Response of various target genes to diet-delivered dsRNA mediated RNA interference in the cotton bollworm, Helicoverpa armigera
Response of various target genes to diet-delivered dsRNA mediated RNA interference in the cotton bollworm, Helicoverpa armigera
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DOI：
10.1007/s10340-013-0541-7
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发表时间：
2014-03-01
影响因子：
4.8
通讯作者：
Sita, T.
中科院分区：
文献类型：
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作者：
Asokan, R.;Chandra, G. Sharath;Sita, T.
棉铃虫Helicoverpa armigera是一种高度多食性害虫，每年危害许多重要经济作物，造成巨大的作物损失。多年来，由于各种因素，包括对广谱化学杀虫剂产生耐药性，这种害虫的管理具有挑战性。RNA干扰（RNAi）在防治害虫方面具有巨大的潜力。然而，RNAi介导的沉默效率因基因而异，因此成功的RNAi介导的害虫防治需要选择合适的靶基因，所述靶基因通过中肠外源递送同源双链RNA（dsRNA）而有效沉默。因此，我们评估了通过半合成饮食递送的两种浓度的dsRNA在沉默五个重要基因中的作用，即谷胱甘肽-S-转移酶、细胞色素P450（两者都参与宿主化感物质的解毒）;胰蛋白酶、胰凝乳蛋白酶（两者都参与蛋白质的消化）和保幼激素酸甲基转移酶（jhamt）（参与幼虫变态）。通过定量实时PCR（qRT-PCR）评估沉默的程度。结果表明，上述目的基因均被沉默，20 μ g dsRNA处理在降低转录水平、幼虫、蛹重和化蛹方面比10 μ g处理效果更显著。与其他靶基因相比，糜蛋白酶的沉默对幼虫和蛹重的影响更明显，而jhamt严重影响化蛹。本研究表明，靶基因对RNAi的敏感性不同，胰凝乳蛋白酶和jhamt被证明是合适的候选基因，可用于RNAi介导的管理H。armigera。
The cotton bollworm, Helicoverpa armigera is a highly polyphagous pest infesting a number of economically important crops, annually causing enormous crop losses. Management of this pest is challenging over the years due to various factors including development of resistance to wide spectrum of chemical insecticides. RNA interference (RNAi) has tremendous potential to combat insect pests. However, RNAi mediated silencing efficiency varies from gene to gene, hence successful RNAi mediated pest control requires selection of suitable target gene(s), which are effectively silenced by the exogenous delivery of cognate double-stranded RNA (dsRNA) through midgut. Therefore, we have evaluated the effect of two concentrations of dsRNA delivered through semi-synthetic diet in silencing five important genes, viz. glutathione-S-transferase, cytochrome P450 (both involved in detoxification of host allelochemicals); trypsin, chymotrypsin (both involved in digestion of proteins) and juvenile hormone acid methyl transferase (jhamt) (involved in larval metamorphosis). Extent of silencing was assessed by quantitative real-time PCR (qRT-PCR). Results revealed that above target genes were silenced variably, 20 mu g dsRNA treatment having a more pronounced effect than 10 mu g in reducing the transcript levels, larval, pupal weight, and pupation. Silencing of chymotrypsin had a more pronounced effect on larval and pupal weight compared to other target genes, while jhamt severely affected pupation. This study demonstrated that target genes have varied sensitivity to RNAi, chymotrypsin, and jhamt were shown to be suitable candidate genes that could be utilized for RNAi mediated management of H. armigera.