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园艺学报 ›› 2021, Vol. 48 ›› Issue (7): 1317-1328.doi: 10.16420/j.issn.0513-353x.2020-0656

• 研究论文 • 上一篇    下一篇

大白菜抗根肿病基因BraA.Pb.8.4的定位及KASP标记开发

杨双娟, 张晓伟, 魏小春, 赵艳艳, 王志勇, 赵肖斌, 李林, 原玉香**()   

  1. 河南省农业科学院园艺研究所,郑州450002
  • 收稿日期:2020-12-24 修回日期:2021-02-19 出版日期:2021-07-25 发布日期:2021-08-10
  • 通讯作者: 原玉香 E-mail:yuanyuxiang@126.com
  • 基金资助:
    国家自然科学基金项目(31872945);国家自然科学基金项目(31801874);河南省中原千人计划中原学者项目(202101510003);国家现代农业产业技术体系建设专项资金项目(CARS-23-G-16);国家重点研发计划项目(2016YFD0100204-18)

Mapping and KASP Markers Development for Clubroot Resistance Gene BraA.Pb.8.4 in Chinese Cabbage

YANG Shuangjuan, Zhang Xiaowei, WEI Xiaochun, ZHAO Yanyan, WANG Zhiyong, ZHAO Xiaobin, LI Lin, YUAN Yuxiang**()   

  1. Institute of Horticulture,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China
  • Received:2020-12-24 Revised:2021-02-19 Online:2021-07-25 Published:2021-08-10
  • Contact: YUAN Yuxiang E-mail:yuanyuxiang@126.com

摘要:

以大白菜1E519BC1F2和4E511BC1F2两个群体为试材,分别构建根肿病抗病和感病池,进行集团分离群体测序(Bulked Segregant Analysis Sequencing,BSA-seq)和分析。结果发现,两个群体中检测到1个共同的根肿病抗性基因候选区间,位于A08染色体7.40 ~ 8.85 Mb,命名为BraA.Pb.8.4。该基因在物理位置上和已经报道的根肿病抗性基因Crr1Rcr9CRs均不同,并且Crr1Rcr9连锁的标记在抗病和感病材料中不具有多态性,CRs连锁的4个标记中只有1个标记(Probe60)在抗病和感病材料中存在多态性,表明BraA.Pb.8.4不同于已经报道的基因,可能为1个新的根肿病抗性基因。在BraA.Pb.8.4候选区间开发了3个KASP标记(BraA.Pb.8.4-K1、BraA.Pb.8.4-K2和BraA.Pb.8.4-K3),均可以有效将纯合抗病、纯合感病和杂合抗病材料区分开,为共显性标记。利用标记BraA.Pb.8.4-K1在5个群体共257个单株中进行验证,发现各单株的基因型和抗病性表型平均一致率高达96.13%。因此,本研究鉴定的BraA.Pb.8.4基因及开发的KASP标记可以高效用于根肿病的分子标记辅助育种。

关键词: 大白菜, 根肿病, BraA.Pb.8.4基因, 基因定位, KASP标记

Abstract:

In this study,Bulked Segregant Analysis Sequencing(BSA-seq)was conducted on two Chinese cabbage populations,named as 1E519BC1F2 and 4E511BC1F2. We identified a common candidate region from the above two populations,which located on 7.40-8.85 Mb of chromosome A08. Here,we named this region as BraA.Pb.8.4. BraA.Pb.8.4 might be a novel clubroot resistance gene for two reasons. Firstly,in terms of physical position,BraA.Pb.8.4 is distinct from previous reported genes,Crr1,Rcr9 and CRs. Secondly,markers linked to Crr1 and Rcr9 amplified the same product without length polymorphysims between resistant and susceptible materials,and only one of the four CRs linked markers (Probe60)produced polymorphysims. Among the candidate region of BraA.Pb.8.4,three KASP markers,BraA.Pb.8.4-K1,BraA.Pb.8.4-K2 and BraA.Pb.8.4-K3,were developed. These three KASP markers are co-dominant markers which can not only effectively separate homozygous resistant genotypes from the homozygous susceptible genotypes,but also distinguish the heterozygous genotypes. Marker BraA.Pb.8.4-K1 were further genotyped in a total of 257 individuals,which belongs to five populations. The results showed that the average consistency rate between genotypes and phynotypes of the above 257 individuals was high to 96.13%. Thus,the BraA.Pb.8.4 gene and the KASP markers developed here can be efficently used for marker-assisted selection(MAS)of clubroot resistant gene in breeding.

Key words: Chinese cabbage, clubroot, BraA.Pb.8.4 gene, gene mapping, KASP marker

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