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Application of Real-Time and Multiplex Polymerase Chain Reaction Assays to Study Leaf Blotch Epidemics in Barley

March 2007 , Volume 97 , Number  3
Pages  297 - 303

J. M. Fountaine , M. W. Shaw , B. Napier , E. Ward , and B. A. Fraaije

First author: Plant-Pathogen Interactions Division, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, U.K. and School of Plant Sciences, The University of Reading, Reading, Berkshire, RG6 6AS, U.K.; second author: School of Plant Sciences, The University of Reading, U.K.; third author: National Institute of Agricultural Botany, Cambridge, Cambridgeshire, CB3 0LE, U.K.; and fourth and fifth authors: Plant-Pathogen Interactions Division, Rothamsted Research

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Accepted for publication 20 September 2006.

Leaf blotch, caused by Rhynchosporium secalis, was studied in a range of winter barley cultivars using a combination of traditional plant pathological techniques and newly developed multiplex and real-time polymerase chain reaction (PCR) assays. Using PCR, symptomless leaf blotch colonization was shown to occur throughout the growing season in the resistant winter barley cv. Leonie. The dynamics of colonization throughout the growing season were similar in both Leonie and Vertige, a susceptible cultivar. However, pathogen DNA levels were approximately 10-fold higher in the susceptible cultivar, which expressed symptoms throughout the growing season. Visual assessments and PCR also were used to determine levels of R. secalis colonization and infection in samples from a field experiment used to test a range of winter barley cultivars with different levels of leaf blotch resistance. The correlation between the PCR and visual assessment data was better at higher infection levels (R2 = 0.81 for leaf samples with >0.3% disease). Although resistance ratings did not correlate well with levels of disease for all cultivars tested, low levels of infection were observed in the cultivar with the highest resistance rating and high levels of infection in the cultivar with the lowest resistance rating.

Additional keywords: epidemic development.

© 2007 The American Phytopathological Society