Quantitative analysis of plant polyamines including thermospermine during growth and salinity stress
Quantitative analysis of plant polyamines including thermospermine during growth and salinity stress
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DOI：
10.1016/j.plaphy.2010.01.013
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发表时间：
2010-07-01
影响因子：
6.5
通讯作者：
Takahashi, Yoshihiro
中科院分区：
文献类型：
--
作者：
Naka, Yukie;Watanabe, Kanako;Takahashi, Yoshihiro
拟南芥被认为含有两个精胺合成酶基因：acaulis 5(Acculis 5)和spms。然而，最近的研究表明，ACL5基因编码温度敏合酶。在这项研究中，我们建立了一种简单的方法，从小于500 mg的植物组织提取物中分离出四胺的两个异构体：精胺和热敏胺。从植物组织中提取的多胺(PA)被苯甲酰化，在C-18反相柱上，用42%(v/v)的乙腈在30℃的水中以等度方式洗脱，在254 nm处检测，衍生物被完全拆分。对不能合成温敏胺和/或精胺的acl5和spms的单突变体和双突变体进行PA分析，证实了该方法的相关性。此外，利用这种方法，我们监测了海棠不同组织中的温温胺含量，发现茎和花中的温温胺含量是整株幼苗和成熟叶片的两到三倍。在水稻和番茄中证实了温度敏素的存在。最后，我们探讨了盐胁迫是否会改变拟南芥中多胺类物质的含量，其中包括温敏胺。(C)2010年爱思唯尔·马森公司。版权所有。
Arabidopsis thaliana was thought to contain two spermine synthase genes, ACAULIS 5 (ACLS) and SPMS. Recent investigations, however, revealed that the ACL5 gene encodes thermospermine synthase. In this study, we have established a simple method to separate two isomers of tetraamine, spermine and thermospermine, in extracts from plant tissues of less than 500 mg. Polyamines (PAs) extracted from plant tissues were benzoylated, and the derivatives were completely resolved by high-performance liquid chromatography on a C-18 reverse-phase column, by eluting with 42% (v/v) acetonitrile in water in an isocratic manner at 30 degrees C and monitoring at 254 nm. The relevance of the method was confirmed by co-chromatography with respective PAs and by the PA analysis of the single- and double-mutants of acl5 and spms, which could not synthesize thermospermine and/or spermine, respectively. Furthermore, with this method, we monitored the thermospermine contents in various tissues of A. thaliana and found that stems and flowers contain two- to three-fold more thermospermine compared to whole seedlings and mature leaves. The presence of thermospermine was confirmed in Oryza sativa and Lycopersicon pesculentum. Finally we addressed whether salinity stress changes the contents of PAs including thermospermine in Arabidopsis. (C) 2010 Elsevier Masson SAS. All rights reserved.