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    响应槲皮素诱导的美国白蛾GST基因鉴定及表达分析

    Identification and expression analysis of GST genes in response to quercetin induction in Hyphantria cunea

      摘要
    • 摘要:
      目的 谷胱甘肽S-转移酶（GSTs）是昆虫体内一种重要的解毒酶，在植食性昆虫对植物次生物质的解毒适应中发挥重要作用。本研究筛选和鉴定了美国白蛾中肠响应槲皮素诱导的GST基因，分析了槲皮素对GST基因的诱导表达模式，为阐明美国白蛾GST基因在解毒代谢槲皮素中的功能奠定基础。
      方法 基于槲皮素诱导的美国白蛾中肠转录组，筛选和鉴定响应槲皮素诱导的GST基因；通过构建系统发育树分析GST基因的家族类别；采用实时荧光定量PCR技术研究槲皮素对GST基因诱导的剂量效应和时间效应。
      结果 鉴定了美国白蛾响应槲皮素诱导的6个GST基因；系统发育分析显示HcGST-E1、HcGST-E2、HcGST-E3属于GST Epsilon家族，HcGST-S1、HcGST-S2属于GST Sigma家族，HcGST-O1属于GST Omega家族。不同质量分数槲皮素对GST基因的诱导作用不同，本实验质量分数（0.5%、1.0%、2.0%、4.0%）的槲皮素均能诱导HcGST-E1表达水平显著上调，0.5%、1.0%和 2.0%的槲皮素诱导HcGST-E3表达水平显著升高；0.5%的槲皮素诱导HcGST-O1的表达水平显著增加。本实验质量分数的槲皮素均诱导HcGST-S1显著下调表达；0.5%和1.0%的槲皮素诱导HcGST-S2的表达水平显著下调。3个上调GST基因均在24 h或36 h内显著上调表达响应槲皮素的诱导。
      结论 槲皮素能显著诱导美国白蛾中肠6个GST基因的表达水平，但对不同基因诱导的表达模式不同。HcGST-E1、HcGST-E3和HcGST-O1显著上调表达响应不同质量分数槲皮素的诱导，推测这3个基因是参与美国白蛾解毒代谢槲皮素的关键基因。

       

      Abstract:
      Objective Glutathione S-transferases (GSTs) are important detoxification enzymes in insects, playing an important role in the physiologhical adaptation of phytophagous insects to plant secondary substances. In this study, we screened and identified GST genes and analyzed their expression patterns induced by quercetin in Hyphantria cunea midgut, laying the foundation for elucidating the function of GST genes in detoxification and metabolism of quercetin in H. cunea.
      Method Based on the quercetin-induced midgut transcriptome of H. cunea, the GST genes in response to quercetin induction were screened and identified. The family categories of GST genes were analyzed by constructing phylogenetic tree. The dose effect and time effect of quercetin on GST gene induction were studied by real-time fluorescence quantitative PCR.
      Result Six GST genes were identified from the gut in response to quercetin induction. Phylogenetic analysis showed that HcGST-E1, HcGST-E2 and HcGST-E3 belong to the GST Epsilon family, HcGST-S1 and HcGST-S2 belong to the GST Sigma family, and HcGST-O1 belongs to the GST Omega family. Various mass fraction of quercetin had different induction effects on GST genes. The tested mass fraction of quercetin (0.5%, 1.0%, 2.0% and 4.0%) significantly up-regulated HcGST-E1 expression, below 2.0% of quercetin significantly increased HcGST-E3 expression, and 0.5% quercetin significantly enhanced HcGST-O1 expression. By contrast, the tested mass fraction of quercetin significantly induced the down-regulation of the expression level of HcGST-S1 and 0.5% and 1.0% of quercetin significantly down-regulated the expression level of HcGST-S2. Three up-regulated GST genes were significantly up-regulated within 24 h or 36 h by quercetin.
      Conclusion Quercetin could significantly induce the expression levels of six GST genes in the midgut of H. cunea, but the expression patterns are various for each induced gene. HcGST-E1, HcGST-E3 and HcGST-O1 are significantly up-regulated in response to the quercetin induction. It is, therefore, speculated that these 3 genes are the key genes involved in the detoxification of quercetin in H. cunea.

       

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