| 摘要: |
| 【目的】探究南亚果实蝇谷胱甘肽S-转移酶基因BtauGSTd1和BtauGSTd2的分子特征及组织表达模式，揭示BtauGSTd1和BtauGSTd2在南亚果实蝇嗅觉及抗药性方面的潜在生理功能，为南亚果实蝇嗅觉识别及抗药性机制研究提供参考。【方法】基于南亚果实蝇成虫头部的转录组数据，通过逆转录聚合酶链反应（RT-PCR）克隆BtauGSTd1和BtauGSTd2，运用生物信息学分析BtauGSTd1和BtauGSTd2的序列特征，通过同源建模构建BtauGSTd1和BtauGSTd2蛋白的三维模型，并采用实时荧光定量PCR（RT-qPCR）技术检测BtauGSTd1和BtauGSTd2在南亚果实蝇成虫不同组织中的表达水平。【结果】序列分析结果表明，BtauGSTd1开放阅读框为684 bp，编码227个氨基酸残基，BtauGSTd1蛋白的相对分子质量为26.26 ku，等电点为8.43；BtauGSTd2开放阅读框为624 bp，编码207个氨基酸残基，BtauGSTd2蛋白的相对分子质量为23.80 ku，等电点为5.90。BtauGSTd1和BtauGSTd2蛋白均含有羧基端结构域和氨基端结构域，无信号肽，不含跨膜结构域，属于非分泌胞质型谷胱甘肽S-转移酶。系统进化分析结果表明，BtauGSTd1和BtauGSTd2均属于昆虫特有的Delta谷胱甘肽S 转移酶亚家族。构建的三维模型显示，BtauGSTd1和BtauGSTd2氨基端结构域均由3个α螺旋（α1~α3）和4个β折叠（β1~β4）构成，羧基端结构域均由6个α螺旋组成（α4~α9）。RT-qPCR结果显示，BtauGSTd1和BtauGSTd2在南亚果实蝇雌雄成虫各供试组织中均有表达，其中BtauGSTd1在雄虫带触角的头部相对表达量显著高于其他组织，在雌虫各供试组织间相对表达量无显著性差异；BtauGSTd2在雌、雄成虫带触角的头部和去触角的头部相对表达量均显著高于去头部的虫体。【结论】BtauGSTd1和BtauGSTd2在南亚果实蝇体内除了充当解毒酶的角色外，很可能在其嗅觉系统中作为气味降解酶，行使降解气味化合物、保护嗅觉系统免受毒害的功能。 |
| 关键词: 南亚果实蝇；谷胱甘肽S-转移酶；生物信息学分析 表达模式 |
| DOI：10.13207/j.cnki.jnwafu.2025.10.012 |
| 分类号: |
| 基金项目:农业部华中作物有害生物综合治理重点实验室/农作物重大病虫草害防控湖北省重点实验室开放基金项目（2023ZTSJJ5）；国家自然科学基金面上项目（31972274）；湖北省高等学校实验室研究项目（HBSY2023-016）；湖北省高等学校优秀中青年科技创新团队项目（T2022009） |
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| Identification and tissue expression of glutathione S-transferase gene in Bactrocera tau |
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TIAN Xiaoli1, XU Dong2, LIU Yi3, WAN Peng2, MING Xing3, GUI Lianyou3, ZHANG Guohui3
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1.College of Life Science,Yangtze University,Jingzhou,Hubei 434025,China;2.Key Laboratory of Integrated Pests Management on Crops in Central China,Ministry of Agriculture and Rural Affairs/Hubei Key Laboratory of Crop Diseases，Insect Pests and Weeds Control,Institute of Plant Protection,Soil and Fertilizer Research,Hubei Academy of Agricultrual Sciences,Wuhan,Hubei 430064,China;3.College of Agriculture,Yangtze University,Jingzhou,Hubei 434025,China
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| Abstract: |
| 【Objective】This study aimed to explore the potential physiological functions of GST genes in the olfaction and insecticide resistance of Bactrocera tau,and the molecular characteristics and tissue expression patterns of two GST genes (BtauGSTd1 and BtauGSTd2) to provide significant references for the olfaction recognition and insecticide resistance mechanism researches in B.tau.【Method Based on the transcriptome data of the head of B.tau，BtauGSTd1 and BtauGSTd2 were cloned by reverse transcription polymerase chain reaction (RT-PCR).The sequence characteristics of BtauGSTd1 and BtauGSTd2 were analyzed by bioinformatics.Three dimensional (3D) models of BtauGSTd1 and BtauGSTd2 were constructed by homology modeling.Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of BtauGSTd1 and BtauGSTd2 in different tissues of B.tau adults.【Result】BtauGSTd1 contained an open reading frame of 684 bp that encoded 227 amino acids.The putative BtauGSTd1 protein had a relative molecular weight of 26.26 ku with an isoelectric of 8.43.BtauGSTd2 comprised an open reading frame of 624 bp that encoded 207 amino acids,with the deduced molecular weight of 23.80 ku and isoelectric point of 5.90.Both putative BtauGSTd1 and BtauGSTd2 had no signal peptide and transmembrane domain,thus belonging to the cytoplasmic GSTs.The phylogenetic tree analysis indicated that BtauGSTd1 and BtauGSTd2 belonged to Delta GST subfamily,which was unique to insects.The three-dimensional model showed that both of BtauGSTd1 and BtauGSTd2 consisted of a N-terminal domain and a C-terminal domain.The N-terminal domain contained three α helices (α1-α3) and four β strands (β1-β4).The C-terminal domain contained six α helices (α4-α9).The results of RT-qPCR showed that BtauGSTd1 and BtauGSTd2 were expressed in all tissues tested of B.tau adults.The relative expression level of BtauGSTd1 in the head of male insects with antennae was significantly higher than that in other tissues,and no significant difference was observed in the relative expression level among the tested tissues of female insects.The relative expression levels of BtaGSTd2 in the head with antennae and the head without antennae of female and male adults were significantly higher than those in adults without heads.【Conclusion】In addition to acting as detoxifying enzymes,BtauGSTd1 and BtauGSTd2 have high probability to act as odorant degrading enzymes in the olfactory system of B.tau adults,which can degrade odorant compounds and protect the olfactory system from toxic substances. |
| Key words: Bactrocera tau glutathione S-transferase bioinformatics analysis expression pattern |
