Improved accuracy of evapotranspiration (ET) estimation, including its partitioning between transpiration (T) and surface evaporation (E), is key to monitor agricultural water use in vineyards, especially to enhance water use efficiency in semi-arid regions such as California, USA. Remote-sensing methods have shown great utility in retrieving ET from surface energy balance models based on thermal infrared data. Notably, the two-source energy balance (TSEB) has been widely and robustly applied in numerous landscapes, including vineyards. However, vineyards add an additional complexity where the landscape is essentially made up of two distinct zones: the grapevine and the interrow, which is often seasonally covered by an herbaceous cover crop. Therefore, it becomes more complex to disentangle the various contributions of the different vegetation elements to total ET, especially through TSEB, which assumes a single vegetation source over a soil layer. As such, a remote-sensing-based three-source energy balance (3SEB) model, which essentially adds a vegetation source to TSEB, was applied in an experimental vineyard located in California’s Central Valley to investigate whether it improves the depiction of the grapevine-interrow system. The model was applied in four different blocks in 2019 and 2020, where each block had an eddy-covariance (EC) tower collecting continuous flux, radiometric, and meteorological measurements. 3SEB’s latent and sensible heat flux retrievals were accurate with an overall RMSD ~ 50 W/m2 compared to EC measurements. 3SEB improved upon TSEB simulations, with the largest differences being concentrated in the spring season, when there is greater mixing between grapevine foliage and the cover crop. Additionally, 3SEB’s modeled ET partitioning (T/ET) compared well against an EC T/ET retrieval method, being only slightly underestimated. Overall, these promising results indicate 3SEB can be of great utility to vineyard irrigation management, especially to improve T/ET estimations and to quantify the contribution of the cover crop to ET. Improved knowledge of T/ET can enhance grapevine water stress detection to support irrigation and water resource management.