Agrobacterium tumefaciens-mediated transformation of Guignardia citricarpa

Guignardia citricarpa, the causal agent of Citrus Black Spot, was successfully transformed via Agrobacterium turnefaciens with cassettes for gfp and bar expression. Transformation is essential to understand the role of genes during interaction between plants and its pathogens. Using a binary plasmid vector based in the pPZP201BK, both germinated conidia and physically fragmented hyphae of G. citricarpa were transformed. Eight independent transformants of G. citricarpa resistant to ammonium glifosinate displayed GFP fluorescence. The majority (93.75%) of the G. citricarpa transformants was mitotically stable and contained a single T-DNA copy ectopically integrated to the chromosome. This is the first report of G. citricarpa transformation and will allow future work on virulence determinants of the fungus and possibly its control. (C) 2009 Elsevier B.V. All rights reserved.