title: Mushroom (Agaricus bisporus).
abstract: We have devised an easy and effective genetic transformation method for the preeminent edible mushroom, Agaricus bisporus. Our method exploits the T-DNA transfer mechanism in Agrobacterium tumefaciens and relies on the reproductive fruiting body as the recipient tissue. The use of fruiting body explants, particularly the gill, provided high-frequency transformation, overcoming the inefficacy of Agrobacterium-based methods targeting fungal spores or vegetative mycelium. The protocol entails incubation of A. tumefaciens for 3 h with acetosyringone, a signaling molecule that launches the gene transfer mechanism, co-cultivation of the induced bacterium and gill explants for 3 d, and selection for transformants based on an inherited resistance to the antibiotic hygromycin. Between 7 and 28 d on the selection medium, upwards of 95% of the gill explants generate hygromycin-resistant colonies. About 75% of the mushroom transformants show a single-copy of the hygromycin-resistant gene integrated at random sites in the genome.
authors.affiliation: Department of Plant Pathology, The Pennsylvania State University, University Park, PA, USA.
authors.forename: C Peter
authors.initials: CP
authors.lastname: Romaine
authors.forename: Carl
authors.initials: C
authors.lastname: Schlagnhaufer
mesh_terms: D000364:Agaricus / Q000166:cytology / Q000235:genetics* / Q000502:physiology; D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D015139:Blotting, Southern; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D003470:Culture Media; D005822:Genetic Vectors; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media