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Agrobacterium tumefaciens-mediated transformation of Lasiodiplodia theobromae, the causal agent of gummosis in cashew nut plants

Agrobacterium tumefaciens-mediated transformation of Lasiodiplodia theobromae, the causal agent of gummosis in cashew nut plants

Authors :
Maria Izabel Florindo Guedes
Manoel Souza
G. F. da Silva
Gert H. J. Kema
Celli Rodrigues Muniz
Francisco das Chagas Oliveira Freire
Source :
Genetics and Molecular Research, 13(2), 2906-2913, Genetics and Molecular Research 13 (2014) 2
Publication Year :
2014

Abstract

Lasiodiplodia theobromae is a major pathogen of many different crop cultures, including cashew nut plants. This paper describes an efficient Agrobacterium tumefaciens-mediated transformation (ATMT) system for the successful delivery of T-DNA, transferring the genes of green fluorescent protein (gfp) and hygromycin B phosphotransferase (hph) to L. theobromae. When the fungal pycnidiospores were co-cultured with A. tumefaciens harboring the binary vector with hph-gfp gene, hygromycin-resistant fungus only developed with acetosyringone supplementation. The cashew plants inoculated with the fungus expressing GFP revealed characteristic pathogen colonization by epifluorescence microscopy. Intense and bright green hyphae were observed for transformants in all extensions of mycelium cultures. The penetration of parenchyma cells near to the inoculation site, beneath the epicuticle surface, was observed prior to 25 dpi. Penetration was followed by the development of hyphae within invaded host cells. These findings provide a rapid and reproducible ATMT method for L. theobromae transformation.

Details

ISSN :
16765680
Volume :
13
Issue :
2
Database :
OpenAIRE
Journal :
Genetics and molecular research : GMR
Accession number :
edsair.doi.dedup.....7231f2f41b573bcaf99f2b87d46ab989