Water supply and valorization are two urgent issues in the utilization of maize biomass in the context of climate change and replacement of fossil resources. Maximizing maize biomass valorization is of interest to make biofuel conversion competitive, and to increase forage energetic value for animal fodder. One way to estimate biomass valorization is to quantify cell wall degradability. In this study, we evaluated the impact of water supply on cell wall degradability, cell wall contents and structure, and distribution of lignified cell types in maize internodes using dedicated high-throughput tools to effectively phenotype maize internodes from 11 inbred lines under two contrasting irrigation scenarios in field trials over three years. Overall, our results clearly showed that water deficit induced significant changes in lignin content and distribution along with a reduction in lignin p-coumaroylation, thereby impacting cell wall degradability. Additionally, we also observed that responses to a water deficit vaned between the lines examined, underscoring biochemical and histological target traits for plant breeding.