Mycobiology

The Korean Society of Mycology (한국균학회)
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Molecular Analysis of Intraspecific Variations of the Indonesian Cochliobolus heterostrophus

  • Gafur, Abdul (Department of Plant Protection, University of Lampung) ;
  • Mujim, Subli (Department of Plant Protection, University of Lampung) ;
  • Aeny, Titik Nur (Department of Plant Protection, University of Lampung) ;
  • Tjahjono, Budi (Department of Plant Pests and Diseases, Bogor Agricultural University) ;
  • Suwanto, Antonius (Department of Biology, Bogor Agricultural University)
  • Published : 2003.03.31
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Abstract

The primary objective of the current research was to detect genetic variations within the Indonesian isolates of Cochliobolus heterostrophus collected from ecologically different places of the country at molecular level using PCR-RFLP analyses. The primer pair of NS3 and NS6 produced amplification fragment in all of the isolates tested. A single fragment of estimated 907 bp was observed in the PCR product pattern. RFLP analysis of the PCR product employing three restriction enzymes, HaeIII, HhaI, and RsaI, respectively, did not reveal intraspecific variations within the fungus. Similarly, nucleotide sequences of portion of small subunit of the ribosomal DNA gene of two of the isolates collected showed no appreciable differences, indicating the absence of genetic diversities among the isolates tested. A phylogenetic tree was constructed and the Indonesian C. heterostrophus, represented by SM-1 isolate, was found to be phylogenetically located near C. sativus, a closely related species.

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References

  1. Alcorn, J. L. 1983. Generic concepts in Drechslera, Bipolaris and Exserohilum. Mycotaxon 17: 1-86
  2. Bronson, C. R, Taga, M. and Yoder, O. C. 1990. Genetic control and distorted segregation of the T-toxin production in field isolates of Cochliobolus heterostrophus. Phytopathology 80: 819-823 https://doi.org/10.1094/Phyto-80-819
  3. Bruns, T. D., White, J. and Taylor, J. W. 1991. Fungal molecular systematics. Ann. Rev. Ecol. Syst. 22: 525-564 https://doi.org/10.1146/annurev.es.22.110191.002521
  4. Chee, H. Y. and Kim, Y. K. 2002. Molecular analysis of Exophiala species using molecular markers. Mycobiology 30: 1-4 https://doi.org/10.4489/MYCO.2002.30.1.001
  5. Gafur, A., Tanaka, C., Shimizu, K., Ouchi, S. and Tsuda, M. 1998. Molecular analysis and characterization of the Cochliobolus heterostrophus $\beta$-tubulin gene and its possible role in conferring resistance to benomyl. I. Gen. Appl. Microbiol. 44: 217-223 https://doi.org/10.2323/jgam.44.217
  6. Gafur, A., Tanaka, C., Ouchi, S. and Tsuda, M. 1997. A PCR-based method for mating type determination in Cochliobolus heterostrophus. Mycoscience 38: 455-458 https://doi.org/10.1007/BF02461689
  7. Gardes, M. and Bruns, T. D. 1993. ITS primers with enhanced specificity for basidiomycetes - application to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118 https://doi.org/10.1111/j.1365-294X.1993.tb00005.x
  8. Jones, M. J. and Dunkle, L. D. 1993. Analysis of Cochliobolus carbonum races by PCR with arbitratory and gene-specific primers. Phytopathology 83: 366-370 https://doi.org/10.1094/Phyto-83-366
  9. Landvik, S., Kristiansen, R and Schumacher, T. 1998. Phylogenetic and structural studies in the Thelebolaceae (Ascomycota). Mycoscience 39: 49-56 https://doi.org/10.1007/BF02461578
  10. Nakada, M., Tanaka, C., Tsunewaki, K and Tsuda, M. 1994. RFLP analysis for species separation in the genera Bipolaris and Curvularia. Mycoscience 35: 271-278 https://doi.org/10.1007/BF02268449
  11. Nakamura, H., Kaneko, S., Yamaoka, Y. and Kakishima, M. 1998. Differentiation of Melampsora rust species on willows in Japan using PCR-RFLP analysis of ITS regions of ribosomal DNA. Mycoscience 39: 105-113 https://doi.org/10.1007/BF02464048
  12. Sambrook, J., Fritsch, E. E and Maniatis, T. 1989. Molecular Cloning - A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press. Cold Spring Harbor
  13. Sanger, E, Nicklen, S. and Coulson, A. R 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A. 74: 5463-5467 https://doi.org/10.1073/pnas.74.12.5463
  14. Sivanesan,A 1987. Graminicolous species of Bipolaris, Curvularia, Exserohilum and their te1eomorphs. CAB Mycol. Pap. 158: 1-261
  15. Tae, M. S., Eom, A. H. and Lee, S, S. 2002. Sequence analyses of PCR amplified partial SSU of ribosomal DNA for identifying arbuscular mycorrhizal fungi in plant roots. Mycobiology 30: 13-17
  16. Tanaka, C., Kubo, Y. and Tsuda, M. 1991. Genetic analysis and characterization of Cochliobolus heterostrophus color mutants. Mycol. Res. 92: 49-56
  17. Tanaka, C., Nakada, M. and Tsuda, M. 1992. Electrophoretic separation of chromosomes of some graminicolous fungi. Trans. Mycol. Soc. lpn. 33: 95-102
  18. White, T. J., Bruns, T., Lee, S. and Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Pp. 315-322 In: PCR Protocols. Innis, M. A, Gelfrand, D. H., Sninsky, J. J. and White, J. (Eds.). Academic Press. San Diego
  19. Yoder, O. C. 1988. Cochliobolus heterostrophus, cause of southern com leaf blight. pp. 93-112. In: Genetics of plant pathogenic fungi (Advances in Plant Pathology 6). Ingram, D. D. and Williams, P. H. (Eds.). Academic Press. London