ABSTRACT: Osmotic stress is a major physiologic dysfunction that alters the water movement across the cell membrane. Soil salinity and water stress are major causal factors of osmotic stress that severely affect agricultural productivity and sustainability. Herein, we suggested and evaluated the impact of integrated biochar-based soil amendment and exogenous glycine betaine application on the growth, physiology, productivity, grain quality, and osmotic stress tolerance of rice (Oryza sativa L., cv. Sakha 105) grown in salt-affected soil under three irrigation intervals (6, 9, or 12 days), as well as soil properties and nutrient uptake under field conditions during the 2019 and 2020 seasons. Our findings showed that dual application of biochar and glycine betaine (biochar + glycine betaine) reduced the soil pH, electrical conductivity, and exchangeable sodium percentage. However, it enhanced the K+ uptake which increased in the leaves of treated-rice plants. Additionally, biochar and glycine betaine supplementation enhanced the photosynthetic pigments (chlorophyll a, b, and carotenoids) and physiological attributes (net photosynthetic rate, stomatal conductance, relative water content, and electrolyte leakage) of osmotic-stressed rice plants. Biochar + glycine betaine altered the activity of antioxidant-related enzymes (catalase, ascorbate peroxide, and peroxidase). Moreover, it improved the yield components, biological yield, and harvest index, as well as the nutrient value of rice grains of osmotic-stressed rice plants. Collectively, these findings underline the potential application of biochar and glycine betaine as a sustainable eco-friendly strategy to improve plant resilience, not only rice, but other plant species in general and other cereal crops in particular, to abiotic stress, particularly those growing in salt-affected soil.