Abstract
Introns are generally highly polymorphic regions within genes and were proven to be of great interest for discriminating among phylogenetically-close Phytophthora species. Phytophthora ramorum and P. fragariae are considered as quarantine pathogens by the European Union and accurate detection tools are therefore necessary for their monitoring. From introns located in different single copy genes (GPA1, RAS-like, and TRP1), we developed a series of PCR primers specific to P. ramorum and P. fragariae. The specificity of these primers was successfully checked with a wide collection of Phytophthora isolates and a protocol was developed to detect both pathogens directly in infected plant tissues. These genes should be of particular interest for the development of additional species-specific detection tools within the Phytophthora genus.
References
Anon. (2000) Council directive 2000/29/EC of 8 May 2000 on protective measures against the introduction into the community of organisms harmful to plants or plant products and against their spread within the community. OJ No L169, 2000.7.10, 1–112
Anon. (2002) Commission decision (2002/757/EC) of 19 September 2002 on provisional emergency phytosanitary measures to prevent the introduction into and the spread within the Community of Phytophthora ramorum Werres, De Cock & Man in ‘t Veld sp. nov. OJ No L252, 2002-09-20, 3739
Bonants PJM, van Gent-Pelzer MPE, Hooftman R, Cooke DEL, Guy DC, Duncan JM (2004) A combination of baiting and different PCR formats, including measurement of real-time quantitative fluorescence, for the detection of Phytophthora fragariae in strawberry plants. European Journal of Plant Pathology 110:689–702
Brasier CM, Kirk SA, Delcan J, Cooke DEL, Jung T, Man in’t Veld WA (2004) Phytophthora alni sp. nov. and its variants: Designation of emerging heteroploid hybrid pathogens spreading on Alnus trees. Mycological Research 108:1172–1184
Cooke DEL, Drenth A, Duncan JM, Wagels G, Brasier CM (2000) A molecular phylogeny of Phytophthora and related oomycetes. Fungal Genetics and Biology 30:17–32
Garbelotto M (2003) Molecular diagnostics of Phytophthora ramorum, causal agent of Sudden Oak Death. Sudden Oak Death Online Symposium. www.apsnet.org/online/SOD
Hayden KJ, Rizzo D, Tse J, Garbelotto M (2004) Detection and quantification of Phytophthora ramorum from California forests using a real-time polymerase chain reaction assay. Phytopathology 94:1075–1083
Ioos R, Husson C, Andrieux A, Frey P (2005) SCAR-based PCR primers to detect the hybrid pathogen Phytophthora alni and its subspecies causing alder disease in Europe. European Journal of Plant Pathology 112:323–335
Ioos R, Andrieux A, Marçais B, Frey P (2006) Genetic characterization of the natural hybrid species Phytophthora alni as inferred from nuclear and mitochondrial DNA analyses. Fungal Genetics and Biology 43:511–529
Kroon LPNM, Verstappen ECP, Kox LFF, Flier WG, Bonants PJM (2004) A rapid diagnostic test to distinguish between American and European populations of Phytophthora ramorum. Phytopathology 94:613–620
Martin FN, Tooley PW, Blomquist C (2004) Molecular detection of Phytophthora ramorum, the causal agent of Sudden Oak Death in California, and two additional species commonly recovered from diseased plant material. Phytopathology 94:621–631
Rizzo DM, Garbelotto M, Davidson JM, Slaughter GW, Koike ST (2002) Phytophthora ramorum as the cause of extensive mortality in Quercus spp. and Lithocarpus densiflorus in California. Plant Disease 86:205–214
Robin C, Desprez-Loustau ML, Capron G, Delatour C (1998) First record of Phytophthora cinnamomi on cork and holm oak in France and evidence of pathogenicity. Annals of Forest Science 55:869–883
Tomlinson JA, Boonham N, Hughes KJD, Griffin RL, Barker I (2005) On-site extraction and Real-time PCR for detection of Phytophthora ramorum in the field. Applied and Environmental Microbiology 11:6702–6710
Acknowledgements
The authors are very grateful to Drs A. Chandelier, S. Prospero, E.M. Hansen and K.J.D. Hughes for providing P. ramorum and P. fragariae isolates, and to Mrs D. Lesgourgues and SRPV Objat for providing strawberry and raspberry plants naturally infected by P. fragariae var. fragariae and P.␣fragariae var. rubi, respectively.
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Ioos, R., Laugustin, L., Schenck, N. et al. Usefulness of single copy genes containing introns in Phytophthora for the development of detection tools for the regulated species P. ramorum and P. fragariae . Eur J Plant Pathol 116, 171–176 (2006). https://doi.org/10.1007/s10658-006-9051-2
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DOI: https://doi.org/10.1007/s10658-006-9051-2