Researching rapid multiplication techniques for mosaic-resistant cassava in tunnel greenhouses is crucial for ensuring food security and sustainable agriculture. Our previous findings indicated that a combination of 50% shading and 70-75% humidity significantly enhanced the formation of shoots and growth of cassava seedlings in a tunnel greenhouse. The current study examines the impact of different substrate compositions on cassava propagation. The experiments were designed as split plots with three replicates and four formulas including CTRL: 100% sand with a thickness of 40 cm; CT-1: 100% sand with a thickness of 30 cm; CT-2: 60% sand and 40% gravel with a thickness of 40 cm; CT-3: 70% sand and 30% gravel with a thickness of 40 cm; and CT-4: 50% sand and 50% gravel with a thickness of 40 cm. The results demonstrated that CT-3 yielded the highest shoot formation rate and growth of cassava seedlings, suggesting that the combination of sand (70%) and gravel (30%) at a thickness of 40 cm provide the most favorable conditions for cassava propagation in tunnel greenhouse conditions.