昆虫学报 ›› 2022, Vol. 65 ›› Issue (11): 1459-1468.doi: 10.16380/j.kcxb.2022.11.007

• 研究论文 • 上一篇    下一篇

禾谷缢管蚜钠通道辅助亚基对钠通道基因表达水平及杀虫剂敏感性的影响

张存环, 刘朗, 彭雄, 郄杏桃, 陈茂华*   

  1. (西北农林科技大学植物保护学院, 农业农村部西北黄土高原作物有害生物综合治理重点实验室, 旱区作物逆境生物学国家重点实验室, 陕西杨凌 712100)
  • 出版日期:2022-11-20 发布日期:2022-12-02

Effects of sodium channel auxiliary subunits on the expression levels of sodium channel genes and insecticide susceptibility in Rhopalosiphum padi (Hemiptera: Aphididae)

ZHANG Cun-Huan, LIU Lang, PENG Xiong, QIE Xing-Tao, CHEN Mao-Hua*   

  1.  (State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture and Rural Affairs, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China)
  • Online:2022-11-20 Published:2022-12-02

摘要: 【目的】通过分析禾谷缢管蚜Rhopalosiphum padi钠通道辅助亚基对钠通道功能的影响,探究辅助亚基在钠离子通道的门控性质中的作用。【方法】分别显微注射dsRpNavH1和dsRpNavH2对钠通道基因RpNavH1和RpNavH2进行RNAi后,采用实时定量PCR(qRT-PCR)技术测定禾谷缢管蚜成蚜5个钠通道辅助亚基基因(RpTEH1, RpTEH2, RpTEH3, RpTEH4和RpTipE)的表达量;利用qRT-PCR技术和杀虫剂生物测定分别测定RNAi干扰RpTipE对禾谷缢管蚜成蚜钠通道及其辅助亚基基因表达量以及LC50浓度高效氯氟氰菊酯敏感性的影响;利用双电压钳技术检测非洲爪蟾Xenopus laevis卵母细胞单独注射果蝇Drosophila钠通道基因DmNav22 cRNA及DmNav22 cRNA分别与果蝇钠通道辅助亚基基因DmTipE及禾谷缢管蚜钠通道辅助亚基基因RpTEH2, RpTEH3, RpTEH4和RpTipE的cRNA共注射时的Na+电流,分析禾谷缢管蚜钠通道辅助亚基的功能。【结果】RNAi敲降禾谷缢管蚜成虫钠通道基因RpNavH1后,禾谷缢管蚜成蚜4个钠通道辅助亚基基因RpTEH1, RpTEH2, RpTEH3以及RpTipE的表达量显著降低,RpTEH4的表达量没有显著变化;RNAi敲降禾谷缢管蚜钠通道基因RpNavH2后,钠通道辅助亚基基因RpTEH1, RpTEH2, RpTEH3, RpTEH4和RpTipE的表达量都显著降低。RNAi敲降禾谷缢管蚜钠通道辅助亚基基因RpTipE后,钠通道基因RpNavH1和RpNavH2及钠通道辅助亚基基因RpTEH1, RpTEH2, RpTEH3和RpTEH4的表达量都显著降低。RNAi干扰RpTipE后48 h时,禾谷缢管蚜成蚜对LC50浓度的高效氯氰菊酯的敏感性降低。双电压钳技术检测结果表明,将果蝇钠通道基因DmNav22 cRNA分别与禾谷缢管蚜钠通道辅助亚基基因RpTEH2, RpTEH3, RpTEH4和RpTipE cRNA共注射到非洲爪蟾卵母细胞中时均提高果蝇钠通道DmNav22的Na+电流。【结论】禾谷缢管蚜钠通道辅助亚基可能参与调节该虫钠离子通道的门控性质。

关键词: 禾谷缢管蚜, 钠通道, 钠通道辅助亚基, 电生理实验, RNAi, 基因表达

Abstract: 【Aim】 This study aims to analyze the effect of sodium channel auxiliary subunits on the sodium channel function in Rhopalosiphum padi, and to explore the roles of auxiliary subunits in the gating properties of sodium channel. 【Methods】 The expression levels of five sodium channel auxiliary subunit genes (RpTEH1, RpTEH2, RpTEH3, RpTEH4 and RpTipE) in R. padi adults after RNAi of sodium channel genes RpNavH1 and RpNavH2 by microinjection of dsRpNavH1 and dsRpNavH2, respectively, were detected by real-time quantitative PCR (qRT-PCR), the effects of RNAi of RpTipE on the expression levels of genes of sodium channels and their auxiliary subunits in R. padi adults and the susceptibility of these adults to LC50 of beta-cypermethrin were determined by qRT-PCR and insecticide bioassay, respectively, and the Na+ currents in Xenopus laevis oocytes evoked by the Drosophila sodium channel gene DmNav22 cRNA injected alone and DmNav22 cRNA co-injected with cRNAs of the sodium channel auxiliary subunit genes DmTipE of Drosophila, and RpTEH2, RpTEH3, RpTEH4 and RpTipE of R. padi, respectively, were detected by the two-electrode voltage clamp technology to analyze the function of sodium channel auxiliary subunits of R. padi. 【Results】 Knockdown of the sodium channel gene RpNavH1 of R. padi by RNAi significantly reduced the expression levels of four sodium channel auxiliary subunit genes (RpTEH1, RpTEH2, RpTEH3 and RpTipE) in R. padi adults, whereas showed no significant influence on the expression level of RpTEH4. Knockdown of the sodium channel gene RpNavH2  of R. padi by RNAi significantly reduced the expression levels of the sodium channel auxiliary subunit genes RpTEH1, RpTEH2, RpTEH3, RpTEH4 and RpTipE. The expression levels of the sodium channel genes RpNavH1 and RpNavH2 and the sodium channel auxiliary subunit genes RpTEH1, RpTEH2, RpTEH3 and RpTEH4 were significantly reduced after knockdown of the sodium channel auxiliary subunit gene RpTipE of R. padi by RNAi. The susceptibility of R. padi adults to LC50 of beta-cypermethrin was significantly decreased at 48 h after RNAi of RpTipE. The two-electrode voltage clamp recording showed that co-injection of DmNav22 cRNA with cRNAs of the sodium channel auxiliary subunit genes RpTEH2, RpTEH3, RpTEH4 and RpTipE of R. padi, respectively, in X. laevis oocytes increased the Na+ current of Drosophila sodium channel DmNav22. 【Conclusion】 Sodium channel auxiliary subunits may be involved in regulating the gating properties of sodium channels in R. padi.

Key words: Rhopalosiphum padi, sodium channel, sodium channel auxiliary subunits, electrophysiological experiments, RNAi, gene expression