Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Wester
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ORIGINAL PAPER
Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte) Pamela M. Bachman • Renata Bolognesi • William J. Moar • Geoffrey M. Mueller • Mark S. Paradise • Parthasarathy Ramaseshadri • Jianguo Tan • Joshua P. Uffman JoAnne Warren • B. Elizabeth Wiggins • Steven L. Levine
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Received: 10 December 2012 / Accepted: 26 May 2013 Ó The Author(s) 2013. This article is published with open access at Springerlink.com
Abstract The sequence specificity of the endogenous RNA interference pathway allows targeted suppression of genes essential for insect survival and enables the development of durable and efficacious insecticidal products having a low likelihood to adversely impact non-target organisms. The spectrum of insecticidal activity of a 240 nucleotide (nt) dsRNA targeting the Snf7 ortholog in Western Corn Rootworm (WCR; Diabrotica virgifera virgifera) was characterized by selecting and testing insects based upon their phylogenetic relatedness to WCR. Insect species, representing 10 families and 4 Orders, were evaluated in subchronic or chronic diet bioassays that measured potential lethal and sublethal effects. When a specific species could not be tested in diet bioassays, the ortholog to the WCR Snf7 gene (DvSnf7) was cloned and corresponding dsRNAs were tested against WCR and Colorado potato beetle (Leptinotarsa
decemlineata); model systems known to be sensitive to ingested dsRNA. Bioassay results demonstrate that the spectrum of activity for DvSnf7 is narrow and activity is only evident in a subset of beetles within the Galerucinae subfamily of Chrysomelidae ([90 % identity with WCR Snf7 240 nt). This approach allowed for evaluating the relationship between minimum shared nt sequence length and activity. A shared sequence length of C21 nt was required for efficacy against WCR (containing 221 potential 21-nt matches) and all active orthologs contained at least three 21 nt matches. These results also suggest that WCR resistance to DvSnf7 dsRNA due to single nucleotide polymorphisms in the target sequence of 240 nt is highly unlikely.
Electronic supplementary material The online version of this article (doi:10.1007/s11248-013-9716-5) contains supplementary material, which is available to authorized users.
Introduction
P. M. Bachman (&) W. J. Moar G. M. Mueller M. S. Paradise J. Tan J. P. Uffman J. Warren S. L. Levine Monsanto Company, 800 N Lindbergh Blvd., St. Louis, MO 63167, USA e-mail: [email protected] R. Bolognesi P. Ramaseshadri B. E. Wiggins Monsanto Company, 700 Chesterfield Parkway W, Chesterfield, MO 63017, USA
Keywords RNAi Specificity Coleoptera Activity spectrum Non-target organism
RNA interference (RNAi) technology can achieve sequence-specific gene silencing in some insects by feeding double-stranded RNAs (dsRNAs) (Baum et al. 2007; Whyard et al. 2009; Swevers and Smagghe 2012). Sequence-specific gene silencing combined with th
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