title: Genetic basis for mycophenolic acid production and strain-dependent production variability in Penicillium roqueforti.
abstract: Mycophenolic acid (MPA) is a secondary metabolite produced by various Penicillium species including Penicillium roqueforti. The MPA biosynthetic pathway was recently described in Penicillium brevicompactum. In this study, an in silico analysis of the P. roqueforti FM164 genome sequence localized a 23.5-kb putative MPA gene cluster. The cluster contains seven genes putatively coding seven proteins (MpaA, MpaB, MpaC, MpaDE, MpaF, MpaG, MpaH) and is highly similar (i.e. gene synteny, sequence homology) to the P. brevicompactum cluster. To confirm the involvement of this gene cluster in MPA biosynthesis, gene silencing using RNA interference targeting mpaC, encoding a putative polyketide synthase, was performed in a high MPA-producing P. roqueforti strain (F43-1). In the obtained transformants, decreased MPA production (measured by LC-Q-TOF/MS) was correlated to reduced mpaC gene expression by Q-RT-PCR. In parallel, mycotoxin quantification on multiple P. roqueforti strains suggested strain-dependent MPA-production. Thus, the entire MPA cluster was sequenced for P. roqueforti strains with contrasted MPA production and a 174bp deletion in mpaC was observed in low MPA-producers. PCRs directed towards the deleted region among 55 strains showed an excellent correlation with MPA quantification. Our results indicated the clear involvement of mpaC gene as well as surrounding cluster in P. roqueforti MPA biosynthesis.
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Gillot
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Jany
authors.affiliation: Área de Microbiología, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, 24071 León, Spain; Instituto de Biotecnología de León (INBIOTEC), Avenida Real n°1, Parque Científico de León, 24006 León, Spain.
authors.forename: Rebeca
authors.initials: R
authors.lastname: Dominguez-Santos
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Poirier
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Stella
authors.initials: S
authors.lastname: Debaets
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Pedro I
authors.initials: PI
authors.lastname: Hidalgo
authors.affiliation: mAbxience, Upstream Production, Parque Tecnológico de León, Julia Morros s/n, Armunia, 24009, León, Spain.
authors.forename: Ricardo V
authors.initials: RV
authors.lastname: Ullán
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Coton
authors.affiliation: Université de Brest, EA 3882 Laboratoire Universitaire de Biodiversité et d'Ecologie Microbienne, IBSAM ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France. Electronic address: monika.coton@univ-brest.fr.
authors.forename: Monika
authors.initials: M
authors.lastname: Coton
mesh_terms: D002611:Cheese / Q000382:microbiology; D003198:Computer Simulation; D015870:Gene Expression; D020868:Gene Silencing; D005800:Genes, Fungal; D016681:Genome, Fungal; D005810:Multigene Family; D009173:Mycophenolic Acid / Q000378:metabolism*; D010407:Penicillium / Q000235:genetics* / Q000378:metabolism*; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis
chemical_list: D048630:Polyketide Synthases; D009173:Mycophenolic Acid
keywords: Gene silencing; Mycophenolic acid; Mycotoxins; Penicillium roqueforti