The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains

  • Kim, Ki-Woo (National Instrumentation Center for Environmental Management, Seoul National University) ;
  • Lee, In-Jung (Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Hyun, Jae-Wook (Citrus Experiment Station, National Institute of Subtropical Agriculture, Rural Development Administration) ;
  • Lee, Yong-Hoon (Department of Biotechnology, Chonbuk National University) ;
  • Park, Eun-Woo (Department of Agricultural Biotechnology, Seoul National University)
  • Received : 2009.09.01
  • Accepted : 2010.02.04
  • Published : 2010.03.01
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Abstract

Staining profiles and bacterial morphology were compared in Xanthomonas citri pv. citri by a transmission electron microscopy. Four types of negative staining regimes were employed depending on culture media and heavy metal stains. The bacterial cells grown on LB agar media often appeared clustered on the supporting film. Meanwhile, individual bacterial cells could be readily found on the preparations from LB broth media. Typical rod-shaped cells (ca. $1\;{\mu}m$ in length) and their flagella were observed in either 2% uranyl acetate (UA) or 2% neutralized potassium phosphotungstate (PTA) staining. The UA-stained bacteria often showed relatively intact cell morphology and rather positively stained cells with a thin electron-dense stain depth around bacteria. The PTA-stained bacteria were characterized by the wrinkled cell surface where the stain was entrapped in grooves. In addition, distinct electron-dense stain depth was evident around the PTA-stained preparations. Numerous fimbriae could be mostly observed from the PTA-stained preparations of the two culture media, but not from the UA-stained preparations.

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References

  1. Bozzola, J. J. and Russell, L. D. 1999. Electron Microscopy: Principles and Techniques for Biologists, 2nd Ed. Jones and Bartlett Publishers, Boston, NY. 670 pp.
  2. Glasauer, S., Langley, S. and Beveridge, T. J. 2001. Sorption of Fe (hydr)oxides to the surface of Shewanella putrefaciens: Cellbound fine-grained minerals are not always formed de novo. Appl. Environ. Microbiol. 67:5544-5550. https://doi.org/10.1128/AEM.67.12.5544-5550.2001
  3. Hayat, M. A. 2000. Principles and Techniques of Electron Microscopy: Biological Applications, 4th Ed. Cambridge University Press, Cambridge, UK. 543 pp.
  4. Hoppert, M. and Holzenburg, A. 1998. Electron Microscopy in Microbiology. BIOS Scientific Publishers, Oxford, UK. 112pp.
  5. Kim, K. W., Jung, W. K. and Park, Y. H. 2008. Comparison of pre-stain suspension liquids in the contrasting ability of neutralized potassium phosphotungstate for negative staining of bacteria. J. Microbiol. Biotech. 18:1762-1767.
  6. Lee, I. J., Kim, K. W., Hyun, J. W., Lee, Y. H. and Park, E. W. 2009. Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri. Microsc. Res. Tech. 72:507-516.
  7. Moreira, L. M., De Souza, R. F., Almeida, N. F. Jr., Setubal, J. C., Oliveira, J. C. F., Furlan, L. R., Ferro, J. A. and Da Silva, A. C. R. 2004. Comparative genomics analyses of citrus-associated bacteria. Annu. Rev. Phytopathol. 42:163-184. https://doi.org/10.1146/annurev.phyto.42.040803.140310
  8. Ohi, M., Li, Y., Cheng, Y. and Walz, T. 2004. Negative staining and image classification-powerful tools in modern electron microscopy. Biol. Proced. Online 6:23-34. https://doi.org/10.1251/bpo70
  9. Schaad, N. W., Postnikova, E., Lacy, G. H., Sechler, A., Agarkova, I., Stromberg, P. E., Stromberg, V. K. and Vidaver, A. K. 2005. Reclassification of Xanthomonas species pathogenic on citrus. Sys. Appl. Microbiol. 28:494-518. https://doi.org/10.1016/j.syapm.2005.03.017
  10. Stone, K. D., Zhang, H.-Z., Carlson, L. K. and Donnenberg, M. S. 1996. A cluster of fourteen genes from enteropathogenic Escherichia coli is sufficient for the biogenesis of a type IV pilus. Mol. Microbiol. 20:325-337. https://doi.org/10.1111/j.1365-2958.1996.tb02620.x
  11. Van Doorn, J., Hollinger, T. C. and Oudega, B. 2001. Analysis of the type IV fimbrial-subunit gene fimA of Xanthomonas hyacinthi: Application in PCR-mediated detection of yellow disease in hyacinths. Appl. Environ. Microbiol. 67:598-607. https://doi.org/10.1128/AEM.67.2.598-607.2001
  12. Walz, T., Häner, M., Wu, X.-R., Henn, C., Engel, A., Sun, T.-T. and Aebi, U. 1995. Towards the molecular architecture of the asymmetric unit membrane of the mammalian urinary bladder epithelium: a closed “twisted ribbon” structure. J. Mol. Biol. 248:887-900.

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