The number of
cycles in the PCR was set to 40, as the 40th cycle represents the extrapolated threshold cycle for a reaction with a theoretical single copy of the template DNA. A total of six 10-fold dilution steps of plasmid standard (10–106 gene copies) were run in triplicateon every wellplate as well as a no-template control (water instead of sample) and a no-amplification control (containing plasmid standard and 0.01% SDS). PCR efficiency was calculated from threshold cycles of these standard dilution steps. As a positive control the DNA extracts from all used isolates were
also tested with a the beta-tubulin PCR assay targeting all Fusarium species described in Yli-Mattila et al. (2004) GenBank accession numbers for beta-tubulin
sequences obtained from 16 Austrian Fusarium iso- lates used in this study are included inTable 1.