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麦长管蚜唾液蛋白C002的基因克隆与
Cloning and RNA interference analysis of the Sitobion avenae salivary protein C002 gene
李雪峰1** 范 佳2 孙永伟1 王根平1 陈红梅1 闫 婷1 杜文明1 夏兰琴1
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DOI:10.7679/j.issn.2095-1353.2014.175
作者单位:(1. 中国农业科学院作物科学研究所/国家农作物基因与改良重大科学工程,北京 100081;2. 中国农业科学院植物保护研究所,北京 100193
中文关键词:麦长管蚜,蚜虫唾液蛋白,C002,RNA干扰,荧光定量PCR
英文关键词: Sitobion avenae, aphid saliva protein, C002, RNA interference, real-time quantitative PCR
中文摘要:

      【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 avenaeF.)取食过程中的作用和功能。方法】根据豌豆蚜基因C002列,同源克隆了麦长管蚜C002基因,并应用荧光定量PCRRNA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cDNA开放阅读框长663 bp,编码220个氨基酸,命名为SvC002。转录水平的表达时序分析发现,SvC002在蚜虫各个时期都有表达,在2龄若蚜时期表达量最高。sRNA饲喂结果显示,麦长管蚜取食dsC0024d6d后,C002基因的表达量分别下降54%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%53.3%。将饲喂dsC002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%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。

英文摘要:

      [Objectives]  To study the function of the C002 salivary protein gene in the aphid Sitobion avenae (F.). [Methods]  A C002 gene was cloned based on the C002 gene of A. pisum. Real-time quantitative PCR and RNAi experiments were then conducted to explore the expression patterns and the function of this gene. [Results]  Bioinformatics analysis shows that the cloned sequence contained a 663 bp open reading frame encoding a protein of 220 amino acid residues. It was therefore named SaC002. Transcripts of SaC002 were detected in all stages of S. avenae, and its expression reached the highest level in 2nd instar nymphs. RNAi analysis revealed that S. avenae fed with dsC002 were disturbed, and that the expression levels of C002 decreased by 54% and 69% after continuous feeding for 4 d and 6 d, respectively; significantly lower than in the control group (P<0.01). Larval survival rates decreased by 32.2% and 53.3% compared to the control group after continuous feeding for 4 d and 6 d. When S. avenae were fed aphid-susceptible wheat (Beijing837) after feeding on dsC002 for 4 d, its survival rate decreased rapidly within a short time. On day 6 and 8, the survival rate dropped to 44.4% and 35.6%, respectively, significantly lower than that of the control group. [Conclusion]  The results indicate that silencing the C002 gene can be lethal to aphids feeding on specific host plants. SaC002 may serve as a potential target gene for controlling S. avenae.

 

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