title: Several Grassland Soil Nematode Species Are Insensitive to RNA-Mediated Interference.
abstract: Phenotypic analysis of defects caused by RNA mediated interference (RNAi) in Caenorhabditis elegans has proven to be a powerful tool for determining gene function. In this study we investigated the effectiveness of RNAi in four non-model grassland soil nematodes, Oscheius sp FVV-2., Rhabditis sp, Mesorhabditis sp., and Acrobeloides sp. In contrast to reference experiments performed using C. elegans and Caenorhabditis briggsae, feeding bacteria expressing dsRNA and injecting dsRNA into the gonad did not produce the expected RNAi knockdown phenotypes in any of the grassland nematodes. Quantitative reverse-transcribed PCR (qRT-PCR) assays did not detect a statistically significant reduction in the mRNA levels of endogenous genes targeted by RNAi in Oscheius sp., and Mesorhabditis sp. From these studies we conclude that due to low effectiveness and inconsistent reproducibility, RNAi knockdown phenotypes in non-Caenorhabditis nematodes should be interpreted cautiously.
authors.affiliation: Ecological Genomics Institute, Kansas State University, Manhattan, KS 66506. ; Division of Biology, Kansas State University, Manhattan, KS 66506. ; Present address: Department of Biology, University of Rochester, Rochester, NY 14627.
authors.forename: David
authors.initials: D
authors.lastname: Wheeler
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Darby
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Todd
authors.forename: Michael A
authors.initials: MA
authors.lastname: Herman
keywords: Konza prairie; RNAi; molecular biology; soil nematode