Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

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• 作者：Pamela M. Bachman (1)
  Renata Bolognesi (2)
  William J. Moar (1)
  Geoffrey M. Mueller (1)
  Mark S. Paradise (1)
  Parthasarathy Ramaseshadri (2)
  Jianguo Tan (1)
  Joshua P. Uffman (1)
  JoAnne Warren (1)
  B. Elizabeth Wiggins (2)
  Steven L. Levine (1)
  
• 关键词：RNAi ; Specificity ; Coleoptera ; Activity spectrum ; Non ; target organism
• 刊名：Transgenic Research
• 出版年：2013
• 出版时间：December 2013
• 年：2013
• 卷：22
• 期：6
• 页码：1207-1222
• 全文大小：
• 作者单位：Pamela M. Bachman (1)
  Renata Bolognesi (2)
  William J. Moar (1)
  Geoffrey M. Mueller (1)
  Mark S. Paradise (1)
  Parthasarathy Ramaseshadri (2)
  Jianguo Tan (1)
  Joshua P. Uffman (1)
  JoAnne Warren (1)
  B. Elizabeth Wiggins (2)
  Steven L. Levine (1)
  
  1. Monsanto Company, 800?N Lindbergh Blvd., St. Louis, MO, 63167, USA
  2. Monsanto Company, 700 Chesterfield Parkway W, Chesterfield, MO, 63017, USA
  
• ISSN：1573-9368

文摘

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 ?1 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.