TY - JOUR
T1 - Stable genetic transformation of the ectomycorrhizal fungus Pisolithus tinctorius
AU - Rodríguez-Tovar, Aída V.
AU - Ruiz-Medrano, Roberto
AU - Herrera-Martínez, Aseneth
AU - Barrera-Figueroa, Blanca E.
AU - Hidalgo-Lara, M. Eugenia
AU - Reyes-Márquez, Blanca E.
AU - Cabrera-Ponce, José Luis
AU - Valdés, María
AU - Xoconostle-Cázares, Beatriz
N1 - Funding Information:
We wish to thank Dr. José Ruiz Herrera for critical review of the manuscript. Thanks are also given to the members of our laboratory for useful discussions. AVR-T, AH-M and BEB-F were CONACyT fellows. The present work was supported by grants from CONACyT México B-5327 and 39961 (to BXC), COSNET (DAAE 09/2001).
PY - 2005/10
Y1 - 2005/10
N2 - 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.
AB - 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.
KW - Agro-transformation
KW - GFP expression
KW - Microprojectile bombardment
UR - http://www.scopus.com/inward/record.url?scp=24644499922&partnerID=8YFLogxK
U2 - 10.1016/j.mimet.2005.02.016
DO - 10.1016/j.mimet.2005.02.016
M3 - Artículo
C2 - 15936101
SN - 0167-7012
VL - 63
SP - 45
EP - 54
JO - Journal of Microbiological Methods
JF - Journal of Microbiological Methods
IS - 1
ER -