Gene knockdown by intro-thoracic injection of double-stranded RNA in the brown planthopper,Nilaparvata lugens |
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| Authors: | Shuhua Liu Zhiping Ding Chengwei Zhang Baojun Yang Zewen Liu |
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| Institution: | 1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2. Scientific Observing and Experimental Station of Crop Pests in Guilin, Ministry of Agriculture, Guilin 541399, China;3. Southern Regional Collaborative Innovation Center for Grain and Oil Crops in China, Changsha 410128, China |
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| Abstract: | RNA interference (RNAi) is a powerful strategy for gene function study in insects. Here, we described the development of a RNAi technique by microinjection of double-stranded RNA (dsRNA) in the brown planthopper Nilaparvata lugens. Based on the mortality and RNAi efficiency criteria, the conjunctive between prothorax and mesothorax was selected as the injection site and 50 nl as injection volume. Three genes with different expression patterns were selected to evaluate the RNAi efficiency. A comparable 40% decrease of gene expression was observed at the 4th day after injection for the ubiquitously expressed calreticulin and the gut specific cathepsin-B genes, but only 25% decrease at the 5th day for the central nervous system specific Nlβ2 gene. Double injection could increase the RNAi efficiency, such as from 25% to 53% for Nlβ2 gene. The gene knockdown technique developed in this study will be an essential post-genomic tool for further investigations in N. lugens. |
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