大麦(Hordeum vulgare Linn.)α-淀粉酶基因Amy6-4表达模式的半定量RT-PCR分析

A Semi-quantitative RT-PCR Method to Measure α-amylase Amy6-4 Gene Expression in Barley(Hordeum vulgare Linn.)

大麦α-淀粉酶编码基因的表达对α-淀粉酶的活性有重要影响。以α-淀粉酶活性具有显著差异的两个大麦品种为试材,研究不同发芽时期大麦种子中Amy6-4基因的表达模式。利用Amy6-4基因的特异性引物,通过半定量RT-PCR,对发芽0h、12h、24h、48h、72h、96h和120h的大麦种子的表达模式进行分析。结果表明,两品种在发芽的0时期基因表达量为零;随着发芽时期的延长两品种的Amy6-4基因表达水平呈现增加的趋势,且两品种在发芽第5天,该基因表达水平均达到最高;在发芽后的各个时期,品种ZDM7825的Amy6-4基因的表达量均高于ZDM5271。

The expression of coding gene,Amy6-4 plays an important role in α-amylase activity in the barley.The expression pattern of Amy6-4 gene in germinated seed during different germinating period was investigated with two different barley cultivars,ZDM5271 and ZDM7825.Semi-quantitative RT-PCR method was used to investigate expression patterns of Amy6-4 gene in 0,12,24,48,72,96 and 120 h post-treatment of germinated seed for two cultivars with different α-amylase activity.The results showed that the Amy6-4 gene didn＇t express at 0h,but after that the gene expressing level was increased in both varieties,and it was higher in the ZDM7825 than that in the ZDM5271 during different period of germinated times.The expression level of Amy6-4 gene reached the highest point at 120 h post-treatment in both cultivars.It concluded that the Amy6-4 gene could not express in ungerminated seed and the expression level of Amy6-4 gene increased with the extension of the germination time.