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Background  The plant pathogenic fungus,  Sphaeropsis visci  a dark-spored species of Botryosphaeriaceae, which causes the leaf spot disease of the European mistletoe ( Viscum album ). This species seems to have potential as a tool for biological control of the hemiparasite. For the rapid detection of  S. visci  haplotypes we tested a direct PCR assay without prior DNA purification. This approach was based on a polymerase enzyme from the crenarchaeon  Sulfolobus solfataricus  engineered by fusion protein technology, which linked the polymerase domain to a sequence non-specific DNA binding protein (Sso7d) .Findings  Most isolates of  Sphaeropsis visci  grouped together in our phylogenetic analyses, indicating that isolates had a previously reported haplotype sequence, which is commonly found in the analyzed Hungarian population. This haplotype was also reported from diseased mistletoe bushes from other European countries. We further identified unique single nucleotide polymorphisms (SNPs) in the ITS region, which were specific to the only well resolved clade in the phylogenetic analysis.    Conclusions  The diPCR approach allowed amplification of ITS rRNA gene directly from small amounts of fungal samples without prior DNA extraction. This simple bioassay in plant disease management enables collection of genomic data from fungal plant pathogen populations.

Application of direct PCR in rapid rDNA ITS haplotype determination of the hyperparasitic fungus Sphaeropsis visci (Botryosphaeriaceae)