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中国鲎(Tachypleus tridentatus)微卫星标记筛选及种群遗传多样性和遗传结构分析
翁朝红, 谢仰杰, 肖志群, 王志勇
集美大学水产学院 农业农村部东海海水健康养殖重点实验室 厦门 361021
摘要:
采用FIASCO方法构建了中国鲎(Tachypleus tridentatus)(CA)n微卫星富集文库,对214个阳性克隆进行测序,筛选获得60条含有微卫星的序列,其中可设计引物的序列共38条。38对微卫星引物在37个中国鲎个体中PCR检测发现,只有9个位点具有多态性,其等位基因数为5-14个,每个位点平均具有8.1个等位基因。9个位点之间不存在连锁。利用该9个具有多态性微卫星标记分析了中国沿海9个中国鲎地理群体的种群遗传多样性和遗传结构。结果表明:中国沿海的中国鲎种群遗传多态性水平仍较高,9个地方群体间无遗传差异,它们之间未见显著分化,且有着较高的基因流;推测人为的迁移是造成遗传无分化的主要原因。我们建议中国鲎已绝迹的海域可从其他海域引入中国鲎个体来恢复地方群体资源。
关键词:  中国鲎  微卫星标记  FIASCO  遗传多样性  遗传结构
DOI:10.11693/hyhz20191100204
分类号:Q958.2
基金项目:国家水产种质资源平台项目,2019DKA30470号。
ISOLATION OF MICROSATELLITE MARKERS IN TACHYPLEUS TRIDENTATUS AND ANALYSIS OF THEIR GENETIC POLYMORPHISM AND GENETIC STRUCTURE
WENG Zhao-Hong, XIE Yang-Jie, XIAO Zhi-Qun, WANG Zhi-Yong
Fisheries College, Jimei University, Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Xiamen 361021, China
Abstract:
Microsatellite-enriched libraries of (CA)n in Chinese horseshoe crab (Tachypleus tridentatus) were constructed using FIASCO (Fast Isolation by AFLP of Sequences Containing repeats) method, from which 214 positive clones were selected to sequence. After screening, 60 sequences containing microsatellite loci were obtained, in which 38 sequences could be used to design primers. Thirty eight pairs of primers were used to PCR in 37 horseshoe crabs. Results show that only nine microsatellite loci were polymorphism with the allele number ranging from 5 to 14, in average of 8.1. There was no genetic linkage between the nine loci. The genetic diversity and structure were analyzed using these polymorphic microsatellite markers. The results indicate that the polymorphism of the nine Chinese horseshoe crab populations along the southeast coast of China remained high, and there was no genetic difference among nice local populations of Chinese horseshoe crab. Therefore, no significant differentiation was found in high gene flow among the populations. It was speculated that human migration was the main cause of non-genetic differentiation. To restore the local population resources, the horseshoe crabs should be introduced to those sea areas where the horseshoe crab has been extinct.
Key words:  Chinese horseshoe crab Tachypleus tridentatus  microsatellite markers  FIASCO  genetic polymorphism  genetic structure
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