DOI QR코드

DOI QR Code

Genetic Differentiation of Phytoplasma Isolates by DNA Heteroduplex Mobility Assay and Single-Strand Conformation Polymorphism Analysis

  • Cha, Byeongjin (Department of Agricultural Biology, Chungbuk National University) ;
  • Han, Sangsub (Department of Agricultural Biology, Chungbuk National University)
  • 발행 : 2002.12.01

초록

Heteroduplex mobility assay (HMA) and single-strand conformation polymorphism (SSCP) analyses combined with PCR were developed for genetic differentiation of various phytoplasma isolates. In the HMA and SSCP analyses, differences in the mobility shifts and the SSCP band patterns identified three distinct types of phyto-plasmas: Type Ⅰ, jujube witches'-broom (JWB) and ligustrum witches'-broom (LiWB); Type Ⅱ, mulberry dwarf(MD) and sumac witches'-broom (SuWB); and Type Ⅲ, paulownia witches'-broom (PaWB). Results of the sequence analyses revealed that phytoplasmas of JWB and MD had 100% homology with LiWB and SuWB, respectively. On the other hand, PaWB phyto-plasma had 97.8% homology with MD phytoplasma. The PCR-HMA and SSCP techniques were very useful in determining variations in sequence among several isolates of phytoplasmas. Furthermore, the methods were rapid, economical, highly sensitive, and easy to handle with the gels.

키워드

참고문헌

  1. Ahrens, U. and Seemuller, E. 1992. Detection of DNA of plantpathogenic mycoplasma-like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathotogy 82:828-832 https://doi.org/10.1094/Phyto-82-828
  2. Chai, J. K. and Kim, Y. H. 1989. Studies on witches' broom of Ligustrum ovatifoIium Hasskarl caused by mycoplasma-likeorganism (MLO). J. Korean For. Soc. 78:103-118
  3. Chang, B. H. and Kim, C. J. 1971. Studies on the dwarf disease ofmulberry tree. Seri. J. Korea 13:17-21
  4. Cousin, M.-T., Roux, J., Boudon-Padieu, E., Berges, R., Seemuller,E. and Hiruki, C. 1998. Use of heteroduplex mobility analysisfor differentiating phytoplasma isolates causing witches'-broom disease on Poputus nigra cv. itatica and stolbur or bigbud symptoms on tomato. J. Phytopathot. 146:97-102 https://doi.org/10.1111/j.1439-0434.1998.tb04664.x
  5. Delwart, E. L., Shpaer, E. G., Louwagie, J., McCutchan, F. E.,Grez, M., Waigmann, H. R. and Mullins, J. I. 1993. Geneticrelationships determined by a DNA heteroduplex mobilityassay: Analysis ofHIV-l genes. Science 262:1257-1261 https://doi.org/10.1126/science.8235655
  6. Deng, S. J. and Hiruki, C. 1990. The use of cloned DNA probes for diagnosis of non-cultivable plant mollicutes. Proc. Japan Acad. 668:58-61 https://doi.org/10.2183/pjab.66.58
  7. Deng, S. J. and Hiruld, C. 1991a. Amplification of 16S rRNA genes from culturable and non-culturable mollicutes. J. Microbiot. Methods 14:53-61 https://doi.org/10.1016/0167-7012(91)90007-D
  8. Deng, S. J. and Hiruki, C. 1991b. Genetic relatedness between two nonculturable mycoplasmalike organisms revealed by nucleic acid hybridization and polymerase chain reaction. Phytopathology 81:1475-1479 https://doi.org/10.1094/Phyto-81-1475
  9. Doi, Y., Teranaka, M., Yora, K. and Asuyama, H. 1967. Myco-plasma or P. L. T. group-like microorganisms found in the phloem elements of plants infected with mulberry dwarf, potato witches'-broom, aster yellows or paulownia witches'- broom. Ann. phytopath. Soc. Japan 33:259-266 https://doi.org/10.3186/jjphytopath.33.259
  10. Han, S. S., Hiruki, C. and Kim, S. M. 1998. Rapid analysis of genetic relationship of phytoplasma isolates by a DNA heteroduplex mobility assay. Korean J. Ptant PathoI. 45:382-385
  11. Kim, C. J. 1968. Witches' broom ofjujube tree (Zizyphus jujuba Mill. var. inermis Rehd.). Res. Butt. Choonchun Agric. CotIege 2:47-53
  12. Kim, Y. H. 1980. Studies on mycoplasma-like organism associated with witches' broom of Rhus javanica. Korean J. For-estry 47:1-15
  13. Kirkpatrick, B. C., Smart, C. D., Gardner, S. L., Gao, J.-L., Ahrens, U., Maurer, R., Schneider, B., Lorenz, K.-H., Seemuller, E., Harrison, N. A., Namba, S. and Daire, X. 1994. Phylogenetic relationships of plant pathogenic MLOs established by 16/23S rDNA spacer sequences. IOM Lett. 3:228-229
  14. Kollar, A., Seemuller, E., Bonet, F., Saillaird, S. and Bove, J. M. 1990. Isolation of the DNA of various plant pathogenic mycoplasmalike organisms from infected plants. Phytopathotogy 80:233-3237 https://doi.org/10.1094/Phyto-80-233
  15. La, Y. J. 1968. Insect transmission of paulownia witches'-broom disease in Korea. Korea Observer 8:55-64
  16. Lee, I.-M., Gundersen-Rindal, D. E., Davis, R. E. and Bartoszyk, I. M. 1998. Revised classification scheme of phytoplasmas based on RFLP analysis of 16S rRNA and ribosomal protein gene sequences, Int. J. Syst. Bacteriat. 48:1153-1169 https://doi.org/10.1099/00207713-48-4-1153
  17. Orita, M., Iwahana, H., Kanazawa, H., Hayashi, K. and Sekiya, T. 1989. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc. Natt. Acad. Sci. USA 86:2766-2770 https://doi.org/10.1073/pnas.86.8.2766
  18. Prosser, J. 1993. Detecting single-base mutations. TIBTECH 11:224. https://doi.org/10.1016/0167-7799(93)90135-V
  19. Schneider, B., Seemuller, E., Smart, C. D. and K.irkpatrick, B. C. 1995. Phylogenetic classification of plant pathogenic mycoplasmalike organisms or phytoplasmas. In: Motecutar and Diagnostic Procedures in Mycoptasmotogy, ed. by R. Razin and J. G. Tully, PP. 1:369-380. San Diego, CA: Academic
  20. Sheffield, V. C., Beck, J. S., Kwitex, A. E., Sabdstrom, D. W. and Stone, E. M. 1993. The sensitivity of single-strand conformation polymorphism analysis for the detection of single base substitutions. Genomics 16:325-332 https://doi.org/10.1006/geno.1993.1193
  21. Wang, K. and Hiruki, C. 1999. Rapid detection of a single-basepair mutation in phytoplasmal conserved genes by DNA heteroduplex mobility assay (HMA). Proc. Japan Acad. 75B:259-262 https://doi.org/10.2183/pjab.75.259

피인용 문헌

  1. Multiple Infection of Apple Trees by Distinct Strains of ‘CandidatusPhytoplasma mali’ and Its Pathological Relevance vol.100, pp.9, 2010, https://doi.org/10.1094/PHYTO-100-9-0863
  2. Diagnostic techniques for detection of phytoplasma diseases: past and present vol.120, pp.1, 2013, https://doi.org/10.1007/BF03356449