Prediction and early detection of mycotoxigenic Fusarium culmorum in wheat by direct PCR-based procedure

Cereals contamination with mycotoxigenic species of Fusarium is considered as a major source of trichothecenes and other mycotoxin groups which cause severe yield losses and serious diseases in human and animal health. Early detection of Fusarium species could be for a great interest to prevent mycotoxin contaminating agro-products. We have established for the first time a direct polymerase chain reaction (DPCR) protocol to detect contamination with trichothecene-producing E culmorum in wheat samples. We have successfully amplified fungal genomic DNA using specific primers targeting the trichothecenes biosynthetic Tri5 gene. We further investigated a versatile multiplex-DPCR on the basis of Tri5 gene and IGS (Intergenic Spacer of rDNA) specific sequence of E culmorum for its identification at specie level and prediction of its potential trichothecenes production simultaneously. Our protocol allowed amplification directly from crude templates with no need of DNA extraction or purification methods and did not require any culture-based approach. These DPCR assays represent a reliable tool for high throughput screening, detection and rapid characterization of mycotoxigenic isolates as well as diverse applications in food industry. (C) 2011 Elsevier Ltd. All rights reserved.