2021年1月14日
Effectiveness of orally-delivered double-stranded RNA on gene silencing in the stinkbug Plautia stali
PLOS ONE
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- 巻
- 16
- 号
- 1
- 開始ページ
- e0245081
- 終了ページ
- e0245081
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1371/journal.pone.0245081
- 出版者・発行元
- Public Library of Science (PLoS)
Development of a reliable method for RNA interference (RNAi) by orally-delivered double-stranded RNA (dsRNA) is potentially promising for crop protection. Considering that RNAi efficiency considerably varies among different insect species, it is important to seek for the practical conditions under which dsRNA-mediated RNAi effectively works against each pest insect. Here we investigated RNAi efficiency in the brown-winged green stinkbug <italic>Plautia stali</italic>, which is notorious for infesting various fruits and crop plants. Microinjection of dsRNA into <italic>P</italic>. <italic>stali</italic> revealed high RNAi efficiency–injection of only 30 ng dsRNA into last-instar nymphs was sufficient to knockdown target genes as manifested by their phenotypes, and injection of 300 ng dsRNA suppressed the gene expression levels by 80% to 99.9%. Knockdown experiments by dsRNA injection showed that multicopper oxidase 2 (MCO2), vacuolar ATPase (vATPase), inhibitor of apoptosis (IAP), and vacuolar-sorting protein Snf7 are essential for survival of <italic>P</italic>. <italic>stali</italic>, as has been demonstrated in other insects. By contrast, <italic>P</italic>. <italic>stali</italic> exhibited very low RNAi efficiency when dsRNA was orally administered. When 1000 ng/μL of dsRNA solution was orally provided to first-instar nymphs, no obvious phenotypes were observed. Consistent with this, RT-qPCR showed that the gene expression levels were not affected. A higher concentration of dsRNA (5000 ng/μL) induced mortality in some cohorts, and the gene expression levels were reduced to nearly 50%. Simultaneous oral administration of dsRNA against potential RNAi blocker genes did not improve the RNAi efficiency of the target genes. In conclusion, <italic>P</italic>. <italic>stali</italic> shows high sensitivity to RNAi with injected dsRNA but, unlike the allied pest stinkbugs <italic>Halyomorpha halys</italic> and <italic>Nezara viridula</italic>, very low sensitivity to RNAi with orally-delivered dsRNA, which highlights the varied sensitivity to RNAi across different species and limits the applicability of the molecular tool for controlling this specific insect pest.
- リンク情報
- ID情報
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- DOI : 10.1371/journal.pone.0245081
- eISSN : 1932-6203
- PubMed ID : 33444324
- PubMed Central 記事ID : PMC7808618