Browse > Article
http://dx.doi.org/10.5423/PPJ.2002.18.6.308

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)
Publication Information
The Plant Pathology Journal / v.18, no.6, 2002 , pp. 308-312 More about this Journal
Abstract
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.
Keywords
heteroduplex; PCR; phytoplasma; polymorphism; SSCP;
Citations & Related Records
연도 인용수 순위
  • Reference
1 La, Y. J. 1968. Insect transmission of paulownia witches'-broom disease in Korea. Korea Observer 8:55-64
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 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   DOI   ScienceOn
4 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   DOI
5 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
6 Kim, C. J. 1968. Witches' broom ofjujube tree (Zizyphus jujuba Mill. var. inermis Rehd.). Res. Butt. Choonchun Agric. CotIege 2:47-53
7 Kim, Y. H. 1980. Studies on mycoplasma-like organism associated with witches' broom of Rhus javanica. Korean J. For-estry 47:1-15
8 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
9 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   DOI
10 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   DOI
11 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   DOI   ScienceOn
12 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   DOI
13 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   DOI
14 Deng, S. J. and Hiruld, C. 1991a. Amplification of 16S rRNA genes from culturable and non-culturable mollicutes. J. Microbiot. Methods 14:53-61   DOI   ScienceOn
15 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   DOI
16 Prosser, J. 1993. Detecting single-base mutations. TIBTECH 11:224.   DOI   ScienceOn
17 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
18 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   DOI
19 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   DOI
20 Chang, B. H. and Kim, C. J. 1971. Studies on the dwarf disease ofmulberry tree. Seri. J. Korea 13:17-21
21 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   DOI