Drought is a common abiotic stress that considerably limits crop production. The objective of this study is to explore the influence of water deficiency on the yield and physiologic-biochemical in upland cotton cultivars (Gossypium hirsutum L). Cotton cultivars, 'Ishonch', 'Navbakhor-2','C-6524' and 'Tashkent-6' were selected to study the relationships among their physiologic, biochemical and yield attributes during water deficiency. Deficit irrigation was designed by modifying the traditional watering protocol to reduce water use. Results indicate that cotton cultivars respond differently to water deficit stress. Water deficit significantly influenced plant valuable economic traits and yield in four cultivars. However, yield components such as the number of bolls per plant, seed number per boll, mass of boll per plant and productivity plant yield were significantly reduced only in and C-6524 and Tashkent-6. The malondialdehyde decreased and total chlorophyll, chlorophyll "a" increased in 'Ishonch' and Navbakhar-2 under deficit irrigation conditions. The direct relationship between physiology, biochemistry, valuable economic traits, and yield may be a useful selection criterion for determining candidate parents for cotton drought tolerance breeding.