Crop wild resources are excellent sources of new genetic variation for resilience against climate extremes. However, detailed characterization of the desirable phenotypes is essential before using these crop wild resources in breeding programs. This current study was, therefore, conducted to investigate the water stress responses of eight wild species and two wheat cultivars. The experiment was carried out under varying levels of osmotic stress induced by polyethylene glycol and progressive water stress through different field capacities. Water stress significantly reduced both physiological and biochemical traits compared to control, ranging from 7.1% (protein content) to 34.5% (chlorophyll) under moderate stress and 9.1-45.8% under severe stress. The anatomical features were also affected under progressive water stress, including a reduction in xylem vessel diameter (7.92 and 16.50%), phloem length (4.36 and 7.18%), vascular bundle length (3.09 and 6.04%), and ground tissue thickness (2.36 and 5.52%), respectively. Conclusively, (endemic to Swat, Pakistan). . . , and . outperformed the check wheat cultivar, Pirsabak 2005, which is a rainfed variety. The results revealed that species belonging to the tertiary gene pool of bread wheat could be an excellent drought tolerance source for use in a breeding program.