Real-time PCR detection and quantification of <i>Fusarium poae, F. graminearum, F. sporotrichioides</i> and <i>F. langsethiae</i> in cereal grains in Finland and Russia

Abstract

Abstract TaqMan real-time quantitative PCR assays were developed for the accurate detection and quantification of DNA from Fusarium poae and F. graminearum species, which are able to produce trichothecenes. These and other PCR assays were used for the quantification of trichothecene-producing Fusarium fungi in cereal grains. A correlation was found between the levels of F. poae DNA and nivalenol and enniatins in barley and between the levels of F. graminearum DNA and deoxynivalenol in oats. The correlations between F. poae DNA and nivalenol and F. graminearum DNA and deoxynivalenol levels were higher than those between these mycotoxins and morphologically determined F. poae and F. graminearum/F. culmorum contamination levels. The use of F. poae specific primers and probe together with F. sporotrichioides/F. langsethiae specific primers and probe in a multiplex qPCR assay yielded results in accordance with those obtained using these primers and probes separately. Keywords: BarleydeoxynivalenolenniatinsnivalenoloatsTaqMan PCR Acknowledgements This study was supported by grants from the National Technology Agency of Finland (No. 40168/03), and the Academy of Finland (No. 52104) and the Nordic Research Board (No. 040291). The authors want to thank Nina Painilainen, Taina Lahtinen, Teija Koivula and Tarja Nordenstedt for their assistance with DNA extraction and qPCR analyses, Kati Lehti for skilful assistance with mycotoxin analyses, Dr Sonja Klemsdal for designing TMTRI and TMLAN primers and probe, Dr Jouni Ahlholm for help in statistical analyses and Dr Cees Waalwijk for critically reading the manuscript. The English was checked by Ellen Valle of the Department of English, University of Turku.