Functional analysis of arabidopsis CYP714A1 and CYP714A2 reveals that they are distinct gibberellin modification enzymes

Takahito Nomura, Hiroshi Magome, Atsushi Hanada, Noriko Takeda-Kamiya, Lewis N. Mander, Yuji Kamiya, Shinjiro Yamaguchi

    Research output: Contribution to journalArticlepeer-review

    40 Citations (Scopus)

    Abstract

    Endogenous levels of bioactive gibberellins (GAs) are controlled by both biosynthetic and inactivation processes, and some cytochrome P450s are involved in this control mechanism. We have previously reported that CYP714B1 and CYP714B2 encode the enzyme GA 13-oxidase, which is required for GA1 biosynthesis, and that CYP714D1 encodes GA 16a, 17-epoxidase, which inactivates the non-13-hydroxy GAs in rice. Arabidopsis has two CYP714 members, CYP714A1 and CYP714A2. To clarify the possible role of these genes in GA metabolism, enzymatic activities of their recombinant proteins were analyzed using a yeast expression system. We found that the recombinant CYP714A1 protein catalyzes the conversion of GA12 to 16-carboxylated GA12 (16-carboxy-16b, 17-dihydro GA12), a previously unidentified GA metabolite. Bioassays of this GA product showed that CYP714A1 is an inactivation enzyme in Arabidopsis. This was confirmed by the extreme GA-deficient dwarf phenotype shown by CYP714A1-overexpres-sing plants. Intriguingly, the recombinant CYP714A2 protein catalyzed the conversion of ent-kaurenoic acid into steviol (ent-13-hydroxy kaurenoic acid). When GA12 was used as a substrate for CYP714A2, 12a-hydroxy GA12 (GA111) was produced as a major product and 13-hydroxy GA12 (GA53) as a minor product. Transgenic Arabidopsis plants overexpres-sing the CYP714A2 gene showed semi-dwarfism. GA analysis showed that the levels of non-13-hydroxy GAs, including GA4, were decreased, whereas those of 13-hydroxy GAs, including GA1 (which is less active than GA4), were increased in the transgenic plants. Our results suggest that the CYP714 family proteins contribute to the production of diverse GA compounds through various oxidations of C and D rings in both monocots and eudicots.

    Original languageEnglish
    Pages (from-to)1837-1851
    Number of pages15
    JournalPlant and Cell Physiology
    Volume54
    Issue number11
    DOIs
    Publication statusPublished - 2013 Nov

    Keywords

    • Arabidopsis
    • Biosynthesis
    • Cytochrome P450
    • Gibberellin
    • Inactivation

    ASJC Scopus subject areas

    • Physiology
    • Plant Science
    • Cell Biology

    Fingerprint Dive into the research topics of 'Functional analysis of arabidopsis CYP714A1 and CYP714A2 reveals that they are distinct gibberellin modification enzymes'. Together they form a unique fingerprint.

    Cite this