小麦脱水素基因wzy2表达量实时荧光定量PCR方法的建立和应用

以陕合6号小麦品种为材料,用SYBR GreenⅡ染料,参照小麦脱水素wzy2和-βactin基因序列设计引物,建立小麦wzy2基因的实时荧光定量PCR分析方法.结果表明:通过对标准品标准曲线和熔解曲线分析,其标准曲线的Ct值检测范围为12~30,PCR扩增效率高达111.3%;不同的标准品扩增曲线的间距为3.078,接近于理想值3.32,且可显示产物特异的单峰,引物的特异性扩增强.小麦脱水素基因wzy2表达量实时荧光定量PCR的测定结果表明,小麦脱水素基因wzy2在胁迫24 h的表达量明显高于胁迫12 h的表达量.

Establishment and Application of Real-Time Quantitative PCR to Detect the Expression of Wheat Dehydrin Gene wzy2

In oder to establish the real-time quantitative PCR,we used the primers designed according to the sequence of wheat dehydrin gene wzy2 and β-actin,to detect the expression of wzy2 in wheat variety ’Shanhe 6’ using SYBR GreenⅡ.The results showed that the Ct values ranged from 12 to 30,the amplification efficiency of PCR was 111.3%,which indicated that the method was practicable.The distance between amplification curves of different standard samples was 3.078.The products demonstrated that primers were specific having only one peak.Then we detected the expression levels of wheat gene wzy2 by real-time quantitative PCR.And the results showed the expression level of wzy2 at 24 h drought stress was significantly higher than that at 12 h drought stress.