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