title: Geneticin (G418) resistance and electroporation-mediated transformation ofFusarium graminearumandF. culmorum.
abstract: Fusarium graminearumandF. culmorumare phytopathogenic species causing scab and root rot diseases in all small grain cereals worldwide including Turkey. In this study, resistance levels to geneticin (G418) of 14F. graminearumand 24F. culmorumisolates collected from cereals were determined. Fungal cultures were grown on potato dextrose agar medium supplemented with 0, 25, 50, 75 and 100 µg/mL of G418. Minimum inhibitory concentration was determined as 25 µg/mL. As a result, it was concluded that all isolates were highly sensitive to G418. Plasmid pFA6-kanmx4 containing geneticin resistance gene (kanmx) was introduced singly or co-electroporated with pEGFP75 plasmid, containing GFP gene, into fungal protoplast cultures obtained with lytic enzyme. Transformants were grown in media including 25 µg/mL G418. Transformation frequencies were 2.8 and 1.8 transformant per µg plasmid forF. graminearumandF. culmorumisolates, respectively. Transformation process was also confirmed by spectrofluorimetric assay. Relative fluorescence unit values in co-transformants were calculated as 1.87 ± 0.04 forF. graminearumand 2.26 ± 0.08 forF. culmorum. The results obtained from the study gave information about antibiotic resistance levels of twoFusariumspecies in Turkey. Moreover, it was shown that pFA6-kanmx4 plasmid was a suitable vector, which can be used in genetic manipulation studies of these two fungal species in particular suppression of endogenous and/or the expression of exogenous genes.
authors.affiliation: Programme of Molecular Biology and Genetics, Institute of Science, Istanbul University , 34134 Vezneciler , Istanbul , Turkey.
authors.forename: Emre
authors.initials: E
authors.lastname: Yörük
authors.affiliation: Department of Molecular Biology and Genetics, Faculty of Science, Istanbul University , 34134 Vezneciler , Istanbul , Turkey.
authors.forename: Gülruh
authors.initials: G
authors.lastname: Albayrak
keywords: Fusarium culmorum; Fusarium graminearum; electroporation; geneticin (G418); pEGFP75; pFA6-kanmx4; transformation