New PCR Assays for the Identification of Fusarium verticillioides, Fusarium subglutinans, and Other Species of the Gibberella fujikuroi Complex

Abstract

Fusarium verticillioides and Fusarium subglutinans are important fungal pathogens of maize and other cereals worldwide. In this study, we developed PCR-based protocols for the identification of these pathogens targeting the gaoB gene, which codes for galactose oxidase. The designed primers recognized isolates of F. verticillioides and F. subglutinans that were obtained from maize seeds from several producing regions of Brazil but did not recognize other Fusarium spp. or other fungal genera that were either obtained from fungal collections or isolated from maize seeds. A multiplex PCR protocol was established to simultaneously detect the genomic DNA from F. verticillioides and F. subglutinans. This protocol could detect the DNA from these fungi growing in artificially or naturally infected maize seeds. Another multiplex reaction with a pair of primers developed in this work combined with a pre-existing pair of primers has allowed identifying F. subglutinans, F. konzum, and F. thapsinum. In addition, the identification of F. nygamai was also possible using a combination of two PCR reactions described in this work, and another described in the literature.

Oxidation with galactose oxidase: Multifunctional enzymatic catalysis

“…Cordeiro subglutinans [20]. GAOB genes were also recently found from F. konzum, F. thapsinum, and F. nygamai [21].…”

The composition of Fusarium species in wheat husks and grains in south-eastern Poland

“…PCR is a commonly used technique for sensitive and rapid identification of Fusarium species. PCR can be an alternative or a complement to the identification based on morphological characteristics (Demeke et al 2005;Spanic et al 2010;Faria et al 2012;Suga et al 2013). The most frequently identified species by PCR are F. graminearum and F. culmorum (Kuzdraliński et al 2014).…”

Fungi Isolated from Maize (Zea mays L.) Grains and Production of Associated Enzyme Activities

“…The economic importance of maize is characterized by its high nutritional value; it is rich in starch, protein, triacylglycerol and fibers. Faria et al [ 20 ] have isolated fungi from maize grains presenting rotten symptoms. In view of the rot symptoms, it was assumed that these isolated fungi might produce hydrolases for degrading maize grain components.…”