title: Genetic and Molecular Basis of Botrydial Biosynthesis: Connecting Cytochrome P450-Encoding Genes to Biosynthetic Intermediates.
abstract: Over two hundred species of plants can be infected by the phytopathogenic fungus Botrytis cinerea under a range of different environmental conditions. In response to these, the fungus produces unique terpenoid and polyketide metabolites. Parts of the plants may be killed by the phytotoxin botrydial, enabling the fungus to feed on the dead cells. In this paper, we describe the genetic and molecular basis of botrydial biosynthesis and the function of the five genes of the genome of B. cinerea that together constitute the botrydial biosynthetic gene cluster. Genes BcBOT3 and BcBOT4, encoding two cytochrome P450 monooxygenases, were inactivated by homologous recombination and were shown to catalyze regio- and stereospecific hydroxylations at the carbons C-10 and C-4, respectively, of the presilphiperfolan-8β-ol skeleton. The null mutants, bcbot3Δ and bcbot4Δ, accumulated key intermediates in the botrydial biosynthesis enabling the complete genetic and molecular basis of the botrydial biosynthetic pathway to be established. Furthermore, the bcbot4Δ mutant overproduced a significant number of polyketides, which included, in addition to known botcinins, botrylactones and cinbotolide A, two new botrylactones and two new cinbotolides, cinbotolides B and C.
authors.affiliation: Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Puerto Real, 11510 Cádiz, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Moraga
authors.affiliation: UMR BIOGER, INRA, AgroParisTech, Université Paris-Saclay , 78850 Thiverval-Grignon, France.
authors.forename: Bérengère
authors.initials: B
authors.lastname: Dalmais
authors.affiliation: Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Puerto Real, 11510 Cádiz, Spain.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Izquierdo-Bueno
authors.affiliation: Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Puerto Real, 11510 Cádiz, Spain.
authors.forename: Josefina
authors.initials: J
authors.lastname: Aleu
authors.affiliation: Department of Organic Chemistry, University of Sussex , Brighton, Sussex BN1 9QJ, United Kingdom.
authors.forename: James R
authors.initials: JR
authors.lastname: Hanson
authors.affiliation: Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Puerto Real, 11510 Cádiz, Spain.
authors.forename: Rosario
authors.initials: R
authors.lastname: Hernández-Galán
authors.affiliation: UMR BIOGER, INRA, AgroParisTech, Université Paris-Saclay , 78850 Thiverval-Grignon, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Viaud
authors.affiliation: Departamento de Química Orgánica, Facultad de Ciencias, Universidad de Cádiz , Puerto Real, 11510 Cádiz, Spain.
authors.forename: Isidro G
authors.initials: IG
authors.lastname: Collado
mesh_terms: D000447:Aldehydes / Q000378:metabolism*; D020171:Botrytis / Q000235:genetics*; D001643:Bridged Bicyclo Compounds / Q000378:metabolism*; D066241:Carbon-13 Magnetic Resonance Spectroscopy; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D008401:Gas Chromatography-Mass Spectrometry; D005800:Genes, Fungal; D061065:Polyketides / Q000378:metabolism; D066244:Proton Magnetic Resonance Spectroscopy
chemical_list: D000447:Aldehydes; D001643:Bridged Bicyclo Compounds; D061065:Polyketides; C432873:botrydial; D003577:Cytochrome P-450 Enzyme System