Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error
2008 | journal article. A publication with affiliation to the University of Göttingen.
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Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error
Brandfass, C. & Karlovsky, P. (2008)
International Journal of Molecular Sciences, 9(11) pp. 2306-2321. DOI: https://doi.org/10.3390/ijms9112306
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- Authors
- Brandfass, Christoph; Karlovsky, Petr
- Abstract
- Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W. G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in plant tissue. We demonstrated that increasing the amount of plant material used for DNA extraction to 0.5-1.0 g considerably reduced sampling error and improved the reproducibility of DNA yield. The costs of DNA extraction at different scales and with different methods (commercial kits versus cetyltrimethylammonium bromide-based protocol) and qPCR systems (doubly labeled hybridization probes versus SYBR Green) were compared. A cost-effective protocol for the quantification of F. graminearum and F. culmorum DNA in wheat grain and maize stalk debris based on DNA extraction from 0.5-1.0 g material and real-time PCR with SYBR Green fluorescence detection was developed.
- Issue Date
- 2008
- Journal
- International Journal of Molecular Sciences
- Organization
- Fakultät für Agrarwissenschaften ; Department für Nutzpflanzenwissenschaften ; Abteilung Molekulare Phytopathologie und Mykotoxinforschung
- ISSN
- 1661-6596