STUDENT ID： 2011308060001

COLLEGE: COLLEGE OF FISHERIES

MAJOR: AQUACULTURE

DEGREE: Ph.D.

RESEARCH AREA: FISH GENETICS AND BREEDING

THESIS TITLE: cDNA LIBRAR CONSTRUCTION!

EST ANALSIS AND E"PRESSION OF IMMUNE#

RELATED GENES IN BLUNT#SNOUT BREAM!

MEGALOBRAMA AMBLYCEPHALA AFTER

BACTERIA INFECTION

AD$ISOR : PROFESSOR %ANG %EIMIN

MEMBERS OF GUIDING GROUP:

DOCTOR GAO &E"IA

PROFESSOR LIU HONG

06'12'2012


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Th( P)*+*,(- R(,()ch %*)/! I, O)45 - C)(((,,1.1 %*)/ +)*+*,(-

The blunt-snout bream ( Megalobrama amblycephala) is herbivorous and distributes in

the middle portion of Yangtze River (Fishbase 2012). This speies is a ma!or fresh"ater

speies and urrentl# ultured in $hina beause of man# valuable harateristis% suh

as omprising of high delia#% high larval survival rate% natural foods feeding% fast

gro"th% tender flesh% high diseases resistane and so on (Tsao 1&'0 hou et al 200*).

The prodution of blunt-snout bream has inreased in reent #ears% "ith ++2&00 tons in

200+ and '2+,*& tons in 200&% raned sith in "hole $hinese fresh"ater fish

prodution (/inistr# of griulture of the eoples Republi of $hina 200' /inistr#

of griulture of the eople3s Republi of $hina 2010). The seletion on this fish

speies "as started in 1&*' from a population eploited from Yuni 4ae% and after '

generations the fish gro"th "ere improved up to 2&5 (4i and $ai 2000 4i and $ai

2006).

7ine 1&'0s% man# studies had been arried out on blunt-snout bream. The previous

researhes mainl# foused on the improvement on the ultured fish biomass% nutrient

re8uirement and geneti variations in ultured and "ild population. 9o"ever% studies

on blunt-snout bream diseases "hih aused great loss in ultured fish are still limited%

espeiall#% on the response of immune-related genes to the pathogens. Therefore% a

detailed stud# on the epression of immune-related genes of blunt-snout bream is

neessar#.

1.2 O)45 - c)(((,,

1.2.1 $onstrution of :; librar# on the immune-related genes based on net

generation se8uene b# using lunt-snout bream ( M. amblycephala)% also alled ?uhang fish% is a member of

finfish $lass of tinopter#gii% @rder of $#priniformes and Famil# of $#prinidae.

ording to The Fish :atabase of Tai"an (2012)% the bream has small% laterall#%

ompressed and pointed head% short snout% large e#es and the mouth "hih is small%

protratile% slightl# obli8ue and arhed. The fish bod# ompressed and high depthdorsal and ventral profile arhed eel on bell# bet"een pelvi fin origin and anal fin


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origin. The sales are moderatel# large% #loid lateral line omplete (4.l.A+0-+*)

dorsal fin ra#s (BBB% ,) petoral fin ra#s (1C1D-1&) pelvi fin ra#s (1C*) anal fin ra#s

(6C2,-62). etoral fin end lose to pelvi fin origin pelvi fin at lo"er side of bod#

anal fin base long audal fin fored. :ar gra#ish dorsall#% side and bell# silver#

"hite. ll fins are gra#ish.

The blunt-snout bream prefers to be benthopelagi and habitat in deep fresh"ater pools

(arrange of +-20 m) "ith slo" running or an be found in laes% impoundments and

ba"aters of rivers. This is a h#poia-sensitive speies (7hen et al 2010). Bn $hina%

the fish ommonl# distribute in the middle portion of Yangtze River found in

;e"shan 4ae (a ba# of 4iangzi 4ae "ith an area of 2+' m2) and Yuli 4ae

(Fishbase 2012).

>lunt-snout bream is basiall# native herbivorous fresh"ater finfish (Tsao 1&'0

@u#ang et al 2001). Bn natural ondition% the fish "as defined mainl# feed on a8uati

plants suh as Vallisneria natans ('*.65)% Hydrilla verticillata (66.,5)% algae and

zooplanton (Tsao 1&'0). The protein re8uirement for fingerling "as 6+5 and lipid D-

,5 (hou et al 1&&, 4i et al 2010). 9o"ever% protein demand "as demonstrated in

orrelation "ith temperature onditions the optimum protein onsumption for fish are

from 2,-60.D5 at 20o$ and from 2+.'-D1.D5 at 2+-60o$ (7hi et al 1&**).

The bream rapidl# gro"th in earl# period and slo"l# later in the life (@u#ang et al

2001)% in "hih gaining of 1'.D m (1 #ear)% 60., m (2 #ears)% 6*.* (6 #ears)% D1.* m

(D #ears)% D1.6 m (+ #ears) and D+.+ m (' #ears) (Tsao 1&'0). The fish are matured

after 2 #ears "ith the relative feundit# average 262 eggEg (@u#ang et al 2001).

1.2 S7-(, * 947#,*7 9)(

Physiology

The o# onsumption of blunt-snout bream "as lo" in bigger bod# "eight and lo"

temperature of the "ater% i.e. the fish "eighs of 10.6 g and 2&+ g onsume of 6'D

mgEgEh and 1&6.** mgEgEh% respetivel# (@u#ang et al 2001). This is a

benthopelagi speies d"elling in +-20 m of "ater depth% ho"ever% the o#gen

onsumption "as relativel# high. Bn the room temperature of 2,o$ !ust for a short time

of h#poia (belo" 0.+ mg @2E4) ould ause the mortalit# of fish (@u#ang et al 2001).

The funtion of h#poia-sensitive of fish "as identified has an involved to the

epression of t"o genes% 9BF-1 and 9BF-2 though% these genes have a different

ph#siologial funtion under normoia and h#poia situations (7hen et al 2010). Thespleen% idne# and liver "ere developmental plaes for red blood ells formation (?u


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1&&0). The ativities of t"o antioidized enz#mes (the glutathione peroidase (


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to the stress b# the liver funtions and non-immunit# (4i et al 2012a). The fish oil

"hih "as supplied into feed formulate "as good for gro"th of bream and the

proportion of D-'5 should be added (hou et al 1&&,). 9o"ever% to the bream

fingerling silver oil and the so#bean oil "ere defined more effetivel# for gro"th than

other oils (4iu et al 1&&, ?ei and $hen 2006).

Replaement of trifolium for 20-D05 in fish diet (1+-605 protein) improved the fish

protein ontent and meat 8ualit# (Gia et al 1&&1). >esides% alfalfa meal "as evaluated to

be suitable for the fish gro"th "hen suppl# of 100 gEg bream feed (9u et al 200+).

The bream diet "hih supplied "ith emodin ('0 mgEg) or Hitamin $ (,00 mgEg)

ma# redue the F$R and enhane the fish gro"th and non-speifi immunit# under the

stress onditions- ro"ded% temperature and baterial infetion (/ing et al 2011a

/ing et al 2011b /ing et al 2012a).

Genetics and reading

From 1&'0s% domestiation of blunt-snout bream "as started. Bn 1&*'% first seleted

generation ahieved aording to the fish origin from 4ae Yuni (the middle of

Yangtze River% $hina) (4i and $ai 2000 4i and $ai 2006). 7urprisingl#% the ultured

bream "ere reorded "ith earlier maturit#% gro"th rate depression thin and longer

bod# and disease suseptibilit# (4i and $ai 2006 esides% the er#thro#te morpholog# in pol#ploid# fish "ere onsidered

and ompletel# studied in general% the differenes of er#thro#te morpholog# in

pol#ploid# fish ma# ause a positive or negative response to the umulative mortalit#

of ultured fish (ou et al 200'). Bn omparison of the fertile autotetraploid (produed

b# preise suppression of the first mitoti leavage of the embr#o b# thermal sho)

and the interploid 6n (produed b# mating of tetraploid females and diploid males) and

the diploid sho"ed differenes in 8uantit# of er#thro#te% the diameter of er#thro#tes

and nulei% the er#thro#te surfae area and the nulear and er#thro#te volume (ouet al 200'). /oreover% interploid h#brids "ere spa"ned from the allotetraploid males


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matted to blunt snout bream females "hih "ere high in fertilization and stable

survival after '0 da#s the inrease of gro"th rate of ,.+5 in omparison bet"een

interploid 6n and diploid blunt-snout bream during 1*0 da#s (ou et al 2011).

/an# studies on the geneti struture b# using of moleular marers on blunt-snout

bream (mt:;% R:% 7R% mirosatellite 77% 7;) "ere onduted (hang%

2001 4i% 200* Tang et al 200* 4i% 2010


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/oreover% the diet for ultured fish "ere supplied "ith emodin or Hitamin $ ma#

enhane for higher ativities of the serum T% 47% J levels% liver 7@: and $T of

blunt-snout bream after A. hydrophila infetion (/ing et al 2011a) high dietar# lipid

redues the resistane to A. hydrophila of blunt-snout bream in laborator# ondition

(4i et al 2012a).

1.3 S7-(, * -,(,(, )(,,c( *; ;,h c7,(- 95 A# hydro$hila

The baterium A. hydrophila belong to the genus of Aeromonas is onsidered to be

almost s#non#mous "ith "ater and a8uati environments. The baterium "as no"n

as an opportunisti pathogen in fresh"ater a8uati habitats "ith heavil# organi load

(ustin and dams 1&&') and in the stress ondition (eters et al 1&**). A. hydrophila

has been assoiated "ith sin lesions% tail and fin rot% haemorrhagi septiaemia over

the bod# and tissue destrution% epizooti uleration and nerosis in the liver and

idne# (ustin and dams 1&&' Ganda and bbout 2010).

A. hydrophila "ere reorded "idel# infeting to fresh"ater fish and marine fish

speies (ustin and dams 1&&' :ouas et al 1&&*). This baterium "as isolated in

arp% Cyprinus carpio ($hirilK et al 200*)% goldfish% Carrassius auratus and Joi% C.

carpio koi ($itarasu et al 2011)% and determined as a main pathogen infeted to

rainbo" trout% Salmo gairdneri (eters et al 1&**)% "aling atfish% Clarias batrachus

(nga et al 1&&0)% goldfish (B8bal et al 1&&&)% ;ile tilapia% Oreochromis niloticus

(Bbrahem et al 200*).

/an# previous studies have been onduted on the resistane to disease aused b# A.

hydrophilla based on nonspeifi immunit# funtions% inluding the interspeies

variations among silver arp ( Hypophthalmichthys molitrix)% bighead arp

( Aristichthys nobilis)% grass arp (Ctenopharyngodon idellus) and bla arp

( Mylopharyngodon piceus) the intra-speies variations among three strains of

ommon arp ("ild C. carpio% mirror C. carpio specularis and !ian ommon arp C.

carpio var. jian)% three strains of ruian arp (engzhe ruian arp C. auratus var.

pengesis% allog#nogeneti ruian arp C. auratus ibelio and "hite ruian arp C.

auratus cuvieri)% and t"o populations of soft-shelled turtle (!rionyx sinensis) from

Taihu lae of Yangtze River basin and Tai"an ($ai et al 200D).

The liver of Bndian arp% Cirrhinus mrigala% "hih "as infeted to A. hydrophila% "as

defined as a speifi tissue of the transferrin gene epression due to transferrin mR;

"as deteted onl# in liver R; and to lesser etent in brain tissue (7ahoo et al 200&).Bn another stud#% after families of tlanti salmon "ere hallenged b# A. salmonicida %


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in l#mphoid organs the pro-inflammator# genes (T4R+/% T4R+7% mR;

"ere losel# assoiated "ith A. hydrophila infetion b# the inrease of epression in

man# tissues at the earl# stage of baterial infetion ($hen et al 200&).

double intraperitoneal in!etion eperiment of the baterium% A. hydrophila to

gilthead seabream (Sparus aurata 4.)% authors suggested that a relativel# fast

elimination of the bateria and reover# of fish during the seondar# infetion. This

based on the do"n-regulated of transript levels of T4R% ;$$R-1% 9N% T$R% Bg/%

/9$-BB %ɑ $6 and $7F-1R and up-regulated of B4-1L genes during the first infetion%

but mR; epression levels "ere up-regulated during the re-infetion (Re#es->eerril

et al 2011). The prophenoloidase (pro@) gene "as determined to be related to the

immune response in red s"amp ra#fish% %rocambarus clarkia% after hallenge to A.

hydrophila% sho"ing inrease of epression level in tissue samples% espeiall# highest

in haemo#tes% but "ith no epression in ovar# (4i et al 2012b). Furthermore% three 4-

t#pe letin (NR


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bateriidal ativit# is more important in resistane against A. hydrophila infetion

(7ahoo et al 200D). >esides% 7ahoo et al (2011) onfirmed that some of innate immune

parameters (phago#ti funtional ativities viz.% respirator# burst and

m#eloperoidase ativities and serum eruloplasmin level) an be indiretl# used as

marer traits for seletion for improved resistane to aeromoniasis in rohu. Bn a "hile%

the serum l#soz#me ativit#% haemagglutination titre and speifi antibod# titre "ere

raised in the #. tarda-infeted rohu% ". rohita (/ohant# and 7ahoo 2010). /oreover%

phago#ti ativit# of leuo#tes% plasma l#soz#me ativit# and speifi antibod# titre

"ere evaluated as more effetive in the resistant families of ommon arp% C. carpio in

omparison to the sensitive ones (rdO et al 2010). Furthermore% omplement and non-

ɑ2m-antiprotease ativities pla# an important role in resistane against A. salmonicida

of tlanti salmon% S. salar (/arsden et al 1&&' hang et al 2011). Bn addition%

respirator# burst and peroidase ativit# indiated a reover# of the immune s#stem

against A. hydrophila of gilthead seabream (S. aurata 4.) (Re#es->eerril et al 2011).

7tudiese on the geneti variation in the immune response of fish "ere onduted. Bn

;ile tilapia% O. niloticus% "hen a full# inbred lone suseptible to A. hydrophila "as

rossed "ith a resistant full inbred lone% the resulting progen# ehibited intermediate

levels of resistane to that of their parents (7arder et al 2001). Bn addition% h#brid

(female ;ile tilapiaPmale blue tilapia) sho"ed the highest resistane to the disease

aused b# A. sobria in omparison to ;ile tilapia% O. niloticus% and blue tilapia% O.

aureus ($ai et al 200D).

1 O9


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- >ateriumA A. hydrophila

- 9ealth# blunt-snout bream fingerlings

- JitsA TRBzol Reagent (Bnvitrogen)% Turbo :;-freeT/ (mbion)% R;eas# /ini it

(Qiagen)% QB8ui $R urifiation Jit (Qiagen)% Trimmer-2 it (Nvrogen)% Bon

Total R;-7e8 Jit v2 (Bnvitrogen)% Bon @neTouhT/ 200 Template it

(Bnvitrogen)% Bon 7e8uening it (Bnvitrogen)% Bon 61*T/ $hip (Bnvitrogen)%

Ta8/an Sniversal /aster /i BB "ith S;< (pplied >ios#stems).

=.2 M(h*-,

=.2.1 E>+()(4 ;,h - 9c() ,)

9ealth# fish fingerlings of blunt-snout bream "as olleted from the ultured pond in

N hou% 9ubei provine% $hina. Fish "as ultured t"o "ees "ith the temperature of the "ater about 2,o$ before hallenge eperiment. total of 2+0 fish "ere randoml#

divided to t"o groups (ontrol and in!eted eperiment) of 2 tans for eah.The baterial strain isolated from septiemia blunt-snout bream "as ultured on the

tr#ptone so#a broth% entrifuged% rinsed and diluted in sterile ph#siologial saline. The

onentration of suspension "as spetrophotometeriall# determined at '10 nm.Nperimental fish "ere intraperitoneall# in!eted "ith 0.1 m4 a suspension of 10 , fu

m4-1 live ell of A. hydrophila. The same volume of sterile ph#siologial saline "as

in!eted into ontrol fish. The umulative mortalit# of eperimental fish "as observed

during + da#s post-in!etion. /oribund fish "ith linial signs "ere re-isolated and re-

identifiated the bateria.=.2.2 S+4 - RNA (>)c*

Tissues of 60 fish (musle% hearth% idne#% spleen and liver) separatel# set into 6 pools

"ith 10 fish per pool from ontrol and hallenged fish "ere olleted in 0h% 2Dh% 6 and

+ da#s post in!etion "ith baterium. ll of the samples "ere immediatel# stored into

sample protetion solution (Taara) and stored at -*0o$ until total R; etration.

Total R; of these samples "as etrated using TRBzol Reagent (Bnvitrogen% S7)

aording to the manufaturer3s instrutions% follo"ing b# R; preipitation% R;

"ashing and R; resuspension.The 8ualit# and 8uantit# of total R; "as determined using a ;anodrop 2000

(Thermo 7ientifi% S7). @nl# R; samples "ith a ratio of 2'0 of 2*0 from 1.& to

2.1 and a ratio 2'0 of 260 from 2.0 to 2.+ "ere used for follo"ing anal#sis. The

samples "ere standardized to 200ng m4-1 and then e8ual volumes from eah tissue

samples of different individuals "ere ombined into a pool. The pools "ere :;ase-treated "ith Turbo :;-freeT/ (mbion) and purified "ith R;eas# /ini Jit (Qiagen%


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S7) follo"ing the manufaturer3s instrutions. R; 8ualit# and 8uantit# "as

determined again at the end of the proess.

=.2.3 cDNA ,5h(,, - I* PGM? ,(@7(c

The R; pools "ere used for double-strand :; s#nthesis using 7/RT

tehnolog# (>: >iosienes $lonteh)% and then the :; "as purified using the

QB8ui $R urifiation Jit (Qiagen).Full-length :; sample ontaining e8ual

amounts of :; from eah librar# "as normalized using the Trimmer-2 it

(Nvrogen% Russia) aording to the manufaturer3s instrutions% follo"ing b# step to

step of :; preipitation% :; denaturation and h#bridization and amplifiation of

normalized :;. The normalized :; "as se8uened using the Bon


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Sampling and RNA extractioncDNA synthesisIon PGM™ sequencingS! analysis

!arget gene determinationGene expression le"el estimation #y qR!-P$RGene expression

1 min at '0o$. The ra" 8$R data "ere anal#sed "ith 7tep@ne 7oft"are v2.0

(pplied >ios#stems) (9olopainen et al 2012).D.6 Tehnial approahes

D.6 itfalls and alternatives

D.6.1 Nperimental fish is not enough for sampling% so need to introdue more fish for hallenge test eperiment.D.6.2 $onstrution of :; librar# is not ompleted need to prepare for more

samples.

. Th( *4(-( - T(ch*4*c4 B,(, ;*) h( P)*+*,(- R(,()chB learned and pratied for the moleular tehnolog#% espeiall# in the $R tehni8ue

"hen B "as in undergraduate and graduate stud# program.

6. %*)/ c*+4((- *) P)*)(,,(, * -(B have finished for the hallenged test and sampling tissues.

. E>+(c(- O7c*(,.1 Researh ahievements (


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6. ustin >% dams $. Fish pathogens% &n ustin >% lt"egg /%


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1D.


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2+. 4i 7F% $ai ?Q. % % 4u J4% Mu ?;% ?ang Y. :ietar# arboh#drateElipid ratios affet stress%

oidative status and non-speifi immune responses of fingerling blunt snout bream%

Megalobrama amblycephala. 'ish ( Shell)ish &mmunology% 2012a% 66A61'-626

2&. 4i Y. % hou Q4% 4iu >% Mi >?. Nffets of emodin% vitamin $

and their ombination on biohemial parameters and t"o 97,0s mR; epressions of ?uhang bream ( Megalobrama amblycephala) infeted "ith Aeromonas hydrophil .


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+ournal o) 'ishery Sciences o) China% 2011b% 1*(6)A+**-'01

6,. /inistr# of griulture of the eople3s Republi of $hina. $hinese fisheries #earboo.

$hinese griultural ress% >ei!ing% 200'

6*. /inistr# of griulture of the eople3s Republi of $hina. $hinese fisheries #earboo.

$hinese griultural ress% >ei!ing% 2010

6&. /ohant# >R% 7ahoo J. Bmmune responses and epression profiles of some immune-

related genes in Bndian ma!or arp% "abeo rohita to #d$ardsiella tarda infetion. 'ish (

Shell)ish &mmunology% 2010% 2*A '16-'21

D0. ;ielsen /N% 9Xi 4% 7hmidt 7% Qian :% 7himada T% 7hen GY% 4arsen G4. Bs Aeromonas

hydrophila the dominant motile Aeromonas speies that auses disease outbreas in

a8uaulture prodution in the he!iang rovine of $hina 0is A*uat Org % 2001% D'A26-

2&

D1. @u#ang /% $hen :Y% Yu M. 7tud# on biolog# of Megalobrama amblycephala in o#ang

4ae. Acta Agriculturae +iangxi% 2001% 16(1)AD,-+0

D2. @u#ang /. 7tud# on the ar#ot#pe of Megalobrama amblycephala from o#ang 4ae.

Acta Agriculturae +iangxi% 2000% 2(2)A'1-'D

D6. eters


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D*. 7ahoo J% Rauta R% /ohant# >R% /ahapatra J:% 7aha G;% R#e /% Nnath N.

7eletion for improved resistane to Aeromonas hydrophila in Bndian ma!or arp "abeo

rohitaA 7urvival and innate immune responses in first generation of resistant and

suseptible lines. 'ish ( Shell)ish &mmunology% 2011% 61AD62-D6*

D&. 7arderb /RB% Thompsona J:% enmana :G% /ndre"a >G. Bmmune responses of ;ile

tilapia (Oreochromis niloticus 4.) lonesA B. ;on-speifi responses. 0evelopmental and

Comparative &mmunology% 2001% 2+A6,-D'

+0. 7hen RG% Giang MY% u G?% ou 7/. 9BF-1and -2 genes in a h#poia-sensitive teleost

speies Megalobrama amblycephalaA :; loning% epression and different responses

to h#poia. Comparative ,iochemistry and %hysiology %art ,% 2010% 1+,A2,6-2*0

+1. 7hi ?4% 4iu /% 4u /Y% 9uang FQ. 7tudies on re8uirement of Megalobrama

amblycephala for several main inorgani salts. +ournal o) 'isheries o) China% 1&&,%

21(D)AD+*-D'1

+2. 7hi ?4% 7han G% 4iu /% Yan 9% 9uang FQ% hou M?% 7hen 4. stud# of the optimum

demand of protein b# blunt snout bream ( Megalobrama amblycephala). F@ librar#%

1&**% ession noA 2*&'11

+6. 7u G


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'0. Yin 9>% Fan T% 7un ?% an F% /eng FR. The Jar#ot#pe and :; ontent anal#sis of

blunt snout bream ( Megalobrama amblycephala). Chinese +ournal o) 'isheries% 1&&+%

*(1)A22-2'

'1. hang :. 7tud# on geneti diversit# of bluntnose bla bream from Yunihu and 4iangzi

laes. + China !hree 8orges -niv.% 2001% 6A2*2-2*D

'2. hang ream ( Megalobrama

amblycephala). Anat. Histol. #mbryol % 200&% 6*AD01-D0+

'6. hang 9% eatman N% 4iu 9% Feng TT% $hen 4Q% 4iu G. /oleular haraterization of

three 4-t#pe letin genes from hannel atfish% &ctalurus punctatus and their responses to

#d$ardsiella ictaluri hallenge. 'ish ( Shell)ish &mmunology% 2012% 62A +&*-'0*

'D. hang >% ;iu $G% 7torset % >Xg"ald G% :almo R. $omparison of Aeromonas

salmonicida resistant and suseptible salmon familiesA high immune response is

benefiial for the survival against Aeromonas salmonicida hallenge. 'ish ( Shell)ish

&mmunology % 2011% 61A1-&

'+. hou ?Y% Yu $Y% 4iu G% Yang


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10. P)*+*,(- B7-(

E>+(-7)(, A*7 7 R(,*,

TRBzol Reagent 6%000 R; etration

Turbo :;-freeT/ Jit 1%D00 Removing ontaminated :;

R;eas# /ini Jit 6%000 mR; purifiation

QB8ui $R

urifiation Jit1%000 :; purifiation

Trimmer-2 Jit +%000 :; normalization

Bon Total R;-7e8 Jit v2 1+%000 :; librar# onstrution

Bon @neTouhT/ 200

Template Jit

20%000 Nnrihment of :; librar#

Bon 7e8uening Jit 1+%000 7hotgun se8uening

Bon 61*T/ $hip 16%000 7e8uening 600 /b to 1


20/20

C*(, - R(c*(-* *; A-,*):

- The proposal ontents are realisti and should be studied

- The epeted results "ill provide more information for subse8uent studies on

diseases resistane seletion for blunt-snout bream families in future

A-,*) S7)(:

D(:

C*(, - )(c*(-* *; h( G7- G)*7+:

- The proposal plan "ill be performed basing on appl#ing ne" gene tehnologies.

- The proposal "ors an be arried out and ompleted.

- The proposal plan should be remained and performed.

L(-() *; G7- G)*7+ S7)(：

D(:

M(9(), *; G7- G)*7+

N( T4( Mangi rofessor 8uaulture$ollege of Fisheries%

9uazhong griultural Sniversit#

$hen Miaouan rofessor 8uaulture$ollege of Fisheries%

9uazhong griultural Sniversit#

Documents

Degree Research Plan and Proposal- Tran Ngoc Tuan(1) (1)