病害频发严重制约着水产养殖业的发展，抗病养殖品种的培育对促进水产养殖业的健康发展具有重要意义。本项目将基于前期获得的稀有鮈鲫P0-F2代抗出血病CRISPR/Cas9诱变家系，以F2代抗病个体为亲本，构建F3代全同胞群体；通过GCRV攻毒实验，获得子代存活和死亡样本；采用HRM突变分析系统，对实验样本indel突变位点进行基因分型；通过突变位点的基因型/表型关联分析和F4代群体水平的功能验证，筛选鉴定CRISPR/Cas9诱变导致的功能性突变位点，获得高抗病性稀有鮈鲫家系；利用已有的稀有鮈鲫基因组序列，克隆抗病相关基因；通过突变型抗病家系与野生型易感家系的转录组比较分析，初步解析抗病基因功能及其网络调控机制。本项目获得的GCRV抗性相关突变位点和突变基因将为草鱼抗病育种奠定基础。

The frequent occurrence of diseases seriously restricts the development of aquaculture. The cultivation of disease-resistant varieties is of great significance to promote the healthy development of aquaculture. This project will construct F3 full-sib families of Rare Minnow based on the previously acquired anti-hemorrhagic CRISPR/Cas9 mutant family (P0-F2 generation), using F2 resistant individuals as parents. Survival and death samples of offspring will be obtained by GCRV infection. HRM mutation analysis system will be used to type the indel mutation sites in the experimental samples. Through genotype/phenotype correlation analysis and functional verification at population level, functional mutation sites caused by CRISPR/Cas9 gene editing will be screened and identified, and a high disease-resistant Rare Minnow family will be obtained. Then, the genes related to disease resistance will be cloned based on the genomic sequence of Rare Minnow. Through the comparative analysis of transcriptome between mutant resistant and wild susceptible families, the function of disease-resistant genes and their regulation network will be preliminarily analyzed, which will lay a foundation for grass carp disease-resistant breeding.