Abstract
Primer pairs were designed from a cloned DNA probe of a strain of flavescence dorée (FD) phytoplasma and from a cloned DNA probe of a strain of stolbur phytoplasma. Among an array of reference phytoplasma strains maintained in periwinkle, pair FD9f/r amplified a 1.3 kb DNA fragment only with phytoplasma strains of elm yellows (EY) group, i.e. two strains of FD and two strains of EY. Tru9I restriction analysis of the fragment amplified by FD9f/r revealed a diversity among EY-group phytoplasmas. The FD strains differed from the strains isolated from elm. The profile of the phytoplasmas infecting the grapevine samples from Catalonia and most of the samples from Northern Italy were identical to that of a FD strain. Three other profiles were detected in grapevine from Palatinate, in Germany.
The two primer pairs derived from a stolbur strain, STOL4f/r and STOL11f2/r1, specifically amplified a 1.7 kb and a 0.9 kb DNA fragment, respectively, with all strains in the stolbur subgroup. However, the pair STOL4f/r did not recognise strain MOL. Both pairs allowed to detect phytoplasmas in diseased grapevines from France, Italy, Spain and Israel. Attempts to differentiate between phytoplasmas in the stolbur subgroup by restriction analyses failed. The pairs FD9f/r and STOL11f2/r1 could be used in the same reaction (multiplex PCR) to detect EY-group phytoplasmas, stolbur-subgroup phytoplasmas or both phytoplasmas simultaneously when template DNAs were mixed.
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References
Bertaccini A, Davis RE, Vibio M, Prince JP and Credi R (1993) Detection and characterisation of mycoplasmalike organism (MLO)DNA in naturally infected grapevine cultivars in Emilia-Romagna, Italy: polymerase chain reaction and restriction analysis. Extended abstracts 11th meeting ICVG, Montreux (CH) (pp 88–89)
Boudon-Padieu E, Larrue J and Caudwell A (1989) ELISA and Dotblot detection of Flavescence dorée-MLO in individual leafhopper vectors during latency and inoculative state. Current Microbiology 19: 357–364
Bertaccini A, Vibio M and Stephani E (1995) Detection and molecular characterisation of phytoplasmas infecting grapevine in Liguria (Italy). Phytopathologia mediterranea 34: 137–141
Bianco PA, Davis RE, Prince JP, Lee IM, Gundersen DE, Fortusini A and Belli G (1993) Double and single infections by aster yellows and elm yellows MLOs in grapevines with symptoms characteristic of flavescence dorée. Rivista di Patologia Vegetale 3: 69–82
Caudwell A (1957) Deux années d'études sur la flavescence dorée, nouvelle maladie grave de la vigne. Annales d0Amélioration des Plantes 4: 359–393
Caudwell A (1961) Etude sur la maladie du bois noir de la vigne: ses rapports avec la flavescence dorée. Annales des Epiphyties 12: 241–262
Caudwell A, Gianotti J, Kuszala C and Larrue J (1971) FrEtude du rôle de particules de type “mycoplasme” dans l'étiologie de la flavescence dorée de la vigne. Examen cytologique des plantes malades et des cicadelles infectieuses. Annales de Phytopathologie 3: 109–125
Daire X (1994) FrDetection et differenciation de mycoplasma-like organisms (MLO) associés aux maladies de la vigne de type jaunisse. Thèse des Sciences de la Vie de l'Université de Bourgogne, 24 juin 1994
Daire X, Boudon-Padieu E, Bervillé A, Schneider B and Caudwell A (1992a) Cloned DNA probes for detection of grapevine flavescence dorée mycoplasma-like organism (MLO). Annals of Applied Biology 121: 95–103
Daire X, Boudon-Padieu E, Grenier E, Bervillé A and Caudwell A (1992b) Characterisation of flavescence dorée MLO using cloned DNA probes. IOM Letters 2: 151
Daire X, Clair D, Larrue J, Boudon-Padieu E and Caudwell A (1993a) Diversity among mycoplasma-like organisms inducing grapevine yellows in France. Vitis 32: 159–163
Daire X, Clair D, Larrue J, Boudon-Padieu E, Alma A, Arzone A, Carraro L, Osler R, Refatti E, Granata G, Credi R, Tanne E, Pearson R and Caudwell A (1993b) Occurrence of diverse MLOs in tissues of grapevine affected by grapevine yellows in different countries. Vitis 32: 247–248
Daire X, Clair D, Larrue J and Boudon-Padieu E (1997) Survey for grapevine yellows phytoplasmas in diverse European countries and Israel. Vitis, 36: 53–54
Davis R E, Dally E L, Bertaccini A, Credi R, Lee IM, Osler R, Carraro L and Barba M (1992) Cloned DNA probes for specific detection of italian periwinkle virescence mycoplasmalike organism (MLO) and investigation of genetic relatedness with other MLOs. Phythopathologia mediterranea 31: 5–12
Fos A, Danet JL, Zreik L, Garnier M and Bové JM (1992) Use of monoclonal antibody to detect the stolbur mycoplasmalike organism in plants and insects and to identify a vector in France. Plant disease 76: 1092–1096
Gärtel W (1965) Untersuchungen über das Auftreten und das Verhalten des flavescence dorée in den Weinbaugebieten am Mosel und Rhein. Weinberg und Keller 12: 347–376
Laviña A, Batlle A, Larrue J, Daire X, Clair D and Boudon-Padieu E (1995) First report of grapevine bois noir phytoplasma in Spain. Plant disease 79: 1075
Lee IM, Hammond W, Davis RE and Gundersen DE (1993) Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms. Phytopathology 83: 834–842
Lorenz KH, Schneider B, Ahrens U and Seemüller E (1995) Detection of the apple proliferation and pear decline phytoplasmas by PCR amplification of ribosomal and nonribosomal DNA. Phytopathology 85: 771–776
Maixner M, Rüdel M, Daire X, Boudon-Padieu E (1995a) Diversity of grapevine yellows in Germany. Vitis 34: 235–236
Maixner M, Ahrens U and Seemüller E (1995b) Detection of the german grapevine yellows (Vergilbungskrankheit) MLO in grapevine, alternative hosts and a vector by a specific PCR procedure. European Journal of Plant Pathology 101: 241–250
Mäurer R, Seemüller E and Sinclair WA (1993) Genetic relatedness of mycoplasmalike organisms affecting elm, alder, and ash in Europe and North America. Phytopathology 83: 971–975
Osler R, Boudon-Padieu E, Carraro L, Caudwell A and Reffatti A (1992) First results of the trials in progress to identify the vector of the agent of a grapevine yellows in Italy. Phytopathologia mediterranea 31: 175–181
Poggi Pollini C, Bissani R, Giunchedi L and Vindimian E (1996) First report of phytoplasma in olive trees (Olea europa L.). Journal of Phytopathology 144: 109–111
Prince JP, Davis RE, Wolf TK, Lee IM, Mogen BD, Dally EL, Bertaccini A, Credi R and Barba M (1993) Molecular detection of diverse mycoplasmalike organisms (MLOs) associated with grapevine yellows and their classification with Aster yellows, X-disease and elm yellows MLOs. Phytopathology 83: 1130–1137
Schneider B, Ahrens U, Kirkpatrick BC and Seemüller E (1993) Classification of plant-pathogenic mycoplasma-like organisms using restriction-site analysis of PCR-amplified 16S rDNA. Journal of General Microbiology 139: 519–527
Seemüller E, Schneider B, Mäurer R, Ahrens U, Daire X, Kison H, Lorenz KH and Stackebrandt E (1994) Phylogenetic classi-fication of plant-pathogenic mycoplasmas by sequence analysis of 16S rDNA. International Journal of Systematic Bacteriology 44: 440–446
Sforza R, Clair D, Daire X, Larrue J, and Boudon-Padieu E (1996) Leafhoppers, planthoppers and phytoplasmas. Proceedings of the XX International Congress of Entomology. Firenze, Italy, August 25–31, 1996, p
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Daire, X., Clair, D., Reinert, W. et al. Detection and differentiation of grapevine yellows phytoplasmas belonging to the elm yellows group and to the stolbur subgroup by PCR amplification of non-ribosomal DNA. European Journal of Plant Pathology 103, 507–514 (1997). https://doi.org/10.1023/A:1008641411025
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DOI: https://doi.org/10.1023/A:1008641411025