Wei Xiaochun, Xiao Shixiong, Zhao Yanyan, Zhang Luyue, Nath Ujjal Kumar, Yang Shuangjuan, Su Henan, Zhang Wenjing, Wang Zhiyong, Tian Baoming, Wei Fang, Yuan Yuxiang, Zhang Xiaowei
Institute of Vegetables, Henan Academy of Agricultural Sciences, Graduate T&R Base of Zhengzhou University, Zhengzhou, Henan, China.
School of Agricultural Sciences, Zhengzhou University, Zhengzhou, Henan, China.
Front Plant Sci. 2024 May 17;15:1355090. doi: 10.3389/fpls.2024.1355090. eCollection 2024.
Clubroot disease poses a significant threat to crops, necessitating ongoing updates on resistance gene sources. In F segregants of the clubroot-resistant inbred line BrT18-6-4-3 and susceptible DH line Y510, the genetic analysis identified a single dominant gene responsible for clubroot resistance. Through bulk segregant sequencing analysis and kompetitive allele-specific polymerase chain reaction assays, was mapped within 110 kb (12,255-12,365 Mb) between markers L-CR11 and L-CR12 on chromosome A08. We identified as the candidate gene of . Upon comparison with the sequence of disease-resistant material BrT18-6-4-3, we found 249 single-nucleotide polymorphisms, seven insertions, six deletions, and a long terminal repeat (LTR) retrotransposon (5,310 bp) at 909 bp of the first intron. However, the LTR retrotransposon was absent in the coding sequence of the susceptible DH line Y510. Given the presence of a non-functional LTR insertion in other materials, it showed that the LTR insertion might not be associated with susceptibility. Sequence alignment analysis revealed that the fourth exon of the susceptible line harbored two deletions and an insertion, resulting in a frameshift mutation at 8,551 bp, leading to translation termination at the leucine-rich repeat domain's C-terminal in susceptible material. Sequence alignment of the CDS revealed a 99.4% similarity to , which indicate that is likely an allele of the gene. Two functional markers, CRA08-InDel and CRA08-KASP1, have been developed for marker-assisted selection in CR turnip cultivars. Our findings could facilitate the development of clubroot-resistance turnip cultivars through marker-assisted selection.
根肿病对作物构成重大威胁,因此需要不断更新抗性基因来源。在抗根肿病自交系BrT18 - 6 - 4 - 3和感病双单倍体系Y510的F分离群体中,遗传分析确定了一个负责根肿病抗性的显性单基因。通过混合分组分析法测序分析和竞争性等位基因特异性聚合酶链反应分析,该基因被定位在A08染色体上标记L - CR11和L - CR12之间110 kb(12,255 - 12,365 Mb)范围内。我们确定该基因是该抗性基因的候选基因。与抗病材料BrT18 - 6 - 4 - 3的序列比较后,我们在第一个内含子的909 bp处发现了249个单核苷酸多态性、7个插入、6个缺失和一个长末端重复(LTR)反转录转座子(5,310 bp)。然而,感病双单倍体系Y510的编码序列中没有该LTR反转录转座子。鉴于其他材料中存在无功能的LTR插入,表明该LTR插入可能与感病性无关。序列比对分析表明,感病品系的第四个外显子有两个缺失和一个插入,导致在8,551 bp处发生移码突变,从而使感病材料中富含亮氨酸重复结构域的C末端翻译终止。编码序列的比对显示与该基因有99.4%的相似性,这表明该基因可能是该基因的一个等位基因。已经开发了两个功能性标记CRA08 - InDel和CRA08 - KASP1,用于抗根肿病芜菁品种的标记辅助选择。我们的研究结果有助于通过标记辅助选择培育抗根肿病的芜菁品种。