马尾松谷胱甘肽过氧化物酶PmGPX基因的克隆及表达分析

发布时间：2018-03-26 00:09

本文选题：林木育种学　切入点：马尾松　出处：《浙江农林大学学报》2017年05期

【摘要】：克隆获得马尾松Pinus massoniana PmGPX基因,进行生物信息学分析以及在不同抗虫材料表达模式分析。在前期马尾松转录组测序获得该基因片段基础上,用cDNA末端快速扩增(RACE)技术获得基因全长,实时荧光定量聚合酶链式反应(qRT-PCR)技术分析该基因在不同抗虫材料中的表达模式。克隆获得基因cDNA全长序列,命名为PmGPX。该基因包含741 bp开放阅读框,编码246个氨基酸。序列分析表明:该基因具有GPX家族典型的保守结构域,等电点PI 8.29,蛋白分子量27.08 kDa,与针叶植物北美云杉Picea sitchensis的同源性较高,为91%。与单子叶和双子叶植物同源性较差,大部分在64%~80%,而在GPX家族典型保守结构域的同源性较高,为82%~100%。qRT-PCR技术分析表明:PmGPX在所有组织中均有表达,在针叶中表达量最高,嫩叶的表达量是根的33.45倍,在根、花和球果中的表达量较低,不同材料日表达模式相似,总体上随时间出现"升—降—升"模式。该基因在抗虫材料中启动较早,且表达量高于对照。推测该基因参与了马尾松抗虫防御体系。
[Abstract]:The Pinus massoniana PmGPX gene of Pinus massoniana was cloned and analyzed by bioinformatics and expression patterns of different insect-resistant materials. The full length of the gene was obtained by rapid amplification of the cDNA terminal and the expression pattern of the gene in different insect-resistant materials was analyzed by real-time fluorescence quantitative polymerase chain reaction (QRT-PCR). The full-length sequence of the gene was cloned and obtained. The gene is named PmGPX. the gene contains 741bp open reading frame and encodes 246 amino acids. Sequence analysis shows that the gene has a typical conserved domain of GPX family. Pi 8.29, protein molecular weight 27.08 kDa. the homology with coniferous Picea sitchensis is 91.The homology with monocotyledonous and dicotyledonous plants is poor, most of them are in the position of 80th, but the homology is higher in the typical conserved domain of GPX family. The results of 82%~100%.qRT-PCR analysis showed that: PmGPX was expressed in all tissues, the highest in needles, 33.45 times in young leaves, lower in roots, flowers and cones, and similar in daily expression patterns of different materials. As a whole, the "up-down-ascending" pattern appeared over time, and the gene started earlier in the insect-resistant materials, and its expression level was higher than that of the control. It was speculated that the gene was involved in the anti-insect defense system of Pinus massoniana (Pinus massoniana).
【作者单位】：广西壮族自治区林业科学研究院国家林业局马尾松工程技术研究中心/广西马尾松工程技术研究中心;
【基金】：国家自然科学基金资助项目(31660219) 广西“八桂学者”专项经费资助项目(2011A015) 广西自然科学基金资助项目(2014GXNSFBA118104)
【分类号】：S791.248

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