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Evapotranspiration (ET) represents the largest water loss flux in drylands, but ET and its partition into plant transpiration (T) and soil evaporation (E) are poorly quantified, especially at fine temporal scales. Physically-based remote sensing models relying on sensible heat flux estimates, like the two-source energy balance model, could benefit from considering more explicitly the key effect of stomatal regulation on dryland ET. The objective of this study is to assess the value of solar-induced chlorophyll fluorescence (SIF), a proxy for photosynthesis, to constrain the canopy conductance (Gc) of an optimal stomatal model within a two-source energy balance model in drylands. We assessed our ET estimation using in situ eddy covariance GPP as a benchmark, and compared with results from using the Contiguous solar-induced chlorophyll fluorescence (CSIF) remote sensing product instead of GPP, with and without the effect of root-zone soil moisture on the Gc. The estimated ET was robust across four steppes and two tree-grass dryland ecosystem. Comparison of ET simulated against in situ GPP yielded an average R2 of 0.73 (0.86) and RMSE of 0.031 (0.36) mm at half-hourly (daily) timescale. Including explicitly the soil moisture effect on Gc, increased the R2 to 0.76 (0.89). For the CSIF model, the average R2 for ET estimates also improved when including the effect of soil moisture: from 0.65 (0.79) to 0.71 (0.84), with RMSE ranging between 0.023 (0.22) and 0.043 (0.54) mm depending on the site. Our results demonstrate the capacity of SIF to estimate subdaily and daily ET fluxes under very low ET conditions. SIF can provide effective vegetation signals to constrain stomatal conductance and partition ET into T and E in drylands. This approach could be extended for regional estimates using remote sensing SIF estimates such as CSIF, TROPOMI-SIF, or the upcoming FLEX mission, among others.