昆虫学报 ›› 2022, Vol. 65 ›› Issue (1): 10-20.doi: 10.16380/j.kcxb.2022.01.002

• 研究论文 • 上一篇    下一篇

SfHMGR在白背飞虱生殖调控中的功能分析

周操1, 杨熙彬2, 龚明富2, 杨洪2,3,*, 龙贵云2,  贾泽艳2, 曾庆会2, 金道超2   

  1.  (1. 重庆师范大学生命科学学院, 重庆 401331; 2. 贵州大学昆虫研究所, 贵州山地农业病虫害重点实验室, 农业农村部贵阳作物有害生物科学观测试验站, 贵阳 550025; 3. 贵州大学烟草学院, 贵阳 550025)
  • 出版日期:2022-01-20 发布日期:2022-01-17

Functional analysis of SfHMGR in reproduction regulation of the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)

ZHOU Cao1, YANG Xi-Bin2, GONG Ming-Fu2, YANG Hong2,3,*, LONG Gui-Yun2, JIA Ze-Yan2, ZENG Qing-Hui2, JIN Dao-Chao2   

  1.  (1. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China; 2. Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Scientific Observing and Experimental Station of Crop Pests in Guiyang, Ministry of Agriculture and Rural Affairs, Institute of Entomology of Guizhou University, Guiyang 550025, China; 3. College of Tobacco Science, Guizhou University, Guiyang 550025, China)
  • Online:2022-01-20 Published:2022-01-17

摘要:

【目的】为探究保幼激素(juvenile hormone, JH)生物合成关键基因3-羟基-3-甲基戊二酰辅酶A还原酶基因(HMGR)在白背飞虱Sogatella furcifera生殖中的作用。【方法】基于已公布的白背飞虱基因组和转录组数据,结合RT-PCR技术获得SfHMGR全长cDNA序列;采用RT-qPCR技术分析SfHMGR在白背飞虱不同发育阶段(1-5龄若虫和雌成虫)和雌成虫不同组织(头、肠道、脂肪体、卵巢和体壁)中的表达谱;通过RNAi技术靶向沉默SfHMGR后观察白背飞虱雌成虫卵巢的发育情况,统计单雌产卵量,用RT-qPCR测定JH生物合成下游基因SfJHAMTSfFAMeT、JH信号传导相关基因SfMetSfKr-h1以及卵巢发育关键基因SfVgSfVgR的转录水平。【结果】克隆获得了白背飞虱SfHMGR(GenBank登录号:MW883397)的全长cDNA序列,开放阅读框长2 724 bp,编码907个氨基酸,其编码蛋白预测的分子量为98.96 kD,理论等电点为6.15。序列分析表明,SfHMGR含有典型的HMG-CoA还原酶的保守结构域、C端3个HMG-CoA结合位点、N端7个典型的跨膜结构域。系统进化分析发现,SfHMGR与灰飞虱Laodelphax striatella和褐飞虱Nilaparvata lugens的HMGR亲缘关系最近,氨基酸序列一致性分别达92.40%和89.25%。发育阶段表达谱结果表明,SfHMGR在各阶段发育中期以及初羽化雌虫中高表达;组织表达谱结果显示,SfHMGR在雌成虫各组织中均有表达,在肠道中具有最高的转录水平。通过显微注射dsRNA抑制SfHMGR表达后,显著抑制了雌虫的单雌产卵量和卵巢的发育,并且SfJHAMT, SfFAMeT, SfKr-h1, SfVgSfVgR的转录水平也被显著抑制。【结论】SfHMGR通过影响JH生物合成过程及其JH的信号传导来调控SfVg的转录水平,进而影响白背飞虱雌虫的卵巢发育和产卵量。

关键词: 白背飞虱, 3-羟基-3-甲基戊二酰辅酶A还原酶, 保幼激素, 繁殖, 卵巢发育

Abstract: 【Aim】To explore the role of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene (HMGR), a key gene of juvenile hormone (JH) biosynthesis, in the reproduction of the white-backed planthopper, Sogatella furcifera.【Methods】Based on the published genome and transcriptome data of S. furcifera, the full-length cDNA sequence of SfHMGR was obtained by RT-PCR. The expression profiles of SfHMGR in different developmental stages (1st-5th instar nymph and female adult) and different tissues (head, gut, fat body, ovaries and integument) of female adults of S. furcifera were detected by RT-qPCR. After targeted silencing of SfHMGR by RNAi technology, the ovarian development of female adults was observed, the number of eggs laid by per female adult of S. furcifera was counted, and the transcription levels of downstream genes of JH biosynthesis (SfJHAMT and SfFAMeT), JH signal transduction related genes (SfMet and SfKr-h1), and the key genes of ovarian development (SfVg and SfVgR) were determined by RT-qPCR. 【Results】The full-length cDNA sequence of SfHMGR (GenBank accession number: MW883397) of S. furcifera was cloned, with an open reading frame of 2 724 bp in length, encoding 907 amino acids. The predicted molecular weight of the protein is 98.96 kD, and the theoretical isoelectric point is 6.15. Sequence analysis showed that SfHMGR contains typical HMG-CoA reductase conserved domain, three HMG-CoA binding sites at the C-terminus and seven transmembrane domains at the N-terminus. Phylogenetic analysis showed that SfHMGR has the closest genetic relationship with HMGR of Laodelphax striatellus and Nilaparvata lugens, showing 92.40% and 89.25% amino acid sequence identities with them, respectively. The developmental expression profiles showed that SfHMGR was highly expressed in the middle of various developmental stages and newly emerged female adults. The tissue expression profiles showed that SfHMGR was expressed in various tissues of female adults, and had the highest transcription level in gut. After the inhibition of SfHMGR expression by microinjection of dsRNA, the number of eggs laid by per female and ovarian development of female adults, and the transcription levels of SfJHAMT, SfFAMeT, SfKr-h1, SfVg and SfVgR were significantly inhibited. 【Conclusion】SfHMGR regulates the transcription level of SfVg by affecting the JH biosynthesis process and JH signal transduction, and then affects the ovarian development and the fecundity of female adults of S. furcifera.

Key words: Sogatella furcifera, 3-hydroxy-3-methylglutaryl coenzyme A reductase, juvenile hormone, reproduction, ovarian development