Chlorophyll meters are widely used to guide nitrogen (N) management by monitoring leaf N status in agricultural systems, but the effects of environmental factors and leaf characteristics on leaf N estimations are still unclear. In the present study, we estimated the relationships among SPAD readings, chlorophyll content and leaf N content per leaf area for seven species grown in multiple environments. There were similar relationships between SPAD readings and chlorophyll content per leaf area for the species groups, but the relationship between chlorophyll content and leaf N content per leaf area and the relationship between SPAD readings and leaf N content per leaf area varied widely among the species groups. A significant impact of light-dependent chloroplast movement on SPAD readings was observed under low leaf N supplementation in both rice and soybean but not under high N supplementation. Furthermore, the allocation of leaf N to chlorophyll was strongly influenced by short-term changes in growth light. We demonstrate that the relationship between SPAD readings and leaf N content per leaf area is profoundly affected by environmental factors and leaf features of crop species, which should be accounted for when using a chlorophyll meter to guide N management in agricultural systems.

Stomatal conductance (g_s) and mesophyll conductance (g_m) represent major constraints to photosynthetic rate (A), and these traits are expected to coordinate with leaf hydraulic conductance (K_leaf) across species, under both steady‐state and dynamic conditions. However, empirical information about their coordination is scarce. In this study, K_leaf, gas exchange, stomatal kinetics, and leaf anatomy in 10 species including ferns, gymnosperms, and angiosperms were investigated to elucidate the correlation of H₂O and CO₂ diffusion inside leaves under varying light conditions. Gas exchange, K_leaf, and anatomical traits varied widely across species. Under light‐saturated conditions, the A, g_s, g_m, and K_leaf were strongly correlated across species. However, the response patterns of A, g_s, g_m, and K_leaf to varying light intensities were highly species dependent. Moreover, stomatal opening upon light exposure of dark‐adapted leaves in the studied ferns and gymnosperms was generally faster than in the angiosperms; however, stomatal closing in light‐adapted leaves after darkening was faster in angiosperms. The present results show that there is a large variability in the coordination of leaf hydraulic and gas exchange parameters across terrestrial plant species, as well as in their responses to changing light.

Summary: Leaf hydraulic conductance (K_leaf) and mesophyll conductance (g_m) both represent major constraints to photosynthetic rate (A), and previous studies have suggested that K_leaf and g_m is correlated in leaves. However, there is scarce empirical information about their correlation. In this study, K_leaf, leaf hydraulic conductance inside xylem (K_x), leaf hydraulic conductance outside xylem (K_ox), A, stomatal conductance (g_s), g_m, and anatomical and structural leaf traits in 11 Oryza genotypes were investigated to elucidate the correlation of H₂O and CO₂ diffusion inside leaves. All of the leaf functional and anatomical traits varied significantly among genotypes. K_leaf was not correlated with the maximum theoretical stomatal conductance calculated from stomatal dimensions (g_smax), and neither g_s nor g_smax were correlated with K_x. Moreover, K_ox was linearly correlated with g_m and both were closely related to mesophyll structural traits. These results suggest that K_leaf and g_m are related to leaf anatomical and structural features, which may explain the mechanism for correlation between g_m and K_leaf.

Understanding the physiological responses of crops to drought is important for ensuring sustained crop productivity under climate change, which is expected to exacerbate the frequency and intensity of periods of drought. Drought responses involve multiple traits, and the correlations between these traits are poorly understood. Using a variety of techniques, we estimated the changes in gas exchange, leaf hydraulic conductance, and leaf turgor in rice (Oryza sativa) in response to both short- and long-term soil drought. We performed a photosynthetic limitation analysis to quantify the contributions of each limiting factor to the resultant overall decrease in photosynthesis during drought. Biomass, leaf area, and leaf width significantly decreased during the 2-week drought treatment, but leaf mass per area and leaf vein density increased. Light-saturated photosynthetic rate declined dramatically during soil drought, mainly due to the decrease in stomatal conductance (g_s) and mesophyll conductance (g_m). Stomatal modeling suggested that the decline in leaf hydraulic conductance explained most of the decrease in stomatal closure during the drought treatment, and may also trigger the drought-related decrease of stomatal conductance and mesophyll conductance. The results of this study provide insight into the regulation of carbon assimilation under drought conditions.