基于CO Ⅰ的中国西施舌DNA条形码

以我国大连(DL)、日照(RZ)、启东(QD)、长乐(CL)、漳州(ZZ)和北海(BH)6个野生群体西施舌为研究对象,用引物LCO1490和HCO2198扩增线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因片段序列,PCR产物双向测序,用ClustalX进行DNA序列比对,用MEGA4.1和PhyML软件构建NJ树和ML树,...

DNA Barcodes of Clam Coelomactata antiquata (Bivalvia: Veneroida)in China Based on CO Ⅰ Gene

The population identification barcodes were constructed in six wild stocks of clam Coelomactata antiquata collected from coasts in Dalian in Liaoning(DL),Rizhao in Shandong(RZ),Qidong in Jiangsu(QD),Changle(CL) and Zhangzhou(ZZ) in Fujian,Beihai in Guangxi(BH) provinces in China based on COⅠ nucleotide sequences by PCR amplification with the primers LCO1490(5′-GGT CAA CAA ATC ATA AAG ATA TTG G-3′)and HCO2198(5′-TAA ACT TCA GGG TGA CCA AAA AAT CA-3′).The PCR products were sequenced directly using DNAStar,and aligned in ClustalX.The phylogenetic trees were constructed using Neighbor-Joining method(on MEGA 4.1) and Maximum likelihood method(PHYML) and Analysis of molecular variance(AMOVA) was performed by Arlequin3.11.The COⅠ fregments were found 658 bp in size,and the COⅠ alignmet for 64 individuals contained 658 bp with 101 variable sites(15.3%),with 91 parsimony informative sites(14.1%).The intrastock genetic distance was found to be 0.0020～0.0043 whereas interstock genetic distance 0.0024～0.0883.Amino acid sequence variations were not observed.Further,AMOVA analysis revealed a substantial genetic differentiation between the group DH(included the stocks of CL,ZZ)and group HB(included the stocks of DL,RZ,QD and BH)(FST =0.956～0.970,P﹤0.01).NJ and ML trees all showed that a monophyletic clade was observed in the group DH whereas another separate clade was found in the group HB.There was genetic distance of 0.0837 between the group DL and group BH,with 17.8～23.5 folds in the interstocks.The genetic structure and phylogenetic relationship among six wild stocks of C.antiquata in typical coastal regions provides the molecular basis of DNA barcodes for the identification of CL and ZZ(Fujian province) C.antiquata.