Studies on the Genetic Diversity of Farmed and Wild Populations of Coilia nasus by Analysing Mitochondrial DNA Cyt b genes

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1 - http / /xuebao. jxau. edu. cn Acta Agriculturae Universitatis Jiangxiensis E - mail ndxb7775@ sina. com mtdna Cytb *.. / Coilia nasus DNA b F DNA b bp h. 9 S 9 9 π. K. 7. b S95. 7 A Studies on the Genetic Diversity of Farmed and Wild Populations of Coilia nasus by Analysing Mitochondrial DNA Cyt b genes WEI Guang-lian XU Gang-chun GU Ruo-bo LI Jian-lin XU Pao *. College of Fisheries Nanjing Agriculture University Wuxi China. Key Open Laboratory of Genetic Breeding and Aquaculture Biology of Freshwater Fishes Ministry of Agriculture Freshwater Fisheries Research Center Chinese Academy of Fishery Sciences Wuxi China Abstract To provide a system analysis of differences in genetic diversity and scientific evidences of conservation of the germ plasm resources and industrialized artifical culture of Coilia nasus the genetic diversity and genetic structure of Coilia nasus were assessed from three populations by using mitochondrial Cyt b sequences. The complete mitochondrial Cyt b gene sequence is bp. Twelve haplotypes were detected from the individuals and the haplotype diversity h was. 9. And variation sites were defined of which 9 were singleton polymorphic sites and the other 9 were parsimony informative polymorphic sites. The nucleotide diversity π was. the average nucleotide difference K was. 7 and the average genetic distance was.. The result showed that genetic parameters and heredity diversities of farmed Coilia nasus BADB5 JBFA7 5 DZ- BE 97 E - mail w59577@. com * E - mail xup@ ffrc. cn

2 mtdna Cytb 7 and wild Coilia nasus were higher than those of Coilia nasus taihuensis which meant farmed Coilia nasus populations still had a high genetic diversity. Key words Coilia nasus farmed population wild population Cyt b genetic diversity Coilia nasus Clupeiformes Engraulidae Coilia b cytochrome b Cyt b mtdna 7-9 b.. ~. g. 9 g. ~. g. g 7 9. ~. g. g eppendorf -. DNA. 5 ml eppendorf 7 μl SET. 5 μl % SDS μl mg /ml K 55 / DNA TE DNA DNA DNA DNA 5 ng /μl. PCR L7 H595 Cyt b L7 5'-GACTT- GAAAAACCACCGTTG-' H595 5'-CTCCGATCTCCGGATTACAAGAC-' 5 μl DNA 5 ~ ng reaction buffer. 5 μl μmol /L dntp. μmol / L Taq U PCR 9 min 9 s 5 min 7 min 5 7 min PCR g /L 5'-GCGGATTCTCCGTAGACAAC-' 5'-GTAAAATTGTCTGGGTCTCC-'

3 . Clustal W. DnaSP version. S h π K MEGA version. Kimura NJ Neighbor - joining Bootstrap. Cyt b PCR bp PCR GenBank mtdna Cyt b NC _ 9579 Clastal X 99% M DL mtdna Cyt b Cyt b PCR Fig. Electrophoresis map of Cyt b gene PCR products farmed C. nasus Cyt b bp b A C G T. % 7. 9%. % 7. % A T 5. 5% C + G. 5% b A C G T G Population Cyt b Tab. The Nucleotide Frequencies of Cyt b gene from three populations % Base A G C T migratory C. nasus farmed C. nasus C. nasus taihuensis S. 5% 9 9 h. 9 π. K. 7 Kumar. ~ Cyt b Kumar. Engraulis japonicus Engraulis encrasicolus mtdna Cytb HM_9 NC_9579 MEGA. bootstrap analysis BP

4 mtdna Cytb 9 Tab. DNA Cyt b Variation sites of Cyt b gene of mitochondrial DNA from three populations Variationsite C C G G A T C G G A G A G A T T T G cy9 T... G. T A.. A. T G.... cy T... G. T G.... cy7 T... G. T G.... cy T... G. T G.... cy7 T... G. T G.... cy T.. A G. T A.. A.. G.... cy T.. A G C T A.. A.. G.... cy T T.. G. T G... T cy T... G. T. T.... G.... cy T.. A G. T A..... G.... cy5 T.. A G. T A..... G.... hj..... C hj..... C... G hj hj C... G hj7.. A hj..... C... G hj C ys C ys T ys..... C ys..... C A A A. cy hj ys cy farmed C. nasus hj C. nasus taihuensis ys migratory C. nasus. Tab. Parameter Populational genetic diversity in three populations Migratory C. nasus Farmed C. nasus C. nasus taihuensis Numberofsamples n 7 Numberofhaplotype h / Numberofvariablesites S singleton polymorphic sites / parsimony informative polymorphic sites. 5 5 / 5 / / Haplotypediversity Hd... Nucleotidediversity π... Average nucleotide difference K Kumar Kumar genetic distance. ~.. ~.. ~.

5 Tab. The average genetic distance among three populations DNA DNA Population Farmed C. nasus C. nasus taihuensis C. nasus taihuensis. Migratory C. nasus Cyt b DNA Cyt b b bp S. 5% π cy hj ys fj cy farmed C. nasus hj C. nasus taihuensis ys migratory C. nasus fj Coilia mystus. 5 Cyt b Fig. Molecular phylogenetic tree based on the Cyt b sequences construced by NJ method π... Founder effect F F Billington % Cyt b ~. % % %. 7%. %

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