昆虫学报 ›› 2023, Vol. 66 ›› Issue (10): 1302-1310.doi: 10.16380/j.kcxb.2023.10.004

• 研究论文 • 上一篇    下一篇

烟粉虱卵黄原蛋白通过影响自噬反应调控共生细菌Rickettsia的丰度

孙想, 刘炳奇, 陈展博, 李欢, 李楚翘, 洪继生, 栾军波*   

  1. (沈阳农业大学植物保护学院, 沈阳 110866)
  • 出版日期:2023-10-20 发布日期:2023-11-27

Vitellogenin determines the abundance of the symbiotic bacterium Rickettsia in Bemisia tabaci (Hemiptera: Aleyrodidae) by regulating autophagy

SUN Xiang, LIU Bing-Qi, CHEN Zhan-Bo, LI Huan, LI Chu-Qiao, HONG Ji-Sheng, LUAN Jun-Bo*   

  1.  (College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China)
  • Online:2023-10-20 Published:2023-11-27

摘要: 【目的】探明烟粉虱Bemisia tabaci卵黄原蛋白(vitellogenin, BtVg)调控共生细菌Rickettsia丰度的分子机制。【方法】通过显微注射烟粉虱MEAM1隐种雌成虫dsRNA对BtVg进行RNAi,采用qRT-PCR检测烟粉虱MEAM1隐种雌成虫BtVg和自噬基因BtAtg8的表达量,统计雌成虫死亡率,采用qPCR检测雌成虫中Rickettsia的丰度,采用免疫荧光标记显微镜技术检测BtVgBtAtg8在雌成虫卵巢管中的定位和表达以及Rickettsia在卵巢管和中肠中的丰度。饲喂烟粉虱MEAM1隐种雌成虫含雷帕霉素(10 μmol/L)的人工饲料诱导自噬后,采用免疫荧光标记显微镜技术检测BtAtg8在卵巢管中的表达和定位,采用qPCR检测卵巢中Rickettsia丰度。【结果】与显微注射dsGFP的对照相比,显微注射dsBtVg后3 d时烟粉虱MEAM1隐种雌成虫BtVg的表达量显著降低,BtAtg8的表达量显著升高,死亡率显著升高,Rickettsia丰度显著降低,卵巢管中BtVg表达量明显降低,BtAtg8表达量明显升高,Rickettsia在卵巢管和中肠中的丰度明显减少。与饲喂含DMSO的人工饲料的对照相比,饲喂含雷帕霉素(10 μmol/L)的人工饲料后5 d时烟粉虱MEAM1隐种雌成虫卵巢管中BtAtg8表达量显著升高;饲喂含雷帕霉素(10 μmol/L)的人工饲料后3和5 d时,烟粉虱MEAM1隐种雌成虫卵巢中Rickettsia的丰度显著降低。【结论】结果提示,BtVg能够保护Rickettsia免受自噬的降解,明确了BtVg能够调控Rickettsia在烟粉虱体内的丰度。

关键词: 烟粉虱, 卵黄原蛋白, 自噬; Rickettsia, 自噬基因, 卵巢

Abstract: 【Aim】 To explore the molecular mechanism by which vitellogenin (Vg) regulates the abundance of the symbiotic bacterium Rickettsia in Bemisia tabaci. 【Methods】 Female adult of B. tabaci MEAM1 was microinjected with dsRNA to conduct RNAi of BtVg, the expression levels of BtVg and the autophagy gene BtAtg8 in the female adult were detected by qRT-PCR, the mortality rate of female adults was recorded, and the abundance of Rickettsia in the female adult was quantified by qPCR. The localization and expression of BtVg and BtAtg8 in ovarioles and the abundance of Rickettsia in the ovarioles and midguts of the female adult were examined by immunofluorescence microscopy. After the autophagy induction by feeding the female adult of B. tabaci MEAM1 with the artificial diet containing rapamycin (10 μmol/L), the expression and localization of BtAtg8 in ovarioles were investigated by immunofluorescence microscopy and the abundance of Rickettsia in ovaries was detected by qPCR. 【Results】 The expression levels of BtVg and BtAtg8 in the female adult of B. tabaci MEAM1 were significantly decreased and significantly increased, respectively, the mortality rate of female adults of B. tabaci MEAM1 was significantly increased, the abundance of Rickettsia in the female adult of B. tabaci MEAM1 was significantly decreased, the expression levels of BtVg and BtAtg8 in ovarioles were obviously decreased and obviously increased, respectively, and the abundance of Rickettsia in the ovarioles and midgut obviously decreased at 3 d after microinjection of dsBtVg compared to the control (microinjected with dsGFP). The expression level of BtAtg8 in the ovarioles of the female adult of B. tabaci MEAM1 was increased significantly at 5 d after feeding with the artificial diet containing rapamycin (10 μmol/L), and the abundance of Rickettsia in the ovarioles of the female adult of B. tabaci MEAM1 was significantly decreased at 3 and 5 d after feeding with the artificial diet containing rapamycin (10 μmol/L) compared with the control fed with the artificial diet containing DMSO. 【Conclusion】 These results suggest that BtVg can protect Rickettsia from degradation by autophagy, and demonstrate the role of BtVg in regulating the abundance of Rickettsia in B. tabaci.

Key words: Bemisia tabaci, vitellogenin, autophagy; Rickettsia, autophagy gene, ovary