Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Agrobacterium tumefaciens-mediated Transformation in Colletotrichum falcatum and C. acutatum

  • Maruthachalam, Karunakaran (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials and Center for Fungal Genetic Resources, Seoul National University) ;
  • Nair, Vijayan (Biotechnology Laboratory, Sugarcane Breeding Institute) ;
  • Rho, Hee-Sool (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials and Center for Fungal Genetic Resources, Seoul National University) ;
  • Choi, Jae-Hyuk (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials and Center for Fungal Genetic Resources, Seoul National University) ;
  • Kim, Soon-Ok (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials and Center for Fungal Genetic Resources, Seoul National University) ;
  • Lee, Yong-Hwan (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials and Center for Fungal Genetic Resources, Seoul National University)
  • Published : 2008.02.29
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Abstract

Agrobacterum tumefaciens-mediated transformation (ATMT) is becoming an effective system as an insertional mutagenesis tool in filamentous fungi. We developed and optimized ATMT for two Colletotrichum species, C. falcatum and C. acutatum, which are the causal agents of sugarcane red rot and pepper anthracnose, respectively. A. tumefaciens strain SK1044, carrying a hygromycin phosphotransferase gene (hph) and a green fluorescent protein (GFP) gene, was used to transform the conidia of these two Colletotrichum species. Transformation efficiency was correlated with co-cultivation time and bacterial cell concentration and was higher in C. falcatum than in C. acutatum. Southern blot analysis indicated that about 65% of the transformants had a single copy of the T-DNA in both C. falcatum and C. acutatum and that T-DNA integrated randomly in both fungal genomes. T-DNA insertions were identified in transformants through thermal asymmetrical interlaced PCR (TAIL-PCR) followed by sequencing. Our results suggested that ATMT can be used as a molecular tool to identify and characterize pathogenicity-related genes in these two economically important Colletotrichum species.

Keywords

References

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