首页 >  水产科学 >  基于COI序列绒螯蟹属DNA条形码和遗传多样性研究

2017, 36(4): 480-487. doi: 10.16378/j.cnki.1003-1111.2017.04.013

基于COI序列绒螯蟹属DNA条形码和遗传多样性研究

1. 天津农学院 水产学院, 天津市水产生态及养殖重点实验室, 天津 300384;

2. 天津市水生动物疫病预防控制中心, 天津 300402

收稿日期:2016-05-31
修回日期:2016-08-19

基金项目:   天津市水产局科技发展计划项目(J2013-21). 

关键词: 绒螯蟹 , COI基因 , DNA条形码 , 物种鉴定 , 遗传多样性

DNA Barcoding and Genetic Diversity in Genus Eriocheir Based on COI Gene Sequences

1. Tianjin Key Laboratory of Aqua-Ecology & Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300384, China;

2. Tianjin Diseases Prevention and Control Centre of Aquatic Animals, Tianjin 300221, China

Received Date:2016-05-31
Accepted Date:2016-08-19

Keywords: Eriocheir , COI gene , DNA barcode , species identification , genetic diversity

对绒螯蟹属的中华绒螯蟹如东和七里海群体、日本绒螯蟹、合浦绒螯蟹及狭颚绒螯蟹共80条线粒体COI片段进行扩增和测序,并与GenBank中绒螯蟹属的台湾绒螯蟹2条和近方蟹属的绒毛近方蟹19条COI 基因序列进行联配分析。结果显示,101条序列包含44种单倍型,序列组成表现明显的碱基偏倚性。中华绒螯蟹如东、七里海群体与日本绒螯蟹间的遗传距离分别为1.210%和1.078%,明显低于COI基因DNA条形码鉴别种的遗传距离为2%的阈值,表明中华绒螯蟹和日本绒螯蟹为同一物种;而合浦绒螯蟹与中华绒螯蟹如东和七里海群体及与日本绒螯蟹的遗传距离分别为4.823%、5.101%以及5.011%,明显大于2%的鉴别阈值,说明合浦绒螯蟹为独立的种。以绒毛近方蟹为外群,基于群体内及群体间的遗传距离构建的邻接树显示,中华绒螯蟹与日本绒螯蟹聚在一起,合浦绒螯蟹则聚成单系。本文测序的5个群体除狭颚绒螯蟹外,其余均具有遗传多样性,单倍型多样性为0.593±0.144~0.779±0.068,核苷酸多样性为0.00156~0.01336;此外,中华绒螯蟹如东群体与日本绒螯蟹、合浦绒螯蟹和中华绒螯蟹七里海群体分别共享单倍型H1、H2和H3,说明这些蟹类可能有种质资源混杂或是遗传污染的现象。

A total of 80 mitochondrial COI gene fragments from five Eriocheir populations including E. sinensis Rudong and Qilihai populations, E. japonica,E. hepuensis and E. leptognathus were amplified and sequenced, and then the 80 sequences were compared with that of 2 COI gene sequences of E. formosa and 19 COI sequences of Hemigrapsus penicillatus recorded in GenBank. In this study, 44 haplotypes were detected from 101 individuals, and the base composition of sequences showed a significant bias. Based on the analysis of COI gene sequences, the K2P distances between E. sinensis Rudong population, Qilihai population and E. japonica, being 1.210% and 1.078%, respectively, were significant lower than 2%, which is the threshold of the DNA barcoding species identification, indicating that E. sinensis and E. japonica were the same species. However, the K2P distances between E. hepuensis and the two E. sinensis populations as well as E. japonica were 4.823%, 5.101% and 5.011%, respectively, which were far beyond 2%, indicating that E. hepuensis was an independent species. Using H. penicillatus as out group, based on K2P distances, neighbor-joining (NJ) method was employed for phylogenetic analysis. NJ trees revealed that E. sinensis and E. japonica did not form monophyletic clades, but disturbed, disorderly, whereas E. hepuensis was clustered into a single branch. Five populations sequenced in our study, with the exception of E. leptognathus, the rest 4 populations had genetic diversity, the haplotype diversity ranged from 0.593±0.144 to 0.779±0.068 and the nucleotide diversity ranged from 0.00156 to 0.01336. The E. sinensis Rudong population shared the haplotypes H1, H2 and H3 with E. japonica,E. hepuensis and E. sinensis Qilihai population, respectively, which suggested that the germplasm resources of the economic crabs were mixed or the phenomenon might be due to the genetic pollution.

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基于COI序列绒螯蟹属DNA条形码和遗传多样性研究