Responses of Antioxidative Systems to Drought Stress in Pendunculate Oak and Maritime Pine as Modulated by Elevated CO2.

The aim of the present study was to investigate the effects of an enhanced CO2 concentration alone or in combination with drought stress on antioxidative systems of a deciduous (oak; Quercus robur) and an evergreen (pine; Pinus pinaster) tree species. The seedlings were grown for one season in a greenhouse in tunnels supplied with 350 or 700 [mu]L L-1 CO2. The experiment was repeated in a second year. Antioxidants, protective enzymes, soluble protein, and pigments showed considerable fluctuations in different years. Elevated CO2 caused significant reductions in the activities of superoxide dismutases in both oak and pine. The activities of ascorbate peroxidase and catalase were also reduced in most cases. The activities of dehydroascorbate reductase, monodehydroascorbate radical reductase, glutathione reductase, and guaiacol peroxidase were affected little or not at all by elevated CO2. When the trees were subjected to drought stress by withholding water, the activities of antioxidative enzymes decreased in leaves of pine and oak grown at ambient CO2 and increased in plants grown at elevated CO2 concentrations. The present results suggest that growth in elevated CO2 might reduce oxidative stress to which leaf tissues are normally exposed and enhance metabolic flexibility to encounter increased stress by increases in antioxidative capacity.

Antioxidative systems, pigment and protein contents in leaves of adult mediterranean oak species (Quercus pubescens and Q. ilex) with lifetime exposure to elevated CO2.

The aim of the present study was to investigate the effects of elevated CO2 on the antioxidative systems and the contents of pigments, soluble protein and lipid peroxidation in leaves of adult oaks, Quercus pubescens and Quercus ilex, grown at naturally enriched CO2 concentrations. For this purpose, a field study was conducted at two CO2 springs in Central Italy. Measurements of the pre-dawn water potentials indicated less drought stress in trees close to CO2 springs than in those grown at ambient CO2 concentrations. Most leaf constituents investigated showed significant variability between sampling dates, species and sites. The foliar contents of protein and chlorophylls were not affected in trees grown close to the CO2 vents compared with those in ambient conditions. Increases in glutathione and other soluble thiols were observed, but these responses might have been caused by a low pollution of the vents with sulphurous gases. At CO2 vents, glutathione reductase was unaffected, and superoxide dismutase activity was significantly diminished, in both species. Generally, the activities of catalase, guaiacol peroxidase and ascorbate peroxidase as well as the sum of dehydroascorbate and ascorbate were decreased in leaves from trees grown in naturally CO2 -enriched environments compared with those grown at ambient CO2 concentrations. The reduction in protective enzymes did not result in increased lipid peroxidation, but increased monodehydroascorbate radical reductase and dehydroascorbate reductase activities found in leaves of Q. pubescens suggest that the smaller pool of ascorbate was subjected to higher turnover rates. These data show that changes in leaf physiology persist, even after lifetime exposure to enhanced atmospheric CO2 . The results suggest that the down-regulation of protective systems, which has also previously been found in young trees or seedlings under controlled exposure to elevated CO2 concentrations, might reflect a realistic response of antioxidative defences in mature trees in a future high-CO2 world.

Preliminary studies of ascorbate metabolism in green and albino regions of variegated leaves of Coleus blumei, Benth.

Green and white variegated leaves of Coleus blumei, Benth. were separated into albino and green sections and used to determine the distribution of vitamin C and L-galactose dehydrogenase activity, an enzyme supposed to be involved in ascorbate metabolism, in heterotrophic and autotrophic foliar fractions. Both green and white sections contained vitamin C and activity of L-galactose dehydrogenase. However, in the white parts mainly dehydroascorbate was found, whereas in the green parts the redox state of the ascorbate system varied with light or dark conditions. Characterisation of L-galactose dehydrogenase from illuminated green leaf sections showed increasing activity with increasingly alkaline pH-values and a temperature optimum of 25 degrees C. Since these properties were slightly different than those of L-galactose dehydrogenase activities obtained from albino or darkened green leaf sections, we suggest that the enzyme may be light-modulated.

Growth under elevated CO(2) ameliorates defenses against photo-oxidative stress in poplar (Populus alba x tremula).

To test the hypothesis that growth-CO(2) concentrations affect stress susceptibility, leaves of poplar trees (Populus alba x tremula) grown under ambient or about twofold ambient CO(2) concentrations were subjected to chilling temperatures at high light intensities or were exposed to paraquat. Photosynthesis was less diminished and electrolyte leakage was lower in stressed leaves from poplar trees grown under elevated [CO(2)] as compared with those from ambient [CO(2)]. Severe stress caused pigment and protein degradation but to a lower extent in leaves from elevated as compared with those from ambient [CO(2)]. The protection was accompanied by rapid induction of superoxide dismutase activities (EC 1.15.1.1). Ascorbate and glutathione-related detoxification systems as well as catalase (EC 1.11.1.6) activities were less resistant than superoxide dismutases and declined in stress-exposed leaves from poplars grown under elevated [CO(2)] to a similar extent as in those from trees grown under ambient [CO(2)]. These results suggest that the CO(2)-mediated amelioration of stress was confined to SOD and limited since the destruction of H(2)O(2)-degrading systems was not prevented.

Differential stress responses of antioxidative systems to drought in pendunculate oak (Quercus robur) and maritime pine (Pinus pinaster) grown under high CO(2) concentrations.

Maritime pine (Pinus pinaster), a drought-avoiding species, contained 2--4-fold lower activities of superoxide dismutase, ascorbate peroxidase, catalase, dehydroascorbate reductase, and glutathione reductase than pendunculate oak (Quercus robur), a drought-tolerant species. The levels of ascorbate, monodehydroascorbate radical reductase activity, and glutathione in pine needles were similar to those in oak leaves. In both species the development of drought stress, characterized by decreasing predawn water potentials, caused gradual reductions in antioxidant protection, increased lipid peroxidation, increased oxidation of ascorbate and glutathione and in pine also significant loss in soluble proteins and carotenoids. These results support the idea that increased drought-tolerance in oak as compared with pine is related to increased biochemical protection at the tissue level. To test the hypothesis that elevated CO(2) ameliorated drought-induced injury, young oak and pine trees acclimated to high CO(2) were subjected to drought stress. Analysis of plots of enzymatic activities and metabolites against predawn water potentials revealed that the drought stress-induced decreases in antioxidant protection and increases in lipid peroxidation were dampened at high CO(2). In pine, protein and pigment degradation were also slowed down. At high CO(2), superoxide dismutase activities increased transiently in drought-stressed trees, but collapsed in pine faster than in oak. These observations suggest that the alleviation of drought-induced injury under elevated CO(2) is related to a higher stability of antioxidative enzymes and an increased responsiveness of SOD to stressful conditions. This ameliorating mechanism existed independently from the effects of elevated CO(2) on plant water relations and is limited within a species-specific metabolic window.

Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots.

To investigate whether Cd induces common plant defense pathways or unspecific necrosis, the temporal sequence of physiological reactions, including hydrogen peroxide (H(2)O(2)) production, changes in ascorbate-glutathione-related antioxidant systems, secondary metabolism (peroxidases, phenolics, and lignification), and developmental changes, was characterized in roots of hydroponically grown Scots pine (Pinus sylvestris) seedlings. Cd (50 microM, 6 h) initially increased superoxide dismutase, inhibited the systems involved in H(2)O(2) removal (glutathione/glutathione reductase, catalase [CAT], and ascorbate peroxidase [APX]), and caused H(2)O(2) accumulation. Elongation of the roots was completely inhibited within 12 h. After 24 h, glutathione reductase activities recovered to control levels; APX and CAT were stimulated by factors of 5.5 and 1.5. Cell death was increased. After 48 h, nonspecific peroxidases and lignification were increased, and APX and CAT activities were decreased. Histochemical analysis showed that soluble phenolics accumulated in the cytosol of Cd-treated roots but lignification was confined to newly formed protoxylem elements, which were found in the region of the root tip that normally constitutes the elongation zone. Roots exposed to 5 microM Cd showed less pronounced responses and only a small decrease in the elongation rate. These results suggest that in cells challenged by Cd at concentrations exceeding the detoxification capacity, H(2)O(2) accumulated because of an imbalance of redox systems. This, in turn, may have triggered the developmental program leading to xylogenesis. In conclusion, Cd did not cause necrotic injury in root tips but appeared to expedite differentiation, thus leading to accelerated aging.