Response of various target genes to diet-delivered dsRNA mediated RNA interference in the cotton bollworm, Helicoverpa armigera

Abstract

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 μg dsRNA treatment having a more pronounced effect than 10 μg 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.