Isotope-Based Techniques to Investigate Factors Influencing Water Use Efficiency in Leaves during Plant Growth.

Plant water use efficiency (WUE) is a comprehensive physiological indicator of plant growth and ability to adapt to drought. However, research on the mechanisms controlling WUE during plant growth and development remains weak. Here, we studied as a typical evergreen conifer species in Northeast China. After collecting 80 tree samples with varying diameters at breast height (DBH), we measured δC and δO as an indicator of WUE, leaf morphology (volume, dry weight, and total epidermal area), ecological stoichiometry (carbon, nitrogen, and phosphorus content), and abiotic factors (light environment, soil pH, soil water content, and soil nutrient content). Correlational analysis of these variables revealed distinct differences between smaller/younger and larger/older plants: (1) In plants with DBH less than 52 cm, δC was positively related to DBH, and δO was negatively related to DBH. Plants with DBH greater than 52 cm showed no relationship between δC and DBH, and δO was positively related to DBH. (2) In plants with DBH less than 52 cm, there was a negative correlation between δC and δO and between δC and leaf phosphorus content (LP), but a positive correlation between δC and DBH, leaf mass per area (LMA), and leaf density (LD). The slopes of DBH-δC, δO-δC, leaf nitrogen content (LN)-δC, and LMA-δC correlations were greater in smaller plants than large plants. (3) Structural equation modelling showed that in smaller plants, DBH had a direct positive effect on δC content and a direct negative effect on δO, and there was a direct positive effect of light environment on δO. In larger plants, there was a direct negative effect of light environment on δC and a direct positive effect of DBH on light environment, as well as a negative effect of soil nitrogen content on leaf nitrogen. In smaller plants, DBH was the most important factor influencing δC, followed by δO and soil moisture, with light and soil pH showing minimal influence. In larger plants, light environment influenced δC the most, followed by soil nitrogen content and soil moisture content, with leaf nitrogen and DBH contributing little. The results suggest that water use efficiency strategies of vary according to growth stage, and the effects of abiotic factors and functional traits vary at different growth stages.