Effects of long-term exposure to elevated CO(2) conditions in slow-growing plants using a (12)C-enriched CO(2)-labelling technique.

Despite their relevancy, long-term studies analyzing elevated CO(2) effect in plant production and carbon (C) management on slow-growing plants are scarce. A special chamber was designed to perform whole-plant above-ground gas-exchange measurements in two slow-growing plants (Chamaerops humilis and Cycas revoluta) exposed to ambient (ca. 400 micromol mol(-1)) and elevated (ca. 800 micromol mol(-1)) CO(2) conditions over a long-term period (20 months). The ambient isotopic (13)C/(12)C composition (delta(13)C) of plants exposed to elevated CO(2) conditions was modified (from ca. -12.8 per thousand to ca. -19.2 per thousand) in order to study carbon allocation in leaf, shoot and root tissues. Elevated CO(2) increased plant growth by ca. 45% and 60% in Chamaerops and Cycas, respectively. The whole-plant above-ground gas-exchange determinations revealed that, in the case of Chamaerops, elevated CO(2) decreased the photosynthetic activity (determined on leaf area basis) as a consequence of the limited ability to increase C sink strength. On the other hand, the larger C sink strength (reflected by their larger CO(2) stimulatory effect on dry mass) in Cycas plants exposed to elevated CO(2) enabled the enhancement of their photosynthetic capacity. The delta(13)C values determined in the different plant tissues (leaf, shoot and root) suggest that Cycas plants grown under elevated CO(2) had a larger ability to export the excess leaf C, probably to the main root. The results obtained highlighted the different C management strategies of both plants and offered relevant information about the potential response of two slow-growing plants under global climate change conditions.

Assessing the stable carbon isotopic composition of intercellular CO2 in a CAM plant using gas chromatography-combustion-isotope ratio mass spectrometry.

Most of the literature focused on internal CO(2) (Ci) determinations in plants has used indirect methods based on gas-exchange estimations. We have developed a new method based on the capture of internal air gas samples and their analysis by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). This method provided a direct measure of intercellular CO(2) concentrations combined with stable carbon isotopic composition in O. ficus-indica plants. Plants were grown at both ambient and elevated CO(2) concentration. During the day period, when the stomata are closed, the Ci was high and was very (13)C-enriched in both ambient and elevated CO(2)-grown plants, reflecting Rubisco's fractionation (this plant enzyme has been shown to discriminate by 29 per thousand, in vitro, against (13)CO(2)). Other enzyme fractionations involved in C metabolism in plants, such as carbonic anhydrase, could also be playing an important role in the diurnal delta(13)C enrichment of the Ci. During the night, when stomata are open, Ci concentrations were higher in elevated (and the corresponding delta(13)C values were more (13)C-depleted) than in ambient CO(2)-grown plants.

Changes in respiratory mitochondrial machinery and cytochrome and alternative pathway activities in response to energy demand underlie the acclimation of respiration to elevated CO2 in the invasive Opuntia ficus-indica.

Studies on long-term effects of plants grown at elevated CO(2) are scarce and mechanisms of such responses are largely unknown. To gain mechanistic understanding on respiratory acclimation to elevated CO(2), the Crassulacean acid metabolism Mediterranean invasive Opuntia ficus-indica Miller was grown at various CO(2) concentrations. Respiration rates, maximum activity of cytochrome c oxidase, and active mitochondrial number consistently decreased in plants grown at elevated CO(2) during the 9 months of the study when compared to ambient plants. Plant growth at elevated CO(2) also reduced cytochrome pathway activity, but increased the activity of the alternative pathway. Despite all these effects seen in plants grown at high CO(2), the specific oxygen uptake rate per unit of active mitochondria was the same for plants grown at ambient and elevated CO(2). Although decreases in photorespiration activity have been pointed out as a factor contributing to the long-term acclimation of plant respiration to growth at elevated CO(2), the homeostatic maintenance of specific respiratory rate per unit of mitochondria in response to high CO(2) suggests that photorespiratory activity may play a small role on the long-term acclimation of respiration to elevated CO(2). However, despite growth enhancement and as a result of the inhibition in cytochrome pathway activity by elevated CO(2), total mitochondrial ATP production was decreased by plant growth at elevated CO(2) when compared to ambient-grown plants. Because plant growth at elevated CO(2) increased biomass but reduced respiratory machinery, activity, and ATP yields while maintaining O(2) consumption rates per unit of mitochondria, we suggest that acclimation to elevated CO(2) results from physiological adjustment of respiration to tissue ATP demand, which may not be entirely driven by nitrogen metabolism as previously suggested.

Tumour necrosis factor-alpha uncouples respiration in isolated rat mitochondria.

Recent studies have demonstrated the existence of an intracellular (associated with mitochondria) tumour necrosis factor-alpha (TNF) binding protein. In an attempt to elucidate if this receptor could be involved in TNF action, we have incubated liver isolated mitochondria in the presence of recombinant murine TNF. The results show that the addition of TNF at concentrations as low as 10(-6) U/microl resulted in a clear uncoupling respiration of liver isolated mitochondria, therefore suggesting that TNF can indeed exert intracellular effects, which are possibly linked with its cytotoxic mechanism.