褐飞虱RNAi显微注射中麻醉方法的比较

doi:10.16409/j.cnki.2095-039x.2022.05.009

摘要/Abstract

摘要： 显微注射dsRNA以实现系统性干扰，是一种有效且能准确评估基因功能的常用方法。试验靶标昆虫由于个体较小、活力强，注射前需要先适当麻醉。待显微注射dsRNA完成后，麻醉效应解除，方进一步开展表型测定试验。不同麻醉方法可对昆虫的麻醉效果、麻醉后生理和行为等产生不同程度的影响。因此，选用合适的麻醉方法对显著减少RNAi显微注射试验误差，提升干扰效率具有重要意义。本文以褐飞虱为评估对象，比较应用不同配比的乙醚与乙酸乙酯混合液麻醉，以及应用低温麻醉对褐飞虱的麻醉效应，并测定不同的麻醉方法对RNAi显微注射试验中褐飞虱存活率的影响。麻醉剂处理随乙酸乙酯的浓度升高，褐飞虱的苏醒时间延长。应用不同麻醉方法对RNAi显微注射试验中褐飞虱存活率的影响程度从小到大依次为乙醚:乙酸乙酯（1:2），乙醚:乙酸乙酯（1:1），乙醚，乙醚:乙酸乙酯（2:1），乙醚:乙酸乙酯（3:1），冰上20 min ，冰上40 min。推荐乙醚:乙酸乙酯（1:2）混合液作为最优麻醉处理应用于褐飞虱RNAi显微注射试验。

关键词: 褐飞虱, 液体麻醉, 低温麻醉, 显微注射, RNA干扰

Abstract: Microinjection is an effective and accurate quantitative method to inhibit gene expression, which has a wide range of applications in insect gene function research and pest control. Usually, the tested insects are small and vigorous, and they need to be anesthetized before injection for immobilization. After the anesthesia effect is relieved, further phenotypic determination experiments can be performed. Different anesthesia methods have different effects on the anesthesia effect, physiology, and behavior of insects. Therefore, the determination of insect anesthesia methods is of great significance to reduce errors in microinjection dsRNA interference experiments. In this study, the brown planthopper was used as the experimental subject, and the anesthetic effects of different proportions of ether and ethyl acetate mixed liquid anesthetics and low temperature anesthesia on the brown planthopper were compared, and the effects on the survival rate of brown planthopper were further determined. The results showed that the recovery time of brown planthopper was prolonged with the increase of the concentration of ethyl acetate in anesthetic treatment. The effects of different anesthesia methods on the survival rate of brown planthopper in the RNAi microinjection experiment ranked from high to low were ether:ethyl acetate (1:2), ether:ethyl acetate (1:1), ether, ether:ethyl acetate (2:1), ether:ethyl acetate (3:1), 20 min on ice, and 40 min on ice. The comprehensive evaluation showed that the mixture of ether:ethyl acetate (1:2) could be used as the optimal anesthesia treatment for the RNAi microinjection experiment of brown planthopper.

Key words: brown planthopper, liquid anesthesia, hypothermic anesthesia, microinjection, RNA interference

中图分类号:

S476

张朋稳, 段至柔, 刘常权, 彭英传, 张万娜, 肖海军, 张晶. 褐飞虱RNAi显微注射中麻醉方法的比较[J]. 中国生物防治学报, 2023, 39(5): 1148-1155.

ZHANG Pengwen, DUAN Zhirou, LIU Changquan, PENG Yinchuan, ZHANG Wanna, XIAO Haijun, ZHANG Jing. Comparison of Anesthesia Methods in RNAi Microinjection of Brown Planthopper[J]. Chinese Journal of Biological Control, 2023, 39(5): 1148-1155.

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