PCR Identification of Fusarium graminearum Isolated from Wheat Grain
Species-specific PCR assay was used for the identification of Hungarian Fusarium graminearum isolates in pure mycelial culture. The Fg16F/Fg16R primer pair of the three known species-specific primers appeared to be the most appropriate one to identify F. graminearum.
Two methods were used for comparative determination of the amplicon size of F. graminearum strains: traditional agarose gel electrophoresis, and chip electrophoresis. Our results have shown that the chip electrophoresis is an easy-to-use, time-efficient substitute for conventional agarose gel electrophoresis; moreover it provides a more precise size determination of amplicons. Amplicon size ranging from 415 bp to 421 bp in tested isolates may be associated with genetic diversity in the Hungarian population of F. graminearum.
The PCR assay described in this study can be used for the routine detection and identification of F. graminearum without isolation and morphological investigation of this fungus.
KeywordsFusarium graminearum PCR species-specific primers chip electrophoresis wheat grain
Unable to display preview. Download preview PDF.
- Agilent 2100 Bioanalyzer. Application Compendium. 2005. Agilent Technologies. Printed in Germany, 95 pp.Google Scholar
- Burgess, L.W., Summerell, B.A., Bullock, S., Backhouse, D. 1994. Laboratory Manual for Fusarium Research. Third Edition, Sydney. 133 pp.Google Scholar
- Desjardins, A.E. 2006. Fusarium Mycotoxins. Chemistry, Genetics and Biology. APS Press, St. Paul, Minesota, USA. 260 pp.Google Scholar
- Miller, J.D., Trenholm, H.L. 1994. Mycotoxins in Grain. Compounds other than Aflatoxin. Eagon Press, St. Paul, Minesota, USA. 552 pp.Google Scholar
- O’Donnell, K., Ward, T.J., Geiser, D.M., Kistler, H.C., Aoki, T. 2004. Genealogical concordance between the mating type locus and seven other nuclear genes supports formal recognition of nine phylogenetically distinct species within the Fusarium graminearum clade. Fungal Genetics and Biology 41:600–623.CrossRefGoogle Scholar
- Szécsi, Á., Mesterházy, Á. 1998. A medium for selective isolation and identification of Fusarium spp. from cereal grains and maize kernels. Acta Phytopathologica et Entomologica Hungarica 33:79–87.Google Scholar