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Chinese Journal of Biological Control ›› 2023, Vol. 39 ›› Issue (5): 1148-1155.DOI: 10.16409/j.cnki.2095-039x.2022.05.009

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Comparison of Anesthesia Methods in RNAi Microinjection of Brown Planthopper

ZHANG Pengwen1, DUAN Zhirou1, LIU Changquan1, PENG Yinchuan1, ZHANG Wanna1, XIAO Haijun2, ZHANG Jing1   

  1. 1. Institute of Entomology, Jiangxi Agricultural University, Nanchang 330045, China;
    2. School of Grassland Science, Beijing Forestry University, Beijing 100083, China
  • Received:2022-06-03 Online:2023-10-08 Published:2023-10-25

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

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