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Agrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestyles

  • Prasad Sreenivasaprasad
    ,
  • Pedro Talhinhas
    ,
  • S. Muthumeenakshi
    ,
  • João Neves-Martins
    ,
  • Helena Oliveira
    ,
  • Surapareddy Sreenivasaprasad
  • University of Lisbon
    ,
  • University of Warwick
Research Output: Contribution to journal Article Peer-review

Abstract

Colletotrichum acutatum is a cosmopolitan pathogen causing economically important diseases known as anthracnose on a wide range of hosts. This fungus exhibits varied pathogenicity lifestyles and the tools essential to understand the molecular mechanisms are still being developed. The transformation methods currently available for this species for gene discovery and functional analysis involve protoplast transformation and are laborious and inefficient. We have developed a protocol for efficient Agrobacterium tumefaciens-mediated transformation (ATMT) of C. acutatum. Using this protocol we were able to transform C. acutatum isolates belonging to different genetic groups and originating from different hosts. The transformation efficiency was up to 156 transformants per 10(4) conidia, with >70% transformants showing single location/single copy integration of T-DNA. Binary vector pBHt2-GFP was constructed, enabling green fluorescence protein tagging of C. acutatum strains, which will be a useful tool for epidemiology and histopathology studies. The ATMT protocol developed was used to identify putative pathogenicity mutants, suggesting the applicability of this technique for rapid generation of a large panel of insertional mutants of C. acutatum leading to the identification of the genes associated with the varied lifestyles.

Publication Information

Output type

Research Output: Contribution to journal Article Peer-review

Original language

English

Pages from-to (Number of pages)

Pages 57-67

Journal (Volume, Issue Number)

Molecular Biotechnology (Volume 39, Issue 1)

Publication milestones

  • Published - 09/01/2008

Publication status

Published - 09/01/2008

ISSN

1073-6085

External Publication IDs

  • handle.net: 10547/302152
  • Scopus: 52049106458

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