The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

Simple and Reliable DNA Extraction Method for the Dark Pigmented Fungus, Cercospora sojina

  • Kim, Ji-Seong (Department of Environmental Horticulture, The University of Seoul) ;
  • Seo, Sang-Gyu (Department of Environmental Horticulture, The University of Seoul) ;
  • Jun, Byung-Ki (Department of Environmental Horticulture, The University of Seoul) ;
  • Kim, Jin-Won (Department of Environmental Horticulture, The University of Seoul) ;
  • Kim, Sun-Hyung (Department of Environmental Horticulture, The University of Seoul)
  • Received : 2010.07.18
  • Accepted : 2010.08.08
  • Published : 2010.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

This study used a modified cetyltrimethylammonium bromide (CTAB) method to efficiently extract DNA from the plant pathogenic fungus Cercospora sojina. Total DNA yield obtained by this method was approximately 1 mg/g of mycelia (fresh weight), and the mean ratio of A260/A280 and A260/A230 were 2.04 and 2.1, respectively. The results of random amplified polymorphic DNA (RAPD) analysis, digestion with restriction enzymes, and Southern hybridization indicated that polysaccharides were effectively removed by this method, and the resulting DNA was sufficient for use in subsequent molecular analysis.

Keywords

References

  1. Asakura, T., Adachi, K. and Schwartz, E. 1978. Stabilizing effectof various organic solvents on protein. J. Biol. Chem. 25:6423-6425.
  2. Chi, M. W., Park, S. Y. and Lee, Y. H. 2009. A quick and safemethod for fungal DNA extraction. Plant Pathol. J. 25:108-111. https://doi.org/10.5423/PPJ.2009.25.1.108
  3. Daub, M. E. and Chung, K. R. 2007. Cercosporin: a photoactivatedtoxin in plant disease. APSnet Feature Story. Februarywww.apsnet.org
  4. Farkas, V. 1979. Biosynthesis of cell walls of fungi. Microbiol.Reviews 43:117-144.
  5. Goodwin, S. B., Dunkle, L. D. and Zismann, V. L. 2001. Phylogeneticanalysis of Cercospora and Mycosphaerella based on theinternal transcribed spacer region of ribosomal DNA. Phytopathology91:648-658. https://doi.org/10.1094/PHYTO.2001.91.7.648
  6. Hartman, G. L., Sinclair, J. B. and Rupe, J. C. 1999. Frogeye leafspot. In: Compendium of soybean diseases, 4rd edn. TheAmerican Phytopathological Society Press. St Paul. pp. 20-21.
  7. Karakousis, A., Tan, L., Ellis, D., Alexiou, H. and Wormald, P. J.2006. An assessment of the efficiency of fungal DNA extractionmethods for maximizing the detection of medicallyimportant fungi using PCR. J. Microbiol. Methods 65:38-48. https://doi.org/10.1016/j.mimet.2005.06.008
  8. Kim, S. H. and Hamada, T. 2005. Rapid and reliable method ofextracting DNA and RNA from sweetpotato, Ipomoea batatas(L). Lam. Biotechnol. Lett. 27:1841-1845. https://doi.org/10.1007/s10529-005-3891-2
  9. Kuhad, R. C., Kapoor, R. K. and Lal, R. 2004. Improving theyield and quality of DNA isolated from white-rot fungi. FoliaMicrobiol. 49:112-116. https://doi.org/10.1007/BF02931383
  10. Loeffler, J., Hebart, H., Shumacher, U., Reitze, H. and Einsele, H.1997. Comparison of different methods for extraction of DNAof fungal pathogens from cultures and blood. J. Clin. Microbiol.35: 3311-3312.
  11. Mahuku, G. S. 2004. A simple extraction method suitable for PCRbased analysis of plant, fungal, and bacterial DNA. Plant Mol.Biol. Rep. 22:71-81. https://doi.org/10.1007/BF02773351
  12. Manna, B., Gambhir, A. and Ghosh, P. 1996. Production andrheological characteristics of the microbial polysaccharide gellan.Lett. Applied Microbiol. 23:141-145.
  13. Moller, E. M., Bahnweg, G., Sandermann, H. and Geiger, H. H.1992. A simple and efficient protocol for isolation of highmolecular weight DNA from filamentous fungi, fruit bodies,and infected plant tissues. Nucleic Acids Res. 20:6115-6116. https://doi.org/10.1093/nar/20.22.6115
  14. Pandey, R. N., Adams, R. P. and Flournoy, L. E. 1996. Inhibitionof random amplified polymorphic DNAs (RAPDs) by plantpolysaccharides. Plant Mol. Biol. Rep. 14:17-22. https://doi.org/10.1007/BF02671898
  15. Sreenivasaprasad, S. 1999. Isolation of fungal nucleic acids. In R.Rapley (ed) The Nucleic Acid Protocols Handbook, HumanaPress. Totowa. pp 37-45.

Cited by

  1. Application of Bioassay-Guided Fractionation Coupled with a Molecular Approach for the Dereplication of Antimicrobial Metabolites vol.79, pp.23-24, 2016, https://doi.org/10.1007/s10337-016-3188-8
  2. Molecular characterization of an endophytic Phomopsisliquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites vol.5, pp.2, 2015, https://doi.org/10.1007/s13205-014-0204-2
  3. Genome sequencing and comparative genomics reveal the potential pathogenic mechanism of Cercospora sojina Hara on soybean 2018, https://doi.org/10.1093/dnares/dsx035
  4. Molecular profiling and antimicrobial potential of endophyticGliomastix polychromaCLB32 inhabitingCombretum latifoliumBlume vol.6, pp.3-4, 2015, https://doi.org/10.1080/21501203.2015.1113207
  5. Chemogenomics driven discovery of endogenous polyketide anti-infective compounds from endosymbiotic Emericella variecolor CLB38 and their RNA secondary structure analysis vol.12, pp.2, 2017, https://doi.org/10.1371/journal.pone.0172848
  6. Isolation and characterization of antimicrobial metabolite producing endophytic Phomopsis sp. from Ficus pumila Linn. (Moraceae) vol.4, pp.3, 2013, https://doi.org/10.1016/j.ijcas.2013.08.006
  7. Genomic and chromatographic approach for the discovery of polyketide antimicrobial metabolites from an endophyticPhomopsis liquidambarisCBR-18 vol.8, pp.2, 2015, https://doi.org/10.1080/21553769.2015.1033768
  8. Antimicrobial Profiling and Molecular Identification of Alternaria arborescens CLB12, A Myco-endosymbiont Inhabiting Combretum latifolium Blume vol.7, pp.1, 2017, https://doi.org/10.1080/22311866.2017.1287592