Microwave observations are sensitive to vegetation water content (VWC). Consequently, the increasing temporal and spatial resolution of spaceborne microwave observations creates a unique opportunity to study vegetation water dynamics and its role in the diurnal water cycle. However, we currently have a limited understanding of sub-daily variations in the VWC and how they affect microwave observations. This is partly due to the challenges associated with measuring internal VWC for validation, particularly non-destructively, and at timescales of less than a day. In this study, we aimed to (1) use field sensors to reconstruct diurnal and continuous records of internal VWC of corn and (2) use these records to interpret the sub-daily behaviour of a 10 d time series of polarimetric L-band backscatter with high temporal resolution. Sub-daily variations in internal VWC were calculated based on the cumulative difference between estimated transpiration and sap flow rates at the base of the stems. Destructive samples were used to constrain the estimates and for validation. The inclusion of continuous surface canopy water estimates (dew or interception) and surface soil moisture allowed us to attribute hour-to-hour backscatter dynamics either to internal VWC, surface canopy water, or soil moisture variations. Our results showed that internal VWC varied by 10 %–20 % during the day in non-stressed conditions, and the effect on backscatter was significant. Diurnal variations in internal VWC and nocturnal dew formation affected vertically polarized backscatter most. Moreover, multiple linear regression suggested that the diurnal cycle of VWC on a typical dry day leads to a 2 (HH, horizontally, and cross-polarized) to almost 4 (VV, vertically, polarized) times higher diurnal backscatter variation than the soil moisture drydown does. These results demonstrate that radar observations have the potential to provide unprecedented insight into the role of vegetation water dynamics in land–atmosphere interactions at sub-daily timescales.