Functional expression of the green fluorescent protein in the ectomycorrhizal model fungus Hebeloma cylindrosporum.
Hebeloma cylindrosporum is a model fungus for mycorrhizal studies because of its fast growth rate, simple nutritional requirements, and completion of its life cycle in vitro, and because it is amenable to transformation. To advance cell biological research during establishment of symbiosis, a tool that would enable the direct visualisation of fusion proteins in the different symbiotic tissues [namely, the expression of reporter genes such as Green Fluorescent Protein (GFP)] was still a missing tool. In the present study, H. cylindrosporum was transformed using Agrobacterium carrying the binary plasmid pBGgHg containing the Escherichia coli hygromycin B phosphotransferase (hph) and the EGFP genes, both under the control of the Agaricus bisporus glyceraldehyde-3-phosphate dehydrogenase promoter. EGFP expression was successfully detected in transformants. The fluorescence was uniformly distributed in the hyphae, while no significant background signal was detected in control hyphae. The suitability of EGFP for reporter gene studies in Hebeloma cylindrosporum was demonstrated opening up new perspectives in the Hebeloma genetics.
Müller
Tobias
T
Transport in Ectomycorrhiza, Institute of Cellular and Molecular Botany (IZMB), University Bonn, 53115, Bonn, Germany.
Benjdia
Mariam
M
Plant Physiology, Center for Plant Molecular Biology (ZMBP), Auf der Morgenstelle 1, 72076, Tübingen, Germany.
Avolio
Meghan
M
Transport in Ectomycorrhiza, Institute of Cellular and Molecular Botany (IZMB), University Bonn, 53115, Bonn, Germany.
Voigt
Boris
B
IZMB, Department of Plant Cell Biology, University of Bonn, 53115, Bonn, Germany.
Menzel
Diedrik
D
IZMB, Department of Plant Cell Biology, University of Bonn, 53115, Bonn, Germany.
Pardo
Alejandro
A
Programa de Investigacion en Interacciones Biologicas, Universidad Nacional de Quilmes, Roque Saenz Peña 180, B1876BXD, Bernal, Provincia de Buenos Aires, Argentina.
Frommer
Wolf B
WB
Plant Physiology, Center for Plant Molecular Biology (ZMBP), Auf der Morgenstelle 1, 72076, Tübingen, Germany.
Wipf
Daniel
D
Transport in Ectomycorrhiza, Institute of Cellular and Molecular Botany (IZMB), University Bonn, 53115, Bonn, Germany. dwipf@uni-bonn.de.
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism; D001487:Basidiomycota / Q000235:genetics / Q000378:metabolism*; D015139:Blotting, Southern; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D038821:Mycorrhizae / Q000235:genetics / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
D012329:RNA, Bacterial; D049452:Green Fluorescent Proteins