RESUMO
The role of ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco) in the regulation of photosynthesis in the field was assessed by following changes in the amount and activity of Rubisco and in the amounts of chlorophyll and total nitrogen in a willow (Salix cv. Aquatica gigantea) canopy during two growing seasons. An additional aim of this study was to determine how nitrogen is partitioned in the canopy and whether Rubisco and chlorophyll serve as sources of nitrogen. During both growing seasons, the total activity and the amount of Rubisco were high in young, sun-exposed leaves and decreased as the leaves aged and became shaded. In 1985, the specific activity was low (on average 0.8 micromol mm(-1) mg(-1) Rubisco protein) compared with the values obtained in 1986 (on average 1.8 micromol min(-1) mg(-1) Rubisco protein). These differences in the specific activity of Rubisco between the two years might be associated with changes in partitioning of nitrogen. During the cool summer of 1985, about 20% of the total nitrogen was partitioned to Rubisco compared with about 13% during the warm summer of 1986. Rubisco comprised a high proportion (about 60%) of the total soluble protein throughout the 1985 growing season, whereas during the 1986 growing season, the proportion of Rubisco in the total soluble protein fraction was lower and decreased markedly as the leaves became shaded. Chlorophyll did not serve as a source of nitrogen in the canopy. On the contrary, the proportion of total nitrogen in chlorophyll increased with time, which indicates acclimation of the thylakoid membranes to decreasing irradiance. The degree of activation of Rubisco in samples collected in 1986 was high in young, sun-exposed leaves, which suggests that in these leaves, Rubisco may limit the rate of CO(2) uptake. In the shaded leaves of the lower canopy, the degree of activation of Rubisco was low, which indicates that in these leaves, photosynthesis is limited by factors other than Rubisco.
RESUMO
Development of leaf chloroplast ultrastructure at five levels in a willow (Salix cv. Aquatica gigantea) canopy was followed during one growing season in the field. Changes in chloroplast ultrastructure were compared with the rate of CO(2) uptake of the same leaves. The highest rates of CO(2) uptake were recorded in young leaves exposed to full available sunlight. In these leaves, the area of the grana stacks was less than 20% of the total chloroplast area and the degree of thylakoid stacking was less than 1.5. The chloroplasts of these leaves contained large amounts of starch and small amounts of plastoglobuli. As the canopy grew and the leaves in the lower parts of the canopy became shaded, the structure of the chloroplast thylakoids gradually changed. In leaves at the two lowest levels of the canopy, the degree of stacking at the end of the growing season was close to 2 and correspondingly the rate of CO(2) uptake was low. The areas of grana stacks and plastoglobuli in these chloroplasts increased and were about 30 and 10% of the chloroplast area, respectively, by the end of the growing season. The increase in the degree of thylakoid stacking was caused by increased biosynthesis of grana lamellae, which in general were thinner than the lamellae of young leaves. The length of the stroma lamellae did not change with leaf age. Morphometric measurements showed that the structure of the chloroplasts in leaves 160 cm above ground was dynamic and responsive to environmental conditions so that photosynthetic capacity remained high for 7 weeks despite an increase in leaf shading.
