AbstractThe last few years have been a huge challenge for every farmer in Europe and Hungary because of the increase in hot days and the decrease in precipitation. These facts induced the farmer’s interest in sorghum because it has better stress tolerance than many other cereales, but little is known about mycotoxin-producing fungi, which can infect this crop. Mycotoxins are secondary metabolites of filamentous fungi, and they are not only phytotoxic but also harmful to humans and animals. This study aimed to determine the internal infection caused by Alternaria spp., Fusarium spp., and Aspergillus spp., which are known as mycotoxin-producing fungi in food raw material. In our study in the case of sorghum, the presence of various mycotoxin-producing fungi was detected on Fusarium selective media (Nash and Snyder media), and the results indicate that these genera are present in sorghum grains as potential mycotoxin producers. To determine which Fusarium species is occurring in our sorghum grain samples, a molecular genetic study was performed on isolated fungi using the TEF region primer pairs to identify the occurring Fusarium species. We mainly identified Fusarium proliferatum in our conventional sorghum samples. The results show that the mentioned mycotoxin-producing fungi are in the sorghum grains and may pose a risk to the safety of feed and food because they may produce mycotoxins in the field or while being stored.