To narrow the gap of agricultural water insufficiency in the Lam Takong River Basin, Thailand, we conducted an assessment of water availability and agricultural water demand under climate and land use changes. The water availability was estimated by SWAT, which was calibrated and validated during 2008–2012 and 2013–2018 against the observed daily discharge at the M.164 station. Measured and simulated discharges showed good agreement during calibration and validation, as indicated by values of 0.75 and 0.69 for R2 and 0.74 and 0.63 for Nash–Sutcliffe Efficiency, respectively. The results of GCMs (IPSL-CM5-MR, NorESM1-M, and CanESM2) under RCPs 4.5 and 8.5 were calculated to investigate changes in rainfall and temperature during 2020–2099. The warming tendencies of future maximum and minimum temperatures were projected as 0.018 and 0.022 °C/year and 0.038 and 0.045 °C/year under RCPs 4.5 and 8.5, respectively. The future rainfall was found to increase by 0.34 and 1.06 mm/year under RCPs 4.5 and 8.5, respectively. As compared to the 2017 baseline, the future planted areas of rice, maize, and cassava were projected to decrease during 2020–2099, while the sugarcane plantation area was expected to increase until 2079 and then decline. The top three greatest increases in future land use area were identified as residential and built-up land (in 2099), water bodies (in 2099), and other agricultural land (in 2059), while the three largest decrease rates were paddy fields (in 2099), forest land (in 2099), and orchards (during 2059–2079). Under the increased reservoir storage and future climate and land use changes, the maximum and minimum increases in annual discharge were 1.4 (RCP 8.5) and 0.1 million m3 (RCP 4.5) during 2060–2079. The sugarcane water demand calculated by CROPWAT was solely projected to increase from baseline to 2099 under RCP 4.5, while the increase for sugarcane and cassava was found for RCP 8.5. The future unmet water demand was found to increase under RCPs 4.5 and 8.5, and the highest deficits would take place in June and March during 2020–2039 and 2040–2099, respectively. In this context, it is remarkable that the obtained results are able to capture the continued and growing imbalance between water supply and agricultural demand exacerbated by future climatic and anthropogenic land use changes. This research contributes new insight for compiling a comprehensive set of actions to effectively build resilience and ensure future water sufficiency in the Lam Takong River Basin.