Jie Li1, Qiuhong Chen, Yongjun Lin, Tingru Jiang, Gang Wu, Hongxia Hua. 1. Hubei Insect Resources Utilisation and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.
Abstract
BACKGROUND: An efficient and convenient RNA interference (RNAi) technique involving double-stranded RNA (dsRNA) ingestion is useful for gene function studies of non-model insects. RESULTS: Three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E (V-ATPase-E, 21E01) were synthesised for RNAi in Nilaparvata lugens. dsRNA was found to be stable in 0.1 g mL(-1) sucrose solution, but unstable in artificial fodder. Therefore, dsRNAs were orally delivered into N. lugens in 0.1 g mL(-1) sucrose solution. RNAi was induced by all three of the dsRNAs at 0.05 µg µL(-1) in N. lugens. Time dynamics analysis of gene silencing indicated that significant suppression of the target gene began as early as 2 days after ingestion of ds2-21E01 and ds3-21E01. However, significant repressive effects were recorded up to 10 days after exposure to ds1-21E01. The maximum reduction in target gene mRNA was observed after 10 days of treatment, with suppression ratios induced by ds1-21E01, ds2-21E01 and ds3-21E01 of 41, 55 and 48% respectively. CONCLUSION: An efficient and convenient RNAi technique involving dsRNA ingestion has been successfully developed for N. lugens. This will be a useful tool for further functional genomic investigation in this organism.
BACKGROUND: An efficient and convenient RNA interference (RNAi) technique involving double-stranded RNA (dsRNA) ingestion is useful for gene function studies of non-model insects. RESULTS: Three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E (V-ATPase-E, 21E01) were synthesised for RNAi in Nilaparvata lugens. dsRNA was found to be stable in 0.1 g mL(-1) sucrose solution, but unstable in artificial fodder. Therefore, dsRNAs were orally delivered into N. lugens in 0.1 g mL(-1) sucrose solution. RNAi was induced by all three of the dsRNAs at 0.05 µg µL(-1) in N. lugens. Time dynamics analysis of gene silencing indicated that significant suppression of the target gene began as early as 2 days after ingestion of ds2-21E01 and ds3-21E01. However, significant repressive effects were recorded up to 10 days after exposure to ds1-21E01. The maximum reduction in target gene mRNA was observed after 10 days of treatment, with suppression ratios induced by ds1-21E01, ds2-21E01 and ds3-21E01 of 41, 55 and 48% respectively. CONCLUSION: An efficient and convenient RNAi technique involving dsRNA ingestion has been successfully developed for N. lugens. This will be a useful tool for further functional genomic investigation in this organism.
Authors: Freja Scheys; Els J M Van Damme; Jarne Pauwels; An Staes; Kris Gevaert; Guy Smagghe Journal: Mol Cell Proteomics Date: 2020-01-10 Impact factor: 5.911