Development of PCR primers for detection of Cylindrocladium pauciramosum

Cylindrocladium pauciramosum, hypothesized to benative to South or Central America, has recently beenreported from Italy, where it was identified on the basisof morphological characters as well as mating type studies.PCR oligonucleotide primers named CPAUCIR/CPAUCIF were designed from b-tubulin gene sequencesto improve detection. These primers amplifieda 161 bp fragment of the b-tubulin gene from all C. pauciramosumisolates tested but failed to amplify DNAfrom other four Cylindrocladium species (C. scoparium,C. candelabrum, C. insulare and C. mexicanum) and toisolates of several other fungi or stramenopiles. In nestedPCR asssays, these primers efficiently amplified C.pauciramosum DNA sequences from infected plants,thus proving to be a suitable diagnostic tool.