title: A new PCR approach for the identification of Fusarium graminearum.
abstract: The main objective of this work was to develop a PCR protocol for the identification of Fusarium graminearum, based on a pair of primers targeted to a segment of the 3´coding region of the gaoA gene that codes for the enzyme galactose oxidase (GO). This region has low homology with the same region of GO genes from other fungi. Genomic DNA from 17 strains of Fusarium spp. isolated from diseased cereals, from several other Fusarium species, and from other fungi genera was analyzed in a PCR assay using this primer set. The 17 strains of Fusarium spp. were also analyzed for the GO enzyme production in submerse fermentation in a new formulated liquid medium. All strains that were morphologically and molecularly identified as F. graminearum were able to secrete the enzyme and had a positive result in the used PCR protocol. No DNA fragment was amplified using genomic DNA from other Fusarium species and species of other fungi genera. The results suggest that the proposed PCR protocol is specific and can be considered as a new molecular tool for the identification of F. graminearum. In addition, the new formulated medium is a cheap alternative for screening for GO screening production by F. graminearum.
authors.affiliation: Departamento de Bioquímica, Universidade Estadual de Maringá , Maringá, PR , Brasil.
authors.forename: Gleison Ricardo
authors.initials: GR
authors.lastname: de Biazio
authors.forename: Gabriela Guimarães Sousa
authors.initials: GG
authors.lastname: Leite
authors.forename: Dauri José
authors.initials: DJ
authors.lastname: Tessmann
authors.forename: Ione Parra
authors.initials: IP
authors.lastname: Barbosa-Tessmann
keywords: Fusarium graminearum; galactose oxidase; gaoA gene; molecular identification
references.citation: Curr Genet. 1992 Sep;22(3):213-20
references.pmid: 1525873
references.citation: J Bacteriol. 1977 Apr;130(1):455-63
references.pmid: 15985
references.citation: J Histochem Cytochem. 1983 Mar;31(3):391-403
references.pmid: 6186733
references.citation: Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):12932-7
references.pmid: 11698678
references.citation: Appl Microbiol. 1965 Sep;13(5):686-93
references.pmid: 5867649
references.citation: Methods Enzymol. 1982;89 Pt D:163-71
references.pmid: 6890617
references.citation: Phytopathology. 1997 Sep;87(9):915-23
references.pmid: 18945062
references.citation: J Basic Microbiol. 2001;41(3-4):143-8
references.pmid: 11512446
references.citation: Adv Protein Chem. 2002;60:1-49
references.pmid: 12418174
references.citation: Curr Opin Chem Biol. 2003 Apr;7(2):189-96
references.pmid: 12714051
references.citation: Ann N Y Acad Sci. 1990;613:385-9
references.pmid: 2075984
references.citation: Plant Dis. 1997 Dec;81(12):1340-1348
references.pmid: 30861784
references.citation: Anticancer Res. 1996 Jul-Aug;16(4B):2193-9
references.pmid: 8694542
references.citation: J Biol Chem. 1992 Apr 25;267(12):8146-52
references.pmid: 1569070