Purification and Characterization of Isocitrate Lyase from Ethanol-grown Euglena gracilis

Isocitrate lyase was purified to homogeneity from ethanol-grown Euglena gracilis. The specific activity was 0.26 μmol/min/mg protein. The molecular mass of the enzyme was calculated to be 380 kDa by gel filtration on a Superose 6 column. The subunit molecular mass of the enzyme was 116 kDa as determined by SDS-polyacrylamide gel electrophoresis. These results showed that the native form of this enzyme was a trimer composed of three identical subunits. The pH optimum for cleavage and condensation reactions was 6.5 and 7.0, respectively. The K_m values for isocitrate, glyoxylate and succinate were 3.8, 1.3 and 7.7 mM, respectively. Isocitrate lyase absolutely required Mg for enzymatic activity. This is the first report of the purification of isocitrate lyase to homogeneity from Euglena gracilis.     

Does the photosynthetic light‐acclimation need change in leaf anatomy?

There is a strong correlation between leaf thickness and the light‐saturated rate of photosynthesis per unit leaf area (P_max). However, when leaves are exposed to higher light intensities after maturation, P_max often increases without increasing leaf thickness. To elucidate the mechanism with which mature leaves increase P_max, the change in anatomical and physiological characteristics of mature leaves of Chenopodium album, which was transferred from low to high light condition, were examined. When compared with leaves subjected to low light continuously (LL leaves), the leaves transferred from low to high light (LH leaves) significantly increased P_max. The transfer also increased the area of chloroplasts facing the intercellular space (S_c) and maintained a strong correlation between P_max and S_c. The mesophyll cells of LL leaves had open spaces along cell walls where chloroplasts were absent, which enabled the leaves to increase P_max when they were exposed to high light (LH). However, the LH leaves were not thick enough to allow further increase in P_max to the level in HH leaves. Thus leaf thickness determines an upper limit of P_max of leaves subjected to a change from low to high light conditions. Shade leaves would only increase P_max when they have open space to accommodate chloroplasts which elongate after light conditions improve.     

The effect of the water-lily beetle, Galerucella nymphaeae, on leaf production and leaf longevity of the yellow water-lily, Nuphar lutea

SUMMARY. 1. The effect of the water-lily beetle, Galerucella (= Pyrrhalta ) nymphaeae (L.), (Coleoptera, Chrysomelidae), on the leaf production and leaf longevity of the yellow water-lily, Nuphar lutea (= N. luteum ) (L.) Sibth. & Sm., was studied experimentally in southern Finland during one growth season.
2. Twelve cages, each containing a single rosette of the water-lily, were established in the lake during May. Two treatments were randomly applied to cages: (i) control (herbivory not allowed), and (ii) fifty beetles added at the beginning of the season. Leaves were marked individually and the production of new leaves was followed until October.
3. In both treatments the production of floating leaves continued until the end of August but declined towards the end of summer. Herbivory did not affect the number of leaves produced, i.e. the plants did not compensate for the loss of leaves.
4. Leaf longevity was reduced dramatically (up to —60%) when herbivores were present. Leaves that emerged during the most intense period of larval growth had a life-span one-third of that of the control group.     

Photosynthesis or persistence: nitrogen allocation in leaves of evergreen and deciduous Quercus species

Photosynthetic nitrogen use efficiency (PNUE, photosynthetic capacity per unit leaf nitrogen) is one of the most important factors for the interspecific variation in photosynthetic capacity. PNUE was analysed in two evergreen and two deciduous species of the genus Quercus. PNUE was lower in evergreen than in deciduous species, which was primarily ascribed to a smaller fraction of nitrogen allocated to the photosynthetic apparatus in evergreen species. Leaf nitrogen was further analysed into proteins in the water‐soluble, the detergent‐soluble, and the detergent‐insoluble fractions. It was assumed that the detergent‐insoluble protein represented the cell wall proteins. The fraction of nitrogen allocated to the detergent‐insoluble protein was greater in evergreen than in deciduous leaves. Thus the smaller allocation of nitrogen to the photosynthetic apparatus in evergreen species was associated with the greater allocation to cell walls. Across species, the fraction of nitrogen in detergent‐insoluble proteins was positively correlated with leaf mass per area, whereas that in the photosynthetic proteins was negatively correlated. There may be a trade‐off in nitrogen partitioning between components pertaining to productivity (photosynthetic proteins) and those pertaining to persistence (structural proteins). This trade‐off may result in the convergence of leaf traits, where species with a longer leaf life‐span have a greater leaf mass per area, lower photosynthetic capacity, and lower PNUE regardless of life form, phyllogeny, and biome.     

Effects of elevated CO2 on the size structure in even-aged monospecific stands of Chenopodium album

To investigate the effect of elevated CO₂ on the size inequality and size structure, even‐aged monospecific stands of an annual, Chenopodium album, were established at ambient and doubled CO₂ with high and low nutrient availabilities in open top chambers. The growth of individual plants was monitored non‐destructively every week until flowering. Elevated CO₂ significantly enhanced plant growth at high nutrients, but did not at low nutrients. The size inequality expressed as the coefficient of variation tended to increase at elevated CO₂. Size structure of the stands was analyzed by the cumulative frequency distribution of plant size. At early stages of plant growth, CO₂ elevation benefited all individuals and shifted the whole size distribution of the stand to large size classes. At later stages, dominant individuals were still larger at elevated than at ambient CO₂, but the difference in small subordinate individuals between two CO₂ levels became smaller. Although these tendencies were found at both nutrient availabilities, difference in size distribution between CO₂ levels was larger at high nutrients. The CO₂ elevation did not significantly enhance the growth rate as a function of plant size except for the high nutrient stand at the earliest stage, indicating that the higher biomass at elevated CO₂ at later stages in the high nutrient stand was caused by the larger size of individuals at the earliest stage. Thus the effect of elevated CO₂ on stand structure and size inequality strongly depended on the growth stage and nutrient availabilities.     

A model of the acclimation of photosynthesis in the leaves of C3 plants to sun and shade with respect to nitrogen use

A model of leaf photosynthesis of C₃, plants has been developed to describe their nitrogen economy. In this model, photosynthetic proteins are categorized into five groups depending on their functions. The effects of investment of nitrogen in each of these groups on the maximal rate of photosynthesis and/or the initial slope of the light-response curve are described as simple equations. Using this model, the optimal pattern of nitrogen partitioning which maximizes the daily rate of CO₂ exchange is estimated for various light environments and leaf nitrogen contents. When the leaf nitrogen content is fixed, the amount of nitrogen allocated to Calvin cycle enzymes and electron carriers increases with increasing irradiance, while that allocated to chlorophyll-protein complexes increases with decreasing irradiance. For chlorophyll-proteins of photosystem II, the amount of light-harvesting complex II relative to that of the core complex increases with decreasing irradiance. At any irradiance, partitioning into ribulose bisphosphate carboxylase increases with increasing leaf nitrogen content Taking the total leaf nitrogen content and the daily CO₂ exchange rate as ‘cost’ and ‘benefit’, respectively, the optimal amount and partitioning of nitrogen are examined for various conditions of light environment and nitrogen availability. The leaf nitrogen content that maximizes the rate of daily carbon fixation increases with increasing growth irradiance. It is also predicted that, at low nitrogen availabilities, low leaf nitrogen contents are advantageous in terms, of nitrogen use efficiency. These trends predicted by the present model are largely consistent with those reported for actual plants. The differences in the total amount of leaf nitrogen and in the organization of photosynthetic components that have been reported for plants from different environments would therefore be of adaptive significance, because such differences can contribute to realization of efficient photosynthesis. These results are fürther discussed in an ecological context.     

Comparative ecophysiology of leaf and canopy photosynthesis

Leaves and herbaceous leaf canopies photosynthesize efficiently although the distribution of light, the ultimate resource of photosynthesis, is very biased in these systems. As has been suggested in theoretical studies, if a photosynthetic system is organized such that every photosynthetic apparatus photosynthesizes in concert, the system as a whole has the sharpest light response curve and is most adaptive. This condition can be approached by (i) homogenization of the light environment and (ii) acclimation of the photosynthetic properties of leaves or chloroplasts to their local light environments. This review examines these two factors in the herbaceous leaf canopy and in the leaf. Changes in the inclination of leaves in the canopy and differentiation of mesophyll into palisade and spongy tissue contribute to the moderation of the light gradient. Leaf and chloroplast movements in the upper parts of these systems under high irradiances also moderate light gradients. Moreover, acclimation of leaves and chloroplasts to the local light environment is substantial. These factors increase the efficiency of photosynthesis considerably. However, the systems appear to be less efficient than the theoretical optimum. When the systems are optically dense, the light gradients may be too great for leaves or chloroplasts to acclimate. The loss of photosynthetic production attributed to the imperfect adjustment of photosynthetic apparatus to the local light environment is most apparent when the photosynthesis of the system is in the transition between the light-limited and light-saturated phases. Although acclimation of the photosynthetic apparatus and moderation of light gradients are imperfect, these markedly raise the efficiency of photosynthesis. Thus more mechanistic studies on these adaptive attributes are needed. The causes and consequences of imperfect adjustment should also be investigated.     

Elevated CO2 concentration, nitrogen use, and seed production in annual plants

Elevated atmospheric CO₂ concentration ([CO₂]) stimulates seed mass production in many species, but the extent of stimulation shows large variation among species. We examined (1) whether seed production is enhanced more in species with lower seed nitrogen concentrations, and (2) whether seed production is enhanced by elevated [CO₂] when the plant uses more N for seed production. We grew 11 annuals in open top chambers that have different [CO₂] conditions (ambient: 370 μmol mol⁻¹, elevated: 700 μmol mol⁻¹). Elevated [CO₂] significantly increased seed production in six out of 11 species with a large interspecific variation (0.84–2.12, elevated/ambient [CO₂]). Seed nitrogen concentration was not correlated with the enhancement of seed production by elevated [CO₂]. The enhancement of seed production was strongly correlated with the enhancement of seed nitrogen per plant caused by increased N acquisition during the reproductive period. In particular, legume species tended to acquire more N and produced more seeds at elevated [CO₂] than non-nitrogen fixing species. Elevated [CO₂] little affected seed [N] in all species. We conclude that seed production is limited primarily by nitrogen availability and will be enhanced by elevated [CO₂] only when the plant is able to increase nitrogen acquisition.