title: Quantification of Fusarium graminearum and Fusarium culmorum by real-time PCR system and zearalenone assessment in maize.
abstract: Zearalenone (ZEA) is a mycotoxin produced by some species of Fusarium, especially by Fusarium graminearum and F. culmorum. ZEA induces hyperoestrogenic responses in mammals and can result in reproductive disorders in farm animals. In the present study, a real-time PCR (qPCR) assay has been successfully developed for the detection and quantification of Fusarium graminearum based on primers targeting the gene PKS13 involved in ZEA biosynthesis. A standard curve was developed by plotting the logarithm of known concentrations of F. graminearum DNA against the cycle threshold (Ct) value. The developed real time PCR system was also used to analyze the occurrence of zearalenone producing F. graminearum strains on maize. In this context, DNA extractions were performed from thirty-two maize samples, and subjected to real time PCR. Maize samples also were analyzed for zearalenone content by HPLC. F. graminearum DNA content (pg DNA/ mg of maize) was then plotted against ZEA content (ppb) in maize samples. The regression curve showed a positive and good correlation (R²=0.760) allowing for the estimation of the potential risk from ZEA contamination. Consequently, this work offers a quick alternative to conventional methods of ZEA quantification and mycological detection and quantification of F. graminearum in maize.
authors.affiliation: Laboratory of Microorganisms and Food Irradiation, Lebanese Atomic Energy Commission-CNRS, P.O. Box 11-8281, Riad El Solh, 11072260 Beirut, Lebanon. a.atoui@cnrs.edu.lb
authors.forename: Ali
authors.initials: A
authors.lastname: Atoui
authors.forename: André
authors.initials: A
authors.lastname: El Khoury
authors.forename: Mireille
authors.initials: M
authors.lastname: Kallassy
authors.forename: Ahmed
authors.initials: A
authors.lastname: Lebrihi
mesh_terms: D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis* / Q000235:genetics; D005506:Food Contamination / Q000032:analysis; D005670:Fusarium / Q000737:chemistry* / Q000235:genetics; D009183:Mycotoxins / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014255:Trichothecenes / Q000032:analysis / Q000096:biosynthesis; D003313:Zea mays / Q000382:microbiology*; D015025:Zearalenone / Q000032:analysis* / Q000235:genetics
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D009183:Mycotoxins; D014255:Trichothecenes; D015025:Zearalenone