Congresso Brasileiro de Microbiologia 2023

Congresso Brasileiro de Microbiologia 2023

Resumo: 379-1

379-1

TRI12 GENE QUANTIFICATION BY QUANTITATIVE PCR (qPCR) AS A RAPID SCREENING TOOL FOR HIGH LEVELS OF Fusarium TYPE B TRICHOTHECENES IN BARLEY GRAINS

Autores:

Letícia Aliberti Galego Alves da Silva (FEA/UNICAMP - Universidade Estadual de Campinas, Fac. de Eng. de Alimentos) ; Karim Cristina Piacentini (FEA/UNICAMP - Universidade Estadual de Campinas, Fac. de Eng. de Alimentos) ; Elem Tamirys dos Santos Caramês (ICB/USP - Universidade de São Paulo, Inst. de Ciências Biomédicas) ; Simona Wawroszová (UKZUZ - Central Institute for Supervision and Testing in Agriculture) ; Sylvie Běláková (RIBM - Research Institute Of Brewing and Malting) ; Liliana de Oliveira Rocha (FEA/UNICAMP - Universidade Estadual de Campinas, Fac. de Eng. de Alimentos)

Resumo:

Mycotoxins, such as type B trichothecenes (deoxynivalenol/DON and nivalenol/NIV) are known Fusarium sambucinum species complex’s secondary metabolites and food contaminants. Due to their importance, quick methods for their estimation have become relevant over the years. In this regard, mycotoxin biosynthetic pathway genes quantification by qPCR appears to be promising. The aim of this study was to validate a qPCR assay for the screening of DON and NIV contamination as well as the occurrence of their producing fungi (F. graminearum sensu lato (s.l.) and F. poae) in barley grain samples. To this end, 53 barley samples were chosen for mycotoxin quantification by UPLC-MS/MS, mycobiota analysis and the quantification of TRI12/15-ADON (for the 15-ADON genotype) and TRI12/NIV (for the NIV genotype) and EF1-α (grain reference gene) by qPCR. Pearson correlation analysis was carried out using the software Prism version 9 (GraphPad, 2022, v. 9). qPCR validation exhibited results within the expected standards and the matrix effects obtained were considered to be non-significant. The most frequent mycotoxin was DON (64%) followed by NIV (25%). Regarding mycobiota results, 83% of the samples were contaminated by F. graminearum s.l. and 51% by F. poae. TRI12/15-ADON showed significant correlations with F. graminearum s.l. incidence, whereas TRI12/NIV was positively correlated with F. poae’s occurrence. A relationship was also found among TRI12/15-ADON and DON. No significant correlations were observed among TRI12/NIV and nivalenol, possibly due to the lower occurrence and levels of this toxin in barley samples. According to the results, the developed qPCR assay is a possible tool for screening high levels of type B trichothecenes, as well as the occurrence of their producing fungi in barley grains.

Palavras-chave:

alternative analysis method, mycotoxins, real time PCR, screening, toxigenic Fusarium

Agência de fomento:

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, projeto: 2017/04811-4), Fundo de apoio ao Ensino, pesquisa e extensão (FAEPEX, projeto: 2017/04811-42130/22), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, 001)