AbstractAlternaria is an important toxigenic fungal genus endangering the quality of cereal grains. No data are available in the literature regarding the composition of Alternaria species in wheat from the main wheat‐cultivating regions of China. In this study, a total of 282 strains were isolated from wheat kernels collected in various provinces. Phylogenetic characterization was performed using two informative loci, ASA10 and ASA19, demonstrating that all strains belonged to A. alternata. A low molecular diversity was observed, which did not appear to be influenced by geographic origin. The secondary metabolite profile of the strains was evaluated in rice culture. Altertoxin I was synthesized at a high concentration by the majority of the strains, while the accumulation of altenuene was lower in both frequency and level than that of alternariol and its derivative. A. alternata did not cause readily observable symptoms on wheat spikes after spraying of the conidia at anthesis, but a higher quantity of altertoxin I and Alternaria DNA were detected in inoculated grains. Artificial inoculations of apple fruit showed that all examined strains were capable of causing typical rot symptoms and producing all four toxins to a certain degree. The widespread occurrence of A. alternata on wheat grains and its ability to synthesize various toxic metabolites may pose a serious hazard for both growers and consumers.