Quantitative analysis of plant polyamines including thermospermine during growth and salinity stress

Yukie Naka, Kanako Watanabe, G. H.M. Sagor, Masaru Niitsu, M. Arumugam Pillai, Tomonobu Kusano, Yoshihiro Takahashi

    Research output: Contribution to journalArticlepeer-review

    64 Citations (Scopus)

    Abstract

    Arabidopsis thaliana was thought to contain two spermine synthase genes, ACAULIS 5 (ACL5) 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 °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.

    Original languageEnglish
    Pages (from-to)527-533
    Number of pages7
    JournalPlant Physiology and Biochemistry
    Volume48
    Issue number7
    DOIs
    Publication statusPublished - 2010 Jul 1

    Keywords

    • Arabidopsis thaliana
    • HPLC
    • High salinity
    • Polyamine
    • Spermine
    • Thermospermine

    ASJC Scopus subject areas

    • Physiology
    • Genetics
    • Plant Science

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