New PCR assays for the identification of Fusarium verticillioides, Fusarium subglutinans, and other species of the Gibberella fujikuroi complex.
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.
Department of Biochemistry, State University of Maringá, Av. Colombo, 5790, 87020-900, Maringá, PR, Brazil; E-Mails:  cbertechini.faria@gmail.com  (C.B.F.);  camilaagneslumiabe@gmail.com  (C.A.L.A.).
Carla Bertechini
CB
Faria
Camila Agnes Lumi
CA
Abe
Cleiltan Novais
CN
da Silva
Dauri José
DJ
Tessmann
Ione Parra
IP
Barbosa-Tessmann
D017931:DNA Primers / Q000235:genetics / Q000378:metabolism; D004271:DNA, Fungal / Q000032:analysis* / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics*; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics; D005692:Galactose Oxidase / Q000235:genetics*; D016681:Genome, Fungal; D005874:Gibberella / Q000201:enzymology* / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D003313:Zea mays / Q000382:microbiology
D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D005692:Galactose Oxidase