基于全基因组重测序的不结球白菜KASP分子标记开发与应用  

Development and application of KASP molecular markers for Brassica campestris ssp.chinensis based on whole-genome resequencing

作　　者：张艳[1] 沈卓 杨易 周轩 吴增祥 黎庭耀[1] ZHANG Yan;SHEN Zhuo;YANG Yi;ZHOU Xuan;WU Zeng-xiang;LI Ting-yao(Vegetable Research Institute,Guangdong Academy of Agricultural Sciences/Guangdong Key Laboratory for New Technology Research of Vegetables/Lingnan Modern Agricultural Science and Technology Laboratory of Guangdong,Guangzhou,Guangdong 510640,China)

机构地区：[1]广东省农业科学院蔬菜研究所/广东省蔬菜新技术研究重点实验室/岭南现代农业科学与技术广东省实验室,广东广州510640

出　　处：《南方农业学报》2025年第2期613-621,共9页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(41701369);广东省乡村振兴战略专项种业振兴项目(2022-NPY-00-010);广州市科技计划项目(202103000006);广州市基础研究计划项目(SL2022A04J00783)。

摘　　要：【目的】基于全基因组重测序数据开发不结球白菜KASP分子标记,以期丰富该物种的遗传信息,为不结球白菜种质资源鉴定、基因定位及分子标记辅助育种提供技术支持。【方法】对2份不结球白菜高代自交系C40和JY70进行全基因组及DNA重测序及比对分析,挑选出差异SNP位点用于KASP分子标记开发,并对55份不结球白菜种质资源进行遗传多样性分析和DNA指纹图谱构建。【结果】C40和JY70之间基因型不一致、测序深度大于10X且等位基因纯合的SNP位点有460849个。每条染色体上选择分布相对均匀的15个SNP位点设计成KASP引物,利用150对KASP引物对C40和JY70进行基因型分型检测,发现有86对引物成功分型,基于其对55份不结球白菜种质资源的基因型分型结果,筛选获得41个在不结球白菜中扩增质量高的KASP分子标记。41个KASP分子标记对55份不结球白菜种质资源的遗传多样性分析结果显示,其主要等位基因频率为0.5000~0.9273,平均为0.6643;基因多样性指数为0.1349~0.5000,平均为0.4257;多态性信息含量(PIC)为0.1258~0.3750,平均为0.3322,PIC>0.2500的位点占所有位点的94.5%,多态性较高。55份不结球白菜种质资源分为三大类群,相同类型的种质资源聚类在一起,其中地理来源相同的资源又聚类在一起,有效地反映了种质间的亲缘关系。利用PIC最高的15个KASP分子标记构建了55份不结球白菜种质资源的DNA指纹图谱。【结论】开发获得41个不结球白菜KASP分子标记,可用于不结球白菜种质资源鉴定、基因定位及分子标记辅助育种。【Objective】The purpose of the study was to develop KASP molecular markers for Brassica campestris ssp.chinensis based on whole genome resequencing data,in order to accumulate and enrich the genetic information of this species,and provide technical support for the identification of Brassica campestris ssp.chinensis germplasm resource,gene mapping and marker-assisted breeding.【Method】Whole genome resequencing and comparison analysis on 2 Brassica campestris ssp.chinensis high-generation inbred lines C40 and JY70 were conducted.KASP molecular markers were developed by selecting differential SNP loci,and genetic diversity analysis and DNA fingerprint construction were carried out for 55 Brassica campestris ssp.chinensis germplasm resources.【Result】There were 460849 SNP loci with genotype incompatibility,sequencing depth greater than 10X and homozygous alleles between C40 and JY70.A total of 15 SNP loci with relatively uniform distribution on each chromosome were selected to be designed as KASP primers,and 150 pairs of KASP primers were used to detect C40 and JY70 genotype,and 86 pairs of primers were successfully genotyped.Based on the genotyping results of 55 Brassica campestris ssp.chinensis germplasm resources,41 KASP molecular markers with high amplification quality in Brassica campestris ssp.chinensis were selected.The genetic diversity analysis of 55 Brassica campestris ssp.chinensis germplasm resources with 41 KASP markers showed that,the minor allele frequency ranged from 0.5000 to 0.9273,with an average of 0.6643;the gene diversity index was 0.1349-0.5000,with an average of 0.4257;the polymorphism information content(PIC)was 0.1258-0.3750,with an average value of 0.3322;and 94.5%of the loci had PIC>0.2500,with fine polymorphism.The clustering analysis results showed that 55 Brassica campestris ssp.chinensis germplasm resources were divided into 3 major groups,germplasm resources of the same type were clustered together,and resources of the same geographical origin were clustered together,which ef

关 键 词：不结球白菜 KASP分子标记 遗传多样性分析 DNA指纹图谱 

分 类 号：S634.303.6[农业科学—蔬菜学]