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Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants

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An Erratum to this article was published on 14 March 2003

Abstract.

Protoplasts of the pathogenic plant fungus, Crinipellis perniciosa, were transformed to hygromycin B resistance using the pAN7-1 plasmid, which contains the Escherichia coli hph gene under the control of Aspergillus nidulans regulatory sequences. The pAN7-1 plasmid was introduced by PEG/CaCl2 treatment. Transformation frequencies of 1.6–2.5 transformants/μg of DNA were achieved. About 54% of the transformants were abortive and 40 analyzed transformants were mitotically stable and showed different hygromycin B resistance levels. The presence of the hph gene was checked by PCR in five transformants and the integration of multiple plasmid copies into different genome sites was observed by Southern analysis. This is the first report of a C. perniciosa transformation system and represents an important step for further research into genetic manipulation of this fungal plant pathogen.

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References

  • Azevedo AMC, De Marco JL, Felix CR (2000) Characterization of an amylase produced by a Tricoderma harzianium isolate with antagonistic activity against Crinipellis perniciosa, the causal agent of witches' broom of cocoa. FEMS Microbiol Lett 188:171–175

    Article  PubMed  Google Scholar 

  • Ballance DJ, Turner G (1985) Development of a high-frequency transforming vector for Aspergillus nidulans. Gene 36:321–331

    CAS  PubMed  Google Scholar 

  • Barrett V, Dixon RK, Lemke PA (1990) Genetic transformation of a mycorrhizal fungus. Appl Microbiol Biotechnol 33:313–316

    CAS  Google Scholar 

  • Bastos CN (1996) Potencial de Trichoderma viride no controle da vassoura-de-bruxa (Crinipellis perniciosa) do cacaueiro. Fitopatol Bras 21:509–512

    Google Scholar 

  • Bowers JH, Bailey BA, Hebbar PK, Sanogo S, Lumsden RD (2001) The impact of plant diseases on world chocolate production. Plant Health Prog DOI 10.1094/PHP-2001-0709-01-RV

  • Churchill ACL, Ciuffetti LM, Hansen DR, Van Etten HD, Van Alfen NK (1990) Transformation of the fungal pathogen Cryphonectria parasitica with a variety of heterologous plasmids. Curr Genet 17:25–31

    CAS  Google Scholar 

  • Furlaneto MC, Paião FG, Pinto FGS, Fungaro MHP (1999) Transformation of the entomopathogenic fungus Metarhizium flavoviride to high resistance to benomyl. Can J Microbiol 45:875–878

    Article  CAS  Google Scholar 

  • Hirano T, Sato T, Yaegashi K, Enei H (2000) Efficient transformation of the edible basidiomycete Lentinus edodes with a vector using a glyceraldehyde-3-phosphate dehydrogenase promoter to hygromycin B resistance. Mol Gen Genet 263:1047–1052

    CAS  PubMed  Google Scholar 

  • Idnurm A, Howllet BJ (2001) Pathogenicity genes of phytopathogenic fungi. Mol Plant Pathol 2:241–255

    Article  CAS  Google Scholar 

  • Irie T, Honda Y, Watanabe T, Kuwahara M (2001) Efficient transformation of filamentous fungus Pleurotus ostreatus using single-strand carrier DNA. Appl Microbiol Biotechnol 5:563–565

    Article  Google Scholar 

  • Kachroo P, Potnis A, Chattoo BB (1997) Transformation of the rice blast fungus Magnaporthe grisea to benomyl resistance. World J Microbiol Biotechnol 13:185–187

    CAS  Google Scholar 

  • Leung H, Lehtinen U, Karjalainen R, Skinner D, Tooley P, Leong S (1990) Transformation of the rice blast fungus Magnaporthe grisea to hygromycin B resistance. Curr Genet 17:409–411

    CAS  PubMed  Google Scholar 

  • Malonek S, Meinhardt F (2001) Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic ascomycete Calonectria morganii. Curr Genet 40:152–155

    CAS  PubMed  Google Scholar 

  • Marmeisse R, Gay G, Debaud JC, Casselton LA (1992) Genetic transformation of the symbiotic basidiomycete fungus Hebeloma cylindrosporum. Curr Genet 22:41–45

    CAS  PubMed  Google Scholar 

  • Mullins ED, Kang S (2001) Transformation: a tool for studying fungal pathogens of plants. Cell Mol Life Sci 58:2043–2052

    CAS  PubMed  Google Scholar 

  • Mullins ED, Chen X, Romaine P, Raina R, Geiser DM, Kang S (2001) Agrobacterium-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer. Phytopathology 91:173–180

    CAS  Google Scholar 

  • Oliver RP, Roberts IN, Harling R, Kenyon L, Punt PJ, Dingemanse MA, Hondel CAMJJ van den (1987) Transformation of Fulvia fulva, a fungal pathogen of tomato, to hygromycin B resistance. Curr Genet 12:231–233

    CAS  Google Scholar 

  • Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, Hondel CAMJJ van den (1987) Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli. Gene 56:117–124

    CAS  PubMed  Google Scholar 

  • Rohe M, Searle J, Newton AC, Knogge W (1996) Transformation of the plant pathogenic fungus, Rhynchosporium secalis. Curr Genet 29:587–590

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Schiestl RH, Petes TD (1991) Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 88:7585–7589

    CAS  PubMed  Google Scholar 

  • Schuren FHJ, Wessels JGH (1994) Highly-efficient transformation of the homobasidiomycete Schizophyllum commune to phleomycin resistance. Curr Genet 26:179–183

    CAS  PubMed  Google Scholar 

  • Specht CA, DiRusso CC, Novotny CP, Ullrich RC (1982) A method for extracting high-molecular-weight deoxyribonucleic acid from fungi. Anal Biochem 119:158–163

    CAS  PubMed  Google Scholar 

  • Turgeon BG, Garber RC, Yoder OC (1985) Transformation of the fungal maize pathogen Cochliobolus heterostrophus using the Aspergillus nidulans amdS gene. Mol Gen Genet 201:450–453

    CAS  Google Scholar 

  • Valadares-Inglis MC, Inglis PW (1997) Transformation of the entomopathogenic fungus, Metarhizium flavoviride strain CG423, to benomyl resistance. FEMS Microbiol Lett 155:199–202

    Article  CAS  Google Scholar 

  • Van de Rhee MD, Graça PMA, Huizing HJ, Mooibroek H (1996) Transformation of the cultivated mushroom, Agaricus bisporus, to hygromycin B resistance. Mol Gen Genet 250:252–258

    Article  PubMed  Google Scholar 

  • Wang J, Holden DW, Leong SA (1988) Gene transfer system for phytopathogenic fungus Ustilago Maydis. Proc Natl Acad Sci USA 85:865–869

    CAS  PubMed  Google Scholar 

  • Yelton MM, Hamer JE, Timberlake WE (1984) Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad of Sci USA 81:1470–1474

    CAS  Google Scholar 

Download references

Acknowledgements

We thank the Brazilian agencies CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for financial support. We are grateful to Dr. C.A.M.J.J. van den Hondel for the pAN7-1 plasmid.

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Authors and Affiliations

  1. Departamento de Microbiologia/BIOAGRO, Universidade Federal de Viçosa, , 36.570-000, Viçosa, Minas Gerais, Brazil

    Juliana Oliveira Lima, Jildete Karla dos Santos, Jorge Fernando Pereira, Elza Fernandes de Araújo & Marisa Vieira de Queiroz

  2. Departamento de Fitopatologia, Universidade Federal de Lavras, 37.200-000 , Lavras, Minas Gerais, Brazil

    Mário Lúcio Vilela de Resende

Authors
  1. Juliana Oliveira Lima
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  2. Jildete Karla dos Santos
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  3. Jorge Fernando Pereira
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  4. Mário Lúcio Vilela de Resende
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  5. Elza Fernandes de Araújo
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  6. Marisa Vieira de Queiroz
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Corresponding author

Correspondence to Marisa Vieira de Queiroz.

Additional information

Communicated by U. Kück

An erratum to this article is available at http://dx.doi.org/10.1007/s00294-003-0388-1.

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Lima, J.O., dos Santos, J.K., Pereira, J.F. et al. Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants. Curr Genet 42, 236–240 (2003). https://doi.org/10.1007/s00294-002-0358-z

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  • DOI: https://doi.org/10.1007/s00294-002-0358-z

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