| 陈秀琳*, 李琳琳, 陈玉鑫, 李广伟.梨小食心虫气味受体GmolOR10基因克隆及表达谱分析[J].植物保护,2023,49(3):75-84. |
| 梨小食心虫气味受体GmolOR10基因克隆及表达谱分析 |
| Cloning and expression profiling of odorant receptor gene GmolOR10 in Grapholita molesta Busck |
| 投稿时间：2022-02-24 修订日期：2022-05-23 |
| DOI：DOI: 10.16688/j. zwbh. 2022099 |
| 中文关键词: 梨小食心虫 气味受体10 基因克隆 实时定量PCR 表达模式 |
| 英文关键词:Grapholita molesta odorant receptor 10 gene cloning real time quantitative PCR expression profile |
| 基金项目:国家自然科学基金(32160636)；陕西省自然科学基金（基础研究计划）(2021JQ-620)；延安大学博士科研启动项目(YDBK2019-01)；延安大学科研计划项目(YDY2019-26) |
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| 中文摘要: |
| 为研究梨小食心虫Grapholita molesta Busck嗅觉通讯分子机制, 本研究应用RT-PCR克隆获得了梨小食心虫气味受体基因GmolOR10的完整开放阅读框, 并对其序列结构进行了相关生物信息学分析, 最后采用实时荧光定量PCR对GmolOR10在梨小食心虫成虫不同组织(触角、头、胸、腹、足、翅)与不同发育阶段的表达模式进行了定量分析。结果表明, GmolOR10编码区长1 194 bp, 编码397个氨基酸, 等电点和相对分子量分别为8.57和46.14 kD, 有7个跨膜结构域。多序列比对与系统发育树结果显示GmolOR10与卷蛾科苹果蠹蛾Cydia pomonella CpomOR18和山毛榉卷叶蛾Cydia fagiglandana CfagOR18的氨基酸序列一致性较高, 分别为84.38%和83. 38%。GmolOR10在雄成虫触角中表达量较高, 极显著高于雌虫触角(P<0.01), 在雌、雄成虫头部也有较高的表达量, 在成虫其他组织表达量很低。不同发育阶段的表达谱显示, GmolOR10在梨小食心虫的不同发育阶段均有表达, 在成虫期的表达量最高(在雌雄成虫中的表达量均呈现出随着日龄增加逐渐升高然后下降的趋势), 在其他发育阶段的表达量相对较低。GmolOR10具有昆虫气味受体的结构特征, 在雌、雄虫触角高表达, 在各发育阶段均有表达, 据此推测GmolOR10可能与梨小食心虫觅食、感受及识别性信息素的过程有关。 |
| 英文摘要: |
| In order to investigate the molecular mechanism of olfactory communication in Grapholita molesta Busck, GmolOR10 was cloned by RT-PCR in this study. The sequence structure of GmolOR10 was analyzed by using bioinformatics software, and then its expression profiles in different adult tissues (antenna, head, thorax, abdomen, leg and wing) and at different developmental stages were measured by real time quantitative PCR. The results revealed that the CDS of GmolOR10 was 1 194 bp in length, encoding 397 amino acid residues, with a predicted theoretical isoelectric point and a relative molecular mass of 8.57 and 46.14 kD, respectively, and seven transmembrane domains. Multiple sequence alignment and phylogenetic analysis indicated that GmolOR10 was closely related to the odorant receptor CpomOR18 from Cydia pomonella and CfagOR18 from Cydia fagiglandana, with an amino acid sequence identity of 84.38% and 83. 38%, respectively. Tissue specific expression profiling showed that abundant expression of GmolOR10 existed in the antenna of male, extremely higher than that in the antenna of female moth (P<0.01). GmolOR10 also showed higher expression in the head of both sexes, and displayed low expression level in the other tissues. Developmental stage specific expression profiling demonstrated that GmolOR10 was expressed at different developmental stages of G. molesta and the expression level of GmolOR10 at adult stage was higher (the expression levels in male and female moths increased first and then decreased with day age), and relatively lower expression was detected at other developmental stages. Based on the structural characteristics and the expression profiling results of GmolOR10, it is speculated that GmolOR10 may be involved in the process of foraging, feeling and recognition of sex pheromones in G. molesta. |
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