Analyses of air samples for ascospores of Leptosphaeria maculans and L. biglobosa by light microscopy and molecular techniques

A - Papers appearing in refereed journals

Kaczmarek, J., Jedryczka, M., Fitt, B. D. L., Lucas, J. A. and Latunde-Dada, A. O. 2009. Analyses of air samples for ascospores of Leptosphaeria maculans and L. biglobosa by light microscopy and molecular techniques. Journal of Applied Genetics. 50 (4), pp. 411-419.

AuthorsKaczmarek, J., Jedryczka, M., Fitt, B. D. L., Lucas, J. A. and Latunde-Dada, A. O.
Abstract

Spores of many fungal pathogens are dispersed by wind. Detection of these airborne inocula is important in forecasting both the onset and the risk of epiphytotics. Species-specific primers targeted at the internal transcribed spacer (ITS) region of Leptosphaeria maculans and L. biglobosa - the causal organisms of phoma stem canker and stem lesions of Brassica spp., including oilseed rape - were used to detect DNA extracted from particles deposited on tapes obtained from a spore trap operated in Rarwino (northwest Poland) from September to November in 2004 and 2006. The quantities of DNA assessed by traditional end-point PCR and quantitative real-time PCR were compared to microscopic counts of airborne ascospores. Results of this study showed that fluctuations in timing of ascospore release corresponded to the dynamics of combined concentrations of DNA from L. maculans and L. biglobosa, with significant positive correlations between ascospore number and DNA yield. Thus the utilization of PCR-based molecular diagnostic techniques enabled the detection, identification, and accurate quantification of airborne inoculum at the species level. Moreover, real-time PCR was more sensitive than traditional PCR, especially in years with low ascospore numbers.

Keywordsascospore release; disease forecasting; molecular biological diagnostics; oilseed rape; phoma stem canker; polymerase chain reaction; real-time PCR; spore trapping
Year of Publication2009
JournalJournal of Applied Genetics
Journal citation50 (4), pp. 411-419
Web address (URL)http://jag.igr.poznan.pl/2009-Volume-50/4/abstracts/537.html
Open accessPublished as non-open access
Funder project or codePDM
Output statusPublished
Publication dates
Online2009
Copyright licensePublisher copyright
ISSN1234-1983
PublisherSpringer Heidelberg

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