Department of Plant Sciences, University of Saskatchewan, Saskatoon, SK, Canada.
Plant Biotechnol J. 2019 Jan;17(1):275-288. doi: 10.1111/pbi.12964. Epub 2018 Jul 4.
Whole-genome sequencing-based bulked segregant analysis (BSA) for mapping quantitative trait loci (QTL) provides an efficient alternative approach to conventional QTL analysis as it significantly reduces the scale and cost of analysis with comparable power to QTL detection using full mapping population. We tested the application of next-generation sequencing (NGS)-based BSA approach for mapping QTLs for ascochyta blight resistance in chickpea using two recombinant inbred line populations CPR-01 and CPR-02. Eleven QTLs in CPR-01 and six QTLs in CPR-02 populations were mapped on chromosomes Ca1, Ca2, Ca4, Ca6 and Ca7. The QTLs identified in CPR-01 using conventional biparental mapping approach were used to compare the efficiency of NGS-based BSA in detecting QTLs for ascochyta blight resistance. The QTLs on chromosomes Ca1, Ca4, Ca6 and Ca7 overlapped with the QTLs previously detected in CPR-01 using conventional QTL mapping method. The QTLs on chromosome Ca4 were detected in both populations and overlapped with the previously reported QTLs indicating conserved region for ascochyta blight resistance across different chickpea genotypes. Six candidate genes in the QTL regions identified using NGS-based BSA on chromosomes Ca2 and Ca4 were validated for their association with ascochyta blight resistance in the CPR-02 population. This study demonstrated the efficiency of NGS-based BSA as a rapid and cost-effective method to identify QTLs associated with ascochyta blight in chickpea.
基于全基因组测序的 bulked segregant analysis(BSA)用于定位数量性状基因座(QTL),为传统的 QTL 分析提供了一种有效的替代方法,因为它通过使用全映射群体显著降低了分析的规模和成本,同时具有相当的 QTL 检测能力。我们使用两个重组自交系群体 CPR-01 和 CPR-02 测试了基于下一代测序(NGS)的 BSA 方法在定位鹰嘴豆褐斑病抗性 QTL 中的应用。在 CPR-01 群体中定位了 11 个 QTL,在 CPR-02 群体中定位了 6 个 QTL,位于 Ca1、Ca2、Ca4、Ca6 和 Ca7 染色体上。使用常规的双亲作图方法在 CPR-01 中鉴定的 QTL 用于比较 NGS 基于 BSA 检测鹰嘴豆褐斑病抗性 QTL 的效率。在 Ca1、Ca4、Ca6 和 Ca7 染色体上鉴定的 QTL 与之前在 CPR-01 中使用常规 QTL 作图方法检测到的 QTL 重叠。在两个群体中检测到 Ca4 染色体上的 QTL,并与之前报道的不同鹰嘴豆基因型中褐斑病抗性的 QTL 重叠,表明存在保守区域。在 Ca2 和 Ca4 染色体上使用 NGS 基于 BSA 鉴定的 QTL 区域中的 6 个候选基因在 CPR-02 群体中验证了与褐斑病抗性的关联。这项研究证明了基于 NGS 的 BSA 作为一种快速、具有成本效益的方法来鉴定与鹰嘴豆褐斑病相关的 QTL 的效率。