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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。
In order to screen microsatellite markers related to Vibrio viridans disease and locate QTLs, the Cynoglossus semilaevis family F1412 (Family + year + family number), whose survival rate in 2014 was 52.22% A total of 169 microsatellite markers (SSRs) were screened by bulked segregant analysis (BSA), and a microsatellite marker scaffold479_23523 related to Vibrio harveyi was obtained. The genotypes of 94 resistant and susceptible individuals were genotyped by 37 SSRs on linkage group LG18 where scaffold479_23523 was located. Three maps were obtained: LG18, LG18F and LG18M Two different modes for single-marker analysis and QTL mapping. There were 3 significant correlation markers (P <0.05) and 1 highly significant correlation marker (P <0.01) in pattern 1, and one interval qE-F1 was mapped in pattern LG18F. There were 4 significant correlation markers (P <0.05) ) And one highly significant correlation marker (P <0.01). The LG18M map locates two intervals qE-M1 and qE-M2. QE-F1 is more likely to be associated with disease resistance traits in the interval qE-F1 encompassing qE-M1 and qE-M2 over the entire genome sequence. In this study, microsatellite markers and QTL intervals related to Vibrio harveyi were developed, which laid the foundation for the cultivation of new resistant varieties.