Wheat contains various phytochemicals, most importantly, antioxidants and polyphenolic compounds that have a variety of health-promoting effects. Seed material was obtained from 63 wheat genotypes of three species (T. aestivum, T. durum, and T. sphaerococcum) from Pakistan and Syria. This study was initiated to estimate and compare the interspecific and intraspecific diversity for antioxidant activity (AOA) and total phenolic content (TPC) in different species of wheat. The wild relatives and landraces of three wheat species exhibited a highly significant interspecific and intraspecific diversity for both traits. T. aestivum exhibited more intraspecific diversity. The AOA of the genotypes ranged from 7.45% to 41.68%, being maximum in accessions of T. durum 12977 (41.68%). TPC ranged from 10.09 to 39.28 mg GAE g^-1 and was highest in T. durum accession 12999 (39.28 mg GAE g^-1). The lowest AOA (7.45%) and TPC (10.09 mg GAE g^-1) were observed in T. aestivum landrace LR-27. Hence, T. durum accessions 12999 and 12979 could be our desired accessions for future breeding programs having phytochemicals. Cluster analysis distributed the genotypes into four clusters. Genotypes of different origins grouped differently, indicating an environmental influence in the development of their genetic architecture. Cluster analysis indicated that 41.30% of the genotypes were included in the high AOA and TPC group. Members of T. aestivum that grouped in clusters 1 and 2 showed low to moderate AOA and TPC. The accessions of T. durum and T. sphaerococcum performed much better than T. aestivum for both of the biochemical traits.