State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, Guangdong, China.
Insect Mol Biol. 2010 Dec;19(6):777-86. doi: 10.1111/j.1365-2583.2010.01038.x. Epub 2010 Aug 19.
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 µg/µl 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.
褐飞虱,Nilaparvata lugens,是近年来爆发的最具破坏性的水稻虫害。RNA 干扰 (RNAi) 是一项技术突破,已被开发成为研究基因功能和对害虫进行高度靶向控制的有力工具。在这里,我们研究了使用基于喂食的 RNAi 技术靶向 N. lugens 基因海藻糖磷酸合酶 (TPS) 的效果。N. lugens TPS (NlTPS) 的全长 cDNA 为 3235 bp,开放阅读框为 2424 bp,编码 807 个氨基酸的蛋白质。NlTPS 在脂肪体、中肠和卵巢中表达。定量实时 PCR (qRT-PCR) 分析显示,NlTPS mRNA 在昆虫的整个生命周期中持续表达,变化不大。通过双链 RNA (dsRNA) 喂食高效沉默 TPS 基因导致 TPS mRNA 和酶活性的快速显著降低。此外,喂食 dsRNA 的 N. lugens 幼虫的发育受到干扰,导致致死率,连续摄入 0.5 µg/µl dsRNA 2、4、7 和 10 天后的累积存活率分别降至 75.56%、64.44%、55.56%和 40.00%,显著低于对照组昆虫的存活率,表明 NlTPS dsRNA 可能可用作害虫防治手段。