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Rapid Detection and Quantification of Rhynchosporium secalis in Barley Using a Polymerase Chain Reaction

Author

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  • Jozef Gubiš

    (Slovak Agricultural Research Centre, Research Institute of Plant Production, Piešťany, Slovak Republic)

  • Martina Hudcovicová

    (Slovak Agricultural Research Centre, Research Institute of Plant Production, Piešťany, Slovak Republic)

  • Marcela Gubišová

    (Slovak Agricultural Research Centre, Research Institute of Plant Production, Piešťany, Slovak Republic)

Abstract

PCR primers for diagnosis of Rhynchosporium secalis in seed samples of barley were developed. For the quantification of the pathogen in seed samples a real-time PCR with SYBR Green approach was used. Amounts from 1.8 to 419.1 pg of R. secalis DNA per 100 ng of total DNA were detected in 18 samples of barley seeds contaminated by R. secalis in field conditions. The correctness of this quantitative analysis was checked using an artificial infection of seeds with 1, 2, 5 and 20% level of infection by R. secalis. The level of contamination of artificially infected samples decreased with a lowering amount of added seed powder contaminated by the pathogen, the correlation coefficient for this analysis was 0.98. While the primer pair used in these analyses shows cross-reactions with other pathogens (P. teres, Drechslera tritici-repentis, F. culmorum and F. poe), it is recommended to check the products of RT-PCR by agarose-gel electrophoresis, in which these pathogens are easily distinguishable from R. secalis by different lengths of the amplified fragments.

Suggested Citation

  • Jozef Gubiš & Martina Hudcovicová & Marcela Gubišová, 2006. "Rapid Detection and Quantification of Rhynchosporium secalis in Barley Using a Polymerase Chain Reaction," Czech Journal of Genetics and Plant Breeding, Czech Academy of Agricultural Sciences, vol. 42(3), pages 111-114.
    • Handle: RePEc:caa:jnlcjg:v:42:y:2006:i:3:id:3650-cjgpb
    DOI: 10.17221/3650-CJGPB
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    References listed on IDEAS

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    1. J. Gubiš & M. Hudcovicová & L. Klčová & V. Červená & K. Bojnanská & J. Kraic, 2004. "Detection of Leaf Blotches - Causal Agents in Barley Leaves and Grains," Czech Journal of Genetics and Plant Breeding, Czech Academy of Agricultural Sciences, vol. 40(4), pages 111-117.
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