中国水产科学2016，Vol.23Issue(4)：791-799,9.DOI:10.3724/SP.J.1118.2016.15403
坛紫菜谷胱甘肽过氧化物酶基因的克隆及表达特征
Cloning and expression analysis of the glutathione peroxidase gene fromPyropia haitanensis
摘要
Abstract
At normal situation, algae maintain a balance between reactive oxygen species (ROS) production and scavenging states, and a little amount of ROS molecules can be used as intracellular signal transduction molecules involved in biochemical metabolic processes. But under abiotic stress conditions, excessive accumulation of ROS will cause oxidative damage to cell membrane, then resulting in irreparable loss of metabolic function and eventu-ally leading to cell death. Algae evolved a complex antioxidant defense mechanism (including non-enzymatic system of antioxidant enzymes and antioxidant enzymatic systems) to resist the damages of ROS. Glutathione peroxidase (GPX) is an important member of the ROS scavenging enzymatic system, which can clean the lipid peroxide and H2O2, its mechanism is to reduce ROS through catalytic the reactions between glutathione (GSH) and lipid peroxides (ROOH) or free H2O2, thereby blocking the production of free radicals, inhibiting the peroxidation processes of cells and protecting the enzyme activity of some sulfhydryl. Previous studies have concluded that the level of GPX activity has a positive correlation with the ability of plants to adapt to adversity. So GPX synthe-sis-related gene has been considered as an important stress resistance-related gene.Pyropia haitanensis, naturally growth in coastal intertidal zone, is an important economic macroalgae in Fujian, Zhejiang, Guangdong provinces. With tidal changes, the thallus required to undergo periodic dehydration and rehydration process, therefore it should has a very strong adaptability to adversity. Simultaneously, with global warming, the temperature of sea water increases, which causes damages to seedling ofP. haitanensisand lead to big economic loss for aquaculture industry. As for the physiological and molecular response of P. haitanensis to high temperature and dehydration stress conditions, our previous studies found GPX plays an important role in stress resistance. In this study, the full-length ofGPX gene fromPyropia haitanensis was cloned by using rapid amplification of cDNA ends (RACE) technology, which namedPhGPX (GenBank accession: JX673908). The nucleotide sequence ofPhGPX consists of 1027 bp, including an open reading frame (ORF) of 555 bp, which can be translate into a 184-amino-acid putative peptides with a molecular mass of 19.91 kD and a theoretical PI of 8.76. Multiple sequence alignment and Phy-logenetic analysis indicated that the protein belong to the plant GPX enzyme family. Expression patterns analyzed by qPCR revealed that the expression ofPhGPX gene between the thallus phage and conchocelis phage was not significantly different; during high-temperature stress, the expression levels ofPhGPX significantly upregulated firstly and then decreased; However, during desiccation, the expression levels ofPhGPX had no significantly change when the water loss was≤40%, but as the desiccation continue, the expression levels ofPhGPX were significantly downregulated. These results suggested that the expression ofPhGPX was induced by different stress at different levels.关键词
坛紫菜/谷胱甘肽过氧化物酶/基因克隆/表达分析Key words
Pyropia haitanensis/glutathione peroxidase/gene cloning/expression analysis分类
农业科技引用本文复制引用
张晗晗,徐燕,纪德华,陈昌生,许凯,谢潮添..坛紫菜谷胱甘肽过氧化物酶基因的克隆及表达特征[J].中国水产科学,2016,23(4):791-799,9.基金项目
国家863计划项目(2012AA10A411) （2012AA10A411）
国家自然科学基金项目(41176151,41276177) （41176151,41276177）
福建省种业创新与产业化工程资助项目(2014S1477-10) （2014S1477-10）
福建省自然科学基金项目(2014J05041,2014J07006) （2014J05041,2014J07006）
