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利用高通量全基因组测序进行大豆白粉病抗性基因的遗传定位。

Genetic mapping of powdery mildew resistance genes in soybean by high-throughput genome-wide sequencing.

机构信息

The State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, South China Agricultural University, Guangzhou, 510642, Guangdong, People's Republic of China.

The Key Laboratory of Plant Molecular Breeding of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou, 510642, Guangdong, People's Republic of China.

出版信息

Theor Appl Genet. 2019 Jun;132(6):1833-1845. doi: 10.1007/s00122-019-03319-y. Epub 2019 Mar 2.

Abstract

The Mendelian locus conferring resistance to powdery mildew in soybean was precisely mapped using a combination of phenotypic screening, genetic analyses, and high-throughput genome-wide sequencing. Powdery mildew (PMD), caused by the fungus Microsphaera diffusa Cooke & Peck, leads to considerable yield losses in soybean [Glycine max (L.) Merr.] under favourable environmental conditions and can be controlled by identifying germplasm resources with resistance genes. In this study, resistance to M. diffusa among resistant varieties B3, Fudou234, and B13 is mapped as a single Mendelian locus using three mapping populations derived from crossing susceptible with resistant cultivars. The position of the PMD resistance locus in B3 is located between simple sequence repeat (SSR) markers GMES6959 and Satt_393 on chromosome 16, at genetic distances of 7.1 cM and 4.6 cM, respectively. To more finely map the PMD resistance gene, a high-density genetic map was constructed using 248 F recombinant inbred lines derived from a cross of Guizao1 × B13. The final map includes 3748 bins and is 3031.9 cM in length, with an average distance of 0.81 cM between adjacent markers. This genotypic analysis resulted in the precise delineation of the B13 PMD resistance locus to a 188.06-kb genomic region on chromosome 16 that harbours 28 genes, including 17 disease resistance (R)-like genes in the reference Williams 82 genome. Quantitative real-time PCR assays of possible candidate genes revealed differences in the expression levels of 9 R-like genes between the resistant and susceptible parents. These results provide useful information for marker-assisted breeding and gene cloning for PMD resistance.

摘要

大豆抗白粉病的孟德尔基因座是通过表型筛选、遗传分析和高通量全基因组测序相结合精确定位的。白粉病(PMD)由 Microsphaera diffusa Cooke & Peck 引起,在有利的环境条件下会导致大豆 [Glycine max (L.) Merr.] 产量显著损失,可以通过鉴定具有抗性基因的种质资源来控制。在这项研究中,利用三个由感病和抗病品种杂交产生的作图群体,将 B3、Fudou234 和 B13 等抗性品种对 M. diffusa 的抗性定位为一个单一的孟德尔基因座。B3 中 PMD 抗性基因座的位置位于 16 号染色体上的简单重复序列(SSR)标记 GMES6959 和 Satt_393 之间,遗传距离分别为 7.1 cM 和 4.6 cM。为了更精细地定位 PMD 抗性基因,利用 Guizao1×B13 杂交产生的 248 个重组自交系构建了高密度遗传图谱。最终图谱包含 3748 个 bin,长度为 3031.9 cM,相邻标记之间的平均距离为 0.81 cM。这种基因型分析精确地将 B13 的 PMD 抗性基因座定位到 16 号染色体上的一个 188.06-kb 基因组区域,该区域包含 28 个基因,包括参考 Williams 82 基因组中的 17 个抗病(R)样基因。对可能候选基因的定量实时 PCR 分析显示,抗性和感病亲本之间 9 个 R 样基因的表达水平存在差异。这些结果为 PMD 抗性的标记辅助育种和基因克隆提供了有用信息。

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