东海野生灰鲳mtDNA D-loop序列遗传变异分析

以东海区野生灰鲳背部肌肉的线粒体DNA为模板,采用PCR技术对7个个体的D-loop序列进行了扩增,通过对PCR产物进行双向测序,最终得到了471 bp的核苷酸片断（除去两端部分序列）。用DNAMAN6.0软件进行了排序比较发现,东海野生灰鲳的D-loop序列同源性高达99.48%;用DNASP4.0软件分析得知,7个序列共有6个单倍型（在GenBank登录号：GU970085-GU970087,GU970089-GU970091）,在7个个体中,共检测到14个变异位点,包括13个转换和1个颠换位点。运用MEGA4.0软件计算出了不同个体间的遗传距离,并据此构建了MP和NJ系统树。用DNASP4.0软件计算出的多态位点数为14,核苷酸多样性指数和平均核苷酸差异数分别为0.009 71和4.571。研究结果表明,东海野生灰鲳的D-loop序列个体变异程度并不丰富。

Genetic Diversity of Pampus cinereus in the East China Sea Revealed by mtDNA D-loop Sequence

Mitochondrial DNA（mt DNA） of Pampus cinereus was prepared as templates from their back muscle.The PCR technique was used to amplify the mtDNA D-loop sequence in 7 individuals of P.cinereus collected from the Yangtze River estuary fishing ground and Zhouwai fishing ground in the East China Sea.The PCR products were sequenced and 471 bp segment at the 5＇ end of control region were obtained（Remove partial sequences of the two ends）.By using DNAMAN6.0 software to align and compare the sequences of these 7 individuals with each other,the identity of D-loop pairwise sequences was 99.48 % and by using DNASP4.0 to analyze the genetic diversity.The result showed that there were 6 haplotypes（GenBank accession no.GU970085-GU970087,GU970089-GU970091） and 14 variation sites were observed,of which there were 13 transition sites and 1 transversion sites.The pairwise genetic distances were computed by MEGA4.0 software.The MP and NJ phylogenetic trees were obtained through the cluster analysis over the pairwise genetic distance.As estimated by DNASP4.0,the number of polymorphic sites was 14,the nucleotide diversity and the average number of nucleotide defferences were 0.009 71 and 4.571,respectively.It can be concluded that the genetic diversity in D-loop sequence of P.cinereus in the East China Sea was not relatively rich.The results of this study proviede a basic molecular biology clue to the studies on population genetics and population structure.