Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, Jiangsu, China.
Department of Agronomy, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen, 73140, Nakhon Pathom, Thailand.
Theor Appl Genet. 2021 Feb;134(2):701-714. doi: 10.1007/s00122-020-03724-8. Epub 2020 Nov 13.
This paper reports fine mapping of qCLS for resistance to Cercospora leaf spot disease in mungbean and identified LOC106765332encoding TATA-binding-protein-associated factor 5 (TAF5) as the candidate gene for the resistance Cercospora leaf spot (CLS) caused by the fungus Cercospora canescens is an important disease of mungbean. A QTL mapping using mungbean F and BCF populations developed from the "V4718" (resistant) and "Kamphaeng Saen 1" (KPS1; susceptible) has identified a major QTL controlling CLS resistance (qCLS). In this study, we finely mapped the qCLS and identified candidate genes at this locus. A BCF [KPS1 × (KPS1 × V4718)] population developed in this study and the F (KPS1 × V4718) population used in a previous study were genotyped with 16 newly developed SSR markers. QTL analysis in the BCF and F populations consistently showed that the qCLS was mapped to a genomic region of ~ 13 Kb on chromosome 6, which contains only one annotated gene, LOC106765332 (designated "VrTAF5"), encoding TATA-binding-protein-associated factor 5 (TAF5), a subunit of transcription initiation factor IID and Spt-Ada-Gcn5 acetyltransferase complexes. Sequence comparison of VrTAF5 between KPS1 and V4718 revealed many single nucleotide polymorphisms (SNPs) and inserts/deletions (InDels) in which eight SNPs presented in eight different exons, and an SNP (G4,932C) residing in exon 8 causes amino acid change (S250T) in V4718. An InDel marker was developed to detect a 24-bp InDel polymorphism in VrTAF5 between KPS1 and V4718. Analysis by RT-qPCR showed that expression levels of VrTAF5 in KPS1 and V4718 were not statistically different. These results indicated that mutation in VrTAF5 causing an amino acid change in the VrTAF5 protein is responsible for CLS resistance in V4718.
本研究报告了绿豆对叶斑病抗性的 qCLS 的精细定位，并鉴定了 LOC106765332 编码 TATA 结合蛋白相关因子 5（TAF5）为候选基因，该基因与由真菌 Cercospora canescens 引起的叶斑病（CLS）抗性有关。使用来自“V4718”（抗性）和“Kamphaeng Saen 1”（KPS1；感病）的绿豆 F 和 BCF 群体进行的 QTL 作图鉴定了控制 CLS 抗性的主要 QTL（qCLS）。在本研究中，我们精细地定位了 qCLS 并鉴定了该基因座的候选基因。本研究中开发的 BCF [KPS1 × （KPS1 × V4718）] 群体和之前研究中使用的 F （KPS1 × V4718）群体使用 16 个新开发的 SSR 标记进行了基因型分析。BCF 和 F 群体的 QTL 分析一致表明，qCLS 映射到 6 号染色体上的约 13 kb 基因组区域，该区域仅包含一个注释基因 LOC106765332（命名为“VrTAF5”），编码 TATA 结合蛋白相关因子 5（TAF5），是转录起始因子 IID 和 Spt-Ada-Gcn5 乙酰转移酶复合物的一个亚基。在 KPS1 和 V4718 之间比较 VrTAF5 的序列发现了许多单核苷酸多态性（SNP）和插入/缺失（InDels），其中 8 个 SNP 存在于 8 个不同的外显子中，而位于外显子 8 中的 SNP（G4,932C）导致 V4718 中氨基酸变化（S250T）。开发了一个 InDel 标记来检测 KPS1 和 V4718 之间 VrTAF5 的 24-bp InDel 多态性。通过 RT-qPCR 分析表明，KPS1 和 V4718 中 VrTAF5 的表达水平没有统计学差异。这些结果表明，VrTAF5 中的突变导致 VrTAF5 蛋白中的氨基酸变化是 V4718 中 CLS 抗性的原因。
