RNA interference for the control of whiteflies (Bemisia tabaci) by oral route
RNA interference for the control of whiteflies (Bemisia tabaci) by oral route
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DOI：
10.1007/s12038-011-9009-1
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Tuli, Rakesh
中科院分区：
文献类型：
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作者：
Upadhyay, Santosh Kumar;Chandrashekar, K.;Tuli, Rakesh
探讨了RNA干扰(RNAi)介导的基因沉默技术在烟粉虱防治中的应用。从肌动蛋白同源、ADP/ATP转位酶、α-微管蛋白、核糖体蛋白L9(RPL9)和V-ATPase A亚单位5个不同的基因合成了dsRNAs和siRNAs。建立了一种简化的昆虫生物测定方法，用于通过口服途径传递DS/siRNA，并对其效果进行了评价。在饲养6天后，DS/siRNA导致29-97%的死亡率。每只昆虫每天摄入近150 NL的昆虫饲料，其中最多含有6纳克的RNA。抑制RPL9和V-ATPase A的表达可导致较高的死亡率，其致死率分别为11.21和3.08微克/毫升。半定量聚合酶链式反应显示，处理昆虫的RPL9和V-ATPase A转录本水平显著降低。在室温下，昆虫饲料中的siRNAs至少稳定了7天。在转基因植物中表达RPL9和V-ATPase A的韧皮部特异的dsRNAs来保护烟粉虱，可能是这项技术的一个有趣的应用。
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 mu g/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.