麦长管蚜重要基因的RNA干扰

摘要

In this study RNA interference (RNAi) technology was applied to analyze the effect of RNAi silencing of tweleve important genes of wheat aphids by feeding aphids with double stranded RNA (dsRNA) of the target genes.The results showed that the mortality of wheat aphids significantly increased after RNAi silencing of water channel protein gene (Wsa) and ATP synthase gene (vAd1-78);whereas glucose dehydrogenase encoding gene (GlD-17) RNAi led to a significantly reduced mortality.Quantitative analysis of gene expression level during 5 days after RNAi showed that the expression level of gene Wsa and vAd1-78 was decreased by 74.8％ and 44.6％,respectively.Whereas the expression level of gene GlD-17 was increased by up to 209％.Variance on mortality of wheat aphids was in accordance with gene expression level changes after gene silencing by RNAi.Among the twelve targeted genes,RNA interference of Wsa led to most significant gene silence effect,increasing mortality to 37.5％ and down-regulating gene expression level to 74.8％.Thus water channel protein gene (Wsa)could be used as a candidate targeted gene of RNA interference for further development of wheat varieties of aphids resistance.%选用12个蚜虫生长发育的关键基因,利用喂食的方法将其双链RNA(dsRNA)引入麦长管蚜体内,以死亡率为评价指标,对基因RNA干扰(RNAi)沉默效应进行分析.结果表明,水通道蛋白基因(Wsa)、ATP合酶基因(vAdl-78)的RNA干涉能够显著提高蚜虫死亡率,而葡萄糖脱氢酶编码基因(GlD-17)的RNA干涉则使蚜虫死亡率明显降低.基因表达荧光定量PCR分析结果显示,喂食dsRNA后5d内,水通道蛋白基因(Wsa)和ATP合酶基因(vAdl-78)表达水平分别下降了74.8％和44.6％;而葡萄糖脱氢酶编码基因(GlD-17)表达水平则上调了209.0％,基因表达水平变化规律与蚜虫死亡率变化趋势一致.在所有供试基因中,水通道蛋白基因(Wsa)的RNA干涉沉默效应效果最突出,导致蚜虫死亡率提高到37.5％,基因表达量下调74.8％,可以作为利用RNAi技术培育抗蚜虫小麦新品种有效的靶基因.