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基于DNA重测序和辣椒混合分离群体分析法的辣椒白粉病抗性基因定位

Mapping of resistance genes to powdery mildew based on DNA re-sequencing and bulk segregant analysis in Capsicum.

作者信息

Zhang Tao, Bosland Paul W, Ma Yan, Wang Yuhang, Li Wei, Kong Weifu, Wei Min, Duan Panpan, Zhang Gaoyuan, Wei Bingqiang

机构信息

College of Horticulture, Gansu Agricultural University, Lanzhou, 730070, Gansu, China.

Plant and Environmental Sciences Department, New Mexico State University, P.O. Box 30003, Las Cruces, NM, 88001, USA.

出版信息

Protoplasma. 2025 May;262(3):489-500. doi: 10.1007/s00709-024-02013-1. Epub 2024 Dec 2.

Abstract

Powdery mildew caused by Leveillula taurica adversely affects the development and growth of pepper plants. However, there have been few reports on the fine mapping and quantitative trait locus (QTLs) gene cloning of resistance genes to powdery mildew in pepper. Herein, an F segregating population was constructed using the high resistance material "NuMex Suave Red" and the extremely susceptible material "c89" for bulked segregant analysis and DNA re-sequencing (BSA-seq). Molecular markers were used to achieve fine mapping, followed by expression verification. A major QTL located on chromosome 5 (Chr5, 7.20-11.75 Mb) that is associated with resistance to powdery mildew in pepper was mapped using BSA-seq. A narrow interval of 64.86 kb encompassing five genes was refined using InDel and KSAP molecular markers developed from the QTL region. Among them, the expression of the ubiquitin-conjugating enzyme E2 gene, Capana05g000392, was significantly upregulated in multiple resistant materials. In addition, there was a single nucleotide polymorphism (SNP) of A/G in the 241st position of the CDS sequence of Capana05g000392, which in turn leads to an amino acid polymorphism of M/V between susceptible parent and resistant parent. Overall, these results indicate that the Capana05g000392 gene may serve as a robust potential factor against powdery mildew in pepper. These findings further elucidate the genetic mechanism of resistance to powdery mildew in pepper and facilitate molecular marker-assisted breeding.

摘要

由牛舌草内丝白粉菌引起的白粉病对辣椒植株的发育和生长产生不利影响。然而,关于辣椒对白粉病抗性基因的精细定位和数量性状位点(QTL)基因克隆的报道较少。在此,利用高抗材料“NuMex Suave Red”和极感材料“c89”构建了一个F分离群体,用于混合分组分析法和DNA重测序(BSA-seq)。使用分子标记进行精细定位,随后进行表达验证。利用BSA-seq定位到一个位于5号染色体(Chr5,7.20-11.75 Mb)上与辣椒抗白粉病相关的主要QTL。利用从QTL区域开发的InDel和KSAP分子标记,将一个包含5个基因的64.86 kb的狭窄区间进行了精细定位。其中,泛素结合酶E2基因Capana05g000392在多个抗性材料中的表达显著上调。此外,Capana05g000392的CDS序列第241位存在A/G单核苷酸多态性(SNP),这导致了感病亲本和抗病亲本之间M/V氨基酸多态性。总体而言,这些结果表明Capana05g000392基因可能是辣椒抗白粉病的一个强大潜在因子。这些发现进一步阐明了辣椒抗白粉病的遗传机制,并促进了分子标记辅助育种。

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