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.
Highlights
Fusarium head blight (FHB) is a disease of cereal crops with a severe impact on wheat and barley production worldwide
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
We started with samples of 100 mg wheat flour in 1 mL CTAB buffer using a protocol developed for agar plaques [26] and observed a large variation among replicas
Summary
Fusarium head blight (FHB) is a disease of cereal crops with a severe impact on wheat and barley production worldwide. The infection of heads of small grain cereals and maize plants with Fusarium spp. impairs both grain yield and quality [1,2]. Apart from adversely effecting the grain size, weight, protein content, baking quality of the flour and other technological parameters, a serious consequence of FHB is the contamination of grain and cereal products with Fusarium mycotoxins [3]. Because grains of low quality are used in feedstuff production rather than for human foods, health damage in farm animals and losses in meat production caused by mycotoxin contamination of feeds have regularly been reported [4].
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