The effect of silencing 20E biosynthesis relative genes by feeding bacterially expressed dsRNA on the larval development of Chilo suppressalis.
The effect of silencing 20E biosynthesis relative genes by feeding bacterially expressed dsRNA on the larval development of Chilo suppressalis.
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DOI：
10.1038/srep28697
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发表时间：
2016-06-29
影响因子：
4.6
通讯作者：
Niu CY
中科院分区：
文献类型：
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作者：
Zhu J;Dong YC;Li P;Niu CY
RNA干扰(RNAi)是研究基因功能和害虫防治潜力的有力工具。寻找合适的靶标基因是发展有效的RNAi介导的害虫控制技术的关键步骤。根据二化螟的转录组，鉴定了24个与昆虫激素生物合成相关的非基因。其中，与蜕皮激素发生有关的4个基因，即PTTH、THORE、SPOCK和nm-G被评为功能研究的候选靶点。克隆了这四个基因的部分cDNA，并将其在细菌中表达的dsRNA饲喂昆虫。结果显示，3天后，靶基因的mRNA丰度显著降低。此外，这四个基因的缺失导致了异常的表型和高的幼虫死亡率。15天后，dsspok组、dsptth组、dstorso组和dsmm-g组的昆虫存活率分别比对照显著降低32%、38%、56%和67%。此外，在dsRNA处理组中，约80%的存活幼虫表现出发育迟缓。这些结果表明，口服细菌表达的dsRNA可以抑制ptth、躯干、spok和nm-g的表达。口服细菌表达的dsRNA提供了一种简单和潜在的治疗方案。
RNA interference (RNAi) is a robust tool to study gene functions as well as potential for insect pest control. Finding suitable target genes is the key step in the development of an efficient RNAi-mediated pest control technique. Based on the transcriptome of Chilo suppressalis, 24 unigenes which putatively associated with insect hormone biosynthesis were identified. Amongst these, four genes involved in ecdysteroidogenesis i.e., ptth, torso, spook and nm-g were evaluated as candidate targets for function study. The partial cDNA of these four genes were cloned and their bacterially expressed dsRNA were fed to the insects. Results revealed a significant reduction in mRNA abundance of target genes after 3 days. Furthermore, knocked down of these four genes resulted in abnormal phenotypes and high larval mortality. After 15 days, the survival rates of insects in dsspook, dsptth, dstorso, and dsnm-g groups were significantly reduced by 32%, 38%, 56%, and 67% respectively, compared with control. Moreover, about 80% of surviving larvae showed retarded development in dsRNA-treated groups. These results suggest that oral ingestion of bacterially expressed dsRNA in C. suppressalis could silence ptth, torso, spook and nm-g. Oral delivery of bacterially expressed dsRNA provides a simple and potential management scheme against C. suppressalis.