Abstract

RNA interference (RNAi)-based technology shows great potential for use in agriculture, particularly for management of costly insect pests. In the decade since the insecticidal effects of environmentally-introduced RNA were first reported, this treatment has been applied to several types of insect pests. Through the course of those efforts, it has become apparent that different insects exhibit a range of sensitivity to environmentally-introduced RNAs. The variation in responses across insect is not well-understood, with differences in the underlying RNAi mechanisms being one explanation. This study evaluates eight proteins among three agricultural pests whose responses to environmental RNAi are known to differ: western corn rootworm (Diabrotica virgifera virgifera), fall armyworm (Spodoptera frugiperda), and southern green stink bug (Nezara viridula). These proteins have been identified in various organisms as centrally involved in facilitating the microRNA- and small interfering-RNA-mediated interference responses. Various bioinformatics tools, as well as gene expression profiling, were used to identify and evaluate putative homologues for characteristics that may contribute to the differing responses of these insects, such as the absence of critical functional domains within expressed sequences, the absence of entire gene sequences, or unusually low or undetectable expression of critical genes. Though many similarities were observed, the number of isoforms and expression levels of double-stranded RNA-binding and argonaute proteins varied across insect. Differences among key RNAi machinery genes of these three pests may impact the function of their RNAi pathways, and therefore, their respective responses to exogenous RNAs.

Highlights

  • Control of agricultural pests through use of genetically engineered crops is a critical aspect of the integrated approach needed to provide sustainable food supplies for a growing world population [1]

  • Putative homologues of all eight core RNA interference (RNAi) machinery genes were mined from WCR, FAW, and SGSB complementary DNA datasets through a series of iterative searches beginning with Drosophila melanogaster (Dme) query sequences

  • Multiple versions of potential homologues were identified for Pasha, LOQS, R2D2, Argonaute 1 (AGO1), and Argonaute 2 (AGO2), which may represent products of alternative transcriptional start sites, alternative splicing, or duplicated genes

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Summary

Introduction

Control of agricultural pests through use of genetically engineered crops is a critical aspect of the integrated approach needed to provide sustainable food supplies for a growing world population [1]. Utilization of RNA interference (RNAi)-based technology in genetically engineered crops is currently being widely explored for insect pest management [2]. RNAi encompasses three related mechanisms of control at the RNA level, wherein RNA is targeted for repression or degradation through the action of microRNA (miRNA), small interfering RNA (siRNA), or Piwi-interacting RNA (piRNA). This general process, first discovered in Petunia and described in Caenorhabditis elegans (Cel) [3, 4], is highly conserved across plants and animals. The Dme pathways may serve as an appropriate model system for study of RNAi in other insects, though it remains unclear how much mechanistic information may be reliably extrapolated. As the components most directly involved in the interference response, these are considered “core” RNAi machinery and are the focus of the current study

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