罗非鱼性别相关微卫星标记的初步筛选  

Preliminary screening of sex-related microsatellite markers in Oreochromis niloticus and O.aureus

作　　者：陈文伟 曹建萌[1] 刘志刚[1] 杨娜 卢迈新[1] CHEN Wen-wei;CAO Jian-meng;LIU Zhi-gang;YANG Na;LU Mai-xin(Key Lab of Tropical&Subtropical Fishery Resource Application&Cultivation,Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China;College of Fisheries&Life,Shanghai Ocean University,Shanghai 201306,China)

机构地区：[1]中国水产科学研究院珠江水产研究所,农业农村部热带亚热带水产资源利用与养殖重点实验室,广州510380 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《淡水渔业》2020年第1期22-30,共9页Freshwater Fisheries

基　　金：国家重点研发计划(2018YFD0901201);现代农业产业技术体系(CARS-46);国家自然科学基金(31502205);中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2017HY-ZC0601,2019ZX-003)

摘　　要：为分析罗非鱼群体的遗传多样性以及筛选与罗非鱼性别相关的微卫星标记。应用24对微卫星引物,使用常规PCR及聚丙烯酰胺凝胶电泳的方法在两个尼罗罗非鱼(Oreochromis niloticus)群体和两个奥利亚罗非鱼(O.aureus)群体中初步筛选到UNH931、GM128、GM201、GM258、GM597以及UNH898共6个与性别相关的微卫星标记。然后使用降落PCR以及毛细管电泳的方法在两个尼罗罗非鱼群体、两个奥利亚罗非鱼群体以及ZY 1、WY 1、YY 1和YY 2型罗非鱼群体中进一步扩增这6个微卫星标记,统计各群体的遗传多样性参数:6个微卫星标记在上述群体共297个样本中检测到95个等位基因,其大小在97~302 bp之间,各位点在各群体等位基因数在1~13个之间,各群体平均观测等位基因数为2.167~9.333,平均有效等位基因数为1.624~4.966,平均观测杂合度为0.324~0.983,平均期望杂合度为0.329~0.782,平均多态信息含量为0.275~0.753;两个尼罗罗非鱼群体、WY 1和YY 2群体达到高度多态(PIC>0.5),两个奥利亚罗非鱼群体、ZY 1和YY 1群体为中度多态(0.25<PIC<0.5)。聚类分析显示所有群体聚为两大支,两个奥利亚罗非鱼群体以及ZY 1群体聚为一支,两个尼罗罗非鱼群体、WY 1、YY 1以及YY 2群体聚为另一支。进行性别关联分析发现:GM597与无锡尼罗罗非鱼群体、高要尼罗罗非鱼群体以及番禺奥利亚罗非鱼群体的性别显著相关,UNH898与高要尼罗罗非鱼群体的性别极显著相关。利用GM597与UNH898联合分析可100%鉴定两个尼罗罗非鱼群体的性别。利用UNH898可以100%将YY 1群体与两个尼罗罗非鱼群体区分开。A total of 24 pairs of microsatellite primers were used to screen sex-related microsatellite markers in two Oreochromis niloticus populations and two O.aureus populations by conventional PCR and polyacrylamide gel electrophoresis to analyze the genetic diversity of tilapia and to screen microsatellite markers related to tilapia sex.Six microsatellite markers,including UNH931,GM128,GM201,GM258,GM597 and UNH898 were identified related to tilapia sex.These six microsatellite markers were then amplified in the two O.niloticus populations,two O.aureus populations and the ZY 1,WY 1,YY 1,YY 2 tilapia populations using touchdown PCR and capillary electrophoresis.The genetic diversity parameters of each population were counted at the same time.A total of 95 alleles in 297 samples from the above populations were detected by six microsatellite markers,ranging in size from 97 to 302 bp.The number of alleles in each population ranged from 1 to 13,the average number of observed alleles was 2.167-9.333,the average effective allele number was 1.624-4.966,the average observed heterozygosity was 0.324-0.983,the average expected heterozygosity was 0.329-0.782,and the average polymorphic information content was 0.275-0.753.The two O.niloticus populations of WY 1 and YY 2 reached a high polymorphism(PIC>0.5),and the two O.aureus populations of ZY 1 and YY 1 were moderately polymorphic(0.25<PIC<0.5).Cluster analysis showed that all groups were clustered into two branches.Two O.aureus populations and ZY 1 population were clustered together.Two O.niloticus populations of WY 1,YY 1 and YY 2 were clustered into another branch.Sex association analysis revealed that GM597 was significantly associated with the sex of Wuxi O.niloticus population,Gaoyao O.niloticus population and Panyu O.aureus population(P<0.05).UNH898 was significantly correlated with the sex of the Gaoyao O.niloticus population(P<0.01).The combined analysis of GM597 and UNH898 can absolutely identify the sex of two O.niloticus populations.As conclusions,YY 1 population can

关 键 词：尼罗罗非鱼(Oreochromis niloticus) 奥利亚罗非鱼(O.aureus) 微卫星 性别 遗传多样性 

分 类 号：S917.4[农业科学—水产科学]