›› 2018, Vol. 61 ›› Issue (10): 1121-1131.doi: 10.16380/j.kcxb.2018.10.001

• 研究论文 •    下一篇


杨海博, 胡镇杰, 李定旭, 朱品红, 董钧锋*   

  1. (河南科技大学林学院, 河南洛阳 471000)
  • 出版日期:2018-10-20 发布日期:2018-10-20

Identification of odorant binding protein genes in antennae of Corythucha ciliata (Hemiptera: Tingidae)

YANG Hai-Bo, HU Zhen-Jie, LI Ding-Xu, ZHU Pin-Hong, DONG Jun-Feng*   

  1.  (Forestry College, Henan University of Science and Technology, Luoyang, Henan 471000, China)
  • Online:2018-10-20 Published:2018-10-20

摘要: 【目的】悬铃木方翅网蝽Corythucha ciliata是专性危害悬铃木属Platanus植物的外来入侵害虫,本研究旨在获得该害虫触角中气味结合蛋白(OBPs)基因信息,以期寻求有效控制害虫的嗅觉分子靶标。【方法】利用Illumina HiSeqTM 4000高通量测序技术对悬铃木方翅网蝽雌雄成虫触角进行转录组测序并对测序结果进行生物信息学分析;通过实时荧光定量PCR(qPCR)方法,分析OBP基因在悬铃木方翅网蝽雌雄成虫触角中的表达模式。【结果】对悬铃木方翅网蝽雌雄成虫触角6个样品的转录组测序,共获得 40.87 Gb clean reads,各样品的序列长度均达到6.31 Gb。转录组数据分析共鉴定出26个推测的悬铃木方翅网蝽OBP基因,其编码蛋白中24个(CcilOBP1-24)属于Classic OBPs,CcilOBP25/26属于Plus-C OBPs;与半翅目其他昆虫相关OBPs系统发育分析表明,大部分CcilOBPs形成独立一簇,少数与其他半翅目昆虫OBPs直系同源。qPCR分析显示,有11个OBP基因在雌雄成虫触角中表达量差异显著,其中有9个OBP 基因(CcilOBP5/6/9/10/17/18/21/24/25)在雄成虫触角中显著高表达,有2个OBP基因(CcilOBP14/16)在雌成虫触角中显著高表达。【结论】本研究获得了悬铃木方翅网蝽成虫触角气味结合蛋白基因信息,研究结果为生物控制该害虫提供了重要基础数据和候选分子靶标。

关键词: 悬铃木方翅网蝽, 触角转录组, 高通量测序, 气味结合蛋白, 实时荧光定量PCR

Abstract: 【Aim】 The sycamore lace bug, Corythucha ciliata (Hemiptera: Tingidae), is an invasive and specific forestry pest of Platanus spp. The objective of this study is to obtain the gene information of odorant binding proteins (OBPs) in C. ciliata antennae and to seek effective olfactory molecular targets for pest control. 【Methods】 The antennal transcriptomes of male and female adults of C. ciliata were sequenced using Illumina HiSeqTM 4000 platform and subjected to bioinformatics analysis. The expression patterns of OBP genes in female and male antennae of C. ciliata adults were analyzed by real-time quantitative PCR (qPCR). 【Results】 In total, 40.87 Gb clean reads were obtained from six samples of male and female antennal transcripts of C. ciliata adults, the sequences of all of the samples are more than 6.31 Gb in length. By further screening and identification, we obtained 26 putative OBP genes from C. ciliata, whose encoded proteins correspond to 24 Classic OBPs (CcilOBP1-24) and 2 Plus-C OBPs (CcilOBP25/26). Phylogenetic analysis of OBPs from C. ciliata (CcilOBPs) and other closely related Hemiptera species revealed that most CcilOBPs form a unique cluster, and a few form ortholog groups with OBPs from other hemipterous insects. qPCR analysis showed that there were significant differences in the expression levels of 11 OBP genes in the antennae of male and female adults, including 9 OBP genes (CcilOBP5/6/9/10/17/18/21/24/25) highly expressed in the male antennae, and 2 OBP genes (CcilOBP14/16) highly expressed in the female antennae. 【Conclusion】 This study acquired the molecular information of OBP genes from C. ciliata adult antennae. The results provide valuable basic data and candidate molecular targets for biological control of C. ciliata.

Key words: Corythucha ciliata, antennal transcriptome, high-throughput sequencing; odorant binding protein, realtime quantitative PCR