Chang Ya-Wen, Wang Yu-Cheng, Zhang Xiao-Xiang, Iqbal Junaid, Du Yu-Zhou
College of Horticulture and Plant Protection & Institute of Applied Entomology, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China.
Insects. 2021 Jan 6;12(1):41. doi: 10.3390/insects12010041.
The leafminer fly, , is an invasive pest of vegetable and horticultural crops in China. In this study, a microinjection method based on dsRNA was developed for RNA interference (RNAi) in using genes encoding vacuolar-ATPase (). Expression analysis indicated that and were more highly expressed in adults than in larvae or pupae. Microinjection experiments with and were conducted to evaluate the efficacy of RNAi in adults. Expression analysis indicated that microinjection with 100 ng or led to a significant reduction in transcripts as compared to that of the control (dsRNA specific to green fluorescent protein). Furthermore, lower dsRNA concentrations were also effective in reducing the expression of target genes when delivered by microinjection. Mortality was significantly higher in and -treated insects than in controls injected with . The successful deployment of RNAi in will facilitate functional analyses of vital genes in this economically-important pest and may ultimately result in new control strategies.
斑潜蝇是中国蔬菜和园艺作物的一种入侵性害虫。在本研究中，开发了一种基于双链RNA（dsRNA）的显微注射方法，用于在斑潜蝇中利用编码液泡型ATP酶（V-ATPase）的基因进行RNA干扰（RNAi）。表达分析表明，V-ATPase A和V-ATPase B在斑潜蝇成虫中的表达高于幼虫或蛹。对V-ATPase A和V-ATPase B进行显微注射实验，以评估RNAi在斑潜蝇成虫中的效果。表达分析表明，与绿色荧光蛋白特异性双链RNA（dsRNA）对照相比，注射100 ng V-ATPase A或V-ATPase B双链RNA导致V-ATPase转录本显著减少。此外，通过显微注射递送较低浓度的dsRNA也能有效降低靶基因的表达。V-ATPase A和V-ATPase B处理的昆虫死亡率显著高于注射绿色荧光蛋白双链RNA的对照。RNAi在斑潜蝇中的成功应用将有助于对这种经济上重要害虫的关键基因进行功能分析，并最终可能产生新的控制策略。
