Agrobacterium tumefaciens -mediated genetic transformation of Aspergillus carbonarius.

Maria Helena P. Fungaro, Luiz Rodrigo Ito, Rubens Tadeu Duarte, Márcia Cristina Furlaneto.

Author address: 

Centro de Ciëncias Biológicas, Universidade Estadual de Londrina, CP, 6001, CEP 86051-990, Londrina-Paraná. Brazil.

Abstract: 

Aspergillus carbonarius is a potent ochratoxin A (OTA) producer that has been found in cereal and food commodities. OTA has nephrotoxic effect and it has been classified as a possible carcinogenic substance for humans. Genetic studies of Aspergillus carbonarius are poorly developed and here we described a genetic transformation system for this filamentous fungus, providing an important step towards the genetic manipulation of this ochratoxigenic species. The fungus was transformed to hygromycin B resistance using the AGL-1 strain of Agrobacterium tumefaciens harboring the binary vector pPK2. Transformation frequencies ranged from 23 to 101 transformants per 105 conidia. Highly resistant transformants were obtained which showed capacity of growing on increased concentrations of hygromycin B (up to 1200 ug ml-1).High mitotic stability of the transformants (94%) was demonstrated after ten successive transfers on non-selective media.Considering the efficiency and flexibility of A. tumefaciens-mediated transformation, this method appears to be an efficient alternative to other insertional mutagenesis techniques in characterizing genes related to ochratoxin biosynthesis. Acknowledgements CAPES/CNPq/Fundaçí£o Araucí¡ria/CPG-UEL
2005

abstract No: 

560

Full conference title: 

23rd Fungal Genetics Conference
    • Fungal Genetics Conference 23rd (2002)