小菜蛾普通气味受体基因 PxylOR9 的鉴定及功能研究

›› 2015, Vol. 58 ›› Issue (5): 507-515.doi:

小菜蛾普通气味受体基因 PxylOR9 的鉴定及功能研究

刘一鹏^1,2, 刘杨^2,*, 杨婷², 桂富荣^1,*, 王桂荣²

(1. 云南农业大学植物保护学院, 昆明 650201; 2. 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193)

出版日期:2015-05-20 发布日期:2015-05-20
作者简介:刘一鹏, 男, 1989年9月生, 湖南衡阳人, 硕士研究生, 研究方向为昆虫生化与分子生物学, E-mail: liuyipenglyp@126.com

Identification and characterization of a general odorant receptor gene PxylOR9 in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

LIU Yi-Peng^1,2, LIU Yang², YANG Ting^2,*, GUI Fu-Rong^1,*, WANG Gui-Rong^2

(1. College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China; 2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)

Online:2015-05-20 Published:2015-05-20

摘要/Abstract

摘要： 【目的】克隆小菜蛾 Plutella xylostella 一普通气味受体基因，明确其在不同组织中的表达分布，并鉴定其功能，为了解小菜蛾对寄主植物的识别机制提供理论基础。【方法】在小菜蛾成虫触角转录组测序和分析的基础上，通过PCR技术克隆得到一气味受体基因的全长序列，利用半定量 RT-PCR 研究其在雌雄蛾不同组织中的表达，并通过爪蟾卵母细胞体外表达结合双电极电压钳技术测试该受体对59种植物挥发物刺激的反应。【结果】在前期研究中，通过转录组测序和生物信息学分析，预测获得了多条小菜蛾气味受体基因序列。本研究中，我们克隆得到了其中一小菜蛾气味受体基因 PxylOR9 的cDNA全长序列(GenBank登录号: KP757898）。PxylOR9包括6个跨膜结构域，N末端位于胞内，符合昆虫气味受体的典型特征。与其他鳞翅目昆虫气味受体构建的进化树分析结果显示，PxylOR9与性信息素受体具有明显的分离，并与普通气味受体聚集在一起。半定量RT-PCR结果显示，PxylOR9 仅在雌雄触角中表达且无雌雄差异。双电极电压钳记录结果显示，在测试的植物挥发物中PxylOR9只对植物挥发物 β-紫罗兰酮的刺激具有反应。【结论】本研究从小菜蛾中克隆得到 PxylOR9 的全长序列，其编码产物具有气味受体的典型特征并且属于普通气味受体。该基因在仅在小菜蛾成虫触角中高表达。体外功能研究证明PxylOR9对植物挥发物 β-紫罗兰酮具有特异反应，推测其参与了小菜蛾对植物的识别。

关键词: 小菜蛾, 普通气味受体, 半定量RT-PCR, 双电极电压钳, β-紫罗兰酮

Abstract: 【Aim】 The objective of this study is to clone an odorant receptor gene from the diamondback moth, Plutella xylostella, characterize its expression profiles in different tissues and investigate its function. Our study provides a theoretical basis for understanding the mechanisms of host plant recognition of the diamondback moth. 【Methods】 The full-length cDNA sequence of PxylOR9 gene was obtained and cloned based on the transcriptome analysis of adult P. xylostella antennae. The relative expression levels of this gene in different tissues of both adult males and females were detected using semi-quantitative RTPCR. Two-electrode voltage clamp electrophysiological recording was used to test the response of Xenopus oocytes injected with PxylOR9 to 59 plant volatiles in vitro. 【Results】 Based on transcriptome sequencing and analysis in the preliminary study, several sequences of odorant receptors were predicted in P. xylostella. The full length cDNA of one of these genes, PxylOR9 (GenBank accession number: KP757898), was cloned in this study. Sequence analysis revealed that PxylOR9 has six transmembrane domains with an intracellular N-terminus, which is the typical structure of insect odorant receptors. Phylogenetic analysis showed that PxylOR9 is obviously different from sex pheromone receptors of several lepidopteran insects, but cluster together with their other general odorant receptors. The semi-quantitative RT-PCR results showed that the expression level of PxylOR9 was much higher in the antennae of both males and females than in other tissues. Voltage clamp electrophysiological recording indicated that PxylOR9 could only respond to the stimulation with β-ionone among the tested plant volatiles. 【Conclusion】 PxylOR9 of P. xylostella belongs to the general odorant receptor with a typical characteristic. PxylOR9 is highly expressed in adult antennae of P. xylostella. PxylOR9 can be activated by plant volatile β-ionone, implying that PxylOR9 may be involved in the recognition of P. xylostella to plants.

Key words: Plutella xylostella, general odorant receptor, semi-quantitative RT-PCR, two-electrode voltage clamp, β-ionone

刘一鹏, 刘杨, 杨婷, 桂富荣, 王桂荣. 小菜蛾普通气味受体基因 PxylOR9 的鉴定及功能研究[J]. , 2015, 58(5): 507-515.

LIU Yi-Peng, LIU Yang, YANG Ting, GUI Fu-Rong, WANG Gui-Rong. Identification and characterization of a general odorant receptor gene PxylOR9 in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)[J]. , 2015, 58(5): 507-515.

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