坛紫菜谷胱甘肽过氧化物酶基因的克隆及表达特征

发布时间：2019-05-22 22:19

【摘要】：谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)是植物活性氧(ROS)清除酶促系统的重要成员之一,在植物逆境胁迫应答中发挥着重要作用。本研究以坛紫菜(Pyropia haitanensis)为研究材料采用RACE技术克隆获得了一条坛紫菜的GPX全长基因序列,命名为PhGPX(Gen Bank收录号:JX673908)。该基因序列全长1027 bp,包含555 bp的开放阅读框,所编码的多肽包含184个氨基酸,分子量为19.9 k D,等电点为8.76。多序列比对和系统进化树分析结果表明Ph GPX属于植物GPX基因家族成员。基因表达水平的q PCR分析结果表明Ph GPX基因在坛紫菜叶状体和丝状体世代中的表达水平没有显著差异;高温胁迫不同时间水平下,Ph GPX基因的表达水平呈现为先上调后下调的趋势;不同失水胁迫条件下,Ph GPX基因的表达不受低水平的失水胁迫影响,但可被高水平(40%)的失水胁迫所抑制,且在复水30 min后仍然无法恢复失水胁迫前的水平。由此推测坛紫菜体内的GPX也可能存在多种家族成员,不同的逆境胁迫条件、甚至不同的逆境胁迫水平可能需要不同家族成员的GPX参与ROS的清除和防御。
[Abstract]:Glutathione peroxide (glutathione peroxidase,GPX) is one of the important members of active oxygen (ROS) scavenging enzyme system in plants and plays an important role in plant stress response. In this study, a full-length GPX gene sequence of Laver haitanensis was cloned by RACE using (Pyropia haitanensis) as research material, which was named PhGPX (Gen Bank reception number: JX673908). The full length of the gene sequence is 1027 bp, which contains 555 bp open reading frame. The encoded polypeptide contains 184 amino acids, the molecular weight is 19.9 KD, and the isoelectric point is 8.76. The results of multiple sequence alignment and phylogenetic tree analysis showed that Ph GPX belonged to plant GPX gene family. The results of Q PCR analysis of gene expression level showed that there was no significant difference in the expression level of Ph GPX gene between striatum and filamentous generation of Euphorbia pulcherrima. Under different time levels of high temperature stress, the expression level of, Ph GPX gene was up-regulated at first and then down-regulated. The expression of, Ph GPX gene under different water loss stress was not affected by low level of water loss stress, but could be inhibited by high level (40%) of water loss stress, and the level before water loss stress could not be restored after 30 min of rehydration. It is speculated that there may also be many family members of GPX in Laver, and different stress conditions and even different stress levels may require GPX from different family members to participate in the clearance and defense of ROS.
【作者单位】：集美大学水产学院;
【基金】：国家863计划项目(2012AA10A411) 国家自然科学基金项目(41176151,41276177) 福建省种业创新与产业化工程资助项目(2014S1477-10) 福建省自然科学基金项目(2014J05041,2014J07006)
【分类号】：S917.3

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