title: Agrobacterium-mediated insertional mutagenesis in the mycorrhizal fungus Laccaria bicolor.
abstract: Agrobacterium-mediated gene transfer (AMT) is extensively employed as a tool in fungal functional genomics and accordingly, in previous studies we used AMT on a dikaryotic strain of the ectomycorrhizal basidiomycete Laccaria bicolor. The interest in this fungus derives from its capacity to establish a symbiosis with tree roots, thereby playing a major role in nutrient cycling of forest ecosystems. The ectomycorrhizal symbiosis is a highly complex interaction involving many genes from both partners. To advance in the functional characterization of fungal genes, AMT was used on a monokaryotic L. bicolor. A collection of over 1200 transgenic strains was produced, of which 200 randomly selected strains were analyzed for their genomic T-DNA insertion patterns. By means of insertional mutagenesis, a number of transgenic strains were obtained displaying differential growth features. Moreover, mating with a compatible strain resulted in dikaryons that retained altered phenotypic features of the transgenic monokaryon. The analysis of the T-DNA integration pattern revealed mostly similar results to those reported in earlier studies, confirming the usefulness of AMT on different genetic backgrounds of L. bicolor. Taken together, our studies display the great versatility and potentiality of AMT as a tool for the genetic characterization of L. bicolor.
authors.affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes and Consejo Nacional de Investigaciones Científicas y Técnicas, Roque Saenz Peña 352, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
authors.forename: B I
authors.initials: BI
authors.lastname: Stephan
authors.affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes and Consejo Nacional de Investigaciones Científicas y Técnicas, Roque Saenz Peña 352, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
authors.forename: M C
authors.initials: MC
authors.lastname: Alvarez Crespo
authors.affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes and Consejo Nacional de Investigaciones Científicas y Técnicas, Roque Saenz Peña 352, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
authors.forename: M J
authors.initials: MJ
authors.lastname: Kemppainen
authors.affiliation: Laboratorio de Micología Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes and Consejo Nacional de Investigaciones Científicas y Técnicas, Roque Saenz Peña 352, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina. apardo@unq.edu.ar.
authors.forename: A G
authors.initials: AG
authors.lastname: Pardo
mesh_terms: D060054:Agrobacterium / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D016681:Genome, Fungal / Q000235:genetics; D055399:Laccaria / Q000235:genetics*; D016254:Mutagenesis, Insertional; D038821:Mycorrhizae / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013559:Symbiosis; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D005656:Fungal Proteins; C036483:T-DNA
keywords: Agrobacterium-mediated gene transfer; Insertional mutagenesis; Laccaria bicolor; Monokaryon