A PCR-based method for the detection of Phaeomoniella chlamydospora in grapevines
The oligonucleotide primers, PCL1 and PCL2, were synthesized for Phaeomoniella chlamydospora from the variable internal transcribed spacers ITS1 and ITS2 of the ribosomal DNA, respectively. Polymerase chain reaction (PCR amplification) with PCL1 and PCL2 produced a 325-bp fragment from isolates of Pa. chlamydospora. Amplification of this fragment was achieved from as little as 16 pg of fungal DNA. The specific primers also amplified a 325-bp fragment from infected grapevine tissue. Fungal DNA from closely related genera, Phaeoacremonium and Phialophora, as well as several other fungi commonly occurring in grapevine stems, showed no amplification with the species-specific primers. These primers can now be used to detect the presence of Pa. chlamydospora in infected nursery material, and thus form the basis for a phytosanitary certification scheme.