烟粉虱传毒相关蛋白基因的克隆及其表达量分析
Cloning and relative expression of a virus-transmission related gene in Bemisia tabaci (Gennadius)
刘 馨** 闫文茜 张友军 吴青君 谢 文 王少丽***
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DOI：10.7679/j.issn.2095-1353.2015.006
作者单位：中国农业科学院蔬菜花卉研究所 北京 100081
中文关键词：烟粉虱，番茄黄化曲叶病毒，传毒相关蛋白，荧光定量PCR
英文关键词：Bemisia tabaci, tomato yellow leaf curl virus, virus-transmission related protein, qRT-PCR
中文摘要： 【目的】 烟粉虱Bemisia tabaci（Gennadius）是一种世界性的入侵性害虫，其传播的番茄黄化曲叶病毒（Tomato yellow leaf curl virus，TYLCV）给我国番茄生产造成巨大经济损失。为了阐明烟粉虱传播双生病毒的机理，本文拟研究明确B和Q烟粉虱体内传毒相关蛋白GroEL基因及其表达量。【方法】 采用特异性引物克隆了B和Q烟粉虱内共生菌编码的传毒相关蛋白GroEL基因，并进行序列分析；并利用荧光定量PCR技术检测两种生物型及其获取双生病毒前后该基因的相对表达量。【结果】 烟粉虱内共生菌Hamiltonella编码的GroEL基因全长为1 668 bp，编码555个氨基酸；B、Q烟粉虱该基因的核苷酸序列相似性为99.94%，氨基酸同源性为99.82%；带毒烟粉虱中GroEL基因的表达量显著高于未带毒的对应生物型烟粉虱；无论带毒与否，Q烟粉虱该基因的表达量均显著高于B（P< 0.05）。【结论】 烟粉虱携带TYLCV后可诱导GroEL的表达量升高，B和Q烟粉虱中GroEL基因及其表达量均存在差异，这可能是B和Q烟粉虱传毒效率存在显著差异的原因之一。
英文摘要：[Objectives] The sweetpotato whitefly, Bemisia tabaci (Gennaddius), is one of the most invasive insect pests in the world. Tomato yellow leaf curl virus (TYLCV), transmitted by B. tabaci, has caused great losses to tomato production in China. In order to illustrate the mechanism of TYLCV transmission by B. tabaci, we studied the virus transmission-related gene (GroEL) and its expression in the B. tabaci B and Q. [Methods] The virus transmission-related gene, GroEL, encoded by the Hamiltonella endosymbiont of the B. tabaci B and Q, was cloned using specific primers and the resultant sequences were analyzed. The relative expressions of GroEL were measured using the qRT-PCR technique in B. tabaci that had fed on both healthy and viruliferous tomato plants. [Results] The results showed that the GroEL gene was 1 668 bp in full length, encoding 555 amino acids. There was 99.94% nucleotide sequence similarity and 99.82% amino acid homology in GroEL genes between the B. tabaci B and Q. GroEL expressions in viruliferous B. tabaci were significantly higher than in corresponding uninfected whiteflies. Also GroEL expression in the B. tabaci Q was proved to be significantly higher than B(P <0.05), regardless of whether the insects concerned were viruliferous or not. [Conclusion] GroEL expression was significantly higher in viruliferous whiteflies. Obvious differences in GroEL expression were exhibited between the B. tabaci B and Q, which might be one of reasons for their significant difference in efficiency of virus transmission.