Transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. Aeschynomene By electroporation of germinated conidia.
A highly efficient and reproducible procedure for the transformation of the bioherbicide Colletotrichum gloeosporioides f. sp. aeschynomene by electroporation of germinated conidia is reported. Development of the procedure involved a detailed study of the germination process followed by extensive calibration of the electrical-pulse and selection conditions. Optimization of the transformation protocol was facilitated by the use of the green fluorescent protein that helped in the identification of stable transformants and in a fast assessment of transgene expression levels, colony homogeneity and genetic stability. Under optimal conditions, over 80 stable transformants/cuvette were obtained. Plasmid integration was predominantly homologous, but high transformation rates were obtained both with homologous and non-homologous vectors. The method described not only opens up opportunities for the genetic manipulation of C. gloeosporioides f. sp. aeschynomene, but also provides a framework for the development and optimization of transformation in other fungi.
Robinson
M
M
Institute of Cereal Crop Improvement, Department of Plant Sciences, Tel Aviv University, Israel.
Sharon
A
A
D002138:Calibration; D002455:Cell Division; D002467:Cell Nucleus / Q000235:genetics; D048429:Cell Size; D020231:Colletotrichum / Q000166:cytology / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D004553:Electric Conductivity; D018274:Electroporation / Q000379:methods*; D018628:Gene Dosage; D017930:Genes, Reporter / Q000235:genetics; D005818:Genetic Engineering; D006540:Herbicides; D010572:Pest Control, Biological; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D012689:Sequence Homology, Nucleic Acid; D013172:Spores, Fungal / Q000235:genetics / Q000502:physiology; D013997:Time Factors; D014170:Transformation, Genetic
D006540:Herbicides