Near-infrared reflectance spectroscopy was applied to determine nitrogen (N), phosphorus (P) and calcium (Ca) content in leaf samples of 18 woody species. A total of 183 samples from mountain, riparian and dry areas from the Central-Western Iberian Peninsula were collected for this purpose. The wide intervals of variation observed in nutrient concentrations (6.6-45.0 g kg(-1) for N, 0.24-2.97 g kg(-1) for P, and 1.00-20.06 g kg(-1) for Ca) were due to the great heterogeneity of the samples. To develop calibration equations, multiple linear regression, and partial least-squares regression (PLSR) were used. In both cases, three mathematical transformations of the data were applied: log1/R and first and second derivatives. The best calibration statistics were obtained using PLSR and derivative transformations (second derivative for N and first derivative for P and Ca). The following coefficients of multiple determination (R2) and standard errors of cross validation were obtained: 0.99 and 0.93 for N, 0.94 and 0.15 for P, and 0.95 and 0.88 for Ca. In the external validation the standard errors of prediction obtained were 0.76 (N), 0.11 (P) and 0.60 (Ca).

Summary 1.It has been proposed that in longer-living leaves the allocation of biomass to structural components is greater than in shorter-living leaves, leading to a greater leaf mass per area (LMA) and to lower assimilation rates. However, direct evidence in support of this hypothesis is very scarce. 2. In the present work we investigated the relationships between leaf duration and LMA, leaf thickness and fibre concentrations (cellulose, hemicellulose and lignin) in five oak species, five pine species and three additional tree species, differing in leaf life spans. Correlations among leaf life span and the other leaf traits were obtained both across species (TIPs) and as phylogenetically independent contrasts (PICs). 3. Leaf thickness and LMA increased steadily with leaf longevity. No relationship was found between leaf longevity and the lignin concentration per unit leaf mass. Evergreen leaves were found to have higher mean concentrations of cellulose and hemicellulose than deciduous ones. However, no relationship was observed between leaf longevity and the concentration of structural carbohydrates across the set of evergreen species, although PIC correlations revealed increases in cellulose with leaf longevity within particular lineages. 4. Our findings reveal that leaf reinforcement by structural carbohydrates depends on leaf habit (deciduous vs. evergreen) and, within a given lineage, also on leaf longevity. However, among the evergreen species co-occurring in a particular environment, leaf duration may apparently be increased, with no need for increases in the concentration of structural components per unit leaf mass.

The competitive equilibrium between deciduous and evergreen plant species to a large extent depends on the intensity of the reduction in carbon gain undergone by evergreen leaves, associated with the leaf traits that confer resistance to stressful conditions during the unfavourable part of the year. This study explores the effects of winter harshness on the resistance traits of evergreen leaves. Leaf mass per unit area (LMA), leaf thickness and the concentrations of fibre, nitrogen (N), phosphorus (P), soluble protein, chlorophyll and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) were determined in three evergreen and two deciduous species along a winter temperature gradient. In the evergreen species, LMA, thickness, and P and structural carbohydrate concentrations increased with the decrease in winter temperatures. Nitrogen and lignin concentrations did not show definite patterns in this regard. Chlorophyll, soluble proteins and Rubisco decreased with the increase in winter harshness. Our results suggest that an increase in LMA and in the concentration of structural carbohydrates would be a requirement for the leaves to cope with low winter temperatures. The evergreen habit would be associated with higher costs at cooler sites, because the cold resistance traits imply additional maintenance costs and reduced N allocation to the photosynthetic machinery, associated with structural reinforcement at colder sites.

Near-infrared reflectance spectroscopy (NIRS) was used to estimate N, neutral detergent fibre (NDF), acid detergent fibre (ADF), lignin and cellulose contents in leaves of a heterogeneous group of 17 woody species from the Central Western region of the Iberian Peninsula. The sample set consisted of 182 samples of leaves of deciduous and evergreen species, showing a wide range of concentrations determined by reference methods: 6.60-35.2 g kg-1 (N), 15.5-66.0% (NDF), 10.2-57.3% (ADF), 3.45-27.4% (lignin) and 5.79-31.3% (cellulose). Reflectance spectra, obtained for samples of dried and ground leaves, were recorded as log1/R (R=reflectance) from 1,100 to 2,500 nm. NIRS calibrations were developed using multiple linear (MLR) and partial least-squares (PLSR) regressions, and tested by external validation. Spectral data were transformed to the first and second derivative (1D, 2D). The PLSR method and derivative transformations provided the best statistics and showed lower standard errors of calibration (SEC) and higher coefficients of multiple determination (R2). In the external validation the standard errors of prediction (SEP) were 0.76 g kg-1 (N), 2.11% (NDF), 1.47% (ADF), 0.85% (lignin) and 0.86% (cellulose). The results obtained show that NIRS is very effective for the estimation of these organic constituents in leaf tissue of woody species. This technique can be used in ecological or ecophysiological studies as an alternative to the more time-consuming standard methods.

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