麦红吸浆虫,3-羟基-3-甲基戊二酰辅酶A还原酶,基因克隆,滞育,发育,变态," /> 麦红吸浆虫,3-羟基-3-甲基戊二酰辅酶A还原酶,基因克隆,滞育,发育,变态,"/>  Sitodiplosis mosellana,3-hydroxy-3-methylglutaryl coenzyme A reductase,gene cloning,diapause,developmental metamorphosis,"/> <span style="font-size:13.3333px;">麦红吸浆虫3-羟基-3-甲基戊二酰辅酶A还原酶</span><span style="font-size:13.3333px;">基因<em>SmHMGR</em>的克隆及在滞育与发育变态</span><span style="font-size:13.3333px;">过程中的表达动态</span>

昆虫学报 ›› 2019, Vol. 62 ›› Issue (10): 1140-1149.doi: 10.16380/j.kcxb.2019.10.003

• 研究论文 • 上一篇    下一篇

麦红吸浆虫3-羟基-3-甲基戊二酰辅酶A还原酶基因SmHMGR的克隆及在滞育与发育变态过程中的表达动态

刘禹含1, 梁婷婷1, 赵佳佳1, 成卫宁1,*, 朱克岩2   

  1. (1.西北农林科技大学, 农业部西北黄土高原作物有害生物综合治理重点实验室, 陕西杨凌 712100; 2. Department of Entomology, Texas A&M University, Texas 77843, USA)
  • 出版日期:2019-10-20 发布日期:2019-10-14

Cloning and expression dynamics of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene SmHMGR during diapause and metamorphosis in Sitodiplosis mosellana (Diptera: Cecidomyiidae)

LIU Yu-Han1, LIANG Ting-Ting1, ZHAO Jia-Jia1, CHENG Wei-Ning1,*, ZHU Ke-Yan2   

  1.  (1. Key Laboratory of Integrated Pest Management on Crops in Northwestern Loess Plateau, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China; 2. Department of Entomology, Texas A&M University, Texas 77843, USA)
  • Online:2019-10-20 Published:2019-10-14

摘要: 【目的】3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)是保幼激素(JH)合成途径的限速酶。麦红吸浆虫Sitodiplosis mosellana是一种典型的专性幼虫滞育昆虫。本研究旨在探讨HMGR基因在麦红吸浆虫滞育和发育变态过程中的作用。【方法】通过RT-PCR和RACE技术克隆麦红吸浆虫滞育前幼虫HMGR基因全长cDNA序列;利用生物信息学软件分析HMGR基因核苷酸和其编码的蛋白氨基酸序列特性;采用qPCR技术测定其在麦红吸浆虫滞育不同时期3龄幼虫及不同发育阶段(1-2龄幼虫、预蛹、初蛹、中蛹和后蛹以及雌雄成虫)中的mRNA表达水平。【结果】克隆获得一条麦红吸浆虫HMGR基因全长cDNA序列,命名为SmHMGR(GenBank登录号: MG876766)。该基因全长2 548 bp,其中开放阅读框长2 328 bp,编码775个氨基酸,预测的蛋白分子量为84.16 kD,理论等电点为8.29。序列分析发现该基因编码的蛋白具有HMGR蛋白家族典型的HMG-CoA-reductase-classⅠ催化功能域及其他保守功能基序;序列比对和系统发育分析表明,SmHMGR与达氏按蚊Anopheles darling等长角亚目(Nematocera)昆虫HMGR的相似性最高、亲缘关系最近。SmHMGR在麦红吸浆虫滞育前的3龄早期幼虫中表达量显著升高,进入滞育后一直维持较高水平,并在滞育后静息阶段的当年12月至翌年1月达到最高。SmHMGR在蛹期表达量低于幼虫期,预蛹期表达量最低;在雌成虫中表达量显著高于在蛹和雄成虫中的表达量。【结论】SmHMGR的表达与麦红吸浆虫发育密切相关,可能在滞育诱导、维持及滞育后静息状态的维持及生殖中发挥作用,其表达量的降低可能参与了幼虫到蛹的变态。

关键词: 麦红吸浆虫')">麦红吸浆虫, 3-羟基-3-甲基戊二酰辅酶A还原酶, 基因克隆, 滞育, 发育, 变态

Abstract: 【Aim】 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is a rate-limiting enzyme of juvenile hormone (JH) synthesis. This study aims to explore the potential roles of HMGR gene in diapause and metamorphosis of Sitodiplosis mosellana, an insect species undergoing obligatory larval diapause. 【Methods】 The full-length cDNA of HMGR gene was cloned from S. mosellana pre-diapause larvae by RT-PCR and RACE technologies. The sequence characteristics of the nucleotide and amino acid sequences of the cloned HMGR were analyzed by bioinformatics programs. The mRNA expression levels of HMGR in the 3rd instar larva of S. mosellana in different stages of diapause (pre-diapause, diapause, post-diapause quiescent and post-diapause development) and different developmental stages (the 1st-2nd instar larva, pre-pupa, early pupa, middle pupa and late pupa, as well as male adult and female adult) were determined by qPCR. 【Results】 The full-length cDNA sequence of HMGR was obtained from S. mosellana and named SmHMGR, which was deposited at GenBank under the accession number MG876766. SmHMGR is 2 548 bp in length and contains a 2 328 bp ORF, encoding a protein of 775 amino acids with a predicted molecular weight of 84.16 kD and a theoretical pI of 8.29. The deduced amino acid sequence contains the conserved HMG-CoA-reductase-classⅠ domain and other motifs of the HMGR protein family. Multiple sequence alignment and phylogenetic analysis showed that SmHMGR has the highest similarity and the closest relationship to HMGRs from the Nematocera including Anopheles darling. The expression level of SmHMGR increased as the larvae entered the early 3rd instar before diapause, maintained high after diapause, and peaked in the post-diapause quiescence phase (from December to the next January). The expression level of SmHMGR was lower in the pupal stage than in the larval stage, the lowest in the pre-pupal stage, and significantly higher in female adults than in pupae and male adults. 【Conclusion】 SmHMGR expression is related to the development of S. mosellana, and might play key roles in diapause induction, maintenance and post-diapause quiescence as well as in adult reproduction, whereas the metamorphosis from larva to pupa might be correlated with its reduced expression.

Key words:  Sitodiplosis mosellana')"> Sitodiplosis mosellana, 3-hydroxy-3-methylglutaryl coenzyme A reductase, gene cloning, diapause, developmental metamorphosis