摘要
三系配套是辣椒育种的主要研究方向,鉴定并开发辣椒恢复基因连锁标记是利用分子标记辅助选育恢复系的难点。本研究以自主选育的辣椒胞质雄性不育系014A和恢复系014C为亲本构建F2分离群体,经χ2测验可育与不育的分离比例符合3∶1,表明辣椒胞质雄性不育育性恢复性状受1对显性基因控制。采用BSA法构建可育和不育极端DNA混池,全基因组重测序并与参考基因组比对,通过SNP-index法和Fisher检验将辣椒胞质雄性不育恢复基因定位于辣椒第6号染色体顶端1.44~8.28Mb的区域内。根据亲本间SNP/InDel差异设计引物,在亲本和极端池筛选多态性引物,得到能稳定扩增出特异条带的分子标记PP5和OP59。这2个标记在极端群体验证中准确率均达到100%;标记OP59是共显性标记,位于恢复基因的候选基因T459-15819间区17232 bp,该标记在不育群体准确率为100%,在可育群体准确率为97.21%;标记PP5位于恢复基因的候选基因T459-15819下游318bp,该标记在不育群体和纯合可育群体准确率均为100%,不能区分杂合可育类型。该研究获得与恢复基因紧密连锁2个标记为加速选育辣椒CMS恢复系奠定基础。
Three-line matching is the main research direction of pepper breeding,identification and development of re-storer gene linkage markers in pepper is a difficult problem in restorer line breeding.In this study,the F2 segregation population was constructed by using 014A and 014C as parents,the segregation ratio of fertility and sterility was 3∶1 byχ2 test,indicating that CMS fertility recovery trait of capsicum was controlled by one pair of dominant genes.Using the BSA method to construct the mixed pool of fertile and sterile DNA,the whole genome was re-sequenced and com-pared with the reference genome,by SNP-index Method and Fisher Test,the cytoplasmic male sterility restorer gene was mapped in the region of 1.44‒8.28 Mb at the tip of the chromosome 6.The primers were designed according to the SNP/InDel difference between parents,and the polymorphic primers were screened in both parents and extreme pools,and the molecular markers PP5 and OP59 were obtained.The accuracy of the two markers was 100%in extreme popu-lation validation;The marker OP59 was a co-dominant marker,which was located at the candidate gene T459-15819 interval of 17232 bp.The accuracy of the marker was 100%in the sterile population and 97.21%in the fertile popula-tion;PP5 was located 318 bp downstream of the candidate gene T459-15819 of the restorer gene.The accuracy of PP5 was 100%in both sterile and homozygous fertile populations.In this study,we obtained two markers closely linked with the restorer gene,which would lay the foundation for accelerating the selection of CMS restorer lines in pepper.
作者
王萌
赵虎
徐晓美
潘尧铧
赵曾菁
吴星
王日升
WANG Meng;ZHAO Hu;XU Xiaomei;PAN Yaohua;ZHAO Zengjing;WU Xing;WANG Risheng(Vegetable Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,Guangxi 530007,China;Vegetable Re-search Institute,Guangdong Academy of Agricultural Sciences,Guangzhou,Guangdong 510640,China;Nanning Wuxiangling Forest Park,Nanning,Guangxi 530219,China)
出处
《热带作物学报》
CSCD
北大核心
2023年第8期1534-1541,共8页
Chinese Journal of Tropical Crops
基金
广西自然科学基金项目(No.2019GXNSFBA245053)
粤桂联合基金项目(No.2020A1515410005)
广西农业科学院基本科研业务专项(桂农科2021JM94)。
关键词
辣椒
全基因组重测序
胞质雄性不育
恢复基因
分子标记
pepper
wholegenome resequencing
cytoplasmic male sterility(CMS)
restoration gene
molecular markers
