小麦粒重基因TaGW2-6A等位变异的高通量分子检测

为了开发出一种高通量的TaGW2-6A基因等位变异分子检测技术,利用高分辨率熔解曲线分析技术(High resolution melting curve analysis,HRM)检测了316份小麦材料的TaGW2-6A基因编码区T碱基插入等位变异以及启动子区Hap-6A-A(-593A和-739G)和Hap-6A-G(-593G和-739A)等位变异,同时利用桑格测序和Hap-6A-P1/P2分子标记验证HRM的分析结果。结果表明,HRM技术能够准确、高效的检测TaGW2-6A基因的等位变异。在316份材料中,检测到61份T碱基插入等位变异材料(命名为TT基因型)和255份无T碱基插入的等位变异材料(命名为tt基因型);其中,189份为Hap-6A-A单倍型材料,127份为Hap-6A-G单倍型材料。

High-throughput Molecular Detection of Allelic Variation in TaGW2-6A Gene of Wheat (Triticum aestivum L.)

TaGW2 on chromosome 6A is a key gene governing kernel size of wheat (Triticum aestivum L.). A single T-base insertion has been found in the eighth exon of TaGW2 (977 bp) and two SNPs (Hap-6A-A and Hap-6A-G) in the promoter region. In this study,a high-throughput technology using high resolution melting curve (HRM) analysis was established and used to detect the allelic variations in TaGW2-6A allelic gene among 316 wheat materials.Results of HRM analysis were verified by Sanger sequencing and molecular marker for Hap-6A-P1/P2. HRM can detect the allelic variation in TaGW2-6A gene accurately and efficiently. There were totally 61 wheat materials detected as containing T-base insertion (TT genotype) and 255 wheat materials detected as lacking of T-base insertion (tt genotype),meanwhile,189 and 127 wheat materials were detected as Hap-6A-A haplotype and Hap-6A-G haplotype,respectively.