Abstract
Protoplasts were produced by treating the yeast-like cells of O. piceae and O. quercus with high concentrations of Novozym 234 (20 mg ml−1). Transformants with resistance to hygromycin B were obtained by transforming the protoplasts with the plasmids pAN7-1 and pCB1004 containing the hygromycin B phosphotransferase (hph) gene. For all isolates tested, transformation efficiencies ranged between 103 and 105 transformants μg−1 vector DNA. High polyethylene glycol concentration (66%) increased the transformation efficiency. Integration of the hph gene into the transformant genome was verified by a polymerase chain reaction and dot DNA hybridization analysis. There was no evidence of mitotic instability of hygromycin B resistance in the transformants.
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