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【目的】蚜虫唾液相关蛋白C002是一种水溶性唾液蛋白,在蚜虫取食过程中发挥着重要作用。本研究探讨了蚜虫唾液相关蛋白基因C002在麦长管蚜Sitobion avenae(F.)取食过程中的作用和功能。【方法】根据豌豆蚜基因C002序列,同源克隆了麦长管蚜C002基因,并应用荧光定量PCR和RNA干扰(RNAi)技术分别对该基因的表达规律和功能进行了研究。【结果】结果表明:麦长管蚜C002基因c DNA开放阅读框长663 bp,编码220个氨基酸,命名为Sv C002。转录水平的表达时序分析发现,Sv C002在蚜虫各个时期都有表达,在2龄若蚜时期表达量最高。ds RNA饲喂结果显示,麦长管蚜取食ds C002 4 d和6 d后,C002基因的表达量分别下降54%和69%,差异极显著(P<0.01),麦长管蚜存活率较对照组下降了32.2%和53.3%。将饲喂ds C002的麦长管蚜接种到感蚜小麦(北京837),蚜虫在短时期内出现大量死亡现象,第6天和第8天存活率分别为44.4%和35.6%,低于对照组并达到极显著水平。【结论】C002基因在麦长管蚜的取食行为中发挥着重要作用,可以作为麦长管蚜防治的潜在RNAi靶标基因。
【Aim】 Aphid salivary associated protein C002 is a kind of water-soluble salivary protein that plays an important role in the feeding of aphids. This study investigated the role and function of aphid-related protein gene C002 in the feeding of Sitobion avenae (F.). 【Method】 Based on the C002 gene sequence of pea aphid, the C002 gene was homologously cloned and the expression and function of the gene were studied by fluorescence quantitative PCR and RNA interference (RNAi). 【Result】 The results showed that the open reading frame of C002 gene of wheat aphid C002 gene was 663 bp in length and encoded 220 amino acids and named Sv C002. The analysis of transcriptional level showed that Sv C002 was expressed in all stages of aphids, and was the highest in the second instars. The results of dsRNA feeding showed that the expression of C002 gene in Cichlium adenophorum was decreased by 54% and 69%, respectively, on the 4th and 6th day after feeding ds C002, the difference was significant (P <0.01) Compared with the control group decreased by 32.2% and 53.3%. Aphids infected with ds C002 were inoculated into susceptible wheat (Beijing 837). Aphids died in a large amount in a short period of time. The survival rates on day 6 and day 8 were 44.4% and 35.6%, respectively, which were lower than those of control Group and reached a very significant level. 【Conclusion】 C002 gene plays an important role in the feeding behavior of Aphis gossypii and may be used as a potential RNAi target gene for the control of Aphis gossypii.