Fusarium species are common fungal pathogens of maize. Fusarium graminearum and Fusarium verticillioides, among others, can cause maize ear rot, and they are also mycotoxin producers. The aims of this work were to determine the frequency and diversity of Fusarium species in Uruguayan maize kernels, evaluate the toxigenic potential of the isolates, determine toxin contamination levels on freshly harvested grain, and assess the sensitivity of main Fusarium species against fungicides. Fusarium verticillioides was the most frequent species isolated, followed by Fusarium graminearum sensu stricto . Of F. verticillioides isolates studied for fumonisin production, 72% produced fumonisin B1 and 32% fumonisin B2. Considering in vitro toxin production by F. graminearum sensu stricto isolates, deoxynivalenol was the main toxin produced, followed by zearalenone and nivalenol. Fumonisins were the most frequently found toxins on freshly harvested maize samples (98% in 2018 and 86% in 2019), and also, fumonisin B1 was the toxin with highest concentration in both years studied (4860 µg/kg in 2018 and 1453 µg/kg in 2019). Deoxynivalenol and zearalenone were also found as contaminants. Metconazole and epoxiconazole were the most effective fungicides tested on F. verticillioides isolates. Fusarium graminearum sensu stricto isolates also were more sensitive to metconazole compared to other fungicides; nevertheless, epoxiconazole was less efficient in controlling this species. This is the first study that reports Fusarium species and mycotoxin contamination levels associated with maize grain in Uruguay. Its detection is the main step to develop management strategies in order to minimize fungal infection in maize crops. © 2023, The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.