Molecular characterization, expression analysis and RNAi knock-down of elongation factor 1 alpha and 1 gamma from Nilaparvata lugens and its yeast-like symbiont

文献类型： 外文期刊

第一作者： Wang, W. X.

作者： Wang, W. X.;Li, K. L.;Chen, L. F.;Lai, F. X.;Fu, Q.;Zhu, T. H.;Chen, L. F.

作者机构：

关键词： Nilaparvata lugens;yeast-like symbionts;elongation factor 1 gamma;elongation factor 1 alpha;RNAi

期刊名称：BULLETIN OF ENTOMOLOGICAL RESEARCH （ 影响因子：1.75； 五年影响因子：2.007 ）

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收录情况： SCI

摘要： In the present paper, four cDNAs encoding the alpha and gamma subunits of elongation factor 1 (EF-1) were cloned and sequenced from Nilaparvata lugens, named NlEF-1 alpha, NlEF-1 gamma, and its yeast-like symbiont (YLS), named YsEF-1 alpha and YsEF-1 gamma, respectively. Comparisons with sequences from other species indicated a greater conservation for EF-1 alpha than for EF-1 gamma. NlEF-1 alpha has two identical copies. The deduced amino acid sequence homology of NlEF-1 alpha and NlEF-1 gamma is 96 and 64%, respectively, compared with Homalodisca vitripennis and Locusta migratoria. The deduced amino acid sequence homology of YsEF-1 alpha and YsEF-1 gamma is 96 and 74%, respectively, compared with Metarhizium anisopliae and Ophiocordyceps sinensis. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis revealed that the expression level of NlEF-1 alpha and NlEF-1 gamma mRNA in hemolymph, ovary, fat body and salivary glands were higher than the midgut and leg tissue. YsEF-1 alpha and YsEF-1 gamma was highly expressed in fat body. The expression level of NlEF-1 alpha was higher than that of NlEF-1 gamma. Through RNA interference (RNAi) of the two genes, the mortality of nymph reached 92.2% at the 11th day after treatment and the ovarian development was severely hindered. The RT-qPCR analysis verified the correlation between mortality, sterility and the down-regulation of the target genes. The expression and synthesis of vitellogenin (Vg) protein in insects injected with NlEF-1 alpha and NlEF-1 gamma double-stranded RNA (dsRNA) was significantly lower than control groups. Attempts to knockdown the YsEF-1 genes in the YLS was unsuccessful. However, the phenotype of N. lugens injected with YsEF-1 alpha dsRNA was the same as that injected with NlEF-1 alpha dsRNA, possibly due to the high similarity (up to 71.9%) in the nucleotide sequences between NlEF-1 alpha and YsEF-1 alpha. We demonstrated that partial silencing of NlEF-1 alpha and NlEF-1 gamma genes caused lethal and sterility effect on N. lugens. NlEF-1 gamma shares low identity with that of other insects and therefore it could be a potential target for RNAi-based pest management.

分类号： Q96

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