碱蓬PEAMT基因的克隆及表达分析

doi:10.11924/j.issn.1000-6850.casb14110137

中国农学通报 ›› 2015, Vol. 31 ›› Issue (9): 178-183.doi: 10.11924/j.issn.1000-6850.casb14110137

所属专题：生物技术

碱蓬PEAMT基因的克隆及表达分析

金杭霞,董德坤,杨清华,袁凤杰,郁晓敏,傅旭军,朱丹华

浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所,浙江省农业科学院作物与核技术利用研究所

收稿日期:2014-11-21 修回日期:2015-01-14 接受日期:2015-01-23 出版日期:2015-04-07 发布日期:2015-04-07
通讯作者: 朱丹华
基金资助:
中国博士后科学基金面上资助

Cloning and Analysis of PEAMT Gene in Suaeda glauca

Received:2014-11-21 Revised:2015-01-14 Accepted:2015-01-23 Online:2015-04-07 Published:2015-04-07

摘要/Abstract

摘要： 研究旨在克隆碱蓬的耐逆相关基因——磷酸乙醇胺甲基转移酶(phosphoethanolamine N-methyltransferase,PEAMT)的编码基因，分析PEAMT基因调控机制，为进一步研究PEAMT基因的功能和碱蓬的耐盐机制奠定分子基础。依据GenBank数据库发表的其他物种的PEAMT序列设计简并引物，通过降落PCR法获得碱蓬PEAMT基因全长cDNA，命名为SgPEAMT，生物信息学软件分析其序列特点，荧光定量PCR检测其表达特性。序列分析表明SgPEAMT基因开放阅读框为1485bp，编码494个氨基酸，推测其为亲水性蛋白。保守结构域分析表明，SgPEAMT含有2个独立的S-腺苷甲硫氨酸依赖性甲基转移酶的保守结构域，每个结构域含有4个基序。系统进化树分析确认SgPEAMT与同属的碱蓬属植物亲缘关系最近。实时荧光定量PCR分析显示，盐胁迫或ABA胁迫下碱蓬根、茎、叶中SgPEAMT基因的表达上调，特别是叶中表达量最高。研究结果表明，SgPEAMT基因受NaCl和ABA诱导表达，预示SgPEAMT基因可能在碱蓬对盐胁迫的反应中起重要作用，是一种参与碱蓬耐盐反应的有效耐逆基因，为植物基因工程提供有效的耐逆基因。

关键词: 林下经济, 林下经济, 北京, 山区

Abstract: Phosphoethanolamine N-methyltransferase (PEAMT) is the key enzyme for phosphocholine. A new gene, named SgPEAMT, was cloned by RT- PCR from Suaeda glauca, which was a typical halophyte. Bioinformatics analysis showed that SgPEAMT encoded 494 amino acids. Hydrophobic analysis indicated that SgPEAMT was a hydrophilic protein and located in cytoplasm. SgPEAMT contained two Sadenosylmethionine-dependent catalyzing domain which contained four motifs, respectively. Phylogenetic tree analyses revealed that the amino acid sequence encoded by SgPEAMT gene was highly homologous to PEAMTs from other Suaeda species. Furthermore, SgPEAMT gene expression patterns in response to NaCl and abscisic acid (ABA) in roots, stems, and leaves of Suaeda glauca were investigated using real time quantitative PCR, which indicated SgPEAMT gene probably played an important role in response of salt stress in Suaeda glauca. The research laid a foundation to further study the PEAMT function and salt resistance mechanism in Suaeda glauca.

金杭霞,董德坤,杨清华,袁凤杰,郁晓敏,傅旭军,朱丹华. 碱蓬PEAMT基因的克隆及表达分析[J]. 中国农学通报, 2015, 31(9): 178-183.

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碱蓬PEAMT基因的克隆及表达分析

Cloning and Analysis of PEAMT Gene in Suaeda glauca

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