浙江近海三疣梭子蟹4个群体的控制区序列分析

以浙江近海三疣梭子蟹大螯肌肉的线粒体DNA作为模板，对4群体共67个个体的控制区（D-loop）序列进行了PCR扩增。通过对PCR产物进行双向测序，最终得到了控制区3’端的rRNA部分序列（64 bp）、D-loop全序列（1179 bp）及控制区5’端tRNA-Ile部分序列（73 bp）。经DNASP软件分析得知，67个D-loop序列定义了65个单倍型，在65个单倍型中，除去插入/缺失位点，所有单倍型共检测到344个多态位点，主要发生在D-loop序列的中间区域；D-loop序列检测到26个插入或缺失位点，碱基插入主要发生在中央后端区域，以13 bp重复片断插入的形式进行。每个个体含有0～3个连续重复片断。通过对三疣梭子蟹的遗传多样性分析发现，D-loop序列的单倍型多样性为0.982～1.000，核苷酸多样性指数为0.02770～0.03633，平均核苷酸差异数为31.790～43.304。综合分析，三个成蟹群体的遗传多样性相近，都低于仔蟹的遗传多样性，说明经过遗传育种，可以提高三疣梭子蟹的种质情况，真正实现对三疣梭子蟹的科学利用。

Genetic Diversity of Portunus trituberculatus in the Offing of Zhejiang Revealed by D-loop Sequence

Mitochondrial DNA （mt DNA） of Portunus trituberculatus was prepared as templates from their muscle. The PCR technique was used to amplify the D-loop sequence in 67 individuals of 4 population. The PCR products were sequenced and 64 bp rRNA sequence of 3’end and 1 179 bp of complete D-loop sequences and 73 bp tRNA-Ile of 5’end were obtained （Remove partial sequences of the two ends）. By using DNASP soft-ware to analyze the genetic diversity, the result showed that there were 65 haplotypes and 334 variation sites were observed （no inserting/missing sites）. The variation sites occurred in the middle of the D-loop sequences. 26 inserting/missing sites were observed in the end of the D-loop sequences too. Results revealed a 13 bp length repetitive sequence in the sequences and individuals differed from each other in frequency and length of the repetitive sequences. As estimated by DNASP, the haplotypic diversity region was 0.982-1.000, the nu-cleotide diversity and the average number of nucleotide defferences were 0.027 70-0.036 33 in D-loop se-quences. It can be concluded that the genetic diversity of P. trituberculatus in the offing of Zhejiang was rich.