Determination of the Ribosomal DNA Internal Transcribed Spacers and 5.85 rDNA Sequences of Cordyceps Species

  • Bae, Jin-Sik (College of Natural Resources and Life Science, Dong-A University) ;
  • Nam Sook park (College of Natural Resources and Life Science, Dong-A University) ;
  • Jin, Byung-Rae (College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Ho-Oung (Department of Sericultural and Entomological Biology, Faculty of Agriculture, Miryang National University) ;
  • Park, Eun-Ju (Department of Life Science, Kyungnam University) ;
  • B. Tolgor (Mycological Institute, Jilin Agricultural University) ;
  • Yu Li (Mycological Institute, Jilin Agricultural University) ;
  • Lee, Sang-Mong (Department of Sericultural and Entomological Biology, Faculty of Agriculture, Miryang National University)
  • Published : 2002.09.01

Abstract

The sequences of the internal transcribed spacers (ITS1 and ITS2) and 5.8S ribosomal DNA gene from five Cordyceps species and one Paecilomyces japonica were determined. The total length of the ITSI, 5.8S and ITS2 regions ranged from 528 to 549 bp. When the C. militaris collected from Korea was used as a standard genotype, the sequence showed 88.4%, 88.6%, 91.1% and 86.8% identity to C. pruinosa, C. sphecocephala, C. scarabaeucika and R japonica, respectively, while the lowest identity was found with C. sinensis (75.4%). Interestingly, C. sinensis was phylogenetically distant from the other Cordyceps species. To test geographic variation, furthermore, sequences of the ITS regions in the 8 samples of C. militaris collected from two localities in Korea and China analyzed and compared with the GenBank-searched sequences from Japan and China. The total length of the ITS regions of C. militaris from Korea, Japan and China was completely identical to each other with 528 bp, and the sequence divergence among three localities in pairwise comparisons ranged from 0.2% (1 bp) to 0.4% (2 bp).

Keywords

References

  1. Bachellerie, J. P. and L. H. Qu (1993) Ribosomal RNA probe for detection and identification of species; in Protocols in MoIecular Parasitology. Hyde, J. E. (ed.), pp. 249-264, Humana Press Clifton, New Jersey
  2. Chen, Y. Q., N. Wang, L. H, Qu, T. H. Li and W. M. Zhang (2001) Determination of the anamorph of Cordyceps sinen-sis inferred from the analysis of the ribosomal DNA internal transcribed spacers and 5.8S rDNA. Biochem. System. Ecol. 29, 597-607 https://doi.org/10.1016/S0305-1978(00)00100-9
  3. Cho, S. Y. (1999) Cultivation and distribution of silkworm-dongchunghacho (Paecitomyces japonicd). Proc. $1^st$ Int. Sympo. Cordyceps, Seoul. pp. 73-82
  4. Fujita, T., K. Inoue, S. Yamamoto et al. (1994) FungaI metab-olites II. A potent immunosuppressive activity found in Isaria sinclaini metabolite. J. Antibiotics 47, 208-215 https://doi.org/10.7164/antibiotics.47.208
  5. Furuya, T., M. Hirotani and M. Matsuzawa (1983) $N^6$-(2-hydroxyethy1)adenosine, a biologically active compound from cultured mycelia of Cordyceps and Isaria species. Phytochemistry 22, 2509-2512 https://doi.org/10.1016/0031-9422(83)80150-2
  6. Gonzalez, I. L., J. E. Sylvester, T. F. Smith, D. Stambolian and R. D. Schmickel (1990) Ribosomal RNA gene sequences and hominoid phylogeny. Mol. Biol. Evol. 7, 203-219
  7. Gutell, R. R. (1993) Collection of small subunit (16S- and 16S-like) ribosomal RNA structures, Nucleic Acids Res. 21, 3051-3054 https://doi.org/10.1093/nar/21.13.3051
  8. Ito, Y. and T. Hirano (1997) the determination of the partial 18S ribosomal DNA sequences of Cordyceps species. Lett. AppI. Microbiol. 25, 239-242 https://doi.org/10.1046/j.1472-765X.1997.00203.x
  9. Kneifel, H., W. A. Konig, W. Loeffler and R. Muller (1977) Ophiocordin, an antifungal antibiotic of Cordyceps ophio-glossoides. Arch. Microbiol. 113, 121-130 https://doi.org/10.1007/BF00428591
  10. Lee, E. H., N. S. Park, S. B. Park, H. O. Lee, C. S. Lee, B. R. Jin and S. M. Lee (2001a) Effects of photopenods on the growth of the entomopathogenic fungi, Paecilomyces japon-ica, during the production of the silkworm-dongchunghacho, silkworm vegetable wasp and plant worm. Int. J. Indust. Entomol. 2, 83-86
  11. Lee, S. B. and J. W. Taylor (1992) Phylogeny of five fungus-like protoctistan Phytophthora species, inferred from the internal transcribed spacers of ribosomal DNA. MoI. Biol. EvoI. 9, 636-653
  12. Lee, S. M., N. S. Park, S. Y. Cho, J. S. Hwang and B. R. Jin (2001 b) Production of the wild entomopathogenic fungi, Cordyceps militaris, in the silkworm, Bombyx mori. Int. J. Indust. EntomoI. 3, 105-108
  13. Morales, V. M., L. E. Pelcher and J. L. Taylor (1993) Compar-ison of the 5.8S rDNA and internal transcribed spacers sequences of isolates of Leptospaeria macuIans from differ-ent pathogenicity groups. Curr. Genet. 23, 490-495 https://doi.org/10.1007/BF00312640
  14. Nam, S. H., J. S. Hwang, S. Y. Cho and T. W. Goo (1999) Genetic relationship of Cordyceps spp. based on internal transcribed spacer sequences of ribosomal DNA. Korean J. Seric. Sci. 41, 174-179
  15. Nikoh, N. and T. Fukatsu (2000) Interkingdom host jumping underground: Phylogenetic analysis of entomoparasitic fungi of the genus Cordyceps. Mol. Biol. EvoI. 17, 629-638 https://doi.org/10.1093/oxfordjournals.molbev.a026341
  16. Shimizu, D. (1994) Color iconography of vegetable wasps and plant worms. Seibundo Shinkosha, Tokyo
  17. Swofford, D. L. (1990) PAUP: phylogenetic analysis using parimony, ver. 3.0. IllJnois Natural History Survey, Cham-paign (on disk)
  18. Torres, R. A., M. Ganal and V. Hemleben (1990) GC balance in the internal transcribed spacers ITSl and ITS2 of nuclear ribosomal RNA genes. J. MoI. Evol. 30, 171-181
  19. White, T. J., T. D. Bruns, S. B. Lee and J. W. Taylor (1990) Amplification and detection sequencing of fungal ribosomal RNA genes for phylogenetics; in PCR protocols: A guide to methods and applications. Innis, M. A., D. H. Gelfand, J. J. Sninsky and T. J. White (eds.), pp. 315-321, Academic Press, New York
  20. Wilmotte, A., Y. Van de Peer and A. Goris et at. (1993) Evolu-tionary relationships among higher fungi inferred from small ribosomal subunit RNA sequence analysis. System. Appl. Microbiol. 16, 436-444 https://doi.org/10.1016/S0723-2020(11)80277-8
  21. Zhu, J. S., G. M. Halpem and K. Jones (1998a) The scientific rediscovery of a precious ancient Chinese herbal regimen:Cordyceps sinensis: Part I. J. Altem. Complement Med. 4, 289-303 https://doi.org/10.1089/acm.1998.4.3-289
  22. Zhu, J. S., G. M. Halpern and K. Jones (1998b) The scientific rediscovery of a precious ancient Chinese herbal regimen:Cordyceps sinensis: Part II. J. Altern. Complement Med. 4, 429-457 https://doi.org/10.1089/acm.1998.4.429