caa a22 4500 467929351 CHVBK 20180406152938.0 cr unu---uuuuu 170328e20060401xx s 000 0 eng 10.1007/s10658-006-0001-9 doi (NATIONALLICENCE)springer-10.1007/s10658-006-0001-9 Real-time PCR for Quantification of Toxigenic Fusarium Species in Barley and Malt [Elektronische Daten] [Tuija Sarlin, Tapani Yli-Mattila, Marika Jestoi, Aldo Rizzo, Sari Paavanen-Huhtala, Auli Haikara] A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4-5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys. Springer, 2006 Gibberella nationallicence trichothecenes nationallicence TaqMan nationallicence Sarlin Tuija VTT Biotechnology, P.O. Box 1000, FIN-02044, VTT, Finland aut Yli-Mattila Tapani Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014, Turku, Finland aut Jestoi Marika Department of Chemistry, National Veterinary and Food Research Institute (EELA), FIN-00581, Helsinki, Finland aut Rizzo Aldo Department of Chemistry, National Veterinary and Food Research Institute (EELA), FIN-00581, Helsinki, Finland aut Paavanen-Huhtala Sari Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014, Turku, Finland aut Haikara Auli VTT Biotechnology, P.O. Box 1000, FIN-02044, VTT, Finland aut European Journal of Plant Pathology Kluwer Academic Publishers 114/4(2006-04-01), 371-380 0929-1873 114:4<371 2006 114 10658 https://doi.org/10.1007/s10658-006-0001-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10658-006-0001-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sarlin Tuija VTT Biotechnology, P.O. Box 1000, FIN-02044, VTT, Finland aut NATIONALLICENCE 700 1- Yli-Mattila Tapani Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014, Turku, Finland aut NATIONALLICENCE 700 1- Jestoi Marika Department of Chemistry, National Veterinary and Food Research Institute (EELA), FIN-00581, Helsinki, Finland aut NATIONALLICENCE 700 1- Rizzo Aldo Department of Chemistry, National Veterinary and Food Research Institute (EELA), FIN-00581, Helsinki, Finland aut NATIONALLICENCE 700 1- Paavanen-Huhtala Sari Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014, Turku, Finland aut NATIONALLICENCE 700 1- Haikara Auli VTT Biotechnology, P.O. Box 1000, FIN-02044, VTT, Finland aut NATIONALLICENCE 773 0- European Journal of Plant Pathology Kluwer Academic Publishers 114/4(2006-04-01), 371-380 0929-1873 114:4<371 2006 114 10658 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer