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草鱼7个插入/缺失型突变多态性及与幼鱼生长性状关联分析
引用本文:张猛,沈玉帮,徐晓雁,王荣泉,冷向军,李家乐.草鱼7个插入/缺失型突变多态性及与幼鱼生长性状关联分析[J].水产学报,2019,43(8):1706-1713.
作者姓名:张猛  沈玉帮  徐晓雁  王荣泉  冷向军  李家乐
作者单位:上海海洋大学农业农村部淡水水产种质资源重点实验室, 上海 201306,上海海洋大学农业农村部淡水水产种质资源重点实验室, 上海 201306;上海海洋大学水产养殖工程研究中心, 上海 201306;上海海洋大学水产科学国家级实验教学示范中心, 上海 201306,上海海洋大学农业农村部淡水水产种质资源重点实验室, 上海 201306;上海海洋大学水产养殖工程研究中心, 上海 201306;上海海洋大学水产科学国家级实验教学示范中心, 上海 201306,苏州市申航生态科技发展股份有限公司, 农业农村部大宗淡水鱼繁育与养殖技术重点实验室, 江苏 苏州 215225,上海海洋大学农业农村部淡水水产种质资源重点实验室, 上海 201306;上海海洋大学水产养殖工程研究中心, 上海 201306;上海海洋大学水产科学国家级实验教学示范中心, 上海 201306,上海海洋大学农业农村部淡水水产种质资源重点实验室, 上海 201306;上海海洋大学水产养殖工程研究中心, 上海 201306;上海海洋大学水产科学国家级实验教学示范中心, 上海 201306
基金项目:现代农业产业技术体系专项(CARS-46-04);上海市科委工程技术中心能力提升项目(16DZ2281200);国家自然科学基金青年科学基金(31802285)
摘    要:对草鱼生长性状进行数量性状基因座(quantitative trait locus,QTL)定位过程中,在15号连锁群中定位到一个与体质量相关的QTL,本研究根据已有的草鱼遗传连锁图谱和基因组序列,拟用短片段重复序列(short tandem repeat,STR)分型技术对该连锁群的3个scaffolds中插入/缺失型突变位点进行筛选,以降低分型成本,同时将筛选出的7个多态性位点与草鱼幼鱼生长性状进行关联分析。结果显示,(1)草鱼3个scaffolds中44个插入/缺失型突变位点中有17个简单重复序列(又称微卫星,simple sequence repeats,SSRs),2个位点引物设计不成功,设计的25对引物中仅22对扩增和分型成功;(2) 22个位点中仅有7对引物在4个亲本中存在多态性,直接测序结果发现,STR分型技术不仅可准确对插入/缺失型突变位点进行分型,同时可降低分型成本;(3)将7个插入/缺失型突变位点与323尾选育F2草鱼的生长性状进行关联分析发现,除了位点ID-10H和ID-41F以外,其余5个位点都与草鱼幼鱼的一个或多个生长性状显著相关,其中ID-6H与幼鱼肥满度性状显著相关,位点ID-11F、ID-15F、ID-32F和ID-39F分别与草鱼幼鱼体质量、体长、体宽和体高性状显著相关,可将以上5个与草鱼幼鱼生长性状相关的突变位点用于草鱼生长性状QTL加密和分子标记辅助育种。

关 键 词:草鱼  突变  多态性  生长  关联分析
收稿时间:2018/9/26 0:00:00
修稿时间:2018/11/28 0:00:00

Polymorphism of seven insert/deletion mutations and their connection with growth traits of juvenile grass carp (Ctenopharyngodon idella)
ZHANG Meng,SHEN Yubang,XU Xiaoyan,WANG Rongquan,LENG Xiangjun and LI Jiale.Polymorphism of seven insert/deletion mutations and their connection with growth traits of juvenile grass carp (Ctenopharyngodon idella)[J].Journal of Fisheries of China,2019,43(8):1706-1713.
Authors:ZHANG Meng  SHEN Yubang  XU Xiaoyan  WANG Rongquan  LENG Xiangjun and LI Jiale
Institution:Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China,Key Laboratory of Conventional Freshwater Fish Breeding and Health Culture Technology Germplasm Resources, Ministry of Agriculture and Rural Affairs, Suzhou Shenhang Eco-technology Development Limited Company, Suzhou 215225, China,Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China and Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China
Abstract:A weight-related QTL was found in the 15 linkage group (LG) during mapping the growth traits of grass carp (Ctenopharyngodon idella). And we tried to screen insert/deletion mutations on three scaffolds in the 15 LG, according to the previous consensus linkage map and draft genome of C. idella. At the meantime, the STR genotyping method was uesed in genotyping of insert/deletion mutations. Moreover, the association analysis of selected mutations and growth traits of 323 individuals was conducted. Results found that 17 of the 44 selected insert/deletion mutations were SSRs, and 2 loci failed to design primers; therefore, 25 pairs of primer were successfully designed. PCR amplifications showed that only 22 of the 25 primer pairs were effective. Then STR genotyping method was used for genotyping of those 22 insert/deletion mutations, and 7 polymorphic mutations were found in four parents. Afterwards, PCR products were sequenced to confirm the STR genotyping results. Sequencing results showed that STR genotyping method could not only classify insertion/deletion mutants accurately, but also reduce the genotyping costs. Finally, the general linear model was used to analyze the relevance between those 7 insert/deletion mutations and growth traits of 323 juvenile C. idella. Correlation results found that 2 loci (ID-10H and ID-41F) were not related with growth traits, while the other 5 loci were significantly related with one or more growth traits of juvenile C. idella. According to the 5 loci, locus ID-6H was significantly correlated with fatness traits of juvenile C. idella, while locus ID-11F, ID-15F, ID-32F and ID-39F were significantly correlated with body weight or length or width or height. In summary, the inexpensive STR genotyping method could be used in the development of insert/deletion mutations, and the 5 growth related mutations could be used in QTL density-increasing and molecular marker assisted breeding of C. idella.
Keywords:Ctenopharyngodon idella  mutation  polymorphism  growth traits  connection
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