半滑舌鳎抗哈维氏弧菌病相关微卫星标记筛选及QTL定位  

Detection of SSR and a QTL analysis of Vibrio harveyi resistance in Cynoglossus semilaevis

作　　者：戴欢[1,2,3] 刘洋[2,3] 王文文 位战飞[2,3] 高进 高峰涛[2,3] 李祥孔[1,2,3] 刘洋 陈松林[2,3] 

机构地区：[1]大连海洋大学水产与生命学院,辽宁大连116023 [2]农业部海洋渔业可持续发展重点实验室、中国水产科学研究院黄海水产研究所,山东青岛266071 [3]青岛海洋科学与技术国家实验室、海洋渔业科学与食物产出过程功能实验室,山东青岛266237

出　　处：《中国水产科学》2017年第1期22-30,共9页Journal of Fishery Sciences of China

基　　金：国家自然科学基金项目(31530078);山东省农业良种工程重大课题;山东省泰山学者攀登计划专项

摘　　要：为了筛选出与抗哈维氏弧菌病相关的微卫星标记并进行QTL定位,以2014年感染实验中存活率为52.22%的半滑舌鳎(Cynoglossus semilaevis)家系F1412(Family+年份+家系号)为材料,采用混合分离子分析法(bulked segregant analysis,BSA)对169个微卫星标记(SSR)进行筛选,得到1个可能与哈维氏弧菌病相关的微卫星标记scaffold479_23523。用scaffold479_23523所在的连锁群LG18上的37个SSR对94尾抗病、感病个体进行基因分型,得到3个图谱:整合图谱(LG18)、雌性图谱(LG18F)和雄性图谱(LG18M),并用两种不同的模式进行单标记分析和QTL定位。模式一得到3个显著性相关标记(P<0.05)和1个极显著性相关标记(P<0.01),图谱LG18F定位出一个区间qE-F1;模式二得到4个显著相关标记(P<0.05)和1个极显著相关标记(P<0.01),图谱LG18M定位出两个区间qE-M1、qE-M2。在全基因组序列的分布范围上,区间qE-F1包含了qE-M1及qE-M2,因此qE-F1更可能与抗病性状相关。本研究开发出了抗哈维氏弧菌病性状相关微卫星标记及QTL区间,为抗病新品种的培育奠定了基础。The bacterium Vibrio harveyi has caused tremendous losses in the Cynoglossus semilaevis aquaculture industry. The fundamental approach has been to cultivate a new disease-resistant strain by combining traditional breeding methods with molecular techniques. In this study, bulked segregant analysis and quantitative trait loci (QTL) mapping were used to screen for disease-resistance markers. A total of 100 individuals were selected to form the F1412 family (nomenclature rule:F+year+family number:survival rate, 52.22%), which was challenged with V. harveyi, and 169 microsatellite loci were detected across all chromosomes. Following the genomic scan, the scaffold479_23523 marker in the DNA pool was significantly different between the resistant and susceptible groups (P=0.000006). Ninety-four individuals were genotyped using all 32 simple sequence repeat markers on LG18, where scaffold479_23523 was located. Three new linkage groups (LG18, LG18F, and LG18M) were identified. Furthermore, two different analytical models were applied to perform a single marker analysis and composite interval mapping with different levels of significance in LG18, LG18F, and LG18M, respectively. In model 1, three significant markers (scaffold4475_71287, scx9-1, and cyse80) and one very significant marker (scaffold080437) were identified, and the qE-F1 resistance-related QTL was detected. The scaffold479_23523 marker was the left LG18F marker with a p-value of 0.0516 in model 1. Model 2 detected four significant markers (hncyse110, scaffold414_19940, scaffold4475_71287, and cyse80), one very significant marker (scaffold08043), and the qE-M1 and qE-M2 QTLs. Both scaffold080437 markers were significantly different (P〈0.001) in the two models. Four markers (scaffold080437, scaffold479_23523, scaffold4475_71287, and cyse80) may be closely associated with resistance to V. harveyi infection in C. semilaevis. qE-F1 explained 87.36% of the phenotypic variance and contained G18M qE-M1 and qE-M2. Thus, qE-F1 was considered a major candidate region fo

关 键 词：半滑舌鳎 哈维氏弧菌 微卫星标记SSR 混合分离子分析法BSA 数量性状基因座QTL 

分 类 号：S917[农业科学—水产科学]