Skip to main content
Log in

Specific characters of 16S rRNA gene and 16S–23S rRNA internal transcribed spacer sequences of Xylella fastidiosa pear leaf scorch strains

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Pear leaf scorch, the only Xylella fastidiosa-induced disease reported from Taiwan, was found in area where the variety Hengshan (Pyrus pyrifolia) was grown. Strains of pear leaf scorch Xyl. fastidiosa (XF-PLS) shared similarities to strains of other host origins in the requirement of complex medium and the exhibition of rippled cell walls, however, recent serological and molecular biology studies showed difference among them. Five strains of XF-PLS were compared with 20 other strains originally isolated from almond, oleander, pecan, plum, peach, mulberry, grapes, citrus, coffee, and sycamore by sequence analyses of the 16S rRNA gene and 16S–23S rRNA internal transcribed spacer region (ITS). When sequences of 16S rRNA gene based on fragment size of 1,537–1,540 bp were compared, the similarity index among 5 XF-PLS strains was 99.3–99.8%, whereas it was 97.8–98.6% between XF-PLS strains and strains from other hosts. When sequences of 16S–23S rRNA ITS based on fragment size of 510–540 bp were compared, the similarity index among 5 XF-PLS strains was 99.0–100%, whereas it was 80.7–82% between XF-PLS strains and strains from other hosts. Multiple sequence alignments led to the identification of 5 polymorphic nucleotides in the 16S rRNA gene among the 25 Xyl. fastidiosa strains, and there were considerable variations in the nucleotide sequences of 16S–23S rRNA ITS between XF-PLS and the other 20 Xyl. fastidiosa strains. The phylogenetic trees revealed that XF-PLS strains were separated from strains of other hosts. Strains of other hosts were divided into four subgroups: strains from (1) oleander, (2) grape, almond M23 and mulberry, (3) citrus and coffee, and (4) pecan, peach, plum, sycamore and almond M12. Results indicate that XF-PLS strains were not closely related to the above-mentioned strains from other hosts and could possibly belong to a new subspecies of Xyl. fastidiosa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215, 403–410.

    PubMed  CAS  Google Scholar 

  • Berisha, B., Chen, Y. D., Zhang, G. Y., Xu, B. Y., & Chen, T. A. (1998). Isolation of Pierce’s disease bacteria from grapevines in Europe. European Journal of Plant Pathology, 104, 427–433.

    Article  Google Scholar 

  • Chang, C. J., & Walker, J. T. (1988). Bacterial leaf scorch of northern red oak: isolation, cultivation, and pathogenicity of a xylem-limited bacterium. Plant Disease, 72, 730–733.

    Article  Google Scholar 

  • Chang, C. J., Ganier, M., Zreik, L., Rossetti, V., & Bove, J. M. (1993). Culture and serological detection of xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain of Xylella fastidiosa. Current Microbiology, 27, 137–142.

    Article  CAS  Google Scholar 

  • Chang, C. J., Donaldson, R., Brennen, P. M., Krewer, G., & Boland, B. (2009). Bacterial leaf scorch, a new blueberry disease caused by Xylella fastidiosa. Hortscience, 44, 413–417.

    Google Scholar 

  • Chen, J., Chang, C. J., Jarret, R. L., & Gawel, N. (1992). Genetic variation among Xylella fastidiosa strains. Phytopathology, 82, 973–977.

    Article  CAS  Google Scholar 

  • Chen, J., Bank, D., Jarret, R. L., Chang, C. J., & Smith, B. J. (2000a). Use of 16S rDNA sequences as signature characters to identify Xylella fastidiosa. Current Microbiology, 40, 29–33.

    Article  CAS  Google Scholar 

  • Chen, J., Banks, D., Jarret, R. L., & Jones, J. B. (2000b). Evidence for conserved tRNA genes in the 16S–23S rDNA spacer sequence and two rrn operons of Xylella fastidiosa. Canadian Journal of Microbiology, 46, 1171–1175.

    CAS  Google Scholar 

  • Chen, J., Jarret, R. L., Qin, X., Hartung, J. S., Banks, D., Chang, C. J., et al. (2000c). 16S rDNA sequence analysis of Xylella fastidiosa strains. Systematic and Applied Microbiology, 23, 349–354.

    Article  CAS  Google Scholar 

  • Chen, J., Hartung, J. S., Chang, C. J., & Vidaver, A. K. (2002). An evolutionary perspective of Pierce’s disease of grapevine, citrus variegated chlorosis, and mulberry leaf scorch diseases. Current Microbiology, 45, 423–428.

    Article  PubMed  CAS  Google Scholar 

  • Chen, J., Groves, R., Civerolo, E. L., Viveros, M., Freeman, M., & Zheng, Y. (2005). Two Xylella fastidiosa genotypes associated with almond leaf scorch disease on the same location in California. Phytopathology, 95, 708–714.

    Article  PubMed  CAS  Google Scholar 

  • Chen, J., Xie, G., Han, S., Chertkov, O., Sims, D., & Civerolo, E. L. (2010). Whole genome sequences of two Xylella fastidiosa strains (M12 and M23) causing almond leaf scorch disease in California. Journal of Bacteriology, 192, 4534.

    Article  PubMed  CAS  Google Scholar 

  • Davis, M. J., Purcell, A. H., & Thomson, S. V. (1980). Isolation medium for the Pierce’s disease bacterium. Phytopatholgy, 70, 425–429.

    Article  Google Scholar 

  • Davis, M. J., French, W. J., & Schaad, N. W. (1981). Axenic culture of the bacteria associated with phony disease of peach and plum leaf scald. Current Microbiology, 6, 309–314.

    Article  Google Scholar 

  • Felsenstein, J. (2004). PHYLIP: Phylogeny inference package. Seattle: Department of Genome Sciences and Department of Biology, University of Washington.

    Google Scholar 

  • Gendel, S. M. (1996). Computational analysis of the specificity of 16S rRNA-derived signature sequences for identifying food-related microbes. Food Microbiology, 13, 1–15.

    Article  CAS  Google Scholar 

  • Goncalves, E. R., & Rosato, Y. B. (2002). Phylogenetic analysis of Xanthomonas species based upon 16S–23S rDNA intergenic spacer sequences. International Journal of Systematic and Evolutionary Microbiology, 52, 355–361.

    PubMed  CAS  Google Scholar 

  • Hartung, J. S., Beretta, J., Brlansky, R. H., Spisso, J., & Lee, R. F. (1994). Citrus variegated chlorosis bacterium: axenic culture, pathogenicity, and serological relationships with other strains of Xylella fastidiosa. Phytopathology, 84, 591–597.

    Article  Google Scholar 

  • Hauben, L., Vauterin, L., Swings, J., & Moore, E. R. B. (1997). Comparison of 16S ribosomal DNA sequences of all Xanthomonas species. International Journal of Systematic Bacteriology, 47, 328–335.

    Article  PubMed  CAS  Google Scholar 

  • Hendson, M., Purcell, A. H., Chen, D., Smart, C., Guilhabert, M., & Kirkpatrick, B. (2001). Genetic diversity of Pierce’s disease strains and other pathotypes of Xylella fastidiosa. Applied and Environmental Microbiology, 67, 895–903.

    Article  PubMed  CAS  Google Scholar 

  • Hernandez-Martinez, R., de la Cerda, K., Costa, H. S., Cooksey, D. A., & Wong, F. P. (2007). Phylogenetic relationships of Xylella fastidiosa strains isolated from landscape ornamentals in southern California. Phytopathology, 97, 857–864.

    Article  PubMed  CAS  Google Scholar 

  • Hillis, M. D., & Dixon, M. T. (1991). Ribosomal DNA: molecular evolution and phylogenetic inference. Quarterly Review of Biology, 66, 411–453.

    Article  PubMed  CAS  Google Scholar 

  • Honeycutt, R. J., Sobral, B. W. S., & McClelland, M. (1995). tRNA intergenic spacers reveal polymorphisms diagnostic for Xanthomonas albilineans. Microbiology, 141, 3229–3239.

    Article  PubMed  CAS  Google Scholar 

  • Hopkins, D. L. (1989). Xylella fastidiosa: xylem-limited bacterial pathogen of plants. Annual Review of Phytopathology, 27, 271–290.

    Article  Google Scholar 

  • Hopkins, D. L., & Adlerz, W. C. (1988). Natural hosts of Xylella fastidiosa in Florida. Plant Disease, 72, 429–431.

    Article  Google Scholar 

  • Hopkins, D. L., & Purcell, A. H. (2002). Xylella fastidiosa: cause of Pierce’s disease of grapevine and other emergent diseases. Plant Disease, 86, 1056–1066.

    Article  Google Scholar 

  • Jeannmougin, F., Thompson, J. D., Gouy, M., Higgins, D. G., & Gibson, T. J. (1998). Multiple sequence alignment with Clustal X. Trends in Biochemical Sciences, 23, 403–405.

    Article  Google Scholar 

  • Leu, L. S., & Su, C. C. (1993). Isolation, cultivation, and pathogenicity of Xylella fastidiosa, the causal bacterium of pear leaf scorch disease in Taiwan. Plant Disease, 77, 642–646.

    Article  Google Scholar 

  • Mehta, A., & Rosato, Y. B. (2001). Phylogenetic relationships of Xylella fastidiosa strains from different hosts, based on 16S rDNA and 16–23S intergenic spacer sequences. International Journal of Systematic and Evolutionary Microbiology, 51, 311–318.

    PubMed  CAS  Google Scholar 

  • Nathalie, L. B., Philippe, H., Vangin, I., & Decaris, B. (1996). 16S rRNA and 16S to 23S internal transcribed spacer sequence analyses reveal inter- and intra-specific Bifidobacterium phylogeny. International Journal of Systematic Bacteriology, 46, 102–111.

    Article  Google Scholar 

  • Neefs, J. M., Van de Peer, Y., De Rijk, P., Goris, A., & De Wachter, R. (1991). Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Research, 19(Suppl), 1987–2015.

    PubMed  CAS  Google Scholar 

  • Page, R. D. (1996). TreeView: an application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences, 12, 357–358.

    PubMed  CAS  Google Scholar 

  • Pooler, M. R., & Hartung, J. S. (1995). Genetic relationships among strains of Xylella fastidiosa from RAPD-PCR data. Current Microbiology, 31, 134–137.

    Article  PubMed  CAS  Google Scholar 

  • Purcell, A. H., & Hopkins, D. L. (1996). Fastidious xylem-limited bacterial plant pathogens. Annual Review of Phytopathology, 34, 131–151.

    Article  PubMed  CAS  Google Scholar 

  • Qin, X., Miranda, V. S., Machado, M. A., Lemos, E. G. M., & Hartung, J. S. (2001). An evaluation of the genetic diversity of Xylella fastidiosa isolated from diseased citrus and coffee in São Paulo, Brazil. Phytopathology, 91, 599–605.

    Article  PubMed  CAS  Google Scholar 

  • Randall, J. J., Goldberg, N. P., Kemp, J. D., Radionenko, M., French, J. M., Olsen, M. W., et al. (2009). Genetic analysis of a novel Xylella fastidiosa subspecies found in the southwestern United States. Applied and Environmental Microbiology, 75, 5631–5638.

    Article  PubMed  CAS  Google Scholar 

  • Rosato, Y. B., Neto, J. B., Miranda, V. S., Carlos, E. F., & Manfio, C. P. (1998). Diversity of a Xylella fastidiosa population isolated from Citrus sinensis affected by citrus variegated chlorosis in Brazil. Systematic and Applied Microbiology, 21, 593–598.

    Article  Google Scholar 

  • Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual (2nd ed.). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Schaad, N. W., Postnikova, E., Lacy, G., Fatmi, M., & Chang, C. J. (2004). Xylella fastidiosa subspecies: X. fastidiosa subsp. piercei, subsp. nov., X. fastidiosa subsp. multiplex subsp. nov., X. fastidiosa subsp. multiplex subsp. nov., and X. fastidiosa subsp. pauca subsp. nov. Systematic and Applied Microbiology, 27, 290–300.

    Article  PubMed  CAS  Google Scholar 

  • Schreiber, H. L., IV, Koirala, M., Lara, A., Ojeda, M., Dowd, S. E., Bextine, B., et al. (2010). Unraveling the first Xylella fastidiosa subsp. fastidiosa genome from Texas. Southwestern Entomologist, 35, 479–483.

    Article  Google Scholar 

  • Schuenzel, E. L., Scally, M., Stouthamer, R., & Nunney, L. (2005). A multigene phylogenetic study of clonal diversity and divergence in north American strains of the plant pathogen Xylella fastidiosa. Applied and Environmental Microbiology, 71, 3832–3839.

    Article  PubMed  CAS  Google Scholar 

  • Simpson, A. J., Reinach, F. C., Arruda, P., Abreu, F. A., Acencio, M., Alvarenga, R., et al. (2000). The genome sequence of the plant pathogen Xylella fastidiosa. Nature, 406, 151–157.

    Article  PubMed  CAS  Google Scholar 

  • Smart, C. D., Schneider, B., Biomquist, C. L., Guerra, L. J., Harrison, N. A., Ahrens, U., et al. (1996). Phytoplasma-specific PCR primers based on sequences of the 16S–23S rRNA spacer region. Applied and Environmental Microbiology, 63, 2988–2993.

    Google Scholar 

  • Stackebrandt, E., & Goebel, B. M. (1994). Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA Sequence analysis in the present species definition in bacteriology. International Journal of Systematic Bacteriology, 44, 846–849.

    Article  CAS  Google Scholar 

  • Su, C. C., Yang, W. J., Feng, C. Y., Hsu, S. T., & Tzeng, K. C. (2008). The application of DNA fingerprintings amplified by arbitrary primers in differentiating pear leaf scorch bacterium from other Xylella fastidiosa strains. Plant Pathology Bulletin, 17, 261–269 (Chinese with English abstract).

    CAS  Google Scholar 

  • Toth, I. K., Avrova, A. O., & Hyman, L. J. (2001). Rapid identification and differentiation of soft rot Erwinias by 16S–23S intergenic transcribed spacer-PCR and restriction fragment length polymorphism analyses. Applied and Environmental Microbiology, 67, 4070–4076.

    Article  PubMed  CAS  Google Scholar 

  • Van Sluys, M. A., de Oliveira, M. C., Monteiro-Vitorello, C. B., Miyaki, C. Y., Furlan, L. R., Camargo, L. E., et al. (2003). Comparative analyses of the complete genome sequences of Pierce’s disease and citrus variegated chlorosis strains of Xylella fastidiosa. Journal of Bacteriology, 185, 1018–1026.

    Article  PubMed  Google Scholar 

  • Vandamme, P., Pot, B., Gillis, M., de Vos, P., Kersters, K., & Swings, J. (1996). Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiological Reviews, 60, 407–438.

    PubMed  CAS  Google Scholar 

  • Weisburg, W. G., Barns, S. M., Pelletier, D. A., & Lane, D. J. (1991). 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 173, 697–703.

    PubMed  CAS  Google Scholar 

  • Wells, J. M., Raju, B. C., Hung, H.-Y., Weisburg, W. G., Mandelco-Paul, L., & Brenner, D. J. (1987). Xylella fastidiosa gen. nov., sp. nov: gram-negative, xylem-limited, fastidious plant bacteria related to Xanthomonas spp. International Journal of Systematic Bacteriology, 37, 136–143.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Mr. Che-Ming Chang for his assistance in phylogenetic analysis presented in Figs. 3 and 4. The research was funded by the Council of Agriculture grant 99AS-9.3.1-BQ-B2 to C.C.S. and W.L.D. and the National Science Council grants NSC 98-2811-B-005-044 and NSC100-2811-B-005-001to F.J.J. and C.J.C.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Fuh-Jyh Jan or Wen-Ling Deng.

Additional information

Chiou-Chu Su and Chung-Jan Chang contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Su, CC., Chang, CJ., Yang, WJ. et al. Specific characters of 16S rRNA gene and 16S–23S rRNA internal transcribed spacer sequences of Xylella fastidiosa pear leaf scorch strains. Eur J Plant Pathol 132, 203–216 (2012). https://doi.org/10.1007/s10658-011-9863-6

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-011-9863-6

Keywords

Navigation