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Efficient Genetic Transformation System for the Ochratoxigenic Fungus Aspergillus carbonarius

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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 105 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.

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Acknowledgments

Grants and fellowships from the following Brazilian institutions supported this work: Fundação Araucária and Conselho Nacional de Pesquisa (CNPq).

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Authors and Affiliations

  1. Centro de Ciências Biológicas, Universidade Estadual de Londrina, PO Box 6001, Londrina, PR, 86051-990, Brazil

    Luiz Rodrigo I. Morioka, Márcia C. Furlaneto, Andréa C. Bogas, Patrícia Pompermayer, Rubens T. D. Duarte, Maria Angélica Ehara Watanabe & Maria Helena P. Fungaro

  2. Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, PO Box 83, Piracicaba, SP, 13400-970, Brazil

    Maria Lúcia C. Vieira

Authors
  1. Luiz Rodrigo I. Morioka
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  2. Márcia C. Furlaneto
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  3. Andréa C. Bogas
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  4. Patrícia Pompermayer
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  5. Rubens T. D. Duarte
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  6. Maria Lúcia C. Vieira
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  7. Maria Angélica Ehara Watanabe
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  8. Maria Helena P. Fungaro
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Corresponding author

Correspondence to Maria Helena P. Fungaro.

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Morioka, L.R.I., Furlaneto, M.C., Bogas, A.C. et al. Efficient Genetic Transformation System for the Ochratoxigenic Fungus Aspergillus carbonarius. Curr Microbiol 52, 469–472 (2006). https://doi.org/10.1007/s00284-005-0402-6

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  • DOI: https://doi.org/10.1007/s00284-005-0402-6

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