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Abstract
RNAi promises to reshape pest control by being nontoxic, biodegradable, and species specific. However, due to the plastic nature of RNAi, there is a significant variability in responses. In this study, we investigate small RNA pathways and processing of ingested RNAi trigger molecules in a hemipteran plant pest, the whitefly Bemisia tabaci Unlike Drosophila, where the paradigm for insect RNAi technology was established, whitefly has abundant somatic piwi-associated RNAs (piRNAs). Long regarded as germline restricted, piRNAs are common in the soma of many invertebrates. We sought to exploit this for a novel gene silencing approach. The main principle of piRNA biogenesis is the recruitment of target RNA fragments into the pathway. As such, we designed synthetic RNAs to possess complementarity to the loci we annotated. Following feeding of these exogenous piRNA triggers knockdown as effective as conventional siRNA-only approaches was observed. These results demonstrate a new approach for RNAi technology that could be applicable to dsRNA-recalcitrant pest species and could be fundamental to realizing insecticidal RNAi against pests.
Highlights
RNAi technology has been shown to be applicable as a low-toxicity biopesticide to control agricultural insect pests and vectors of plant pathogens through silencing essential, biologically relevant genes (Zotti & Smagghe, 2015)
To characterize the RNAi pathways of whitefly, we first sought to identify the collection of Ago/Piwi proteins encoded in the whitefly “B biotype” genome (MEAM1v1.2) using existing annotations and BLAST to curate sequences (Fig 1A) (Chen et al, 2016)
We found a significant population of piwi-associated RNAs (piRNAs), which are more abundant than the endogenous siRNAs—the species exploited by existing RNAi approaches
Summary
RNAi technology has been shown to be applicable as a low-toxicity biopesticide to control agricultural insect pests and vectors of plant pathogens through silencing essential, biologically relevant genes (Zotti & Smagghe, 2015). A number of products are available, some arthropod pests exhibit moderate or only minor sensitivity to dsRNA upon ingestion (Yu et al, 2013; Zhu & Palli, 2020) This suggests that to fully realize this strategy across most or all arthropods, RNAi triggers may require unique engineering relevant to each target species (Shukla et al, 2016; Parsons et al, 2018). RNAi pathways were investigated for the whitefly Bemisia tabaci (Genn.) (Aleyrodidae, Hemiptera) to characterize the fundamental features that might be exploited to improve RNAi approach(es). As a group, this whitefly is considered a cryptic or sibling species. The behavior of these molecules is investigated in the context of an extant RNAi mechanism in B. tabaci
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