Feeding-based RNA interference of a trehalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens
Feeding-based RNA interference of a trehalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens
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DOI：
10.1111/j.1365-2583.2010.01038.x
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发表时间：
2010-12-01
影响因子：
2.6
通讯作者：
Zhang, W.
中科院分区：
文献类型：
--
作者：
Chen, J.;Zhang, D.;Zhang, W.
褐飞虱 (Nilaparvata lugens) 是近年来爆发的最具破坏性的水稻害虫。 RNA 干扰 (RNAi) 是一项技术突破，已成为研究基因功能和高度针对性地防治害虫的强大工具。在这里，我们研究了使用基于喂养的 RNAi 技术来靶向褐飞虱中的海藻糖磷酸合酶 (TPS) 基因的效果。褐飞虱TPS(NlTPS)的cDNA全长3235bp，开放阅读框2424bp，编码807个氨基酸的蛋白质。 NlTPS在脂肪体、中肠和卵巢中表达。实时定量PCR (qRT-PCR) 分析表明，NlTPS mRNA 在昆虫的一生中持续表达，变化很小。通过双链 RNA (dsRNA) 喂养有效沉默 TPS 基因，导致 TPS mRNA 和酶活性水平快速显着降低。此外，饲喂dsRNA的褐飞虱幼虫发育受到干扰，导致致死率，连续摄入0.5μg/μl dsRNA 2、4、7和10天后，累积存活率分别降至75.56、64.44、55.56和40.00%。这些值显着低于对照组昆虫的值，表明 NlTPS dsRNA 可用作害虫控制的手段。
The brown planthopper, Nilaparvata lugens, is the most devastating rice insect pest to have given rise to an outbreak in recent years. RNA interference (RNAi) is a technological breakthrough that has been developed as a powerful tool for studying gene function and for the highly targeted control of insect pests. Here, we examined the effects of using a feeding-based RNAi technique to target the gene trehalose phosphate synthase (TPS) in N. lugens. The full-length cDNA of N. lugens TPS (NlTPS) is 3235 bp and has an open reading frame of 2424 bp, encoding a protein of 807 amino acids. NlTPS was expressed in the fat body, midgut and ovary. Quantitative real-time PCR (qRT-PCR) analysis revealed that NlTPS mRNA is expressed continuously with little change during the life of the insect. Efficient silencing of the TPS gene through double-stranded RNA (dsRNA) feeding led to rapid and significant reduction levels of TPS mRNA and enzymatic activity. Additionally, the development of N. lugens larvae that had been fed with the dsRNA was disturbed, resulting in lethality, and the cumulative survival rates dropped to 75.56, 64.44, 55.56 and 40.00% after continuous ingestion of 0.5 mu g/mu l dsRNA for 2, 4, 7 and 10 days, respectively. These values were significantly lower than those of the insects in the control group, suggesting that NlTPS dsRNA may be useful as a means of insect pest control.