Multiplex PCR assay for detection of wheat leaf rust resistance genes Lr9-Lr26 and Lr19-Lr20 in 116 Chinese wheat cultivars（lines) | 116个小麦品种（系）抗叶锈基因Lr9-Lr26、Lr19-Lr20的复合PCR检测

中国人. [目的]建立简单、快速、有效的小麦抗叶锈基因复合PCR体系, 从而提高分子标记辅助选择效率。[方法]以28个‘Thatcher’为背景的近等基因系和16个已知基因载体品系作为试材, 测试了小麦抗叶锈病基因Lr9、Lr26、Lr19和Lr20的STS标记特异性,通过优化PCR反应体系和循环条件, 构建了抗叶锈基因Lr9-Lr26和Lr19-Lr20的复合PCR检测体系。对116个小麦品种（系）所含有的抗叶锈病基因进行了分子检测。[结果] 供试品种均不含有Lr9和Lr20, 47个品种含有Lr26（基因频率为40.5%）,‘中梁22’含有Lr19。经反复验证, Lr9-Lr26和Lr19-Lr20复合PCR技术检测结果可靠, 且与上述单个分子标记检测结果一致。[结论] 建立的Lr9-Lr26和Lr19-Lr20的复合PCR检测体系可以准确、稳定、高效地检测小麦抗叶锈基因Lr9、Lr26、Lr19和Lr20。

英语. [Objective] A simple, fast and effective method of multiplex PCR for wheat leaf rust resistance genes was set up to improve the efficiency of marker-assisted selection. [Method] By testing the specificity of STS markers of Lr9, Lr19, Lr20 and Lr26 based on the near isogenic lines with＇Thacher＇background and known gene lines, multiplex PCR was developed and optimized for detection of wheat leaf rust resistance gene combination of Lr9-Lr26 and Lr19-Lr20. [Result] A total of 116 wheat cultivars were evaluated using the multiplex PCRs, and specific DNA bands of resistance gene Lr26 was present in 47 accessions out of the 116 cultivars or lines tested. Lr9 and Lr20 were not detected and Lr19 may be present in ＇Zhong Liang 22＇. The Lr9-Lr26 and Lr19-Lr20 multiplex PCR results were consistent with those obtained separately by using the STS markers. [Conclusion] The results indicated that the two multiplex PCRs developed could be used to detect the Lr9, Lr26, Lr19 and Lr20 stably and efficiently. The multiplex PCR technique may be efficiently used in marker-assisted selection for wheat breeding and useful for the DNA-based identification of wheat varieties.