Abstract
BackgroundThe western corn rootworm, Diabrotica virgifera virgifera, is a pervasive pest of maize in North America and Europe, which has adapted to current pest management strategies. In advance of an assembled and annotated D. v. virgifera genome, we developed transcriptomic resources to use in identifying candidate genes likely to be involved in the evolution of resistance, starting with members of the ATP-binding cassette (ABC) transporter family.ResultsIn this study, 65 putative D. v. virgifera ABC (DvvABC) transporters were identified within a combined transcriptome assembly generated from embryonic, larval, adult male, and adult female RNA-sequence libraries. Phylogenetic analysis placed the deduced amino-acid sequences of the DvvABC transporters into eight subfamilies (A to H). To supplement our sequence data with functional analysis, we identified orthologs of Tribolium castaneum ABC genes which had previously been shown to exhibit overt RNA interference (RNAi) phenotypes. We identified eight such D. v. virgifera genes, and found that they were functionally similar to their T. castaneum counterparts. Interestingly, depletion of DvvABCB_39715 and DvvABCG_3712 transcripts in adult females produced detrimental reproductive and developmental phenotypes, demonstrating the potential of these genes as targets for RNAi-mediated insect control tactics.ConclusionsBy combining sequence data from four libraries covering three distinct life stages, we have produced a relatively comprehensive de novo transcriptome assembly for D. v. virgifera. Moreover, we have identified 65 members of the ABC transporter family and provided the first insights into the developmental and physiological roles of ABC transporters in this pest species.
Highlights
The western corn rootworm, Diabrotica virgifera virgifera, is a pervasive pest of maize in North America and Europe, which has adapted to current pest management strategies
Due to the relatively conserved sequence of the nucleotide-binding domain (NBD), it has been used for the phylogenetic classification of the ATP-binding cassette (ABC) transporter superfamily into eight subfamilies designated A to H (ABCA to ABCH) [29]
Transcriptome sequencing, assembly, and annotation Over 22 million raw Illumina (MiSeq) sequencing reads were generated across four libraries
Summary
The western corn rootworm, Diabrotica virgifera virgifera, is a pervasive pest of maize in North America and Europe, which has adapted to current pest management strategies. ATP-binding cassette (ABC) proteins comprise one of the largest gene families, and are found across prokaryotic and eukaryotic domains [24]. Most of these proteins function as transmembrane transporters, which actively move a myriad of molecules across cellular membranes [25]. ABC transporter proteins have a two-domain structure: a highly conserved nucleotide-binding domain (NBD) and a variable transmembrane domain (TMD) [26]. Due to the relatively conserved sequence of the NBD, it has been used for the phylogenetic classification of the ABC transporter superfamily into eight subfamilies designated A to H (ABCA to ABCH) [29]
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