1.二つのテトラアミン異性体であるスペルミンとサーモスペルミンのHPLCによる定量分析手法の確立(口頭発表)

概要

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Arabidopsis thaliana was thought to contain two spermine synthase genes, ACAULIS5 (ACL5) and SPMS. Current 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 300mg. 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℃ and monitoring at 254nm. 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 esculentum. Finally we addressed whether salinity stress changes the thermospermine content in Arabidopsis.
植物化学調節学会の論文
2009-10-06