Vegetation Optical Depth (VOD) has emerged as a valuable metric to quantify water stress on vegetation's carbon uptake from a remote sensing perspective. However, existing spaceborne microwave remote sensing platforms face limitations in capturing the diurnal VOD variations and global products lack site‐level validation against plant physiology. To address these challenges, we leveraged the Global Navigation Satellite System (GNSS) L‐band microwave signal, measuring its attenuation by the canopy of a temperate broadleaf forest using a pair of GNSS receivers. This approach allowed us to collect continuous VOD observations at a sub‐hourly scale. We found a significant seasonal‐scale correlation between VOD and leaf water potential. The VOD diurnal amplitude is affected by soil moisture, plant transpiration and leaf surface water. Additionally, VOD can help independently estimate plant transpiration. Our findings pave the way for a deeper understanding of response of the vegetation to water stress at finer temporal scales. Plain Language Summary Microwave satellite measurements offer valuable insights into Vegetation Optical Depth (VOD), which reflects the amount of water present in plants. However, most VOD products currently available only offer observations at two times of the day, which fail to capture the variations of VOD over the course of the day. In this study, we installed Global Navigation Satellite System (GNSS) signal receivers at the top and bottom of a temperate broadleaf forest canopy located at a flux tower in Missouri, USA. This setup allowed us to obtain continuous observations of VOD at sub‐hourly scale. The strong correlation observed between GNSS VOD and local leaf water potential suggests that this robust and cost‐effective technique can complement other labor‐intensive measurements. The VOD diurnal amplitude is influenced by soil water content, evapotranspiration, and leaf surface water in the form of dew and interception. GNSS VOD also exhibits the potential to aid in estimating plant transpiration, which can be further used for partitioning evapotranspiration. The investigation of plant water content dynamics in this study will enhance our understanding of the carbon‐water coupling at a finer temporal scale.