Electroporation and Agrobacterium-mediated spore transformation.
Genetic transformation is a key technology in modern fungal research. Most commonly, protoplasts are transformed using the polyethylene glycol-mediated transformation protocols. Because protoplasts are generated by treatment of mycelia with a crude enzyme preparation, the results tend to be inconsistent. Furthermore, some species cannot be transformed by this method. Electroporation (EP) and Agrobacterium tumefaciens-mediated transformation (AMT) are two alternative methods. These methods allow the transformation of spores or mycelia, they are simple to perform and provide consistent results. In this chapter, we describe EP and AMT protocols for the fungus Colletotrichum gloeosporioides f. sp. aeschynomene (C. gloeosporioides). These protocols can be used as baseline for the calibration of similar transformation protocols in other species.
Minz
Anna
A
Department of Plant Sciences, Tel-Aviv University, Tel Aviv, Israel.
Sharon
Amir
A
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D020231:Colletotrichum / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D013172:Spores, Fungal / Q000502:physiology*; D014170:Transformation, Genetic
D004271:DNA, Fungal