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    王建国, 王建军, 曹传旺, 孙丽丽. 舞毒蛾LdOR2基因克隆及对CO2胁迫下的行为反应[J]. 北京林业大学学报, 2021, 43(9): 101-110. DOI: 10.12171/j.1000-1522.20210184
    引用本文: 王建国, 王建军, 曹传旺, 孙丽丽. 舞毒蛾LdOR2基因克隆及对CO2胁迫下的行为反应[J]. 北京林业大学学报, 2021, 43(9): 101-110. DOI: 10.12171/j.1000-1522.20210184
    Wang Jianguo, Wang Jianjun, Cao Chuanwang, Sun Lili. Cloning of LdOR2 gene in Lymantria dispar and its behavioral response to CO2 stress[J]. Journal of Beijing Forestry University, 2021, 43(9): 101-110. DOI: 10.12171/j.1000-1522.20210184
    Citation: Wang Jianguo, Wang Jianjun, Cao Chuanwang, Sun Lili. Cloning of LdOR2 gene in Lymantria dispar and its behavioral response to CO2 stress[J]. Journal of Beijing Forestry University, 2021, 43(9): 101-110. DOI: 10.12171/j.1000-1522.20210184

    舞毒蛾LdOR2基因克隆及对CO2胁迫下的行为反应

    Cloning of LdOR2 gene in Lymantria dispar and its behavioral response to CO2 stress

    • 摘要:
        目的  本文克隆了舞毒蛾的气味受体基因LdOR2,并阐明该基因在舞毒蛾各发育期和组织中的表达特征及其对CO2浓度胁迫下的行为响应,为进一步研究气候变化下舞毒蛾的嗅觉反应机制提供理论依据。
        方法  通过转录组文库筛选克隆出LdOR2基因,利用生物信息学分析其基因特性,通过实时荧光定量PCR(RT-qPCR)技术检测LdOR2基因在不同发育阶段和组织以及不同CO2浓度(397、550和750 μL/L)下的表达水平,并利用RNA干扰(RNAi)技术研究不同CO2浓度下LdOR2基因沉默后舞毒蛾的行为学反应。
        结果  舞毒蛾LdOR2基因开放阅读框(ORF)为1 203 bp,编码400个氨基酸,蛋白分子量为45.76 kDa,理论等电点为8.22;进化树分析结果表明,舞毒蛾LdOR2与黏虫MsepOR24和双委夜蛾AdisOR21亲缘关系较近,并聚为一类;RT-qPCR结果显示,LdOR2在舞毒蛾各发育阶段均有表达,在雌蛹中表达量最高,雄成虫中表达量最低;在雌、雄成虫不同组织中,雌、雄触角中表达量显著高于其它组织(P < 0.05),但雌、雄虫触角间的表达量差异不明显。高CO2浓度下LdOR2基因表达量降低,其中,550 μL/L和750 μL/L条件下雌虫触角中其表达量与对照组相比分别下降21%和29%(P < 0.05),雄虫触角中其表达量与对照组相比分别下降了43%和7%(P < 0.05)。LdOR2基因沉默后,舞毒蛾雌、雄成虫对丁香酚和顺-3-己烯-1-醇的趋向性减弱,而在高浓度CO2处理条件下,舞毒蛾沉默体对7种挥发物的反应率均有所下降。
        结论  舞毒蛾LdOR2在其气味识别过程中发挥重要作用,CO2浓度变化通过调节舞毒蛾LdOR2基因的表达进而影响其对气味的敏感性。

       

      Abstract:
        Objective  In this study, we cloned the odorant receptor gene (LdOR2) and determined the expression levels of this gene in developmental stages and different tissues of the Lymantria dispar and its behavioral response to CO2 stress. The results will provide a theoretical basis for clarifying the olfactory response mechanism of the L. dispar under climate change.
        Method  The LdOR2 gene was cloned through transcriptome library screening, and its characteristics were analyzed by bioinformatics. The expression levels of LdOR2 gene in different developmental stages and tissues as well as in different CO2 concentrations (397, 550 and 750 μL/L) were determined by real-time fluorescence quantitative PCR technology. In addition, RNA interference (RNAi) technology was used to study the behavioral responses of L. dispar adults silenced by LdOR2 at different CO2 concentrations.
        Result  The open reading frame (ORF) of LdOR2 gene in L. dispar was 1 203 bp, encoding 400 amino acids. The molecular mass of the LdOR2 protein was 45.76 kDa and the theoretical isoelectric point was 8.22. The phylogenetic tree showed that the LdOR2 in L. dispar was closely related to MsepOR24 Mythimna separata and AdisOR21 in Athetis dissimilis, and clustered into one group. RT-qPCR results showed that LdOR2 was expressed at all developmental stages of the L. dispar, with the highest expression level in female pupae and the lowest expression levels in male adults. In different tissues of female and male adults, the expression levels in antennae were significantly higher than those in other tissues (P < 0.05), but showed no difference between the antennae of female and male. The expression of LdOR2 gene decreased under high CO2 concentration. Compared with the control group, the expression of female antennae under 550 μL/L and 750 μL/L conditions decreased by 21% and 29% (P < 0.05), respectively, and the expression levels of antennae of male L. dispar adults decreased by 43% and 7% (P < 0.05). After LdOR2 gene silencing, the tendency of female and male L. dispar adults to eugenol and cis-3-hexene-1-ol was weakened, while the response rates of the L. dispar silencers to seven volatiles decreased under high CO2 concentration.
        Conclusion  LdOR2 plays an important role in the odor recognition of L. dispar. The sensitivity of L. dispar to odor was affected by the expression levels of LdOR2 gene regulated by the changes of CO2 concentration.

       

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