Quantitative PCR (qPCR) for estimating the presence of Fusarium and its mycotoxins in barley grains

Abstract

Members within the Fusarium sambucinum species complex (FSAMSC) are able to produce mycotoxins, such as deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEN) and enniatins (ENNs) in food products. Consequently, alternative methods for assessing the levels of these mycotoxins are relevant for quick decision-making. In this context, qPCR based on key mycotoxin biosynthetic genes could aid in determining the toxigenic fungal biomass, and could therefore infer mycotoxin content. The aim of this study was to verify the use of qPCR as a technique for estimating DON, NIV, ENNs and ZEN, as well as Fusarium graminearum sensu lato (s.l.) and F. poae in barley grains. For this purpose, 53 barley samples were selected for mycobiota, mycotoxin and qPCR analyses. ENNs were the most frequent mycotoxins, followed by DON, ZEN and NIV. 83% of the samples were contaminated by F. graminearum s.l. and 51% by F. poae. Pearson correlation analysis showed significant correlations for TRI12/15-ADON with DON, ESYN1 with ENNs, TRI12/15-ADON and ZEB1 with F. graminearum s.l., as well as ESYN1 and TRI12/NIV with F. poae. Based on the results, qPCR could be useful for the assessment of Fusarium presence, and therefore, provide an estimation of its mycotoxins’ levels from the same sample.

Keywords:

• Mycotoxins
• toxigenic fungi
• analytical methods
• mass spectrometry

Acknowledgements

The authors would like to thank the São Paulo Research Foundation (FAPESP, Project number: 2017/04811-4), Fundo de apoio ao Ensino, pesquisa e extensão (FAEPEX, Project number: 2130/22), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Finance Code 001) and the Ministry of Agriculture of the Czech Republic, institutional support MZE-RO1923.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was financially supported by the São Paulo Research Foundation – FAPESP under Grant [2017/04811-4]; Fundo de apoio ao Ensino, pesquisa e extensão - FAEPEX under Grant [2130/22]; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES under [Finance Code 001]; and the Ministry of Agriculture of the Czech Republic under Grant [MZE-RO1923].