›› 2017, Vol. 60 ›› Issue (12): 1376-1383.doi: 10.16380/j.kcxb.2017.12.003

• 研究论文 • 上一篇    下一篇

萝卜蚜反-β-法尼烯结合蛋白基因的时空表达模式分析(英文)

纪祥龙, 战一迪, 李佩玲, 刘勇*   

  1. (山东农业大学植物保护学院, 昆虫行为与化学生态学研究室, 山东泰安 271018)
  • 出版日期:2017-12-20 发布日期:2017-12-20

Spatio-temporal expression patterns of (E)-β-farnesene-binding protein genes in the turnip aphid, Lipaphis erysimi (Hemiptera: Aphididae) (in English)

JI Xiang-Long, ZHAN Yi-Di, LI Pei-Ling, LIU Yong*   

  1. (Department of Entomology, Shandong Agricultural University, Tai′an, Shandong 271018, China)
  • Online:2017-12-20 Published:2017-12-20

摘要:   【目的】气味结合蛋白(odorant binding proteins, OBPs)有助于提高昆虫嗅觉系统的敏感性,并且在蚜虫报警信息素反-β-法尼烯[(E)-β-farnesene EBF]的通信过程中起重要作用。萝卜蚜Lipaphis erysimi是十字花科作物的主要害虫,为了减少蔬菜田间的农药使用,EBF对蚜虫防治具有很大的潜力。然而,目前萝卜蚜对EBF反应的研究甚少。【方法】本研究利用PCR和RACE技术克隆并鉴定了2种公认的在萝卜蚜中对EBF具有高度亲和力的气味结合蛋白的基因LeryOBP3和LeryOBP7;利用RT-qPCR检测了它们在萝卜蚜不同发育阶段和无翅成蚜不同组织中的表达谱。【结果】克隆获得的萝卜蚜2个基因序列LeryOBP3(GenBank登录号: KJ703012)和LeryOBP7(GenBank登录号: KJ703013)分别与豌豆蚜Acyrthosiphum pisum ApisOBP3和ApisOBP7序列对比有较高的氨基酸序列一致性(分别为94%和88%),基因序列符合气味结合蛋白基因的典型特征。LeryOBP3和LeryOBP7开放阅读框全长分别为357和468 bp,分别编码118和155个氨基酸。发育表达谱表明,LeryOBP3和LeryOBP7的表达量高峰出现于成虫期;组织表达谱表明,LeryOBP3在成蚜足中表达量较高,而LeryOBP7主要表达于成蚜触角组织中。【结论】在萝卜蚜成虫触角中的LeryOBP7结合蛋白可能与其对EBF的响应有关。

关键词: 萝卜蚜, 报警信息素, 气味结合蛋白, 基因表达谱, 实时荧光定量PCR

Abstract:  【Aim】 Odorant binding proteins (OBPs) contribute to the remarkable sensitivity of insect olfactory system and play an important role in aphid alarm pheromone (E)-β-farnesene (EBF) communication. The turnip aphid, Lipaphis erysimi (Kaltenbach) is the major pest of cruciferous crops. To reduce pesticide application in vegetable fields, EBF appears to hold strong potential for aphid control. However, few studies about L. erysimi to EBF are known so far. 【Methods】 Two putative OBP genes, LeryOBP3 and LeryOBP7 encoding proteins with high affinity to EBF were cloned by PCR and RACE and characterized from L. erysimi, and their expression profiles at different developmental stages and in different tissues of wingless adults of L. erysimi were determined by RT-qPCR. 【Results】 The two genes cloned here, LeryOBP3 (GenBank no.: KJ703012) and LeryOBP7 (GenBank no.: KJ703013) share high amino acid sequence identities (94% and 88%, respectively) with ApisOBP3 and ApisOBP7, which encode proteins with affinity to EBF in Acyrthosiphum pisum (Harris). The full length of open reading frames of LeryOBP3 and LeryOBP7 are 357 and 468 bp, encoding 118 and 155 amino acids, respectively. Developmental expression profiles showed that the obvious expression peaks of LeryOBP3 and LeryOBP7 appeared at the adult stage, and tissue expression profiles showed that LeryOBP3 was expressed strongly in legs of adults while LeryOBP7 in adult antennae. 【Conclusion】 These results suggest that LeryOBP7 in the antennae might be responsible for the response of L. erysimi to EBF.

Key words: Lipaphis erysimi; alarm pheromone, odorant-binding protein, gene expression profile, real-time quantitative PCR