Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene

  title={Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene},
  author={Bettina Tudzynski and Hiroshi Kawaide and Yuji Kamiya},
  journal={Current Genetics},
Abstract The gene coding for copalyl diphosphate synthase (CPS), which represents the first gene of the gibberellin pathway, was isolated from the rice pathogen Gibberella fujikuroi. This fungus is used commercially for the production of gibberellic acid and related gibberellins. CPS is a terpene cyclase which catalyzes the first specific step of the gibberellin (GA) pathway as it branches off from the general isoprenoid (biosynthetic) pathway at geranylgeranyl disphosphate (GGDP). A cDNA… 

Gibberellin biosynthetic pathway in Gibberella fujikuroi: evidence for a gene cluster.

Results suggest that at least some of the genes involved in the biosynthesis of gibberellins are closely linked in a gene cluster in G. fujikuroi, as has been recently found for other "dispensable" pathways in fungi.

Isolation and Characterization of the Gibberellin Biosynthetic Gene Cluster in Sphaceloma manihoticola

The close similarity between the two GA gene clusters, the identical gene functions, and the conserved intron positions suggest a common evolutionary origin despite the distant relatedness of the two fungi.

Biosynthesis of gibberellins in Gibberella fujikuroi: biomolecular aspects

  • B. Tudzynski
  • Biology
    Applied Microbiology and Biotechnology
  • 1999
This review gives a short overview of the well-known physiological and biochemical studies and concentrates mainly on the new molecular genetic data from GA research, including new information on the regulation of GA biosynthesis.

The P450-4 Gene of Gibberella fujikuroi Encodes ent-Kaurene Oxidase in the Gibberellin Biosynthesis Pathway

Gas chromatography-mass spectrometry analysis of extracts of mycelia and culture fluid of a P450-4 knockout mutant identified ent-kaurene as the only intermediate of the gibberellin biosynthesis pathway, and data suggest that P 450-4 encodes a multifunctional ent- Kaurene oxidase catalyzing all three oxidation steps betweenent-KAurene and ent-kasurenoic acid.

Characterization of the Final Two Genes of the Gibberellin Biosynthesis Gene Cluster of Gibberella fujikuroi

It is demonstrated by gene disruption that P450-3, one of the four cytochrome P450 monooxygenase genes in the GA gene cluster, encodes the 13-hydroxylase, which catalyzes the conversion of GA7 to GA3, in the last step of the pathway.

Isolation, characterization and disruption of the areA nitrogen regulatory gene of Gibberella fujikuroi

To generate areA-GF − mutants, gene replacement experiments were performed using a linearized replacement vector carrying the hygromycin B phosphotransferase (hph) gene, which resulted in transformants which were unable to utilize nitrogen sources other than ammonium and glutamine, and gave significantly reduced gibberellin production yields.

Regulation of Carotenogenesis and Secondary Metabolism by Nitrogen in Wild-Type Fusarium fujikuroi and Carotenoid-Overproducing Mutants

Results suggest regulatory connections between carotenoid biosynthesis and nitrogen-controlled biosynthetic pathways in this fungus.

Conserved bases for the initial cyclase in gibberellin biosynthesis: from bacteria to plants.

Functional conservation of CPSs from bacteria to plants are suggested, presumably reflecting an early role for derived diterpenoids in both plant development and plant-microbe interactions, eventually leading to GA, and a speculative evolutionary scenario is presented.

Biochemical and Molecular Analyses of Gibberellin Biosynthesis in Fungi

  • H. Kawaide
  • Biology, Chemistry
    Bioscience, biotechnology, and biochemistry
  • 2006
A molecular analysis of GA biosynthesis in Phaeosphaeria sp.



Molecular cloning and characterization of a cDNA encoding the gibberellin biosynthetic enzyme ent-kaurene synthase B from pumpkin (Cucurbita maxima L.).

The first committed step in the formation of diterpenoids leading to gibberellin (GA) biosynthesis is the conversion of geranylgeranyl diphosphate to ent-kaurene, and the deduced amino acid sequence shares significant homology with other terpene cyclases, including the conserved DDXXD motif.

The isoprenoid pathway: cloning and characterization of fungal FPPS genes

Transcription analysis in different light regimes has shown that the FPPS genes in G. fujikuroi and N. crassa are not regulated by light induction, and sequence analysis showed a high degree of similarity between the deduced proteins of all known FPP synthase genes.

Ent-kaurene synthase from the fungus Phaeosphaeria sp. L487. cDNA isolation, characterization, and bacterial expression of a bifunctional diterpene cyclase in fungal gibberellin biosynthesis.

Results indicate that the fungus has a bifunctional diterpene cyclase that can convert geranylgeranyl diphosphate into ent-kaurene and may be separate catalytic sites for the two cyclization reactions.

The Arabidopsis GA1 locus encodes the cyclase ent-kaurene synthetase A of gibberellin biosynthesis.

The isolation of a nearly full-length GA1 cDNA clone from wild-type Arabidopsis encodes an active protein and is able to complement the dwarf phenotype in ga1-3 mutants by Agrobacterium-mediated transformation.

The LS locus of pea encodes the gibberellin biosynthesis enzyme ent-kaurene synthase A.

KSA expression in seeds is developmentally regulated and parallels overall GA biosynthesis, suggesting that KSA expression may play an important role in the regulation ofGA biosynthesis and seed development.

The GA2 locus of Arabidopsis thaliana encodes ent-kaurene synthase of gibberellin biosynthesis.

Results show that the GA2 locus encodes KS, and the recombinant AtKS protein derived from the ga2-1 mutant is truncated by 14 kD at the C-terminal end and does not contain significant KS activity in vitro.

GIBBERELLIN BIOSYNTHESIS: Enzymes, Genes and Their Regulation.

  • P. HeddenY. Kamiya
  • Biology
    Annual review of plant physiology and plant molecular biology
  • 1997
Highlights include the cloning of ent-copalyl diphosphate synthase and ent-kaurene synthase, and the demonstration that the former is targeted to the plastid; the finding that the Dwarf-3 gene of maize encodes a cytochrome P450, although of unknown function; and the clones of GA 20-oxidase and 3beta-hydroxylase genes.

3-Hydroxy-3-methylglutaryl-CoA reductase gene of Gibberella fujikuroi: isolation and characterization

Sequence analysis revealed a high degree of similarity between the deduced amino-acid sequences in the C-terminal catalytic domains of all known HMG-CoA reductases, but the highest degree was found between the sequences of both analysed ascomycetes.

The Gibberella fujikuroi niaD gene encoding nitrate reductase: isolation, sequence, homologous transformation and electrophoretic karyotype location.

A cosmid vector designated pGFniaD was generated based on niaD selection and shown to give comparable transformation efficiencies, giving frequencies of up to 200 transformants per microgram DNA, indicating that the vector may be used reliably for both types of integration.


It is confirmed that a single protein catalyzes the complex, multistep cyclization of geranylgeranyl diphosphate to abietadiene, and an apparent fusion of segments of the two distinct terpenoid cyclase types is consistent with the novel mechanism of the bifunctional abetadiene synthase in catalyzing both protonation-initiated and ionization- initiated cyclization steps.