title: Efficient genetic transformation system for the ochratoxigenic fungus Aspergillus carbonarius.
abstract: Aspergillus carbonarius is a potent ochratoxin A producer that has been found in products such as grapes, coffee, spices, and cocoa. Ochratoxin A has nephrotoxic effect, and it has been classified as a possible carcinogenic substance for humans. Here we describe for the first time a transformation system for A. carbonarius, providing an important step towards its genetic manipulation. Conidia were transformed to acquire hygromycin B resistance using the AGL-1 strain of Agrobacterium tumefaciens. Genetic transformation was evaluated growing A. tumefaciens cells in induction medium containing or not acetosyringone prior to co-cultivation. The mean transforming efficiencies in IM+AS and IM-AS conditions were 62.2 and 44.5 transformants per 10(5) conidia, respectively. The hph gene was random integrated into the genome of A. carbonarius. Fungal sequences flanking the insertion site could be amplified by TAIL-PCR.
authors.affiliation: Centro de Ciências Biológicas, Universidade Estadual de Londrina, PO Box 6001, 86051-990 Londrina, PR, Brazil.
authors.forename: Luiz Rodrigo I
authors.initials: LR
authors.lastname: Morioka
authors.forename: Márcia C
authors.initials: MC
authors.lastname: Furlaneto
authors.forename: Andréa C
authors.initials: AC
authors.lastname: Bogas
authors.forename: Patrícia
authors.initials: P
authors.lastname: Pompermayer
authors.forename: Rubens T D
authors.initials: RT
authors.lastname: Duarte
authors.forename: Maria Lúcia C
authors.initials: ML
authors.lastname: Vieira
authors.forename: Maria Angélica Ehara
authors.initials: MA
authors.lastname: Watanabe
authors.forename: Maria Helena P
authors.initials: MH
authors.lastname: Fungaro
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001230:Aspergillus / Q000235:genetics* / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods; D009793:Ochratoxins / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D009793:Ochratoxins; C025589:ochratoxin A