upscaled ctab-based dna extraction and real-time pcr assays for fusarium culmorum and f. graminearum dna in plant material with reduced sampling error高档ctab-based dna提取和实时pcr检测枯萎culmorum和graminearum dna在植物材料减少抽样误差.pdf

Int. J. Mol. Sci. 2008, 9, 2306-2321; DOI: 10.3390/ijms9112306 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms/ Article Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error Christoph Brandfass and Petr Karlovsky * University of Göttingen, Department of Crop Sciences, Molecular Phytopathology and Mycotoxin Research Division, Grisebachstrasse 6, 37077 Göttingen, Germany * Author to whom correspondence should be addressed; E-Mail: pkarlov@gwdg.de; Tel. +49-551-39-12918; Fax: +49-551-39-12919 Received: 28 August 2008; in revised form: 19 November 2008 / Accepted: 24 November 2008 / Published: 25 November 2008 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