Development of a Highly Sensitive FcMito qPCR Assay for the Quantification of the Toxigenic Fungal Plant Pathogen Fusarium culmorum.

Bilska, Katarzyna; Kulik, Tomasz; Ostrowska-Kołodziejczak, Anna; Buśko, Maciej; Pasquali, Matias; Beyer, Marco; Baturo-Cieśniewska, Anna; Juda, Marcin; Załuski, Dariusz; Treder, Kinga; Denekas, Joerg; Perkowski, Juliusz

<italic>Fusarium culmorum</italic> is a ubiquitous, soil-borne fungus (ascomycete) causing foot and root rot and Fusarium head blight on cereals. It is responsible for yield and quality losses as well as grain contamination with mycotoxins, which are a potential health hazard. An extremely sensitive mitochondrial-based qPCR assay (FcMito qPCR) for quantification of <italic>F. culmorum</italic> was developed in this study. To provide specificity, the FcMito assay was successfully validated against 85 <italic>F. culmorum</italic> strains and 53 isolates of 30 other fungal species. The assay efficiency and sensitivity were evaluated against different <italic>F. culmorum</italic> strains with various amounts of pure fungal DNA and in the presence of background wheat DNA. The results demonstrated the high efficiency of the assay (97.2–106.0%, R2-values > 0.99). It was also shown that, in the presence of background DNA, 0.01 pg of fungal template could be reliably quantified. The FcMito assay was used to quantify <italic>F. culmorum</italic> DNA using 108 grain samples with different trichothecene levels. A significant positive correlation was found between fungal DNA quantity and the total trichothecene content. The obtained results showed that the sensitivity of the FcMito assay was much higher than the nuclear-based qPCR assay for <italic>F. culmorum</italic>.

FUSARIUM culmorum; FUNGAL diseases of plants; FUSARIUM diseases of plants; FUNGAL DNA; MYCOTOXINS

Toxins, 2018, Vol 10, Issue 5, p211

2072-6651

Academic Journal

10.3390/toxins10050211