Molecular diagnostics on the toxigenic potential of Fusarium spp. plant pathogens

• Published in: Journal of applied microbiology 2014-06, Vol.116 (6), p.1607-1620
• Main Authors: Dawidziuk, A, Koczyk, G, Popiel, D, Kaczmarek, J, Buśko, M
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biomarkers; biosynthesis; cost effectiveness; Crop science; diagnostic techniques; early warning systems; essential genes; fumonisins; Fumonisins - analysis; Fundamental and applied biological sciences. Psychology; Fungi; Fusarium; Fusarium - genetics; Fusarium - isolation & purification; Fusarium - pathogenicity; Genes, Fungal; markers; Microbiology; mycotoxins; Original; plant pathogens; Plant pathology; plant protection; Polymerase Chain Reaction; screening; Toxins; trichothecenes; Trichothecenes - analysis; Trichothecenes - genetics; zearalenone; Zearalenone - analysis; Zearalenone - genetics
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.12488

AIMS: We propose and test an efficient and rapid protocol for the detection of toxigenic Fusarium isolates producing three main types of Fusarium‐associated mycotoxins (fumonisins, trichothecenes and zearelanone). METHODS AND RESULTS: The novel approach utilizes partially multiplexed markers based on genes essential for mycotoxin biosynthesis (fumonisin—fum6, fum8; trichothecenes—tri5, tri6; zearalenone, zea2) in Fusarium spp. The protocol has been verified by screening a collection of 96 isolates representing diverse species of filamentous fungi. Each Fusarium isolate was taxonomically identified through both molecular and morphological techniques. The results demonstrate a reliable detection of toxigenic potential for trichothecenes (sensitivity 100%, specificity 95%), zearalenone (sensitivity 100%, specificity 100%) and fumonisins (sensitivity 94%, specificity 88%). Both presence and identity of toxin biosynthetic genes were further confirmed by direct sequencing of amplification products. CONCLUSIONS: The cross‐species‐specific PCR markers for key biosynthetic genes provide a sensitive detection of toxigenic fungal isolates, contaminating biological material derived from agricultural fields. SIGNIFICANCE AND IMPACT OF THE STUDY: The conducted study shows that a PCR‐based assay of biosynthetic genes is a reliable, cost‐effective, early warning system against Fusarium contamination. Its future use as a high‐throughput detection strategy complementing chemical assays enables effective targeted application of crop protection products.