The AquaCrop model was calibrated and validated for canola (Brassica napus) under Moistube irrigation (MTI) and various water regimes [(i) 100%, (ii) 75%, and (iii) 55% of crop water requirement (ETc)] over two seasons, 2019 and 2020. The normalised root mean square (nRMSE), Model Efficiency (EF), R2, and the Willmot’s index of agreement (d) statistics were used to evaluate the model’s efficiency in simulating biomass (B), canopy cover (CC), yield (Y), and harvest index (HI). The calibration results indicated the model simulated with accuracy the CC (under 100% ETcR2 = 0.99, EF = 0.92, nRMSE = 6.4%, d = 0.98) and 75% ETc (R2 = 0.99, EF = 0.92, nRMSE = 10.3%, d = 0.98). The model simulated CC well for validation for 100% ETc (R2 = 0.97, EF = 0.93, nRMSE = 22.5%, d = 0.98) and 75% ETc (R2 = 0.84, EF = 0.45, nRMSE = 59.2%, d = 0.86) irrigation regimes. Final biomass simulations were reasonably good under 100% ETc, 75% ETc, and 55% ETc irrigation regimes (R2 > 0.90, d > 0.65). The study showed the usefulness of AquaCrop for assessing yield response of canola to full and deficit irrigation scenarios under MTI.