While the direct impact of climate change on reference evapotranspiration (ET0) has been extensively studied, there is limited research on the indirect impact resulting from the interaction between climatic variables. This gap hinders a comprehensive understanding of climate change effects on ET0. Additionally, there is scarce exploration into the quantitative effect of freeze-thaw cycles on ET0 variation. In this study, we employed path analysis and dependent variable variance decomposition methods to discern the direct and interactive effects of climatic variables on ET0 in the Tibetan Plateau from 1960 to 2022. Annual ET0 exhibited variation across basins, with the coefficient of variability during the thawed period smaller than that during the non-thawed period. On an annual scale, the largest contribution to ET0 variation came from water vapor pressure deficit (VPD) at 47.7%. This contribution was amplified by its coupled interaction with temperature (T) at 47.1%, although the contribution was partially offset by the interactive effects of VPD with downward shortwave radiation and wind speed at −2.4% and − 27.6%, respectively. During different freezing-thawing periods, VPD primarily controlled ET0 variation, with its interaction with other climatic variables enhancing its impact. Furthermore, soil moisture, influenced by freeze-thaw cycles, exhibited a strong correlation with T and VPD, indicating the significant effect of freeze-thaw cycles on ET0 variation. The weak correlation between ET0 and NDVI suggested that vegetation growth had a limited regulatory effect on ET0. These findings provide valuable insights into the impact of interactions between climatic variables on hydrological processes, enhancing our understanding of the interactive roles of hydrometeorological variables.