National Botanical Research Institute (Council of Scientific and Industrial Research), Rana Pratap Marg, Lucknow 226 001, India.
J Biosci. 2011 Mar;36(1):153-61. doi: 10.1007/s12038-011-9009-1.
RNA interference (RNAi)-mediated gene silencing was explored for the control of sap-sucking pest Bemisia tabaci, commonly known as whitefly. dsRNAs and siRNAs were synthesized from five different genes - actin ortholog, ADP/ATP translocase, alpha-tubulin, ribosomal protein L9 (RPL9) and V-ATPase A subunit. A simplified insect bioassay method was developed for the delivery of ds/siRNA through the oral route, and efficacy was evaluated. ds/siRNA caused 29-97% mortality after 6 days of feeding. Each insect ingested nearly 150 nl of insect diet per day, which contained a maximum of 6 ng of RNA. Knocking down the expression of RPL9 and V-ATPase A caused higher mortality with LC50 11.21 and 3.08 microg/ml, respectively, as compared to other genes. Semi-quantitative PCR of the treated insects showed significant decrease in the level of RPL9 and V-ATPase A transcripts. siRNAs were found stable in the insect diet for at least 7 days at the room temperature. Phloem-specific expression of dsRNAs of RPL9 and V-ATPase A in transgenic plants for the protection against whiteflies might be an interesting application of this technology.
RNA 干扰(RNAi)介导的基因沉默被探索用于控制刺吸式害虫烟粉虱,通常称为粉虱。dsRNA 和 siRNA 是从 5 个不同的基因 - 肌动蛋白同源物、ADP/ATP 易位酶、α-微管蛋白、核糖体蛋白 L9(RPL9)和 V-ATPase A 亚基合成的。开发了一种简化的昆虫生物测定方法,通过口服途径递送 ds/siRNA,并评估了其功效。ds/siRNA 在 6 天的喂养后导致 29-97%的死亡率。每只昆虫每天摄入约 150 毫微升的昆虫饮食,其中含有最多 6 纳克的 RNA。与其他基因相比,RPL9 和 V-ATPase A 的表达下调导致更高的死亡率,LC50 分别为 11.21 和 3.08 微克/毫升。对处理过的昆虫进行半定量 PCR 显示,RPL9 和 V-ATPase A 转录本的水平显著下降。siRNA 在昆虫饮食中在室温下至少稳定 7 天。dsRNA 对 RPL9 和 V-ATPase A 的韧皮部特异性表达可能是这项技术的一个有趣应用。