Phytoplasmas were detected in cactus ( species) plants exhibiting witches'-broom disease symptoms in Yunnan Province, south-western China. Comparative and phylogenetic analyses of 16S rRNA gene sequences indicated that an overwhelming majority of the cactus-infecting phytoplasmas under study belonged to the peanut witches'-broom phytoplasma group (16SrII). Genotyping through use of computer-simulated restriction fragment length polymorphism (RFLP) analysis of 16S rRNA genes revealed a remarkable genetic diversity among these cactus-infecting phytoplasma strains. Based on calculated coefficients of RFLP pattern similarities, seven new 16SrII subgroups were recognized, bringing the total of described group 16SrII subgroups to 12 worldwide. Geographical areas differed from one another in the extent of genetic diversity among cactus-infecting phytoplasma strains. The findings have implications for relationships between ecosystem distribution and the emergence of group 16SrII subgroup diversity.


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  1. Bai, X. D., Zhang, J. H., Ewing, A., Miller, S. A., Radek, A. J., Shevchenko, D. V., Tsukerman, K., Walunas, T., Lapidus, A. & other authors(2006). Living with genome instability: the adaptation of phytoplasmas to diverse environments of their insect and plant hosts. J Bacteriol 188, 3682–3696.[CrossRef] [Google Scholar]
  2. Cai, H., Chen, H. R., Li, F. & Kong, B. H.(2002). First report of a phytoplasma associated with cactus witches'-broom in Yunnan (China). Plant Pathol 51, 394[CrossRef] [Google Scholar]
  3. Choueiri, E., Massad, R., Jreijiri, F., Danet, J. L., Salar, P., Bové, J. M. & Foissac, X.(2005). First report of a 16SrII group phytoplasma associated with shoot proliferation of a cactus (Opuntia monacantha) in Lebanon. Plant Dis 89, 1129 [Google Scholar]
  4. Davis, R. E. & Dally, E. L.(2001). Nonfunctional tRNA gene in an unusual example of rRNA interoperon sequence heterogeneity in phytoplasma. Phytopathology 91, S21 [Google Scholar]
  5. Davis, R. E. & Sinclair, W. A.(1998). Phytoplasma identity and disease etiology. Phytopathology 88, 1372–1376.[CrossRef] [Google Scholar]
  6. Davis, R. E., Jomantiene, R., Kalvelyte, A. & Dally, E. L.(2003). Differential amplification of sequence heterogeneous ribosomal RNA genes and classification of the ‘Fragaria multicipita’ phytoplasma. Microbiol Res 158, 229–236.[CrossRef] [Google Scholar]
  7. Deng, S. & Hiruki, C.(1991). Amplification of 16S rRNA genes from culturable and nonculturable Mollicutes. J Microbiol Methods 14, 53–61.[CrossRef] [Google Scholar]
  8. Firrao, G., Carraro, L., Gobbi, E. & Locci, R.(1996). Molecular characterization of a phytoplasma causing phyllody in clover and other herbaceous hosts in northern Italy. Eur J Plant Pathol 102, 817–822.[CrossRef] [Google Scholar]
  9. Firrao, G., Gibb, K. & Streten, C.(2005). Short taxonomic guide to the genus ‘Candidatus phytoplasma’. J Plant Pathol 87, 249–263. [Google Scholar]
  10. Gundersen, D. E. & Lee, I.-M.(1996). Ultrasensitive detection of phytoplasmas by nested-PCR assays using two universal primer pairs. Phytopathol Mediterr 35, 144–151. [Google Scholar]
  11. Jeanmougin, F., Thompson, J. D., Gouy, M., Higgins, D. G. & Gibson, T. J.(1998). Multiple sequence alignment with clustal_x. Trends Biochem Sci 23, 403–405.[CrossRef] [Google Scholar]
  12. Jomantiene, R., Davis, R. E., Valiunas, D. & Alminaite, A.(2002). New group 16SrIII phytoplasma lineages in Lithuania exhibit interoperon sequence heterogeneity. Eur J Plant Pathol 108, 507–517.[CrossRef] [Google Scholar]
  13. Kumar, S., Tamura, K. & Nei, M.(2004).mega3: integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef] [Google Scholar]
  14. Lauer, U. & Seemüller, E.(2000). Physical map of the chromosome of the apple proliferation phytoplasma. J Bacteriol 182, 1415–1418.[CrossRef] [Google Scholar]
  15. Lee, I.-M., Hammond, R. W., Davis, R. E. & Gundersen, D. E.(1993). Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms. Phytopathology 83, 834–842.[CrossRef] [Google Scholar]
  16. Lee, I.-M., Gundersen-Rindal, D. E., Davis, R. E. & Bartoszyk, I.-M.(1998). Revised classification scheme of phytoplasmas based on RFLP analysis of 16S rRNA and ribosomal protein gene sequences. Int J Syst Bacteriol 48, 1153–1169.[CrossRef] [Google Scholar]
  17. Lee, I.-M., Davis, R. E. & Gundersen-Rindal, D. E.(2000). Phytoplasma: phytopathogenic mollicutes. Annu Rev Microbiol 54, 221–255.[CrossRef] [Google Scholar]
  18. Lee, I.-M., Martini, M., Marcone, C. & Zhu, S. F.(2004). Classification of phytoplasma strains in the elm yellows group (16SrV) and proposal of ‘Candidatus Phytoplasma ulmi’ for the phytoplasma associated with elm yellows. Int J Syst Evol Microbiol 54, 337–347.[CrossRef] [Google Scholar]
  19. Lee, I.-M., Zhao, Y., Davis, R. E., Wei, W. & Martini, M.(2007). Prospects of DNA-based systems for differentiation and classification of phytoplasmas. Bull Insectol 60, 239–244. [Google Scholar]
  20. Leyva-Lopez, N. E., Aguilar-Rojas, O. I., Leal-Klevezas, D. S. & Martínez-Soriano, J. P.(1999). Presence of phytoplasmas in Mexican cacti. Phytopathology 89, S45 [Google Scholar]
  21. Liefting, L. W., Andersen, M. T., Beever, R. E., Gardner, R. C. & Foster, L. S.(1996). Sequence heterogeneity in the two 16S rRNA genes of Phormium yellow leaf phytoplasma. Appl Environ Microbiol 62, 3133–3139. [Google Scholar]
  22. Martini, M.(2004).Ribosomal protein gene-based phylogeny: a basis for phytoplasma classification, p. 103. PhD dissertation, University of Udine, Udine, Italy.
  23. Martini, M., Lee, I.-M., Bottner, K. D., Zhao, Y., Botti, S., Bertaccini, A., Harrison, N. A., Carraro, L., Marcone, C. & other authors(2007). Ribosomal protein gene-based phylogeny for finer differentiation and classification of phytoplasmas. Int J Syst Evol Microbiol 57, 2037–2051.[CrossRef] [Google Scholar]
  24. McCoy, R. E., Caudwell, A., Chang, C. J., Chen, T.-A., Chiykowski, L. N., Cousin, M. T., Dale, J. L., DeLeeuw, G. T. N., Golino, D. A. & other authors(1989). Plant diseases associated with mycoplasma-like organisms. In The Mycoplasmas, vol. v, pp. 545–640. Edited by R. F. Whitcomb & J. G. Tully. New York: Academic Press.
  25. Nei, M. & Li, W.-H.(1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci U S A 76, 5269–5273.[CrossRef] [Google Scholar]
  26. Nyffeler, R.(2002). Phylogenetic relationships in the cactus family (Cactaceae) based on evidence from trnK/matK and trnL-trnF sequences. Am J Bot 89, 312–326.[CrossRef] [Google Scholar]
  27. Oshima, K., Kakizawa, S., Nishigawa, H., Jung, H.-Y., Wei, W., Suzuki, S., Arashida, R., Nakata, D., Miyata, S. & other authors(2004). Reductive evolution suggested from the complete genome sequence of a plant-pathogenic phytoplasma. Nat Genet 36, 27–29.[CrossRef] [Google Scholar]
  28. Padovan, A., Firrao, G., Schneider, B. & Gibb, K.(2000). Chromosome mapping of the sweet potato little leaf phytoplasma reveals genome heterogeneity within the phytoplasmas. Microbiology 146, 893–902. [Google Scholar]
  29. Schneider, B. & Seemüller, E.(1994). Presence of two sets of ribosomal genes in phytopathogenic mollicutes. Appl Environ Microbiol 60, 3409–3412. [Google Scholar]
  30. Shuman, S.(1994). Novel approach to molecular cloning and polynucleotide synthesis using vaccinia DNA Topoisomerase. J Biol Chem 269, 32678–32684. [Google Scholar]
  31. Tamura, K., Dudley, J., Nei, M. & Kumar, S.(2007).mega4: Molecular Evolutionary Genetics Analysis (mega) software version 4.0. Mol Biol Evol 24, 1596–1599.[CrossRef] [Google Scholar]
  32. Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. G.(1997). The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.[CrossRef] [Google Scholar]
  33. Tsai, J. H.(1979). Vector transmission of mycoplasmal agents of plant diseases. In The Mycoplasmas III, Plant and Insect Mycoplasmas, pp. 265–307. Edited by R. F. Whitcomb & J. G. Tully. New York: Academic Press.
  34. Wei, W., Cai, H., Chen, H. R., Davis, R. E. & Zhao, Y.(2007a). First report of a natural infection of Opuntia sp. by a ‘Candidatus phytoplasma asteris’-related phytoplasma in China. Plant Dis 91, 461 [Google Scholar]
  35. Wei, W., Davis, R. E., Lee, I.-M. & Zhao, Y.(2007b). Computer-simulated RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. Int J Syst Evol Microbiol 57, 1855–1867.[CrossRef] [Google Scholar]
  36. Wei, W., Lee, I.-M., Davis, R. E., Suo, X. & Zhao, Y.(2007c). Virtual RFLP analysis of 16S rDNA sequences identifies new subgroups in the clover proliferation phytoplasma group. Bull Insectol 60, 349–350. [Google Scholar]
  37. Weisburg, W. G., Tully, J. G., Rose, D. L., Petzel, J. P., Oyaizu, H., Yang, D., Mandelco, L., Sechrest, J., Lawrence, T. G. & other authors(1989). A phylogenetic analysis of the mycoplasmas: basis for their classification. J Bacteriol 171, 6455–6467. [Google Scholar]
  38. White, D. T., Blackall, L. L., Scott, P. T. & Walsh, K. B.(1998). Phylogenetic positions of phytoplasmas associated with dieback, yellow crinkle and mosaic diseases of papaya, and their proposed inclusion in ‘Candidatus Phytoplasma australiense’ and a new taxon, ‘Candidatus Phytoplasma australasia’. Int J Syst Bacteriol 48, 941–951.[CrossRef] [Google Scholar]
  39. Woese, C. R.(1987). Bacterial evolution. Microbiol Rev 51, 221–271. [Google Scholar]
  40. Zreik, L., Carle, P., Bové, J. M. & Garnier, M.(1995). Characterization of the mycoplasma-like organism associated with witches'-broom disease of lime and proposition of a Candidatus taxon for the organism, ‘Candidatus Phytoplasma aurantifolia’. Int J Syst Bacteriol 45, 449–453.[CrossRef] [Google Scholar]

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