The construction materials are emitting carbon during their production and usage. With an emphasis on the effect of bamboo knot spacing on flexural strength, this study investigates the possibility of employing bamboo as a sustainable substitute for steel reinforcement in concrete slabs. By conducting a comprehensive experimental program, bamboo with varying knot spacings was used to reinforce concrete slabs, and standardized tests were performed to determine their flexural strength. The analysis of test data revealed patterns and correlations between knot spacing and flexural performance. To take into consideration the uncertainties in knot spacing and material properties, sophisticated statistical techniques and reliability analysis were used. The results demonstrated that bamboo-reinforced slabs possess significant flexural strength. The flexural strength increased by 37.086% as the knot spacing increased from 250 mm to 330 mm. The bamboo slab panels considered for this study were found to be 45.012% more energy-efficient and 19.18% more economical compared to conventional RC slabs of similar dimensions.