1887

Abstract

A method has been developed to amplify the 16S rRNA gene of plant-pathogenic mycoplasma-like organisms (MLOs) from infected plant material using the polymerase chain reaction (PCR). The procedure is dependent on the presence of a I restriction site in the 16S rDNA of chloroplasts but not in that of the MLOs. This difference permits the specific amplification of the 16S rDNA of the MLOs from I-digested total DNA from infected plants using primers from conserved regions of this gene. In this study 16S rDNA was obtained from 52 MLO isolates from herbaceous dicots and monocots as well as woody plants. Digestion of the 16S rRNA genes using I endonuclease revealed seven restriction patterns, which were used to group the isolates examined. Group I, which is also characterized by the presence of two I sites, consisted of 31 isolates, most of which are from herbaceous dicots. Isolates assigned to groups II to VI were mostly from woody plants, while the isolates of group VII were from monocots or obtained from a leafhopper. The restriction patterns varied little within groups; however, four group I isolates and one group IV isolate differed slightly from the typical patterns of these groups as a result of a deletion or a slight shift of one restriction site. The groupings uncovered by I restriction were also obtained by digesting the 16S rDNA with I endonuclease. However, some atypical patterns were observed within group V isolates. The groups described on the basis of restriction digest data were supported by sequence analysis. With one exception, the 16S rDNA of isolates within the same group exhibited 97·8 to 99·5% homology while those of different groups showed 89·6 to 92·0% homology.

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1993-03-01
2021-10-27
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References

  1. Ahrens U., Seemüller E. 1992; Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828–832
    [Google Scholar]
  2. Ahrens U., Seemüller E., Lorenz K. H. 1992; Genetic differentiation of the MLOs affecting temperate fruit trees. International Organisation for Mycoplasmology Letters 2:150
    [Google Scholar]
  3. Bertaccini A., Davis R. E., Lee I.-M. 1990; Distinction among mycoplasma-like organisms (MLOs) in Gladiolus, Ranunculus, Brassica, and Hydrangea through detection with nonradioactive cloned DNA probes. Phytopathologia mediterranea 29:107–113
    [Google Scholar]
  4. Bonnet F., Saillard C., Kollar A., Seemüller E., Bovś J. M. 1990; Detection and differentiation of the mycoplasma-like organism associated with apple proliferation disease using cloned DNA probes. Molecular Plant-Microbe Interactions 3:438–443
    [Google Scholar]
  5. Bové J. M., Saillard C. 1979; Cell biology of spiroplasmas. In The Mycoplasmas III pp. 83–153 Whitcomb R. F., Tully J. G. Edited by New York: Academic Press;
    [Google Scholar]
  6. Bouvet A., Grimont F., Grimont P. A. D. 1991; Intraspecific variations in nutritionally variant streptococci: rRNA gene restriction patterns in Streptococcus defectivus and Streptococcus adjacens. International Journal of Systematic Bacteriology 41:483–486
    [Google Scholar]
  7. Carraro L., Osler R., Refatti E., Poggi Pollini C. 1988; Transmission of the possible agent of apple proliferation to Vinca rosea by dodder. Rivista di Patologia Vegetale S.IV:43–52
    [Google Scholar]
  8. Clark M. F., Davies D. L., Buss S. L., Morton A. 1988; Serological discrimination among mycoplasma-like organisms using polyclonal and monoclonal antibodies. Acta Horticulturae 235:107–113
    [Google Scholar]
  9. Davis R. E., Sinclair W. A., Lee I.-M., Dally E. L. 1992; Cloned DNA probes specific for detection of a mycoplasma-like organism associated with ash yellows. Molecular Plant-Microbe Interactions 5:163–169
    [Google Scholar]
  10. Dijkstra J., Lee P. E. 1972; Transmission by dodder of sandal spike disease and the accompanying mycoplasma-like organisms via Vinca rosea. Netherlands Journal of Plant Pathology 78:218–228
    [Google Scholar]
  11. Fox G. E., Wisotzkey J. D., Jurtshuk P. 1992; How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. International Journal of Systematic Bacteriology 42:166–170
    [Google Scholar]
  12. Göbel U. B, Geiser A., Stanbridge E. J. 1987; Oligonucleotide probes complementary to variable regions of ribosomal RNA discriminates between Mycoplasma species. Journal of General Microbiology 133:1969–1974
    [Google Scholar]
  13. Grimont F., Grimont P. A. D. 1986; Ribosomal nucleic acid gene restriction patterns as potential taxonomic tools. Annales de l’Institut Pasteur/Microbiologie 137B:165–175
    [Google Scholar]
  14. Gurtler V., Wilson V. A., Mayall B. C. 1991; Classification of medically important Clostridia using restriction endonuclease site differences of PCR-amplified 16S rDNA. Journal of General Microbiology 137:2673–2679
    [Google Scholar]
  15. Heintz W. 1989; Transmission of a new mycoplasma-like organism (MLO) from Cuscuta odorata (Ruiz et Pav.) to herbaceous plants and attempts to its elimination in the vector. Journal of Phytopathology 125:171–186
    [Google Scholar]
  16. Higgins D. G., Sharp P. M. 1988; Clustal: a package for performing multiple alignments on a microcomputer. Gene 73:237–244
    [Google Scholar]
  17. Jensen D. D. 1956; Insect transmission of virus between tree and herbaceous plants. Virology 2:249–260
    [Google Scholar]
  18. Klein M. 1970; Safflower phyllody - a mycoplasma disease of Carthamus tinctorius in Israel. Plant Disease Reporter 54:735–738
    [Google Scholar]
  19. Kuske C. R., Kirkpatrick B. C. 1992; Phylogenetic relationship between the western aster yellows mycoplasma-like organisms and other prokaryotes established by 16S rRNA gene sequence. International Journal of Systematic Bacteriology 42:226–233
    [Google Scholar]
  20. Kuske C. R., Kirkpatrick B. C., Seemüller E. 1991; Differentiation of virescence MLOs using western aster yellows mycoplasmalike organism chromosomal DNA probes and restriction fragment length polymorphism analysis. Journal of General Microbiology 137:153–159
    [Google Scholar]
  21. Laigret F., Grau O., Bové J. M. 1990; Comparison of 16S rDNA sequences of various mollicutes. Zentralblatt für Bakteriologie Suppl 20:435–440
    [Google Scholar]
  22. Lane D. J., Pace B., Olsen G. J., Stahl D. A., Sogin M. L., Pace N. R. 1985; Rapid determination of 16S ribosomal RNA sequences for phylogenetic analysis. Proceedings of the National Academy of Sciences of the United States of America 826955–6959
    [Google Scholar]
  23. Lee L.-M., Davis R. E. 1988; Detection and investigation of genetic relatedness among aster yellows and other mycoplasma-like organisms by using cloned DNA and RNA probes. Molecular Plant-Microbe Interactions 1:303–310
    [Google Scholar]
  24. Lee I.-M., Davis R. E., Dewitt N. 1990; Nonradioactive screening method for isolation of disease-specific probes to diagnose plant diseases caused by mycoplasma-like organisms. Applied Environmental Microbiology 56:1471–1475
    [Google Scholar]
  25. Lim P.-O., Sears B. B. 1989; 16S rRNA sequence indicates that plant-pathogenic mycoplasma-like organisms are evolutionarily distinct from animal mycoplasmas. Journal of Bacteriology 171:5901–5906
    [Google Scholar]
  26. Lin C.-P., Chen T. A. 1985; Monoclonal antibodies against the aster yellows agent. Science 227:1233–1235
    [Google Scholar]
  27. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual Cold Spring Harbor NY; Cold Spring Harbor Laboratory:
    [Google Scholar]
  28. Marchoux C., Giannotti J. 1971; Interférence entre deux mycoplasmas végétale. Physiologie Végétale 9:595–610
    [Google Scholar]
  29. Marchuk D., Drumm M., Saulino A., Collins F. S. 1991; Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Research 19:54
    [Google Scholar]
  30. Mäurer R., Seemüller E. 1992; Genetic relatedness of European and North American mycoplasma-like organisms from trees. International Organisation for Mycoplasmology Letters 2:149
    [Google Scholar]
  31. Marwitz R. 1990; Diversity of yellows disease agents in plant infections. Zentralblatt für Bakteriologie, Suppl 20:431–434
    [Google Scholar]
  32. Marwitz R., Petzold H. 1976; Elektronenmikroskopischer Nachweis mycoplasmaähnlicher Organismen in Delphinium-Hybriden mit Blütenvergrünung und -verlaubung. Phytopathologische Zeitschrift 87:1–11
    [Google Scholar]
  33. Marwitz R., Petzold H. 1983; Mykoplasmaähnliche Organismen als Krankheitserreger an Primeln. Gesunde Pflanze 35:336–341
    [Google Scholar]
  34. Marwitz R., Petzold H., Özel M. 1974; Untersuchungen zur Übertragbarkeit des möglichen Erregers der Triebsucht des Apfels auf einen krautigen Wirt. Phytopathologische Zeitschrift 81:85–91
    [Google Scholar]
  35. Marwitz R., Petzold H., Roth L. 1979; Eine bisher unbekannte stolburverwandte Krankheit bei feldmäßig angebauten Pflanzen von Solanum marginatum in Ecuador. Phytopathologische Zeitschrift 95:305–317
    [Google Scholar]
  36. Marwitz R., Kuhbandner B., Petzold H. 1987; Übertragung mykoplasmaähnlicher Organismen (MLO) von hexenbesenkranken Heidelbeeren (Vaccinium myrtillus) auf Catharanthus roseus mit Hilfe von Cuscuta. Nachrichtenblatt der Deutschen Pflanzenschutzdienstes 39:129–132
    [Google Scholar]
  37. Mccoy R. E., Caudwell A., Chang C. J., Chen T. A., Chiykowski I. N., Cousin M. T., Dale J. L., De Leeuw G. T. N., Golino D. A., Hackett K. J., Kirkpatrick B. C., Marwitz R., Petzold H., Sinha R. C., Sugiura M., Whitcomb R. F., Young I. L., Zhu B. M., Seemüller E. 1989; Plant diseases associated with mycoplasma-like organisms. In The Mycoplasmas V pp. 545–640 Whitcomb R. F., Tully J. G. Edited by San Diego: Academic Press;
    [Google Scholar]
  38. Moreno P., Llacer G., Medina V. 1985; Descripcion y comparacion de varias micoplasmosis en Vinca rosea L. Anales del Instituto Nacional de Investigaciones Agrarias, Serie Agricola, N. Extr 28:287–309
    [Google Scholar]
  39. Razin S. 1989; Molecular approach to mycoplasma phylogeny. In The Mycoplasmas V pp. 33–69 Whitcomb R. F., Tully J. G. Edited by San Diego: Academic Press;
    [Google Scholar]
  40. Sears B. B., Klomparens K. L. 1989; Leaf tip cultures of the evening primrose allow stable, aseptic culture of mycoplasma-like organisms. Canadian Journal Plant Pathology 11:343–348
    [Google Scholar]
  41. Sinclair W. A., Braun E. J., Larsen A. O. 1976; Update on phloem necrosis of elms. Journal of Arboriculture 2:106–113
    [Google Scholar]
  42. Stackebrandt E. 1991; Unifying phylogeny and phenotypic diversity. In The Prokaryotes I pp. 19–47 Balows A., Trüper H. G., Dworkin M., Harder W., Schleifer K. -H. Edited by New York: Springer Verlag;
    [Google Scholar]
  43. Taschke C., Ruland K., Herrmann R. 1987; Nucleotide sequence of the 16S rRNA of >Mycoplasma hyopneumoniae. Nucleic Acids Research 15:3918
    [Google Scholar]
  44. Taschke C., Wolters J., Herrmann R. 1990; Ribosomal RNA genes of four Mycoplasma species. Zentralblatt für Bakteriologie Suppl 20:329–336
    [Google Scholar]
  45. Weisburg W. G., Tully J. G., Rose D. L., Petzel J. P., Oyaizu H., Yang D., Mandelco L., Sechrest J., Lawrence T. G., Van Etten J., Maniloff J., Woese C. R. 1989; A phylogenetic analysis of the mycoplasmas: basis for their classification. Journal of Bacteriology 171:6455–6467
    [Google Scholar]
  46. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 1991; 16S ribosomal DNA amplification for phylogenetic study. Journal Bacteriology 173:697–703
    [Google Scholar]
  47. Welvaert W., Sakyn G., Lagasse A. 1975; Recherches sur les symptomes de la virescence chez l’Hydrangea macrophylla Thunb. Phytopathologische Zeitschrift 83:152–158
    [Google Scholar]
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