基于微卫星DNA标记的山东微山湖区放养钉螺遗传结构分析

中国寄生虫学与寄生虫病杂志

基于微卫星DNA标记的山东微山湖区放养钉螺遗传结构分析

何健1，缪峰2，杨坤1，赵长磊2，刘新2*

1 江苏省血吸虫病防治研究所，国家卫生计生委寄生虫病预防与控制技术重点实验室，江苏省寄生虫与媒介控制技术重点实验室，无锡 214064；2 山东省医学科学院，山东省寄生虫病防治研究所，济宁 272033

出版日期:2016-06-30 发布日期:2016-10-28

Genetic Variation of Oncomelania hupensis in Weishan Lake of Shandong Province Using Microsatellite DNA Markers

HE Jian1, MIAO Feng2, YANG Kun1, ZHAO Chang-lei2, LIU Xin2*

1 Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases, Wuxi 214064, China；2 Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases, Jining 272033, China

Online:2016-06-30 Published:2016-10-28

摘要/Abstract

摘要： 目的分析在山东微山湖地区长期放养的湖北钉螺（Oncomelania hupensis）群体与其来源地江苏省长江江滩钉螺群体之间的基因差异。方法收集江苏省扬州市江都区、仪征市和丹阳市、南京市六合区的长江钉螺以及山东省济宁市微山湖独山岛的钉螺。微山湖区放养的钉螺来自于扬州市江都区，放养时间为10年（至2014年）。选择A18、C22、T4-33、T6-17等4对微卫星位点对钉螺群体进行PCR扩增，分析各钉螺群体的等位基因数（Na）、期望杂合度（He）、观察杂合度（Ho）和群体间遗传差异平均数等指标，并进行分子变异方差分析（AMOVA），用非加权组平均法（UPGMA）和邻接法（NJ）构建系统进化树描述群体遗传分化及聚类情况。使用Mantel检验探讨遗传距离与地理距离的相关性。结果本研究收集5个钉螺群体共103只钉螺。江都、仪征、丹阳、六合、微山湖群体的Na分别为7.50、12.50、10.00、11.50和12.75；Ho分别为0.16、0.27、0.17、0.30和0.22；He分别为0.81、0.91、0.84、0.90和0.92。微山湖群体的3个遗传多样性指标均在较高水平，提示有较好的遗传多样性，但与其他群体差异无统计学意义（P>0.05）。微山湖与江都钉螺群体间遗传差异平均数最小（0.79），丹阳群体与其他群体之间差异较大，为0.87～0.97。AMOVA分析的遗传变异主要来源于钉螺个体间（占92.50%）。NJ和UPGMA系统进化树显示，江都和微山湖的钉螺群体聚为一支，仪征和六合的钉螺群体聚为一支，丹阳钉螺群体与其他4个群体聚为一支。Mantel检验显示遗传距离与地理距离没有相关性。结论江苏省长江钉螺群体与微山湖钉螺群体基因多样性均较高，微山湖地区放养的钉螺与来源地的长江钉螺群体尚未出现明显遗传分化。

关键词: 微卫星DNA, 湖北钉螺, 微山湖

Abstract: Objective To identify the genetic variation of Oncomelania hupensis between Weishan Lake population and Yangtze River population. Methods O. hupensis snails were collected from Jiangdu District of Yangzhou City, Yizheng City, Danyang City, and Liuhe District of Nanjing City in Jiangsu Province, as well as Weishan Lake of Jining City in Shandong Province. Four polymorphic microsatellite DNA loci（A18, C22, T4-33 and T6-17） were amplified by using microsatellite PCR to analyze the number of alleles （Na）, the expected heterozygosity （He）, the observed heterozygosity （Ho） and the average number of pairwise differences between populations. Analyses of molecular variance（AMOVA） and cluster analysis were performed to find the genetic variation between O. hupensis snail populations. Unweighted pair group method with arithmetic mean （UPGMA） and neighbour joining（NJ） method were used to construct the phylogenetic tree to describe genetic variation and clustering among populations. Mantel test was carried out to detect the correlation between the genetic distance and geographical distance. Results A total of 103 snails were collected. The snail populations of Jiangdu, Yizheng, Danyang, Liuhe, and Weishan Lake had Na of 7.50, 12.50, 10.00, 11.50; Ho of 0.16, 0.27, 0.17, 0.30, 0.22; and He of 0.81, 0.91, 0.84, 0.90, 0.92, respectively. The Weishan Lake population showed relatively higher levels of the three indicators, indicating a higher genetic diversity, but with no significant difference with the other four populations. The average number of pairwise differences was lowest between the Weishan Lake and Jiangdu populations（0.79）, and it was higher between Danyang population and the others（0.87-0.97）. AMOVA result showed the inter-individual genetic variation accounted for 92.50% of the total variations. NJ and UPGMA analysis showed Jiangdu and Weishan Lake populations gathered together in one branch, Yizheng and Liuhe populations formed another relatively independent branch, and then was Danyang population. Genetic distance and geographical distance showed no correlation by Mantel test. Conclusion There is a high genetic diversity of O. hupensis in Weishan Lake and Yangtze River populations, and no significant genetic divergence between Weishan Lake snail population and its original Yangtze River population.

Key words: Microsatellite DNA, Oncomelania hupensis, Weishan Lake region

何健1，缪峰2，杨坤1，赵长磊2，刘新2*. 基于微卫星DNA标记的山东微山湖区放养钉螺遗传结构分析[J]. 中国寄生虫学与寄生虫病杂志.

HE Jian1, MIAO Feng2, YANG Kun1, ZHAO Chang-lei2, LIU Xin2*. Genetic Variation of Oncomelania hupensis in Weishan Lake of Shandong Province Using Microsatellite DNA Markers[J]. Chinese Journal of Parasitology and Parasitic Diseases.

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