02221nas a2200241 4500008004100000245009100041210006900132300001100201490000800212520145200220653001801672653001401690653001801704653002101722100001901743700002201762700002401784700002101808700001801829700002501847700002601872856008101898 2009 eng d00aDNA-based method for rapid identification of the pine pathogen, Phytophthora pinifolia0 aDNAbased method for rapid identification of the pine pathogen Ph a99-1040 v2983 a

Phytophthora pinifolia causes a needle and shoot disease in Pinus radiata, referred to as ‘Daño Foliar del Pino’. This newly discovered disease requires intensive research efforts that necessitate the processing of large numbers of samples for which accurate identification, often by people not experienced in Phytophthora taxonomy, is required. The aim of this study was, therefore, to develop species-specific primers for P. pinifolia that amplify the internal transcribed spacer region of the ribosomal operon and the nuclear Ypt1 gene, respectively. The primers were tested over several Phytophthora spp., as well as fungi isolated from P. radiata. In all cases, only P. pinifolia was amplified. In addition to the species-specific primers, a PCR-restriction fragment length polymorphism protocol using available Phytophthora genus-specific primers was also used to generate a species-specific profile for P. pinifolia. This provided a characteristic profile that allows the identification of P. pinifolia, and it could also discriminate between 27 different species of Phytophthora. Both techniques reported in this study make it possible to identify large numbers of P. pinifolia cultures accurately and efficiently, which will be important for both quarantine work and biological research on this important new pathogen.

10aMonterey pine10aoomycetes10aPinus radiata10atree defoliation1 aDurán, Alvaro1 aSlippers, Bernard1 aGryzenhout, Marieka1 aAhumada, Rodrigo1 aDrenth, Andre1 aWingfield, Brenda, D1 aWingfield, Michael, J uhttp://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2009.01700.x/abstract