Abstract
BackgroundExpression of double strand RNA (dsRNA) designed against important insect genes in transgenic plants have been shown to give protection against pests through RNA interference (RNAi), thus opening the way for a new generation of insect-resistant crops. We have earlier compared the efficacy of dsRNAs/siRNAs, against a number of target genes, for interference in growth of whitefly (Bemisia tabaci) upon oral feeding. The v-ATPase subunit A (v-ATPaseA) coding gene was identified as a crucial target. We now report the effectiveness of transgenic tobacco plants expressing siRNA to silence v-ATPaseA gene expression for the control of whitefly infestation.Methodology/Principal FindingsTransgenic tobacco lines were developed for the expression of long dsRNA precursor to make siRNA and knock down the v-ATPaseA mRNA in whitefly. Molecular analysis and insecticidal properties of the transgenic plants established the formation of siRNA targeting the whitefly v-ATPaseA, in the leaves. The transcript level of v-ATPaseA in whiteflies was reduced up to 62% after feeding on the transgenic plants. Heavy infestation of whiteflies on the control plants caused significant loss of sugar content which led to the drooping of leaves. The transgenic plants did not show drooping effect.Conclusions/SignificanceHost plant derived pest resistance was achieved against whiteflies by genetic transformation of tobacco which generated siRNA against the whitefly v-ATPaseA gene. Transgenic tobacco lines expressing dsRNA of v-ATPaseA, delivered sufficient siRNA to whiteflies feeding on them, mounting a significant silencing response, leading to their mortality. The transcript level of the target gene was reduced in whiteflies feeding on transgenic plants. The strategy can be taken up for genetic engineering of plants to control whiteflies in field crops.
Highlights
Agricultural crops are damaged worldwide by more than 10,000 species of insects [1].To circumvent the yield losses due to insect attack, diverse approaches are evolving for genetic engineering of crops to impart host plant derived pest resistance [2,3]
This study examines if transgenic tobacco plants, designed to express double strand RNA (dsRNA) precursor against the whitefly v-ATPaseA, can form sufficient small interfering RNAs (siRNA) and cause mortality to offer protection against the sucking pest
This is the first report of RNA interference (RNAi) mediated gene silencing in whitefly through feeding on transgenic plants
Summary
Agricultural crops are damaged worldwide by more than 10,000 species of insects [1].To circumvent the yield losses due to insect attack, diverse approaches are evolving for genetic engineering of crops to impart host plant derived pest resistance [2,3]. Sap-sucking pests are emerging as more serious pests in cotton fields in comparison to bollworms [1,9] This group of insect pests (jassid, whitefly, aphid, thrips, mealy bug, mites, mired bug etc.) damage plants by sucking photosynthetic assimilates, and acts as vectors of several plant viruses [10]. They excrete honey dew which promotes fungal infection, causing loss to the quality of farm produce [11,12]. We report the effectiveness of transgenic tobacco plants expressing siRNA to silence vATPaseA gene expression for the control of whitefly infestation
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