Internal conductance to CO2 transfer of adult Fagus sylvatica: Variation between sun and shade leaves and due to free-air ozone fumigation

Two separate objectives were considered in this study. We examined (1) internal conductance to CO₂ (g_i) and photosynthetic limitations in sun and shade leaves of 60-year-old Fagus sylvatica, and (2) whether free-air ozone fumigation affects g_i and photosynthetic limitations. g_i and photosynthetic limitations were estimated in situ from simultaneous measurements of gas exchange and chlorophyll fluorescence on attached sun and shade leaves of F. sylvatica. Trees were exposed to ambient air (1× O₃) and air with twice the ambient ozone concentration (2× O₃) in a free-air ozone canopy fumigation system in southern Germany (Kranzberg Forest). g_i varied between 0.12 and 0.24 mol m⁻² s⁻¹ and decreased CO₂ concentrations from intercellular spaces (C_i) to chloroplastic (C_c) by approximately 55 μmol mol⁻¹. The maximum rate of carboxylation (V_cmax) was 22–39% lower when calculated on a C_i basis compared with a C_c basis. g_i was approximately twice as large in sun leaves compared to shade leaves. Relationships among net photosynthesis, stomatal conductance and g_i were very similar in sun and shade leaves. This proportional scaling meant that neither C_i nor C_c varied between sun and shade leaves. Rates of net photosynthesis and stomatal conductance were about 25% lower in the 2× O₃ treatment compared with 1× O₃, while V_cmax was unaffected. There was no evidence that g_i was affected by ozone.     

Foliage type specific susceptibility to ozone in Picea abies,Pinus cembra and Larix decidua at treeline: A synthesis

Cumulative ozone uptake (COU, mmol m⁻²) and O₃ flux (FO₃, nmol m⁻² s⁻¹) were related to physiological, morphological and biochemical characteristics of field-grown mature evergreen Norway spruce [Picea abies (L.) Karst.], Cembran pine [Pinus cembra L.], and deciduous European larch [Larix decidua Mill.] trees at treeline. The threshold COU causing a statistically significant decline in photosynthetic capacity (A_max) ranged between 19.6 mmol m⁻² in current-year needles of evergreen conifers and 22.0 6 mmol m⁻² in short-shoot needles of deciduous L. decidua subjected to exposure periods of ≥84 and ≥43 days, respectively. The higher O₃ sensitivity of deciduous L. decidua than of evergreen P abies and P. cembra was associated with differences in FO₃ and specific leaf area (SLA), both being significantly higher in L. decidua. FO₃ was 5.9 nmol m⁻² s⁻¹ in L. decidua and 2.7 nmol m⁻² s⁻¹ in evergreen conifers. Species-dependent differences were also related to detoxification capacity expressed through total surface area based concentrations of reduced ascorbate and α-tocopherol that both increased with SLA. Findings suggest that differences in O₃ sensitivity between evergreen and deciduous conifers can be attributed to foliage type specific differences in SLA, the latter determining physiological and biochemical characteristics of the treeline conifers.     

Effects of flooding on photosynthesis and root respiration in saltcedar (Tamarix ramosissima), an invasive riparian shrub

The introduced shrub Tamarix ramosissima invades riparian zones, but loses competitiveness under flooding. Metabolic effects of flooding could be important for T. ramosissima, but have not been previously investigated. Photosynthesis rates, stomatal conductance, internal (intercellular) CO₂, transpiration, and root alcohol dehydrogenase (ADH) activity were compared in T. ramosissima across soil types and under drained and flooded conditions in a greenhouse. Photosynthesis at 1500 μmol quanta m⁻² s⁻¹ (A₁₅₀₀) in flooded plants ranged from 2.3 to 6.2 μmol CO₂ m⁻² s⁻¹ during the first week, but A₁₅₀₀ increased to 6.4–12.7 μmol CO₂ m⁻² s⁻¹ by the third week of flooding. Stomatal conductance (g_s) at 1500 μmol quanta m⁻² s⁻¹ also decreased initially during flooding, where g_s was 0.018 to 0.099 mol H₂O m⁻² s⁻¹ during the first week, but g_s increased to 0.113–0.248 mol H₂O m⁻² s⁻¹ by the third week of flooding. However, photosynthesis in flooded plants was reduced by non-stomatal limitations, and subsequent increases indicate metabolic acclimation to flooding. Root ADH activities were higher in flooded plants compared to drained plants, indicating oxygen stress. Lower photosynthesis and greater oxygen stress could account for the susceptibility of T. ramosissima at the onset of flooding. Soil type had no effect on photosynthesis or on root ADH activity. In the field, stomatal conductance, leaf water potential, transpiration, and leaf δ¹³C were compared between T. ramosissima and other flooded species. T. ramosissima had lower stomatal conductance and water potential compared to Populus deltoides and Phragmites australis. Differences in physiological responses for T. ramosissima could become important for ecological concerns.     

Stomatal,biochemical and morphological factors limiting photosynthetic gas exchange in the mangrove associate Hibiscus tiliaceus under saline and arid environment

The effect of salinity on leaf area and the relative accumulation of Na⁺ and K⁺ in leaves of the mangrove associate Hibiscus tiliaceus were investigated. Photosynthetic gas exchange characteristics were also examined under arid and non-arid leaf conditions at 0, 10, 20 and 30‰ substrate salinity. At salinities ≥ 40‰, plants showed complete defoliation followed by 100% mortality within 1 week. Salinities ≤ 30‰ were negatively correlated with the total leaf area per plant (r² = 0.94). The reduction in the total plant leaf area is attributed to the reduction in the area of individual leaves (r² = 0.94). Selective uptake of K⁺ over Na⁺ declined sharply with increasing salinity, where K⁺/Na⁺ ratio was reduced from 6.37 to 0.69 in plants treated with 0 and 30‰, respectively. Under non-arid leaf condition, increasing salinity from 0 to 30‰ has significantly reduced the values of the intrinsic components of photosynthesis V_c,max (from 50.4 to 18.4 μmol m⁻² s^-1), J_max (from 118.0 to 33.8 μmol photons m⁻² s⁻¹), and V_TPU (from 6.90 to 2.30 μmol m⁻² s⁻¹), while stomatal limitation to gas phase conductance (S_L) increased from 14.6 to 38.4%. Water use efficiency (WUE) has subsequently doubled from 3.20 for the control plants to 8.93 for 30‰ treatment. Under arid leaf conditions, the stomatal factor (S_L) was more limiting to photosynthesis than its biochemical components (73.4 to 26.6%, respectively, at 30‰). It is concluded that salinity causes a drastic decline in photosynthetic gas exchange in H. tiliaceus leaves through its intrinsic and stomatal components, and that the apparent phenotypic plasticity represented by the leaf area modulation is unlikely to be the mechanism by which H. tiliaceus avoids salt stress.     

Stomatal,biochemical and morphological factors limiting photosynthetic gas exchange in the mangrove associate Hibiscus tiliaceus under saline and arid environment

The effect of salinity on leaf area and the relative accumulation of Na⁺ and K⁺ in leaves of the mangrove associate Hibiscus tiliaceus were investigated. Photosynthetic gas exchange characteristics were also examined under arid and non-arid leaf conditions at 0, 10, 20 and 30‰ substrate salinity. At salinities ≥ 40‰, plants showed complete defoliation followed by 100% mortality within 1 week. Salinities ≤ 30‰ were negatively correlated with the total leaf area per plant (r² = 0.94). The reduction in the total plant leaf area is attributed to the reduction in the area of individual leaves (r² = 0.94). Selective uptake of K⁺ over Na⁺ declined sharply with increasing salinity, where K⁺/Na⁺ ratio was reduced from 6.37 to 0.69 in plants treated with 0 and 30‰, respectively. Under non-arid leaf condition, increasing salinity from 0 to 30‰ has significantly reduced the values of the intrinsic components of photosynthesis V_c,max (from 50.4 to 18.4 μmol m⁻² s^-1), J_max (from 118.0 to 33.8 μmol photons m⁻² s⁻¹), and V_TPU (from 6.90 to 2.30 μmol m⁻² s⁻¹), while stomatal limitation to gas phase conductance (S_L) increased from 14.6 to 38.4%. Water use efficiency (WUE) has subsequently doubled from 3.20 for the control plants to 8.93 for 30‰ treatment. Under arid leaf conditions, the stomatal factor (S_L) was more limiting to photosynthesis than its biochemical components (73.4 to 26.6%, respectively, at 30‰). It is concluded that salinity causes a drastic decline in photosynthetic gas exchange in H. tiliaceus leaves through its intrinsic and stomatal components, and that the apparent phenotypic plasticity represented by the leaf area modulation is unlikely to be the mechanism by which H. tiliaceus avoids salt 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.     

Sap flow measurements as a basis for assessing trace-gas exchange of trees

The assessment of trace-gas uptake by plants is of basic interest in plant ecophysiology and atmospheric chemistry. For tall vegetation and extensive canopies micrometeorological methods and modelling of deposition or combinations of both are usually the methods of choice. However, distinguishing between the aerodynamically driven components of deposition and stomatal uptake is difficult and estimates of plant uptake remain uncertain. Canopy conductance derived from sapflow measurements of trees represents an important and highly variable component to determine the uptake of trace gases by trees under free atmospheric conditions. The theory of the assessment of trace-gas uptake by the sapflow-based approach is reviewed and exemplified for the uptake of ozone, nitrogen oxides, and ammonia into coniferous and deciduous tree species. First results on the stomatal ammonia compensation point (χ_s=0.11–0.27 nmol mol⁻¹) of the coniferous tree species Picea abies determined by a bio-assay are reported and compared with published values on herbaceous plants and gas-exchange approaches for P. abies. For a summer period in 2003, the ground-area scaled uptake rate of gaseous NH₃ by P. abies was more than twice as high (1.38 nmol m⁻² s⁻¹) than the uptake of NO_x (0.53 nmol m⁻² s⁻¹). Estimates of ground-area scaled O₃ uptake and phytomedically relevant O₃ doses of Fagus sylvatica were found to be significantly less under dry conditions in August 2003 (cumulative uptake 2.3 mmol m⁻²) than in years with sufficient soil water supply despite higher atmospheric O₃ concentrations in 2003. Cumulative ground-area scaled O₃ uptake of Pinus cembra reached 150 mmol m⁻² during the growing season at an alpine site. Preliminary results and future perspectives are discussed for the transfer of the approach to the uptake of carbon dioxide and hence to determine total net primary production of trees. This novel approach has the potential to reduce uncertainties of C fluxes measured by the eddy-covariance technique and biogeochemical plot studies. It also allows to determine flux components like heterotrophic and autotrophic respiration separately as residuals from budget equations. Overall, it is concluded that the sapflow-based methodology contributes a new quality of flux data significantly improving our current understanding of biospheric aspects of trace-gas fluxes into tall vegetation.     

Physiological control of leaf methane emission from wetland plants

The transport of methane from the rhizosphere to the atmosphere takes place in the intercellular spaces and stomata of wetland plants, and foliar gas exchange is one of the critical steps of the transport process. The objectives of our research were to investigate: (i) variation in foliar gas exchange among four common wetland plant species (i.e., Peltandra virginica L., Orontium aquaticum L., Juncus effusus L., and Taxodium distichum L.), (ii) the role of key environmental factors (i.e., light, temperature, and carbon dioxide concentration) in controlling foliar methane emission, and (iii) physiological mechanisms underlying the variation in methane emission due to species and the environment. Experiments were conducted in an instantaneous, flow-through gas-exchange system that operated on a mass balance approach and concurrently measured foliar fluxes of methane, water vapor, and carbon dioxide. The chamber system allowed for the control of light, temperature, humidity, and carbon dioxide concentration. Diel patterns of methane emission varied among species, with daylight emissions from P. virginica and O. aquaticum 2–4 times those of J. effusus and T. distichum in saturating light. Foliar methane emission from P. virginica (1.80 μmol m⁻² s⁻¹) under ambient daylight conditions was an order of magnitude higher than that of the other three species (∼0.20 μmol m⁻² s⁻¹). As leaf temperature was increased by 10 °C, methane emission increased by a factor of 1.5–2.2, and the temperature effect was independent of stomatal conductance. When data were pooled among the four species, varying the light and carbon dioxide concentrations in a stepwise manner produced changes in foliar methane emission that were associated with stomatal conductance (r² = 0.52). To scale our observations to other wetland plant species, a stepwise multiple regression model is offered that incorporates stomatal conductance and net carbon dioxide assimilation to estimate instantaneous methane emission from foliar surfaces. The model indicates that changes in stomatal conductance affect methane emission three times more than equivalent changes in net carbon dioxide assimilation.     

Decomposition processes in soil of a healthy and a declining Phragmites australis stand

Decomposition processes were investigated in the soil of a declining, more eutrophic and a healthy, less eutrophic freshwater reed (Phragmites australis (Cav.) Trin. ex Steudel) stand in the littoral zone of Rožmberk fishpond, Czech Republic. Soil and pore water were sampled five times from April to October 1998. Chemical properties, CO₂ production in oxic and anoxic conditions, CH₄ production, denitrifying enzyme activity (DEA) and bacterial biomass were measured under laboratory conditions in suspensions prepared from homogenised soil samples. The more eutrophic West stand was more anaerobic than the East stand, with lower redox potential, lower pH and with a higher amount of organic acids, mainly acetic and lactic acid. Mean seasonal concentrations of total nitrogen in pore water, nitrogen of amino acids and proteins, and reducing sugars were all higher in the soil at the more eutrophic stand. Higher nutrient status and more reduced conditions at the more eutrophic stand were accompanied by (i) a limitation of aerobic microbial activities (CO₂ production in oxic conditions: 0.35 versus 0.54 μmol CO₂ cm⁻³ h⁻¹); lower DEA (4.0 versus 20.2 nmol N₂O cm⁻³ h⁻¹) and a lower proportion of bacteria that were active in aerobic conditions; (ii) by a prevalence of anaerobic over aerobic microbial processes; (iii) by a higher rate of methanogenesis (15.0 versus 11.5 nmol CH₄ cm⁻³ h⁻¹) and (iv) by an overall lower rate of microbial processes as compared to less eutrophied stand. The shift from aerobic to anaerobic microbial metabolism, and a coinciding restriction of metabolic activities at the more eutrophic stand are indicative of an elevated oxygen stress in the soil, associated with accumulation of metabolites toxic to both the micro-organisms and the reed. Possible links between eutrophication, decomposition processes in the soil and reed decline are discussed.     

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.     

Effects of overexpressing photosynthetic carbon flux control enzymes in the cyanobacterium Synechocystis PCC 6803

Synechocystis PCC 6803 is a model unicellular cyanobacterium used in e.g. photosynthesis and CO₂ assimilation research. In the present study we examined the effects of overexpressing Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), sedoheptulose 1,7-biphosphatase (SBPase), fructose-bisphosphate aldolase (FBA) and transketolase (TK), confirmed carbon flux control enzymes of the Calvin-Bassham-Benson (CBB) cycle in higher plants, in Synechocystis PCC 6803. Overexpressing RuBisCO, SBPase and FBA resulted in increased in vivo oxygen evolution (maximal 115%), growth rate and biomass accumulation (maximal 52%) under 100 μmol photons m⁻² s⁻¹ light condition. Cells overexpressing TK showed a chlorotic phenotype but increased biomass by approximately 42% under 100 μmol photons m⁻² s⁻¹ light condition. Under 15 μmol photons m⁻² s⁻¹ light condition, cells overexpressing TK showed enhanced in vivo oxygen evolution. This study demonstrates increased growth and biomass accumulation when overexpressing selected enzymes of the CBB cycle. RuBisCO, SBPase, FBA and TK are identified as four potential targets to improve growth and subsequently also yield of valuable products from Synechocystis PCC 6803.     

Differential tolerance to combined salinity and O2 deficiency in the halophytic grasses Puccinellia ciliata and Thinopyrum ponticum: The importance of K+ retention in roots

Saline environments of terrestrial halophytes are often prone to waterlogging, yet the effects on halophytes of combined salinity and waterlogging have rarely been studied. Either salinity or hypoxia (low O₂) alone can interfere with K⁺ homeostasis, therefore the combination of salinity or hypoxia is expected to impact significantly on K⁺ retention in roots. We studied mechanisms of tolerance to the interaction of salinity with hypoxia in Puccinellia ciliata and Thinopyrum ponticum, halophytic grasses that differ in waterlogging tolerance. Plants were exposed to aerated and stagnant saline (250 mM NaCl) treatments with low (0.25 mM) and high (4 mM) K⁺ levels; growth, net ion fluxes and tissue ion concentrations were determined. P. ciliata was more tolerant than T. ponticum to stagnant-saline treatment, producing twice the biomass of adventitious roots, which accumulated high levels of Na⁺, and had lower shoot Na⁺. After 24 h of saline hypoxic treatment, MIFE measurements revealed a net uptake of K⁺ (∼40 nmol m⁻² s⁻¹) for P. ciliata, but a net loss of K⁺ (∼20 nmol m⁻² s⁻¹) for the more waterlogging sensitive T. ponticum. NaCl alone induced K⁺ efflux from roots of both species, with channel blocker tests implicating GORK-like channels. P. ciliata had constitutively a more negative root cell membrane potential than T. ponticum (−150 versus −115 mV). Tolerance to salinity and hypoxia in P. ciliata is related to increased production of adventitious roots, regulation of shoot K⁺/Na⁺, and a superior ability to maintain negative membrane potential in root cells, resulting in greater retention of K⁺.     

Seasonal variations in photosynthetic irradiance response curves of macrophytes from a Mediterranean coastal lagoon

The main photosynthesis and respiration parameters (dark respiration rate, light saturated production rate, saturation irradiance, photosynthetic efficiency) were measured on a total of 23 macrophytes of the Thau lagoon (2 Phanerogams, 5 Chlorophyceae, 10 Rhodophyceae and 6 Phaeophyceae). Those measurements were performed in vitro under controlled conditions, close to the natural ones, and at several seasons. Concomitantly, measurements of pigment concentrations, carbon, phosphorous and nitrogen contents in tissues were performed. Seasonal intra-specific variability of photosynthetic parameters was found very high, enlightening an important acclimatation capacity. The highest photosynthetic capacities were found for Chlorophyceae (e.g. Monostroma obscurum thalli at 17 °C, 982 μmol O₂ g⁻¹ dw h⁻¹ and 9.1 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹, respectively for light saturated net production rate and photosynthetic efficiency) and Phanerogams (e.g. Nanozostera noltii leaves at 25 °C, 583 μmol O₂ g⁻¹ dw h⁻¹ and 2.6 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹ respectively for light saturated net production rate and photosynthetic efficiency). As expected, species with a high surface/volume ratio were found to be more productive than coarsely branched thalli and thick blades shaped species. Contrary to R_d (ranging 6.7–794 μmol O₂ g⁻¹ dw h⁻¹, respectively for Rytiphlaea tinctoria at 7 °C and for Dasya sessilis at 25 °C) for which a positive relationship with water temperature was found whatever the species studied, the evolution of P/I curves with temperature exhibited different responses amongst the species. The results allowed to show summer nitrogen limitation for some species (Gracilaria bursa-pastoris and Ulva spp.) and to propose temperature preferences based on the photosynthetic parameters for some others (N. noltii, Zostera marina, Chaetomorpha linum).     

Partitioning of latent heat flux at a northern peatland

The partitioning of latent heat flux (Q_E) to vascular plant and moss surface components was assessed for a Sphagnum-dominated bog with a hummock–hollow surface having a sparse canopy of low shrubs. Results from porometry and eddy covariance measurements of Q_E showed evaporation from the moss surface ranged from greater than 50% of total Q_E early in the growing season to less than 20% after a dry period toward the end of the growing season. Both soil moisture and vapour pressure deficit (D_a) affected this partitioning with drier moss and peat, lower water table, and smaller D_a all reducing moss Q_E. Daily maximum moss Q_E ranged from greater than 200 W m⁻² early in the growing season to less than 100 W m⁻² during a dry period. In contrast, vascular contribution to total Q_E increased over the season from a daily maximum of about 150 W m⁻² to 250 W m⁻² due to increase in leaf area by leaf replacement and emergence and to drying of the moss surface. Porometry results showed average daily maximum conductance from bog shrubs was near 8 mm s⁻¹. These conductance values were smaller than those reported for vascular plants from more nutrient-rich wetlands. The effect of increases in D_a on vascular Q_E were moderated by decreases in stomatal conductance. At constant available energy, vascular leaf conductance was reduced by as much as 2 mm s⁻¹ and moss surface conductance was enhanced by up to 3 mm s⁻¹ by large D_a. Considering vascular and non-vascular water transport characteristics and frequency of water table position and given the observed variations of Q_E partitioning with water table location and moss and peat water content, it is suggested that modelling efforts focus on how dry hummocks and wet hollows each contribute to Q_E, especially as related to D_a and soil moisture dynamics.     

Effects of ozone fumigation on cotton (Gossypium hirsutum L.) morphology,anatomy, physiology,yield and qualitative characteristics of fibers

This experiment was conducted to study the effect of high ozone concentrations on two cotton (Gossypium hirsutum L.) cultivars. Two cotton cultivars (Romanos and Allegria) were exposed to control (CF < 4 ppb O₃) and 100 ppb O₃. Plant exposure to ozone began eight days after emergence and was interrupted one day before removing the leaves, to calculate the leaf area. Plants were exposed to ozone 7 h/day, in closed and controlled-environment chambers, during their illumination with artificial visible light.In comparison to control plants, plants exposed to O₃ showed chlorotic and necrotic patches on their leaves, increased stomatal or epidermal cell density and yellowness of cotton fibers. Elevated ozone concentration did not have a significant effect on stomatal width, total leaf thickness and thickness of histological components of leaves. Exposure to ozone concentration reduced non-glandular hair density of main leaf veins, plant height, mainstem internode length, chlorophyll content, net photosynthetic rate, stomatal conductance and length and area of bracts and petals. Elevated ozone treatment reduced the maximum length of staminal tube, anther number, pollen grain germination, leaf area, leaf dry weight, boll number, raw cotton weight, total branch length, dry weight of the mainstem–branches–bracts–carpophylls and of root dry weight. Furthermore, exposure to O₃ reduced the seed weight, the lint weight, the yield, the ratio of lint weight to seed weight, the fiber strength, the micronaire, the maturity index and the fiber uniformity index values. This study shows that the exposure to high ozone concentrations mainly affected the rate of photosynthesis, raw cotton weight and strength of cotton fibers.     

Evaluation of the association of mercury(II) with some dicysteinyl tripeptides

The present study was undertaken to gain insight into the associations of mercury(II) with dicysteinyl tripeptides in buffered media at pH 7.4. We investigated the effects of increasing the distance between cysteinyl residues on mercury(II) associations and complex formations. The peptide–mercury(II) formation constants and their associated thermodynamic parameters in 3-(N-morpholino)propanesulfonic acid (MOPS) buffered solutions were evaluated by isothermal titration calorimetry. Complexes formed in different relative ratios of mercury(II) to cysteinyl peptides in ammonium formate buffered solutions were characterized by LTQ Orbitrap mass spectrometry. The results from these studies show that n-alkyl dicysteinyl peptides (CP 1–4), and an aryl dicysteinyl peptide (CP 5) can serve as effective “double anchors” to accommodate the coordination sites of mercury(II) to form predominantly one-to-one Hg(peptide) complexes. The aryl dicysteinyl peptide (CP 5) also forms the two-to-two Hg₂(peptide)₂ complex. In the presence of excess peptide, Hg(peptide)₂ complexes are also detected. Notably, increasing the distance between the ligating groups or “anchor points” in CP 1–5 does not significantly affect their affinity for mercury(II). However, the enthalpy change (ΔH) values (ΔH₁ ∼ −91 kJ mol⁻¹ and ΔH₂ ∼ −66 kJ mol⁻¹) for complex formation between CP 4 and 5 with mercury(II) are about one and a half times larger than the related values for CP 1, 2 and 3 (ΔH₁ ∼ −66 kJ mol⁻¹ and ΔH₂ ∼ 46 kJ mol⁻¹). The corresponding entropy change (ΔS) values (ΔS₁ ∼ −129 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −116 J K⁻¹ mol⁻¹) of the structurally larger dicysteinyl peptides CP 4 and 5 are less entropically favorable than for CP 1, 2 and 3 (ΔS₁ ∼ −48 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −44 J K⁻¹ mol⁻¹). Generally, these associations result in a decrease in entropy, indicating that these peptide–mercury complexes potentially form highly ordered structures. The results from this study show that dicysteinyl tripeptides are effective in binding mercury(II) and they are promising motifs for the design of multi-cysteinyl peptides for binding more than one mercury(II) ion per peptide.     

Improvement of succinate production by overexpression of a cyanobacterial carbonic anhydrase in Escherichia coli

Succinate fermentation was investigated in Escherichia coli strains overexpressing cyanobacterium Anabaena sp. 7120 ecaA gene encoding carbonic anhydrase (CA). In strain BL21 (DE3) bearing ecaA, the activity of CA was 21.8 U mg⁻¹ protein, whereas non-detectable CA activity was observed in the control strain. Meanwhile, the activity of phosphoenolpyruvate carboxylase (PEPC) increased from 0.2 U mg⁻¹ protein to 1.13 U mg⁻¹ protein. The recombinant bearing ecaA reached a succinate yield of 0.39 mol mol⁻¹ glucose at the end of the fermentation. It was 2.1-fold higher than that of control strain which was just 0.19 mol mol⁻¹ glucose. EcaA gene was also introduced into E. coli DC1515, which was deficient in glucose phosphotransferase, lactate dehydrogenase and pyruvate:formate lyase. Succinate yield can be further increased to 1.26 mol mol⁻¹ glucose. It could be concluded that the enhancement of the supply of HCO₃⁻ in vivo by ecaA overexpression is an effective strategy for the improvement of succinate production in E. coli.     

The effects of NH4+ and NO3− on growth,resource allocation and nitrogen uptake kinetics of Phragmites australis and Glyceria maxima

The effects of NH₄⁺ or NO₃⁻ on growth, resource allocation and nitrogen (N) uptake kinetics of two common helophytes Phragmites australis (Cav.) Trin. ex Steudel and Glyceria maxima (Hartm.) Holmb. were studied in semi steady-state hydroponic cultures. At a steady-state nitrogen availability of 34 μM the growth rate of Phragmites was not affected by the N form (mean RGR = 35.4 mg g⁻¹ d⁻¹), whereas the growth rate of Glyceria was 16% higher in NH₄⁺-N cultures than in NO₃⁻-N cultures (mean = 66.7 and 57.4 mg g⁻¹ d⁻¹ of NH₄⁺ and NO₃⁻ treated plants, respectively). Phragmites and Glyceria had higher S/R ratio in NH₄⁺ cultures than in NO₃⁻ cultures, 123.5 and 129.7%, respectively.Species differed in the nitrogen utilisation. In Glyceria, the relative tissue N content was higher than in Phragmites and was increased in NH₄⁺ treated plants by 16%. The tissue NH₄⁺ concentration (mean = 1.6 μmol g fresh wt⁻¹) was not affected by N treatment, whereas NO₃⁻ contents were higher in NO₃⁻ (mean = 1.5 μmol g fresh wt⁻¹) than in NH₄⁺ (mean = 0.4 μmol g fresh wt⁻¹) treated plants. In Phragmites, NH₄⁺ (mean = 1.6 μmol g fresh wt⁻¹) and NO₃⁻ (mean = 0.2 μmol g fresh wt⁻¹) contents were not affected by the N regime. Species did not differ in NH₄⁺ (mean = 56.5 μmol g⁻¹ root dry wt h⁻¹) and NO₃⁻ (mean = 34.5 μmol g⁻¹ root dry wt h⁻¹) maximum uptake rates (V_max), and V_max for NH₄⁺ uptake was not affected by N treatment. The uptake rate of NO₃⁻ was low in NH₄⁺ treated plants, and an induction phase for NO₃⁻ was observed in NH₄⁺ treated Phragmites but not in Glyceria. Phragmites had low K_m (mean = 4.5 μM) and high affinity (10.3 l g⁻¹ root dry wt h⁻¹) for both ions compared to Glyceria (K_m = 6.3 μM, affinity = 8.0 l g⁻¹ root dry wt h⁻¹). The results showed different plasticity of Phragmites and Glyceria toward N source. The positive response to NH₄⁺-N source may participates in the observed success of Glyceria at NH₄⁺ rich sites, although other factors have to be considered. Higher plasticity of Phragmites toward low nutrient availability may favour this species at oligotrophic sites.     

Different O3 response of sensitive and resistant snap bean genotypes (Phaseolus vulgaris L.): The key role of growth stage,stomatal conductance,and PSI activity

The ozone (O₃) biomonitoring system based on sensitive (S156) and resistant (R123) genotypes of snap bean (Phaseolus vulgaris L.), was tested in a fumigation experiment, with the aim of: (i) assessing the performance of the S156/R123 system under simulated climatic conditions and O₃ levels often found in Mediterranean sites; (ii) contributing to identify possible ecophysiological mechanisms determining the different O₃ sensitivity of S156 and R123, along different plant growth stages. In contrast with what reported by other authors, differences in stomatal O₃ uptake were evident between sensitive and resistant plants. In particular, R123 plants showed an O₃-induced stomatal closure (−38.1% than the control) during flowering and the onset of visible injury, a behaviour that can be regarded as an O₃ avoidance mechanism. At the end of the fumigation period (AOT40 = 4.66 ppm h), despite the reduction of assimilation and the photoprotective down regulation of PSII photochemistry, the capacity to reduce the final electron acceptors beyond PSI was up-regulated, particularly in S156, while the PSI activity remained constant in both genotypes, an unusual response to O₃ stress. Increased energy demand for maintenance and repair processes also determined increased dark respiration rates (Rd) in all fumigated plants; nocturnal stomatal conductance (g_sN) was also enhanced, differently in the two genotypes, with possible implication for higher nocturnal stomatal uptake of S156 in field conditions. No clear genotype × O₃ effect was instead detected on pod biomass, although a significant O₃-induced reduction of yield was evident in both genotypes.     

Effects of intraspecific competition on growth and photosynthesis of Atriplex prostrata

We investigated the effect of intraspecific competition on growth parameters and photosynthesis of the salt marsh species Atriplex prostrata Boucher in order to distinguish the effects of density-dependent growth inhibition from salt stress. High plant density caused a reduction of 30% in height, 82% in stem dry mass, 80% in leaf dry mass, and 95% in root dry mass. High density also induced a pronounced 72% reduction in leaf area, 29% decrease in length of mature internodes and 50% decline in net photosynthetic rate. The alteration of net photosynthesis paralleled growth inhibition, decreasing from 7.6 ± 0.9 μmol CO₂ m⁻² s⁻¹ at low density to 3.5 ± 0.4 μmol CO₂ m⁻² s⁻¹ at high density, indicating growth inhibition caused by intraspecific competition is mainly due to a decline in net photosynthesis rate. Plants grown at high density also exhibited a reduction in stomatal conductance from 0.7 ± 0.1 mol H₂O m⁻² s⁻¹ at low density to 0.3 ± 0.1 mol H₂O m⁻² s⁻¹ at high density and a reduction in transpiration rate from 6.0 ± 0.3 mmol H₂O m⁻² s⁻¹ at low density to 4.3 ± 0.3 mmol H₂O m⁻² s⁻¹ at high density. Biomass production was inhibited by an increase in plant density, which reduced the rate of photosynthesis, stomatal conductance and leaf area of plants.