Transformation of the plant pathogenic fungus, Rhynchosporium secalis

The barley leaf scald fungus, Rhynchosporium secalis, was transformed to hygromycin-B and phleomycin resistance using the hph gene from E. coli and the ble gene from Streptoalloteichus hindustanus under the control of Aspergillus nidulans promoter and terminator sequences. Plasmid DNA was introduced into fungal protoplasts by PEG/CaCl2 treatment. Transformation frequencies varied from 59 to 493 transformants per 10 microg of DNA and 5 x 10(7) protoplasts. The antibiotic-resistant phenotype appeared to be stable under selective, as well as under non-selective, conditions for several generations. Co-transformation using the E. coli uidA gene under the control of A. nidulans promoter and terminator sequences on a non-selectable plasmid occurred at frequencies of up to 66%.