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Plant and Cell Physiology 2004 45(9):1129-1138; doi:10.1093/pcp/pch149
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© 2004 Oxford University Press

Rapid Paper

Emission of ent-Kaurene, a Diterpenoid Hydrocarbon Precursor for Gibberellins, into the Headspace from Plants

Minoru Otsuka1,2,4, Hiromichi Kenmoku2, Mikihiro Ogawa2, Kazunori Okada3,5, Wataru Mitsuhashi1, Takeshi Sassa1, Yuji Kamiya2, Tomonobu Toyomasu1,6 and Shinjiro Yamaguchi2

1 Course of the Science of Bioresources, United Graduate School of Agricultural Sciences, Iwate University (Yamagata University), Tsuruoka, Yamagata, 997-8555 Japan
2 Laboratory for Cellular Growth and Development, Plant Science Center, RIKEN (Institute of Physical and Chemical Research), Suehiro-cho 1-7-22, Tsurumi-ku, Yokohama, Kanagawa, 230-0045 Japan
3 Department of Biology, Tokyo Gakugei University, Nukuikitamachi 4-1-1, Koganei, Tokyo, 184-8501 Japan

ent-Kaurene is a tetracyclic hydrocarbon precursor for gibberellins (GAs) in plants and fungi. To address whether fungal GA biosynthesis enzymes function in plants, we generated transgenic Arabidopsis plants overexpressing ent-kaurene synthase (GfCPS/KS) from a GA-producing fungus Gibberella fujikuroi. GfCPS/KS catalyzes a two-step reaction corresponding to ent-copalyl diphosphate synthase (CPS) and ent-kaurene synthase (KS) activities in plants. When GfCPS/KS was overexpressed and targeted to plastids, a range of GA-deficient phenotypes of the ga1-3 and ga2-1 mutants (defective in CPS and KS, respectively) were restored to wild type. Unexpectedly, the transgenic lines overproducing GfCPS/KS emitted the GA precursor ent-kaurene into the headspace besides its accumulation in the plant body. When co-cultivated with the ent-kaurene overproducers in a closed environment, the airborne ent-kaurene was able to fully complement the dwarf phenotype of ga1-3 and ga2-1 mutants, but not that of the ga3-1 mutant (defective in ent-kaurene oxidase). These results suggest that ent-kaurene may be efficiently metabolized into bioactive GAs in Arabidopsis when supplied as a volatile. We also provide evidence that ent-kaurene is released in the headspace of wild-type Chamaecyparis obtusa and Cryptomeria japonica plants, suggesting the occurrence of this hydrocarbon GA precursor as a volatile in nature.

4 Present address: Cryobiosystem Research Center, Faculty of Agriculture, Iwate University, Morioka, Iwate, 020-8550 Japan.

5 Present address: Biotechnology Research Center, University of Tokyo, Bunkyo-ku, Tokyo, 113-8657 Japan.

6 Corresponding author: E-mail, toyomasu{at}tds1.tr.yamagata-u.ac.jp; Fax, +81-235-28-2812.


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