Tree-ring intra-annual stable isotopes (delta C-13 and delta O-18) are powerful tools for revealing plant ecophysiological responses to climatic extremes. We analyzed interannual and fine-scale intra-annual variability of tree-ring delta C-13 and delta O-18 in Chinese red pine (Pinus massoniana) from southeastern China to explore environmental drivers and potential trade-offs between the main physiological controls. We show that wet season relative humidity (May-October RH) drove interannual variability of delta O-18 and intra-annual variability of tree-ring delta O-18. It also drove intra-annual variability of tree-ring delta C-13, whereas interannual variability was mainly controlled by February-May temperature and September-October RH. Furthermore, intra-annual tree-ring delta O-18 variability was larger during wet years compared with dry years, whereas delta C-13 variability was lower during wet years compared with dry years. As a result of these differences in intra-annual variability amplitude, process-based models (we used the Roden model for delta O-18 and the Farquhar model for delta C-13) captured the intra-annual delta O-18 pattern better in wet years compared with dry years, whereas intra-annual delta C-13 pattern was better simulated in dry years compared with wet years. This result suggests a potential asymmetric bias in process-based models in capturing the interplay of the different mechanistic processes (i.e., isotopic source and leaf-level enrichment) operating in dry versus wet years. We therefore propose an intra-annual conceptual model considering a dynamic trade-off between the isotopic source and leaf-level enrichment in different tree-ring parts to understand how climate and ecophysiological processes drive intra-annual tree-ring stable isotopic variability under humid climate conditions.

Understanding varying climate responses in tree-ring data across tree ages is important, but little is known about tree-age effects on climate responses in tree-ring stable isotopes. To detect whether age differences in tree-ring delta C-13 and delta O-18 could lead to differing climate responses, we measured tree-ring cellulose delta C-13 and delta O-18 (1901-2010) from Schrenk spruce (Picea schrenkiana) trees in northwestern China with ages ranging from 110 to 470 years, which we binned into three age groups. Tree-ring delta C-13 (pin-corrected) and delta O-18 exhibited similar year-to-year variability between age groups and did not feature age-related trends. delta C-13 series from old trees (270-470 years) showed stronger legacy effects, reflecting influences from the antecedent period (due to carbohydrate reserves and climate), compared to young trees (110-125 years). Both tree-ring delta C-13 and delta O-18 values decreased with increasing relative humidity (RH) and precipitation and with decreasing mean and maximum temperatures during the main growing season (May-August). delta C-13 and delta O-18 exhibited age-dependent climate responses: Young trees had a stronger climate response in delta C-13 but a weaker or similar climate response in delta O-18 compared to old trees. We developed multiple growing-season RH reconstructions based on composite chronologies using delta C-13 and delta O-18 series from different age groups. In particular, we found that including delta C-13 from young trees improved the skill of RH reconstructions because of the age-specific mechanisms driving the delta C-13-climate relationship, but that caution is warranted with regard to extreme values. We therefore suggest that young trees should be considered when using stable isotopes, particularly in delta C-13, for climate reconstruction.