Imaging mycorrhizal fungal transformants that express EGFP during ericoid endosymbiosis.
Ericoid endomycorrhizal fungi form intracellular associations with the epidermal root cells of plants belonging to Ericales. In natural environments, these fungi increase the ability of their host plants to colonise soils polluted with toxic metals, although the underlying mechanisms are not clearly understood. Genetic transformation is a powerful tool to study the function of specific genes involved in the interaction of symbiotic fungi with the host plants and with the environment. Here, we investigated the possibility to genetically transform an ericoid endomycorrhizal strain. A metal tolerant mycorrhizal Oidiodendron maius strain isolated from a contaminated area was chosen to develop the transformation system. Two different protocols were used: protoplasts and Agrobacterium-mediated transformation. Stable transformants were obtained with both techniques. They remained competent for mycorrhizal formation and GFP-transformed fungi were visualised in planta. This is the first report of stable transformation of an ericoid endomycorrhizal fungus. The protocol set up could represent a good starting point for the identification of genes important in the ericoid mycorrhiza formation and in the understanding of how this symbiosis is established and functions. The success in the genetic transformation of this strain will allow us to better define its potential use in bioremediation strategies.
Martino
Elena
E
Dipartimento di Biologia Vegetale dell'Università di Torino, Centre of Excellence for Plant and Microbial Biosensing (CEBIOVEM) and Istituto per la Protezione delle Piante del CNR, Sezione di Torino, Torino, Italy. elena.martino@unito.it
Murat
Claude
C
Vallino
Marta
M
Bena
Andrea
A
Perotto
Silvia
S
Spanu
Pietro
P
D016960:Agrobacterium tumefaciens / Q000235:genetics; D001203:Ascomycota / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D049452:Green Fluorescent Proteins / Q000235:genetics; D019216:Metals, Heavy / Q000378:metabolism; D038821:Mycorrhizae / Q000235:genetics* / Q000378:metabolism*; D018517:Plant Roots / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012988:Soil Microbiology; D012989:Soil Pollutants / Q000378:metabolism; D013559:Symbiosis / Q000235:genetics; D014170:Transformation, Genetic; D015032:Zinc / Q000378:metabolism
D019216:Metals, Heavy; D011994:Recombinant Proteins; D012989:Soil Pollutants; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D015032:Zinc