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Sensitivity to RNA interference-mediated gene silencing in intact and ligated larvae of the armyworm, Mythimna separata (Lepidoptera: Noctuidae)

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Abstract

RNA interference (RNAi) is a common tool for analysis of gene function in both model and non-model insects, but it is becoming evident that RNAi efficiency varies considerably from species to species. We examined RNAi efficiency in larvae of the armyworm Mythimna separata (Walker) using multiple genes and tissues. First, we showed that five different target genes exhibited distinct tissue distribution patterns by quantitative determination of mRNA in total hemocytes, foregut, midgut, hindgut, Malpighian tubules and fat body: neuroglian mRNA was most abundant in fat body; inhibitor of apoptosis proteins mRNA was found to be ubiquitous; aquaporin 4 mRNA was most enriched in hindgut; cueball and prophenoloxidase 2 were mainly expressed in hemocytes. Second, we assessed sensitivity to gene silencing by double-strand RNA injection of these five genes in the six different tissues. We found that these genes generally showed refractoriness to double-strand RNA-mediated gene knockdown irrespective of the tissue tested. Finally, we demonstrated that appreciable gene knockdown was achieved at least in the adhering hemocyte fraction when larval isolated abdomen was prepared by ligation and subjected to dsRNA injection. Our study thus added detailed information on the refractoriness of larval tissues of a lepidopteran insect to gene silencing through RNAi and provided a new potential approach to improve RNAi efficiency.

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Acknowledgments

This work was supported in part by Grant-in-Aid 23658047 and 25450486, to KM from the Japan Society for the Promotion of Science.

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Author notes

  1. Masashi Kamezaki

    Present address: Biology Group, Discovery Health and Crop Sciences Research Laboratory, Sumitomo Chemical Co., Ltd, 4-2-1 Takatsukasa, Takarazuka, 665-8555, Japan

  2. Tatsuhiro Yoshida

    Present address: Aichi Nishimikawa Office of Agriculture, Forestry and Fisheries, 1-4 Myodaijihonmachi, Okazaki, 444-0860, Japan

Authors and Affiliations

  1. Applied Entomology Laboratory, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa, Nagoya, 464-8601, Japan

    Kakeru Yokoi, Toshiharu Tanaka & Ken Miura

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  1. Kakeru Yokoi
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  2. Masashi Kamezaki
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  4. Toshiharu Tanaka
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Correspondence to Ken Miura.

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Yokoi, K., Kamezaki, M., Yoshida, T. et al. Sensitivity to RNA interference-mediated gene silencing in intact and ligated larvae of the armyworm, Mythimna separata (Lepidoptera: Noctuidae). Appl Entomol Zool 48, 431–439 (2013). https://doi.org/10.1007/s13355-013-0201-7

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  • DOI: https://doi.org/10.1007/s13355-013-0201-7

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