Elevated CO2 and water-availability effect on gas exchange and nodule development in N2-fixing alfalfa plants

N₂-fixing alfalfa plants were grown in controlled conditions at different CO₂ levels (350 μmol mol^?1 versus 700 μmol mol^?1) and water-availability conditions (WW, watered at maximum pot water capacity versus WD, watered at 50% of control treatments) in order to determine the CO₂ effect (and applied at two water regimes) on plant growth and nodule activity in alfalfa plants. The CO₂ stimulatory effect (26% enhancement) on plant growth was limited to WW plants, whereas no CO₂ effect was observed in WD plants. Exposure to elevated CO₂ decreased Rubisco carboxylation capacity of plants, caused by a specific reduction in Rubisco (EC 4.1.1.39) concentration (11% in WW and 43% in WD) probably explained by an increase in the leaf carbohydrate levels. Plants grown at 700 μmol mol^?1 CO₂ maintained control photosynthetic rates (at growth conditions) by diminishing Rubisco content and by increasing nitrogen use efficiency. Interestingly, our data also suggest that reduction in shoot N demand (reflected by the TSP and especially Rubisco depletion) affected negatively nodule activity (malate dehydrogenase, EC 1.1.1.37, and glutamate-oxaloacetate transaminase, EC 2.6.1.1, activities) particularly in water-limited conditions. Furthermore, nodule DM and TSS data revealed that those nodules were not capable to overcome C sink strength limitations.     

Effects of shade treatments on the photosynthetic capacity,chlorophyll fluorescence,and chlorophyll content of Tetrastigma hemsleyanum Diels et Gilg

Tetrastigma hemsleyanum Diels et Gilg was grown under full sunlight and moderate and high levels of shade for one month to evaluate its photosynthetic and chlorophyll fluorescence response to different light conditions. The results showed that T. hemsleyanum attained greatest leaf size and P_n when cultivated with 67% shade. Leaves of seedlings grown with 90% shade were the smallest. Leaf color of plants grown under full sunlight and 50% shade was yellowish-green. The P_n value increased rapidly as PPFD increased to 200 μmol m^?2 s^?1 and then increased slowly to a maximum, followed by a slow decrease as PPFD was increased to 1000 μmol m^?2 s^?1. P_n was highest for the 67% shade treatment and the LSP for this shade treatment was 600 μmol m^?2 s^?1. Full sunlight and 50% shade treatments resulted in significant reduction of ETR and qP and increased NPQ. Chl a, Chl b and total chlorophyll content increased and Chl a/b values decreased with increased shading. Results showed that light intensity greater than that of 50% shade depressed photosynthetic activity and T. hemsleyanum growth. Irradiance less than that of 75% shade limited carbon assimilation and led to decreased plant growth. Approximately 67% shade is suggested to be the optimum light irradiance condition for T. hemsleyanum cultivation.     

Growth and development of cotton (Gossypium hirsutum L.) in response to CO2 enrichment under two different temperature regimes

An increase in atmospheric CO₂ concentration ([CO₂]) together with other climate change factors could greatly affect agricultural productivity. Understanding the impact of the change in atmospheric [CO₂] in conjunction with the ongoing global change is crucial to prepare for mitigation and any adaptation for future agricultural production. The main goal of this project was to study the time-course pattern of cotton plant growth in response to [CO₂] and temperature to investigate the hypothesis that whether response to elevated [CO₂] would change at different temperatures. An experiment was conducted in the controlled-environment chambers of the Georgia Envirotron with two different day/night temperatures levels, e.g., 25/15 °C and 35/25 °C, and three CO₂ concentrations, e.g., 400, 600 and 800 μmol l^?1. The experimental design was completely randomized with four replicates (plastic containers) per treatment. Growth analysis was conducted at bi-weekly intervals during the growing season. In addition, leaf area, leaf dry mass, root dry mass, square dry mass, boll dry mass and total above dry mass per plant were also measured at each sampling. Plant traits, including plant height, number of leaves, number of squares and number of bolls were recorded weekly. The number of days to emergence, squaring, flowering and maturity were also observed. The results showed that by increasing [CO₂] to 600 μmol l^?1 total biomass increased at both temperature levels, but a further increase of [CO₂] up to 800 μmol l^?1 increased total biomass only at the temperature of 35/25 °C. Throughout the growing season, there was no significant effect of [CO₂] levels on LAI. Increasing temperature from 25/15 °C to 35/25 °C had a positive impact on LAI across all CO₂ levels (P < 0.05). Increasing CO₂ from 400 to 600 μmol l^?1 significantly increased the number of squares by 31.4%, but a further increase to 800 μmol l^?1 caused a 6.6% decrease (non-significant) in the number of squares. The interactive effects of [CO₂] and temperature indicated that at a higher temperature, CO₂ would be more beneficial as we proceed towards the end of the growing season. However, further studies are needed to really understand the interaction between higher [CO₂] and temperature levels and cultivar characteristics.     

Gas exchange acclimation to elevated CO2 in upper-sunlit and lower-shaded canopy leaves in relation to nitrogen acquisition and partitioning in wheat grown in field chambers

Growth at elevated CO₂ often decreases photosynthetic capacity (acclimation) and leaf N concentrations. Lower-shaded canopy leaves may undergo both CO₂ and shade acclimation. The relationship of acclimatory responses of flag and lower-shaded canopy leaves of wheat (Triticum aestivum L.) to the N content, and possible factors affecting N gain and distribution within the plant were investigated in a wheat crop growing in field chambers set at ambient (360 μmol mol⁻¹) and elevated (700 μmol mol⁻¹) CO₂, and with two amounts of N fertilizer (none and 70 kg ha⁻¹ applied on 30 April). Photosynthesis, stomatal conductance and transpiration at a common measurement CO₂, chlorophyll and Rubisco levels of upper-sunlit (flag) and lower-shaded canopy leaves were significantly lower in elevated relative to ambient CO₂-grown plants. Both whole shoot N and leaf N per unit area decreased at elevated CO₂, and leaf N declined with canopy position. Acclimatory responses to elevated CO₂ were enhanced in N-deficient plants. With N supply, the acclimatory responses were less pronounced in lower canopy leaves relative to the flag leaf. Additional N did not increase the fraction of shoot N allocated to the flag and penultimate leaves. The decrease in photosynthetic capacity in both upper-sunlit and lower-shaded leaves in elevated CO₂ was associated with a decrease in N contents in above-ground organs and with lower N partitioning to leaves. A single relationship of N per unit leaf area to the transpiration rate accounted for a significant fraction of the variation among sun-lit and shaded leaves, growth CO₂ level and N supply. We conclude that reduced stomatal conductance and transpiration can decrease plant N, leading to acclimation to CO₂ enrichment.     

Acclimation of peanut (Arachis hypogaea L.) leaf photosynthesis to elevated growth CO2 and temperature

Peanut (Arachis hypogaea L. cv. Florunner) was grown from seed sowing to plant maturity under two daytime CO₂ concentrations ([CO₂]) of 360 μmol mol⁻¹ (ambient) and 720 μmol mol⁻¹ (elevated) and at two temperatures of 1.5 and 6.0 °C above ambient temperature. The objectives were to characterize peanut leaf photosynthesis responses to long-term elevated growth [CO₂] and temperature, and to assess whether elevated [CO₂] regulated peanut leaf photosynthetic capacity, in terms of activity and protein content of ribulose bisphosphate carboxylase-oxygenase (Rubisco), Rubisco photosynthetic efficiency, and carbohydrate metabolism. At both growth temperatures, leaves of plants grown under elevated [CO₂] had higher midday photosynthetic CO₂ exchange rate (CER), lower transpiration and stomatal conductance and higher water-use efficiency, compared to those of plants grown at ambient [CO₂]. Both activity and protein content of Rubisco, expressed on a leaf area basis, were reduced at elevated growth [CO₂]. Declines in Rubisco under elevated growth [CO₂] were 27–30% for initial activity, 5–12% for total activity, and 9–20% for protein content. Although Rubisco protein content and activity were down-regulated by elevated [CO₂], Rubisco photosynthetic efficiency, the ratio of midday light-saturated CER to Rubisco initial or total activity, of the elevated-[CO₂] plants was 1.3- to 1.9-fold greater than that of the ambient-[CO₂] plants at both growth temperatures. Leaf soluble sugars and starch of plants grown at elevated [CO₂] were 1.3- and 2-fold higher, respectively, than those of plants grown at ambient [CO₂]. Under elevated [CO₂], leaf soluble sugars and starch, however, were not affected by high growth temperature. In contrast, high temperature reduced leaf soluble sugars and starch of the ambient-[CO₂] plants. Activity of sucrose-P synthase, but not adenosine 5′-diphosphoglucose pyrophosphorylase, was up-regulated under elevated growth [CO₂]. Thus, in the absence of other environmental stresses, peanut leaf photosynthesis would perform well under rising atmospheric [CO₂] and temperature as predicted for this century.     

The harmful alga Aureococcus anophagefferens utilizes 19′-butanoyloxyfucoxanthin as well as xanthophyll cycle carotenoids in acclimating to higher light intensities

Aureococcus anophagefferens is a picoplanktonic microalga that is very well adapted to growth at low nutrient and low light levels, causing devastating blooms (“brown tides”) in estuarine waters. To study the factors involved in long-term acclimation to different light intensities, cells were acclimated for a number of generations to growth under low light (20 μmol photons m^? 2 s^? 1), medium light (60 or 90 μmol photons m^? 2 s^? 1) and high light (200 μmol photons m^? 2 s^? 1), and were analyzed for their contents of xanthophyll cycle carotenoids (the D pool), fucoxanthin and its derivatives (the F pool), Chls c₂ and c₃, and fucoxanthin Chl a/c polypeptides (FCPs). Higher growth light intensities resulted in increased steady state levels of both diadinoxanthin and diatoxanthin. However, it also resulted in the conversion of a significant fraction of fucoxanthin to 19′-butanoyloxyfucoxanthin without a change in the total F pool. The increase in 19′-butanoyloxyfucoxanthin was paralleled by a decrease in the effective antenna size, determined from the slope of the change in F₀ as a function of increasing light intensity. Transfer of acclimated cultures to a higher light intensity showed that the conversion of fucoxanthin to its derivative was a relatively slow process (time-frame of hours). We suggest the replacement of fucoxanthin with the bulkier 19′-butanoyloxyfucoxanthin results in a decrease in the light-harvesting efficiency of the FCP antenna and is part of the long-term acclimative response to growth at higher light intensities.     

Plasticity of rice tiller production is related to genotypic variation in the biomass response to elevated atmospheric CO2 concentration and low temperatures during vegetative growth

Appropriate resource partitioning to either production of new tillers or growth of individual tillers is a critical factor for increasing rice biomass production and facilitating adaptation to climate change. We examined the contributions of genotypic variation to the tiller number and individual tiller growth of 24 rice cultivars in response to an elevated atmospheric CO₂ concentration [CO₂] (control + 191 μmol mol⁻¹) and a low air temperature (control minus 4.7 °C) during 56 days of vegetative growth after transplanting. For all genotypes combined, biomass increased by 27% under elevated [CO₂] and decreased by 34% at low temperature, with a significant genotype × temperature interaction. The increase caused by elevated [CO₂] resulted from increased tiller number, and the decrease caused by low temperature resulted from decreased growth of individual tillers. Despite the different overall responses to elevated [CO₂] and low temperature, most of the genotypic variation in biomass at elevated [CO₂] and low temperature was explained by the responses of tiller number rather than by individual tiller growth. The genotypes with the highest biomass response to elevated [CO₂] had a smaller reduction of biomass under low temperature. These results highlight the greater importance of genotypic variation in tiller number than in individual tiller growth in the response of biomass to environmental change.     

Sex-specific light acclimation of Chara canescens (Charophyta)

Bisexual populations of the charophyte Chara canescens (Desv. et Loisel. in Loisel., 1810) containing male and female individuals are rarely found. Two experiments were carried out to study whether male and female algae from the same site exhibit different physiological capacities, especially with respect to light acclimation.Algae from two different shore levels and from laboratory cultures acclimated to six irradiance conditions (35–500 μmol photons m⁻² s⁻¹) were compared. Field measurements showed that both female and male algae of C. canescens are able to acclimate to daily changes in solar irradiance. The quantum yield of Photosystem II (PSII) decreased with increasing irradiance in the morning and increased with decreasing irradiance in the afternoon. Growth experiments showed increasing growth rates from 35 μmol photons m⁻² s⁻¹ (∼7 mg FW) up to 500 μmol photons m⁻² s⁻¹ (∼27 mg FW) in female and male C. canescens. The irradiance saturation point for photosynthesis (E_k) was about 140 μmol m⁻² s⁻¹ for both sexes within the whole range of acclimation irradiances. The maximum photosynthesis rate at saturating irradiances (P_max) of male algae was highest at E_k, whereas P_max of female algae was highest at 500 μmol photons m⁻² s⁻¹. The photosynthetic efficiency in the light-limited range (α) increased in female C. canescens and decreased in male C. canescens. The ratio of the non-photochemical quenching parameter (NPQ) to the relative electron transport rates rETR_(MT) increased in both sexes with irradiance, but showed a steeper increase in male than in female algae. Pigment analysis showed similar acclimation pattern for male and female C. canescens. Chl a/Chl b ratios of both sexes were constant over the whole range of E_g, whereas Chl a/carotenoid ratios in male and female C. canescens decreased from 70 μmol photons m⁻² s⁻¹ upwards. Pigment analysis pointed out that the carotenes α-, β- and γ-carotene were more prominent in male than in female algae.Our results indicate that female C. canescens are more efficient in light acclimation than male algae from the same site. Nevertheless, further investigations of bisexual C. canescens populations resolving CO₂-uptake mechanisms and/or genetic differences are needed.     

Pea plant responsiveness under elevated [CO2] is conditioned by the N source (N2 fixation versus NO3− fertilization)

The main goal of this study was to test the effect of [CO₂] on C and N management in different plant organs (shoots, roots and nodules) and its implication in the responsiveness of exclusively N₂-fixing and NO₃⁻-fed plants. For this purpose, exclusively N₂-fixing and NO₃⁻-fed (10 mM) pea (Pisum sativum L.) plants were exposed to elevated [CO₂] (1000 μmol mol⁻¹ versus 360 μmol mol⁻¹ CO₂). Gas exchange analyses, together with carbohydrate, nitrogen, total soluble proteins and amino acids were determined in leaves, roots and nodules. The data obtained revealed that although exposure to elevated [CO₂] increased total dry mass (DM) in both N treatments, photosynthetic activity was down-regulated in NO₃⁻-fed plants, whereas N₂-fixing plants were capable of maintaining enhanced photosynthetic rates under elevated [CO₂]. In the case of N₂-fixing plants, the enhanced C sink strength of nodules enabled the avoidance of harmful leaf carbohydrate build up. On the other hand, in NO₃⁻-fed plants, elevated [CO₂] caused a large increase in sucrose and starch. The increase in root DM did not contribute to stimulation of C sinks in these plants. Although N₂ fixation matched plant N requirements with the consequent increase in photosynthetic rates, in NO₃⁻-fed plants, exposure to elevated [CO₂] negatively affected N assimilation with the consequent photosynthetic down-regulation.     

Effects of Zn2+ and niflumic acid on photosynthesis in Glycine soja and Glycine max seedlings under NaCl stress

The effects of two anion/Cl^? channel inhibitors, Zn²⁺ and niflumic acid (NA), on seedling photosynthetic and fluorescent parameters of two Glycine soja populations (salt-tolerant BB52; salt-sensitive N23227) and Glycine max cultivar (salt-tolerant Lee68) were studied and compared under salt stress. Treatments with Zn²⁺ and NA only (10, 20 μmol L^?1) were also imposed for comparisons. Results showed that, there were non-toxic and non-nutritional effects of Zn²⁺ and NA treatments alone on seed germination and seedling growth of soybeans. Under 150 mmol L^?1 NaCl for 6 d, leaf chlorophyll and carotenoid contents, net photosynthetic rate (P_n), stomatal conductance (G_s), intercellular CO₂ concentration (C_i), transpiration rate (Tr), and the maximum photochemical efficiency of photosystem II (PS II) (F_v/F_m) except the stomatal limitation (Ls) significantly decreased in three kinds of soybean seedlings when compared with their control plants. The NaCl stress plus additional 20 μmol L^?1 Zn showed an obvious enhancement of leaf chlorophyll and carotenoid contents, P_n, G_s, C_i and Tr, especially for the G. max cultivar Lee68, but the supplementation of 20 μmol L^?1 NA showed the reverse effects.     

Purification,kinetic and thermodynamic characterization of soluble acid invertase from sugarcane (Saccharum officinarum L.)

We report for the first time kinetic and thermodynamic properties of soluble acid invertase (SAI) of sugarcane (Saccharum officinarum L.) salt sensitive local cultivar CP 77-400 (CP-77). The SAI was purified to apparent homogeneity on FPLC system. The crude enzyme was about 13 fold purified and recovery of SAI was 35%. The invertase was monomeric in nature and its native molecular mass on gel filtration and subunit mass on SDS-PAGE was 28 kDa. SAI was highly acidic having an optimum pH lower than 2. The acidic limb was missing. Proton transfer (donation and receiving) during catalysis was controlled by the basic limb having a pKa of 2.4. Carboxyl groups were involved in proton transfer during catalysis. The kinetic constants for sucrose hydrolysis by SAI were determined to be: k_m = 55 mg ml^?1, k_cat = 21 s^?1, k_cat/k_m = 0.38, while the thermodynamic parameters were: ΔH* = 52.6 kJ mol^?1, ΔG* = 71.2 kJ mol^?1, ΔS* = ?57 J mol^?1 K^?1, ΔG*_E–S = 10.8 kJ mol^?1 and ΔG*_E–T = 2.6 kJ mol^?1. The kinetics and thermodynamics of irreversible thermal denaturation at various temperatures 53–63 °C were also determined. The half -life of SAI at 53 and 63 °C was 112 and 10 min, respectively. At 55 °C, surprisingly the half -life increased to twice that at 53 °C. ΔG*, ΔH* and ΔS* of irreversible thermal stability of SAI at 55 °C were 107.7 kJ mol^?1, 276.04 kJ mol^?1 and 513 J mol^?1K^?1, respectively.     

Elevated CO2 will not select for enhanced growth in white spruce despite genotypic variation in response

The effect of elevated atmospheric CO₂ on plant growth has been well-documented in the literature. However, few studies have quantified intra-specific genetic variation in growth response, and the potential for natural and artificial selection to act upon this variation. This study examined intra-specific variation in growth response to elevated CO₂ in 29 genotypes of white spruce (Picea glauca), a widely distributed and economically important species of the boreal forest region in North America. Trees were exposed to either ambient (370 μL L^?1) or twice-ambient CO₂ (740 μL L^?1). The opportunity for selection (i.e. the relative variation in fitness) was determined at low and high CO₂ levels with size as a measure of fitness and heritability of this variation determined. There was considerable variation among the genotypes in size and response to elevated CO₂. The increase in mass at elevated CO₂ ranged from 23% to 108% depending upon genotype. In spite of this variation, the genetic correlation between the two environments approached unity, as genotype variance was much greater than the genotype×CO₂ variance. Elevated CO₂ had no effect on heritability of the size-related traits we examined, and either had no effect on opportunity for selection, or decreased it. We conclude that selection at elevated atmospheric CO₂ is unlikely to increase mean plant size in white spruce beyond that observed for present day populations grown at elevated CO₂, despite the substantial genetic variation in CO₂ response displayed by this species.     

Effects of gradual versus step increases in carbon dioxide on Plantago photosynthesis and growth in a microcosm study

This study investigated the effects of a gradual versus step increases in carbon dioxide (CO₂) on plant photosynthesis and growth at two nitrogen (N) levels. Plantago lanceolata were grown for 80 days and then treated with the ambient CO₂ (as the control), gradual CO₂ increase and step CO₂ increase as well as low and high N additions for 70 days. While [CO₂] were kept at constant 350 and 700 μmol mol⁻¹ for the ambient and step CO₂ treatments, respectively, [CO₂] in the gradual CO₂ treatment was raised by 5 μmol mol⁻¹ day⁻¹, beginning at 350 μmol mol⁻¹ and reaching 700 μmol mol⁻¹ by the end of experiment. The step CO₂ treatment immediately resulted in an approximate 50% increase in leaf photosynthetic carbon fixation at both the low and high N additions, leading to a 20–24% decrease in leaf N concentration. The CO₂-induced nitrogen stress, in return, resulted in partial photosynthetic downregulation since the third week at the low N level and the fourth week at the high N level after treatments. In comparison, the gradual CO₂ treatment induced a gradual increase in photosynthetic carbon fixation, leading to less reduction in leaf N concentration. In comparison to the ambient CO₂, both the gradual and step CO₂ increases resulted in decreases in specific leaf area, leaf N concentration but an increase in plant biomass. Responses of plant shoot:root ratio to CO₂ treatments varied with N supply. It decreased with low N supply and increased with high N supply under the gradual and step CO₂ treatments relative to that under the ambient CO₂. Degrees of those changes in physiological and growth parameters were usually larger under the step than the gradual CO₂ treatments, largely due to different photosynthetic C influxes under the two CO₂ treatments.     

The mechanism(s) involved in the photoprotection of PSII at elevated CO2 in nodulated alfalfa plants

In a previous study, we found that enhanced CO₂ subjected to nodulated alfalfa plants grown at different temperatures (ambient and ambient + 4 °C) and water availability regimes could protect PSII from photodamage. The main objective of this study was to determine the mechanism(s) involved in the photoprotection of PSII at elevated CO₂ levels in this plant. Elevated CO₂ reduced carboxylation capacity-induced photosynthetic acclimation and reduced enzymatic and/or nonenzymatic antioxidant activities, suggesting that changes in electron flow did not cause any photooxidative damage (which was also confirmed by H₂O₂ and lipid peroxidation analyses). Enhanced nonphotochemical quenching and xanthophyll cycle pigments revealed that plants grown at 700 μmol mol^?1 CO₂ compensated for the reduction in energy sink with a larger capacity for nonphotochemical dissipation of excitation energy as heat, i.e., modulating the status of the VAZ components. Elevated CO₂ induced the de-epoxidation of violaxanthin to zeaxanthin, facilitating thermal dissipation and protecting the photosynthetic apparatus against the deleterious effect of excess excitation energy.     

Response of ecosystem CO2 exchange to biomass productivity in a high yield grassland

We measured the biomass production and ecosystem carbon CO₂ exchange in a high yield grassland dominated by Miscanthus sinensis. The experimental grassland is managed by mowing once a year in winter every year and the harvested biomass on the ground is left to become the humus. The maximum aboveground and belowground biomasses were 1117 and 2803 g d.w. m^?2 in our grassland. Although the high potential of our grassland for biomass production led to higher carbon uptake than with other types of grassland, the large biomass contributed to a higher respired carbon loss. Biomass increase led to a linear increase in ecosystem respiration. Over the 3 years, RE₁₀ increased with increasing aboveground biomass. The potential gross primary production at a photosynthetic photon flux density of 2000 μmol m² s^?1 logarithmic increased with LAI. These responses of CO₂ exchange to biomass production suggest this grassland behaved as weak CO₂ sink or near carbon neutral (?78 and 17 g C m^?2 year^?1) in current management.     

The metabolic cost of bicarbonate use in the submerged plant Elodea nuttallii

The aquatic plant Elodea nuttallii (Planch.) St. John has been shown to express plasticity in the source of inorganic carbon it uses for photosynthesis. An investigation was undertaken to determine what effect the switch from CO₂ to HCO₃⁻ use had on the growth of E. nuttallii. Plants were grown under reduced CO₂ availability that favoured the switch, together with control plants (CO₂ at equilibrium with air) that continued to use CO₂ only. The extent to which both sets of plants could utilise HCO₃⁻ was determined (as the ratio of oxygen evolution at pH 9 and 6.5), and several measures of growth were made. Although reduced CO₂ availability produced an increase in HCO₃⁻ utilisation, no differences were found in the measured growth of the plants. Therefore, it was possible to estimate, from the difference between the estimated rate of photosynthesis of the plants utilising HCO₃⁻ and those using CO₂ only, the approximate cost of constructing, maintaining and running the bicarbonate utilisation mechanism in this species as 69 μmol photons m⁻² s⁻¹. This value can be used to estimate an irradiance of circa 80 μmol m⁻² s⁻¹ below which HCO₃⁻ use would not be expected in this species, an irradiance commonly experienced by submerged macrophytes in the field.     

A show cave management: Anthropogenic CO2 in atmosphere of Výpustek Cave (Moravian Karst,Czech Republic)

Anthropogenic impact on CO₂ levels was studied in the Bear Chamber of the Výpustek Cave, a show cave in the Moravian Karst (Czech Republic), during a period of active ventilation and enhanced attendance. The study showed that the natural CO₂ levels were controlled by (i) the natural CO₂ influxes from soils/epikarst (up to ∼5.64 × 10⁻² mol s⁻¹); and, (ii) the advective CO₂ fluxes out of cave atmosphere (up to 4.66 × 10⁻² mol s⁻¹). During visitor presence, the anthropogenic CO₂ flux into the chamber reached up to ∼0.13 mol s⁻¹ and exceeded all other CO₂ fluxes. The reachable anthropogenic steady states at sufficient duration of stay (up to 2.65 × 10⁻¹ mol m⁻³) could exceed the natural CO₂ levels by factor of more than nine based on the number of visitors. Recession analysis of anthropogenic pulses showed that intervals between individual visitor groups would have to be up to ∼6 h long if the cave environment has to return to natural conditions. As such pauses between individual tours are hardly realizable, a risk analysis was conducted to find the consequences of breaking natural conditions. It showed that the condition under which dripwater becomes aggressive to calcite (i.e., the point when P_CO2 in cave atmosphere exceeds the hypothetical CO₂ concentrations in epikarst that has participated on the water formation, P_CO2(H) = 10^−1.56) is potentially reachable under extreme conditions only (enormous visitor stay period and visitor number). In case of condensed water, however, any increase in CO₂ concentration will cause an increase of water aggressiveness to calcite. Therefore, in the periods and sites of enhanced condensation, it is important to strive for preservation of natural conditions.     

Leaf photosynthesis of Haberlea rhodopensis before and during drought

Haberlea rhodopensis is a homoiochlorophyllous resurrection plant that shows a low rate of leaf net CO₂ uptake (4–6 μmol m^?2 s^?1) under saturating photosynthetic photon flux densities in air (21% O₂ and about 390 ppm CO₂). However, leaf net CO₂ uptake reaches values of 17–18 μmol m^?2 s^?1 under saturating CO₂ and light. H. rhodopensis leaves have a very low mesophyll CO₂ conductance that can partly explain the low rate of leaf net CO₂ uptake in normal air. Experimental evidences suggest that mesophyll conductance is not sensitive to temperature in the 20–35 °C range. In addition, it is shown that the (1) transpiration rate of H. rhodopensis is nearly linearly related to the vapour pressure difference between the leaf and the ambient air within the interval from 0.5 kPa to 2.5 kPa at a leaf temperature of 25 °C and (2) leaf net CO₂ uptake in normal air under saturating light does not change much with leaf temperature (between 20 °C and 30 °C). At a leaf relative water content of between 90% and 30%, the decrease of leaf net CO₂ assimilation during drought can be explained by a decrease of leaf CO₂ diffusional conductance. Accordingly the non-photochemical chlorophyll fluorescence quenching decreases only at relative water contents lower than 20%, indicating that photosynthetic activity maintains a trans-thylakoidal proton gradient over a wide range of leaf water contents. Moreover, PSII photochemistry (as estimated by the Fv/Fm ratio and the thermoluminescence B band intensity) is only affected at leaf relative water contents lower than about 20%, thus confirming that primary photosynthetic reactions are resistant to drought. Interestingly, the effect of leaf desiccation on photosynthetic capacity, measured at very high ambient CO₂ molar ratios under saturating PPFD, is identical to that observed for three non-resurrection C₃ mesophytes. This demonstrates that the photosynthetic apparatus of H. rhodopensis is not more resistant to desiccation when compared to other C₃ plants. Since the leaf area decreases by more than 50% when the leaf relative water content is reduced to about 40% during drought it is supposed, following Farrant et al. [Farrant, J.M., Vander, W.C., Lofell, D.A., Bartsch, S., Whittaker, A., 2003. An investigation into the role of light during desiccation of three angiosperms resurrection plants. Plant Cell Environ. 26, 1275–1286], that H. rhodopensis leaf cells avoid mechanical stress.     

Interspecific variability of plant stomatal response to step changes of [CO2]

The kinetics of a stomatal response to sudden increases or decreases of CO₂ concentrations ([CO₂]) was studied in 13 plant species growing in the field. Plants were well supplied with water. In each plant, gas exchange measurements were made on a fully developed leaf that was first left to achieve steady-state stomatal conductance (g_s) at 400 μmol (CO₂) mol⁻¹) and then exposed to a step change of [CO₂] (to 700 μmol mol⁻¹ in one experiment; and to 700 and back to 400 μmol mol⁻¹ in a second experiment). Porometric data were captured in intervals of 3 s until a new steady state was reached.A comparison of t_1/2, the half-time needed to achieve new g_s, indicates similar responses of stomata in grasses when compared to herbs. The stomata of C₄ plants responded in approximately 5 min, the highest closure rate was detected in Echinochloa crus-galli and Digitaria sanguinalis. Opening rates were similar to closing rates and the response as a whole was rather symmetric. In C₃ plants, the full response of stomata was much slower. Analysis revealed differences in absolute rates of g_s change between C₃ and C₄ plants. These differences can be related to the specificities of the type of photosynthetic metabolism. C₄ photosynthesis enables plants to reduce g_s, which can hasten further changes of diffusivity in response to the environmental signals. A possible coupling of C₄ metabolism to the regulation of guard cells also has to be taken into account when explaining the observed results.