Fusarium head blight (FHB) in wheat and triticale leads to contamination of the grain with the mycotoxin deoxynivalenol (DON) that is harmful to animal and man. A fast, low-cost, and reliable method for quantification of the DON content in the grain is essential for selection. We analysed 113 wheat and 55 triticale genotypes for their symptom development on spikes, Fusarium exoantigen (ExAg) and DON content in the grain after artificial inoculation with a highly aggressive isolate of F. culmorum in three (wheat) and six (triticale) location-by-year combinations. Additionally, in triticale the amount of Fusarium damaged kernels (FDK) was assessed. ExAg content was analysed by a newly developed Fusarium-specific plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) and DON content by an immunoassay. A moderate disease severity resulted in an ExAg content of 0.87 optical density (OD) units in wheat and 1.02 OD in triticale. DON content ranged from 12.0 to 105.2 mg kg −1 in wheat and from 24.2 to 74.0 mg kg −1 in triticale. Genotypic and genotype-by-environment interaction variances were significant (P < 0.01). Coefficient of phenotypic correlation between DON content analysed by the immunoassay and ExAg content was r = 0.86 for wheat and r = 0.60 for triticale. The highest correlation between DON content and symptom rating was found by FHB rating in wheat (r = 0.77) and by FDK rating in triticale (r = 0.71). In conclusion, selection for reduced FHB symptoms should lead to a correlated selection response in low fungal biomass and low DON content in the grain.
Aims: The objective of this study was to evaluate the capability of Fusarium culmorum to produce non-hydrophobin surface-active proteins in vitro, to isolate and characterize such proteins from liquid cultures, to analyse their effect on overfoaming (gushing) of beer and to elucidate their prevalence in pure cultures and infected malt. Methods and Results: A 20 kDa protein was isolated from liquid cultures of F. culmorum BBA 62182 upon enrichment by foaming. BLAST search with Nterminal and internal sequences of the protein revealed high homology with a hypothetical protein predicted within the F. graminearum PH1 genome sequence. Oligonucleotide primers designed to bind 30 nt upstream and downstream of the predicted gene were used to amplify a 695 nt PCR fragment from genomic DNA of F. culmorum BBA 62182. Cloning and sequencing of the product revealed a 635 nt open reading frame which had 98% homology to the predicted F. graminearium PH1 gene code. Removal of a 59 nt intron and translation resulted in a 191 amino acid protein of 20AE754 kDa with a calculated pI of 9AE1. Amino acids obtained by Edman sequencing of fragments within the 20 kDa protein were 100% homologous with the sequence deduced from the DNA sequence. According to its properties, the new protein was termed alkaline foam protein A (AfpA). Sequence comparison revealed some homologies with proteins in Emericella nidulans, which are involved in phialide development and response to antifungal agents. Homologies with other hypothetical fungal proteins suggest a new group of proteins, for which we suggest the name fungispumins. Addition of AfpA to beer showed that overfoaming (gushing) is not induced in stable beer but can significantly enhance this effect in beer showing moderate gushing.
Plant pathogenic fungi of the genusFusarium cause agriculturally important diseases of small grain cereals and maize. Especially the contamination of grains with the mycotoxin deoxynivalenol (DON) is harmful for animals and humans. For quantification of the severity ofFusarium head blight (FHB) in resistance evaluation and selection of cereals a fast, economical and reliable method is essential. Immunological methods appear to be particularly suitable for such an assessment. The results demonstrate that a selected antiserum was appropriate for the detection ofFusarium-exoantigens (ExoAg) in cereal grains by an indirect ELISA-format, although a discrimination between variousFusarium-species was not possible. The polyclonal antibody detection system was optimized for different parameters and a standard test protocol was elaborated.
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