Agrobacterium-mediated transformation of the ectomycorrhizal symbiont Laccaria bicolor S238N.
The development of an efficient transformation system is required to alter the expression of symbiosis-regulated genes and to develop insertional mutagenesis in the ectomycorrhizal basidiomycete Laccaria bicolor S238N. Vegetative mycelium of this fungus was transformed by Agrobacterium tumefaciens-mediated gene transfer. The selection marker was the hygromycin resistance gene of Escherichia coli (hph) under the control of the gpd promoter from Agaricus bisporus and the CaMV 35S terminator as part of the T-DNA. PCR amplification of hph and Southern blot analyses showed that the genome of the hygromycin-resistant transformants contained the cassette. The latter proved mostly single copy and random integration of part of the transgene into the fungal genome. A. tumefaciens-mediated gene transfer should facilitate future development of insertional mutagenesis, targeted gene disruption and RNA interference technology in L. bicolor.
Kemppainen
Minna
M
Programa de Investigación en Interacciones Biológicas, Universidad Nacional de Quilmes, Roque Sáenz Peña 180, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
Circosta
Ariana
A
Programa de Investigación en Interacciones Biológicas, Universidad Nacional de Quilmes, Roque Sáenz Peña 180, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
Tagu
Denis
D
UMR INRA-UHP 1136 Interactions Arbres/Micro-organismes, Centre INRA de Nancy, 54280, Champenoux, France.|UMR BIO3P, Centre INRA de Rennes, BP 35327, 35653, Le Rheu Cedex, France.
Martin
Francis
F
UMR INRA-UHP 1136 Interactions Arbres/Micro-organismes, Centre INRA de Nancy, 54280, Champenoux, France.
Pardo
Alejandro G
AG
Programa de Investigación en Interacciones Biológicas, Universidad Nacional de Quilmes, Roque Sáenz Peña 180, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina. apardo@unq.edu.ar.
D000363:Agaricales / Q000235:genetics* / Q000502:physiology; D000935:Antifungal Agents / Q000494:pharmacology; D015139:Blotting, Southern; D017796:Caulimovirus / Q000235:genetics; D002934:Cinnamates / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D018628:Gene Dosage; D016681:Genome, Fungal; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D016254:Mutagenesis, Insertional; D025282:Mycelium / Q000235:genetics; D038821:Mycorrhizae / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D012231:Rhizobium / Q000235:genetics*; D012641:Selection, Genetic; D013559:Symbiosis; D014170:Transformation, Genetic
D000935:Antifungal Agents; D002934:Cinnamates; D004269:DNA, Bacterial; D004271:DNA, Fungal; D029968:Escherichia coli Proteins; C036483:T-DNA; D006921:Hygromycin B; C026273:hygromycin A