title: Stable genetic transformation of the ectomycorrhizal fungus Pisolithus tinctorius.
abstract: In the present work the genetic transformation and the expression of gene markers in transgenic Pisolithus tinctorius are reported. The ectomycorrhizae are facultative symbionts of plant roots, which are capable of affording mineral nutrients to its co-host in exchange of fixed carbon. Given the importance of this association (more than 80% of gymnosperms are associated with these fungi), its study from both basic and applied viewpoints is relevant. We have transformed this fungus with reporter genes and analyzed their expression in its saprophytic state. Genetic transformation was performed by microprojectile bombardment and Agrobacterium-mediated transformation. This last method proved to be the more efficient. Southern analysis of biolistic-transformed fungi revealed the random integration of the transgene into the genome. The accumulation of the transcript of the reporter gene was demonstrated by RT-PCR. The visualization of GFP-associated fluorescence in saprophytic mycelia confirmed the expression of the reporter gene. This is the first report on the stable transformation and expression of GFP in the ectomycorrhizal fungus P. tinctorius.
authors.affiliation: Departamento de Biotecnología y Bioingeniería, Irapuato, Centro de Investigación y Estudios Avanzados del IPN., Av. IPN. 2508, San Pedro Zacatenco 07360 México, D.F.
authors.forename: Aída V
authors.initials: AV
authors.lastname: Rodríguez-Tovar
authors.forename: Roberto
authors.initials: R
authors.lastname: Ruiz-Medrano
authors.forename: Aseneth
authors.initials: A
authors.lastname: Herrera-Martínez
authors.forename: Blanca E
authors.initials: BE
authors.lastname: Barrera-Figueroa
authors.forename: M Eugenia
authors.initials: ME
authors.lastname: Hidalgo-Lara
authors.forename: Blanca E
authors.initials: BE
authors.lastname: Reyes-Márquez
authors.forename: José Luis
authors.initials: JL
authors.lastname: Cabrera-Ponce
authors.forename: María
authors.initials: M
authors.lastname: Valdés
authors.forename: Beatriz
authors.initials: B
authors.lastname: Xoconostle-Cázares
mesh_terms: D016960:Agrobacterium tumefaciens; D001487:Basidiomycota / Q000378:metabolism* / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D028223:Pinus / Q000502:physiology; D018517:Plant Roots / Q000502:physiology; D013559:Symbiosis; D014170:Transformation, Genetic
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins