利用ARMS-Tm-shift-qPCR技术检测水稻低直链淀粉含量基因Wx-mq

摘要

Wx-mq基因是水稻低直链淀粉性状控制基因,与米饭食味品质密切相关.本研究根据水稻控制低直链淀粉含量Wx-mq基因第497位存在的G＞A单核苷酸变异,将基于SYBR Green Ⅰ的ARMS和Tmshift的两种实时PCR基因分型方法相结合,建立了可准确区分Wx-mq基因第497位的GG纯合、从纯合及GA杂合三种类型的实时荧光定量PCR体系.试验表明这种基于SYBR Green Ⅰ的ARMS-Tm-shift实时PCR基因分型方法无需合成探针,试验设计简单,是一种快速、简便、特异性好的基因分型方法,可用于Wx-mq基因分子标记辅助育种中基因的高通量分型.%Wx-mq gene is a low amylose trait controlled gene and closely relates with the eating quality of rice.According to the rice low amylose starch content control gene Wx-mq No.497 position existing G ＞A single nucleotide variants,two real-time PCR genotyping methods,ARMS and Tm-shift based on SYBR Green Ⅰ,was combined to establish the fluorescence real-time quantitative PCR system,which could accurately distinguish the GG homozygote,AA homozygote and GA heterozygosis of Wx-mq gene No.497 position.The experiment results showed that the ARMS-Tm-shift real-time PCR genotyping method based on SYBR Green Ⅰ did not need probe synthesis with simple designs.It was a rapid,simple and specific genotyping method.It could be used in high-throughput genotyping of Wx-mq gene in molecular marker assisted breeding.