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Ecosystem-scale water-use efficiency (WUE) is an important indicator for understanding the intimately coupled relationship between carbon and water cycles in ecosystems. Previous studies have suggested that both abiotic and biotic factors have significant effects on WUE in forest ecosystems. However, responses of WUE to phenology in the context of climate change remain poorly understood. In this study, we analyzed the sensitivity and response patterns of seasonal WUE to phenology in Zhejiang Province where typical subtropical forest ecosystems are located, and discussed potential causes of the changes of the sensitivity and response patterns along different climate gradient during 2000–2014. The results of interannual partial correlation analysis showed widespread negative correlations between WUE and the start of growing season (SOS) in spring. This is because the increase in gross primary product (GPP) is larger than that of evapotranspiration (ET), resulting from an advanced SOS. The positive correlation between WUE and SOS was widely observed in summer mainly because of water stress and plant ecological strategy. The autumn WUE enhanced with the delay in the end of growing season (EOS) mainly because of the increase in GPP meanwhile the decrease or steadiness in ET, resulting from a delayed EOS. In space, the sensitivity of spring WUE to SOS significantly decreased along the radiation gradient, which might be related to strong soil evaporation in high radiation area; the sensitivity of WUE to SOS in summer showed a positive correlation with precipitation and a negative correlation with temperature, respectively, which might be attributed to the compensation of GPP to the delayed SOS and water stress caused by high temperature. The sensitivity of WUE to EOS increased significantly along the radiation and precipitation gradients in autumn, which may be because the increase of radiation and precipitation provides more water and energy for photosynthesis. [ABSTRACT FROM AUTHOR]