title: Agrobacterium tumefaciens-mediated transformation of Lasiodiplodia theobromae, the causal agent of gummosis in cashew nut plants.
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.
authors.affiliation: Embrapa Agroindústria Tropical, Fortaleza, CE, Brasil celli.muniz@embrapa.br.
authors.forename: C R
authors.initials: CR
authors.lastname: Muniz
authors.affiliation: Laboratório de Biologia Molecular, Embrapa Amazônia Ocidental, Manaus, AM, Brasil.
authors.forename: G F
authors.initials: GF
authors.lastname: da Silva
authors.affiliation: Embrapa Agroenergia, Parque Estação Biológica, Brasília, DF, Brasil.
authors.forename: M T
authors.initials: MT
authors.lastname: Souza
authors.affiliation: Embrapa Agroindústria Tropical, Fortaleza, CE, Brasil.
authors.forename: F C O
authors.initials: FC
authors.lastname: Freire
authors.affiliation: Biointeractions and Plant Health, Plant Research International B.V., Wageningen, The Netherlands.
authors.forename: G H J
authors.initials: GH
authors.lastname: Kema
authors.affiliation: Universidade Estadual do Ceará, Centro de Ciência da Saúde, Fortaleza, CE, Brasil.
authors.forename: M I F
authors.initials: MI
authors.lastname: Guedes
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D031021:Anacardium / Q000235:genetics* / Q000254:growth & development / Q000382:microbiology; D001203:Ascomycota / Q000235:genetics* / Q000472:pathogenicity; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D025301:Hyphae / Q000235:genetics / Q000472:pathogenicity; D009754:Nuts / Q000235:genetics* / Q000254:growth & development; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D049452:Green Fluorescent Proteins