昆虫学报 ›› 2019, Vol. 62 ›› Issue (3): 284-293.doi: 10.16380/j.kcxb.2019.03.002

• 研究论文 • 上一篇    下一篇

异色瓢虫气味结合蛋白基因HaxyOBP1和HaxyOBP6的克隆及表达谱分析

韩世鹏, 梁超, 李新兵, 韩慧, 赵锋, 何运转*   

  1. (河北农业大学植物保护学院, 河北保定 071000)
  • 出版日期:2019-03-20 发布日期:2019-03-14

Cloning and expression profiling of the odorant binding protein genes HaxyOBP1 and HaxyOBP6 in Harmonia axyridis (Coleoptera: Coccinellidae)

HAN Shi-Peng, LIANG Chao, LI Xin-Bing, HAN Hui, ZHAO Feng, HE Yun-Zhuan*   

  1. (College of Plant Protection, Agricultural University of Hebei, Baoding, Hebei 071000, China)
  • Online:2019-03-20 Published:2019-03-14

摘要:

【目的】本研究旨在探索异色瓢虫Harmonia axyridis气味结合蛋白的结构和分布,更好地了解气味结合蛋白在异色瓢虫嗅觉系统中的作用。【方法】利用生物信息学方法克隆异色瓢虫2个气味结合蛋白基因序列并对其蛋白结构进行分析;通过实时荧光定量PCR分析克隆获得的这2个基因在异色瓢虫各个发育阶段和成虫不同组织中的表达水平。【结果】本研究成功克隆得到异色瓢虫两个气味结合蛋白基因HaxyOBP1和HaxyOBP6(GenBank登录号分别为MG757923和MG757927)。HaxyOBP1开放阅读框全长447 bp,编码148个氨基酸,具有6个保守的半胱氨酸,表明HaxyOBP1属于Classic OBPs。HaxyOBP1在异色瓢虫各发育阶段均有表达,在雄性成虫中表达量最高;组织表达谱分析表明HaxyOBP1在雄虫头部表达量最高。HaxyOBP6开放阅读框全长414 bp,编码137个氨基酸,具有4个保守的半胱氨酸,表明HaxyOBP6属于Minus-C OBPs。HaxyOBP6在成虫期的表达量明显高于幼虫阶段,并且在雄成虫中的表达量显著高于雌成虫;组织表达谱分析表明HaxyOBP6在雄成虫头部和雌成虫翅中表达量最高。【结论】本研究克隆得到异色瓢虫两个气味结合蛋白基因,表达谱分析表明HaxyOBP1和HaxyOBP6分别在异色瓢虫成虫头和翅中具有较高的表达水平,说明气味结合蛋白基因可能在异色瓢虫非嗅觉组织中同样具有重要作用。本研究结果为深入研究异色瓢虫气味结合蛋白结构和功能奠定了基础。

关键词: 异色瓢虫, 气味结合蛋白, 基因克隆, 表达谱分析, 实时荧光定量PCR

Abstract: 【Aim】 The study aims to explore the structure and distribution of odorant binding proteins of Harmonia axyridis, so as to provide a basis for further understanding the functions of odorant binding proteins in the olfactory system of H. axyridis. 【Methods】 Two odorant binding protein genes were cloned from H. axyridis, and their protein structures were analyzed by bioinformatics methods. The expression levels of the two genes in different developmental stages and adult tissues of H. axyridis were analyzed by qRT-PCR. 【Results】 Two odorant binding proteins genes of H.axyridis, HaxyOBP1 and HaxyOBP6 (GenBank accession numbers: MG757923 and MG757927, respectively), were successfully cloned and sequenced. The open reading frame of HaxyOBP1 is 447 bp in length. It encodes 148 amino acids with six conserved cysteines, indicating that HaxyOBP1 belongs to Classical OBPs subfamily. HaxyOBP1 was expressed in various developmental stages of H. axyridis and had the highest expression level in male adults. Tissue expression profiles showed that HaxyOBP1 had the highest expression level in the head of male adult. The open reading frame of HaxyOBP6 is 414 bp in length. It encodes 137 amino acids with four conserved cysteines, indicating that HaxyOBP6 belongs to Minus-C OBPs subfamily. The expression level of HaxyOBP6 in adults was higher than that in larvae, and the expression level in male adults was significantly higher than that in female adults. Tissue expression profiles showed that HaxyOBP6 had the highest expression levels in the head of male adult and the wing of female adult. 【Conclusion】 In this study, two odorant binding protein genes were cloned from H. axyridis. The expression profiles showed that HaxyOBP1 and HaxyOBP6 were highly expressed in the head and the wing of adults, respectively, suggesting that the OBPs of H. axyridis may also play important roles in non-olfactory tissues. The results of this study lay a foundation for further study on the structure and function of OBPs of H. axyridis.

Key words: Harmonia axyridis; odorant binding protein, gene cloning, expression profiling, qRT-PCR