Czech J. Genet. Plant Breed., 2004, 40(4):111-117 | DOI: 10.17221/3708-CJGPB

Detection of Leaf Blotches - Causal Agents in Barley Leaves and Grains

J. Gubiš, M. Hudcovicová, L. Klčová, V. Červená, K. Bojnanská, J. Kraic
Division of Applied Genetics and Breeding, Research Institute of Plant Production, Piešťany, Slovak Republic

The objective of this paper was to adapt PCR-based detection method for R. secalis and P. teres DNA isolated from pathogens and also from artificially infected juvenile leaves and seeds using pathogen-specific primers. It has been proven that primers specific to P. teres and R. secalis can reliably diagnose pathogen DNA as well as its presence in the mixture with barley DNA. Two primers set for detection of R. secalis were compared. The intensity of the corresponding DNA band after amplification with primer pair RS1-RS3 was higher than that amplified with RS8-RS9. The primer set RS1-RS3 was also used to detect R. secalis in barley seeds. DNA from infected seeds was isolated by two ways - according to the method of Dellaporta et al. (1983) or by the Adgen DNA Extraction System. The DNA extracted using the Adgen kit showed higher quality, however the amplification of the pathogen DNA was accomplished in both cases.

Keywords: Pyrenophora teres; Rhynchosporium secalis, DNA; PCR; pathogen diagnostics

Published: December 31, 2004  Show citation

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Gubiš J, Hudcovicová M, Klčová L, Červená V, Bojnanská K, Kraic J. Detection of Leaf Blotches - Causal Agents in Barley Leaves and Grains. Czech J. Genet. Plant Breed. 2004;40(4):111-117. doi: 10.17221/3708-CJGPB.
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References

  1. B J.A., T E.J.A., K D.M., T J.E. (2 1): The application of real-time PCR to the identification, detection and quantification of Pyrenophora species in barley seed. Molec. Plant Pathol., 2: 49-57.
  2. B J.J., L J.M. (1995): Polymerase chain reaction assay for the detection of Stagonospora nodorum and Septoria tritici in wheat. Phytopathology, 85: 319-324. Go to original source...
  3. B M.P., S B.J., W R.K. (1993): Host range of Pyrenophora teres isolates from California. Plant Dis., 77: 942-947. Go to original source...
  4. D S.L., W J., H J.B. (1983): A plant DNA minipreparation: version II. Plant Mol. Biol. Rep., 1: 19-21. Go to original source...
  5. D F.M., P D.W., J P., N P. (1998): The use of species-specific PCR-based assays to analyse Fusarium ear blight of wheat. Plant Pathol., 47: 197-205. Go to original source...
  6. F B.A., L D.J., C J.M., B S., H D.W. (2001): PCR-based assays to assess wheat varietal resistance to blotch (Septoria tritici and Stagonospora nodorum) and rust (Puccinia striformis and Puccinia recondita) diseases. Eur. J. Plant Pathol., 107: 905-917.
  7. G P.H., H T., X B.G., L H.W. (1995): Differentiation of Gaemannomyces graminis from other turf-grass fungi by amplification with primers from ribosomal internal transcribed spacers. Plant Pathol., 44: 384-391. Go to original source...
  8. G D.S. (1999): Quantitative real-time polymerase chain reaction for the core facility using TaqMan and the Perkin Elmer/Applied Biosystem Division 7700 sequence detector. J. Biomolec. Technol., 10: 11-16.
  9. H D.R., B T.E., P K.A. (1994): Use of random amplified polymorphic DNA markers for the detection of genetic variation in Magnaporthe poea. Phytopathology, 84: 1312-1316. Go to original source...
  10. K K., O A., H M. (1997): Detection of Pythium ultimum using polymerase chain reaction with species-specific primers. Plant Dis., 81: 1155-1160. Go to original source... Go to PubMed...
  11. K J.G., O H. (1973): Transmission of Rhynchosporium secalis on barley grain. Trans. Brit. Mycolog. Soc., 60: 405-411. Go to original source...
  12. L H.K., T J.P., T T.K. (2001): A PCRbased assay to detect Rhynchosporium secalis in barley seed. Plant Dis., 85: 220-225. Go to original source... Go to PubMed...
  13. M C H.A., F S.J., F B.A., W E. (2003): Molecular diagnostics for fungal plant pathogens. Pest Manag. Sci., 59: 129-142. Go to original source... Go to PubMed...
  14. P D.W., N P. (1996): Development of a PCR assay to detect Fusarium poe in wheat. Plant Pathol., 45: 383-391. Go to original source...
  15. R J.C. (1999): Nucleic acid techniques in testing for seed-borne diseases. In: S J.H., A R. (eds): New Diagnostics in Crop Science. CAB International, Wallingford, UK, 127-149. chosporium secalis from three continents suggests moderate gene flow and regular recombination. Phythopatology, 90: 901-908. Go to original source...
  16. S A.A. (1994): Rynchosporioz jačmenja. Zašč. Rast., 5: 21-22. 49: 623-626. identification of Tilletia indica, causal agent of kernel bunt of wheat. Phytopathology, 86: 115-122. 59: 3681-3685. PCR detection of Pyrenophora graminea from barley seed. Plant Pathol., 50: 347-355. tion of new diagnostic technique for the detection of seed-borne Pyrenophora species (leaf stripe and net blotch) on barley. In: Project No. 244, NIAB, Huntingdon Road, Cambridge CB3 OLE. Application of a real-time PCR method to detect Pyrenophora species in barley seed, and implications for seed treatment strategies. EPPO Bull., 30: 555-558. H. (2001): Development and use of an assay based on the polymerase chain reaction that differentiates the pathogens causing spot form and net form of net blotch of barley. Aust. Plant Pathol., 30: 37-44. transmission of sorghum downy mildew: detection by DNA hybridization. Seed Sci. & Technol., 18: 201-207. of Diaporthe phaseolorum and Phomis longicolla from soybean seeds. Phytopathology, 89: 796-804.

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