Agrobacterium tumefaciens-mediated transformation in Colletotrichum falcatum and C. acutatum

Karunakaran Maruthachalam, Vijayan Nair, Hee Sool Rho, Jaehyuk Choi, Soonok Kim, Yong Hwan Lee

Research output: Contribution to journalArticlepeer-review

Abstract

Agrobacterium 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.

Original languageEnglish (US)
Pages (from-to)234-241
Number of pages8
JournalJournal of Microbiology and Biotechnology
Volume18
Issue number2
StatePublished - Feb 28 2008

Keywords

  • C. acutatum
  • C. falcatum
  • Insertional mutagenesis
  • Pepper anthracnose
  • Sugarcane red rot
  • Transformation

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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