Biological functions of ent- and syn-copalyl diphosphate synthases in rice: key enzymes for the branch point of gibberellin and phytoalexin biosynthesis.
Rice (Oryza sativa L.) produces ent-copalyl diphosphate (ent-CDP) and syn-CDP as precursors for several classes of phytoalexins and the phytohormones, gibberellins (GAs). It has recently been shown that a loss-of-function mutation of OsCPS1, a gene encoding a putative ent-CDP synthase, results in a severely GA-deficient dwarf phenotype in rice. To clarify the biological functions of the ent- and syn-CDP synthases involved in the biosynthesis of phytoalexins and/or GAs, we isolated two cDNAs, OsCyc1 and OsCyc2, encoding putative diterpene cyclases from ultraviolet (UV)-irradiated rice leaves (cv. Nipponbare). The production of phytoalexins in rice leaves is known to be highly induced by UV treatment. Using a bacterial expression system, we demonstrated that OsCyc1 encodes syn-CDP synthase and that OsCyc2 and OsCPS1 encode ent-CDP synthase. The level of expression of the OsCyc1 and OsCyc2 transcripts in rice leaves increased drastically in response to UV treatment, whereas expression of the OsCPS1 transcript was not induced by UV light. These results suggest that OsCyc1, OsCyc2 and OsCPS1 are responsible for the biosynthesis of momilactones A and B and oryzalexin S, oryzalexins A-F and phytocassanes A-E, and GAs, respectively. Our results strongly suggest the presence of two ent-CDP synthase isoforms in rice, one that participates in the biosynthesis of GAs and a second that is involved in the biosynthesis of phytoalexins.
Otomo
Kazuko
K
Course of the Science of Bioresources, The United Graduate School of Agricultural Science, Iwate University (Yamagata University), Tsuruoka, Yamagata 997-8555, Japan.
Kenmoku
Hiromichi
H
Oikawa
Hideaki
H
König
Wilfried A
WA
Toshima
Hiroaki
H
Mitsuhashi
Wataru
W
Yamane
Hisakazu
H
Sassa
Takeshi
T
Toyomasu
Tomonobu
T
D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D005875:Gibberellins / Q000096:biosynthesis*; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012275:Oryza / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D010936:Plant Extracts / Q000096:biosynthesis*; D018515:Plant Leaves / Q000201:enzymology / Q000528:radiation effects; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012717:Sesquiterpenes; D013729:Terpenes; D014466:Ultraviolet Rays; D000094162:Phytoalexins
D017931:DNA Primers; D005875:Gibberellins; D007527:Isoenzymes; D010936:Plant Extracts; D010940:Plant Proteins; D012717:Sesquiterpenes; D013729:Terpenes; D019883:Alkyl and Aryl Transferases; C090963:ent-kaurene synthetase A; D000094162:Phytoalexins