Characterization of high-level deoxynivalenol producer Fusarium graminearum and F-culmorum isolates caused head blight and crown rot diseases in Turkey

Abstract

In vitro and in planta determination and quantification of deoxynivalenol (DON) are a crucial step in food safety. tri13 amplicons of 282 bp and tri3 partial region of 863 bp were obtained from 16 isolates with DON and 15-acetyldeoxynivalenol (15-AcDON) mycotoxin profiles. Similarly, 42 3-acetyldeoxynivalenol (3-AcDON) isolates yielded 282-bp band of tri13 and 583 bp of tri3 gene in multiplex polymerase chain reaction (PCR) assays. None of the isolates were characterized as NIV producers via PCR. All Fusarium isolates have been determined as high-level DON producers (more than 1 ppm) via tri5-tri6 intergenic regions polymorphisms. tri11 expression via reverse transcriptase PCR (RT-PCR) confirmed results obtained at genomic level. High-pressure liquid chromatographic (HPLC) analysis showed that DON and/or acetylated derivatives can be co-produced with NIV. Even if each assay characterized Turkish Fusarium species as high trichothecene producer via PCR assays, HPLC analysis yielded 0-2481 mu g/kg level of trichothecene production including DON, 3-AcDON and NIV. In this study, chemotype determination was carried out via multiplex PCR and tri13, and tri3 genes were co-targeted in a single reaction tube. It is posited that this is the first report on low-and high-level DON production differentiation analysis in Fusarium graminearum isolates via tri5-tri6 intergenic region analysis worldwide. Results showed that high-and low-DON production potential of F. culmorum isolates' from tri5 to tri6 nucleotide data could be useful in other Fusarium sp. characterization. This study covers the widest range of geographical regions in Turkey to be subjected to comprehensive class B-trichothecene analysis.