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Sumikova T., Gabrielova L., Kucera L., Žabka M., Chrpova J. (2013): Mycotoxin production, chemotypes and diversity of Czech Fusarium graminearum isolates on wheat. Czech J. Food Sci., 31: 407–412. Fusarium head blight (FHB) is a serious cereal disease in the Czech Republic. The most important pathogen associ ated with FBH is Fusarium graminearum , which can produce trichothecenes, mainly deoxynivalenol (DON) and its derivates. A set of 103 F. graminearum isolates were isolated from naturally infected wheat ears collected from 20 localities (25 ears from one locality) within the Czech Republic, in the year 2004. The ears were evaluated for DON content by ELISA. Maximum detected value was 30.7 mg/kg. A group of PCR assays targeting the segments of the Tri7 , Tri13 and Tri3 genes were used to determine the chemotypes of F. graminearum isolates. All the isolates belonged to DON producing chemotype. Further discrimination revealed that almost all (99.03%) isolates belonged to 15-ADON chemotype, and only one (0.97%) isolate belonged to 3-ADON chemotype. The genetic variability of the isolates was assessed from their AFLP fingerprints. The populations were highly heterogeneous both within and between locations, and no clear evidence for the association between AFLP profile and geographic origin was found out.