Role of ethylene in alleviation of cadmium-induced photosynthetic capacity inhibition by sulphur in mustard

Sulphur (S) assimilation leads to the formation of glutathione (GSH) and alleviation of cadmium (Cd) stress. GSH is synthesized from its immediate metabolite cysteine, which also serves as a metabolite for ethylene formation through S‐adenosyl methionine. To assess the role of ethylene in S‐induced alleviation of Cd stress on photosynthesis, the effects of S or ethephon (ethylene source) on GSH and ethylene were examined in mustard (Brassica juncea L. cv. Varuna). Sufficient‐S at 100 mg S kg^?1 soil alleviated Cd‐induced photosynthetic inhibition more than excess‐S (200 mg S kg^?1 soil) via ethylene by increased GSH. Under Cd stress, plants were less sensitive to ethylene, despite high ethylene evolution, and showed photosynthetic inhibition. Ethylene sensitivity of plants increased with ethephon or sufficient‐S, triggering the induction of an antioxidant system, and leading to increased photosynthesis even under Cd stress. The effects of ethephon and S under Cd stress were similar. The effects of S were reversed by ethylene biosynthesis inhibitor, aminoethoxyvinylglycine (AVG), suggesting that ethylene plays an important role in S‐induced alleviation of Cd stress on photosynthesis.     

Cross-talk between salicylic acid and NaCl-generated reactive oxygen species and nitric oxide in tomato during acclimation to high salinity

Hydrogen peroxide (H₂O₂) and nitric oxide (NO) generated by salicylic acid (SA) are considered to be functional links of cross‐tolerance to various stressors. SA‐stimulated pre‐adaptation state was beneficial in the acclimation to subsequent salt stress in tomato (Solanum lycopersicum cv. Rio Fuego). At the whole‐plant level, SA‐induced massive H₂O₂ accumulation only at high concentrations (10^?3–10^?2M), which later caused the death of plants. The excess accumulation of H₂O₂ as compared with plants exposed to 100 mM NaCl was not associated with salt stress response after SA pre‐treatments. In the root tips, 10^?3–10^?2M SA triggered the production of reactive oxygen species (ROS) and NO with a concomitant decline in the cell viability. Sublethal concentrations of SA, however, decreased the effect of salt stress on ROS and NO production in the root apex. The attenuation of oxidative stress because of high salinity occurred not only in pre‐adapted plants but also at cell level. When protoplasts prepared from control leaves were exposed to SA in the presence of 100 mM NaCl, the production of NO and ROS was much lower and the viability of the cells was higher than in salt‐treated samples. This suggests that, the cross‐talk of signalling pathways induced by SA and high salinity may occur at the level of ROS and NO production. Abscisic acid (ABA), polyamines and 1‐aminocyclopropane‐1‐carboxylic acid, the compounds accumulating in pre‐treated plants, enhanced the diphenylene iodonium‐sensitive ROS and NO levels, but, in contrast to others, ABA and putrescine preserved the viability of protoplasts.     

Ethylene reverses photosynthetic inhibition by nickel and zinc in mustard through changes in PS II activity,photosynthetic nitrogen use efficiency,and antioxidant metabolism

We investigated the influence of exogenously sourced ethylene (200 μL L^?1 ethephon) in the protection of photosynthesis against 200 mg kg^?1 soil each of nickel (Ni)- and zinc (Zn)-accrued stress in mustard (Brassica juncea L.). Plants grown with Ni or Zn but without ethephon exhibited increased activity of 1-aminocyclopropane carboxylic acid synthase, and ethylene with increased oxidative stress measured as H₂O₂ content and lipid peroxidation compared with control plants. The oxidative stress in Ni-grown plants was higher than Zn-grown plants. Under metal stress, ethylene protected photosynthetic potential by efficient PS II activity and through increased activity of ribulose-1,5-bisphosphate carboxylase and photosynthetic nitrogen use efficiency (P-NUE). Application of 200 μL L^?1 ethephon to Ni- or Zn-grown plants significantly alleviated toxicity and reduced the oxidative stress to a greater extent together with the improved net photosynthesis due to induced activity of ascorbate peroxidase and glutathione (GSH) reductase, resulting in increased production of reduced GSH. Ethylene formation resulting from ethephon application alleviated Ni and Zn stress by reducing oxidative stress caused by stress ethylene production and maintained increased GSH pool. The involvement of ethylene in reversal of photosynthetic inhibition by Ni and Zn stress was related to the changes in PS II activity, P-NUE, and antioxidant capacity was confirmed using ethylene action inhibitor, norbornadiene.     

Chloride and sulphur concentrations in chloroplasts of spinach

Chloride concentrations within individual chloroplasts, the adjacent cytoplasm and nearby vacuoles of spinach mesophyll cells (Spinacea oleracea L. cv. Hybrid 102) were determined by means of electron probe X-ray microanalysis in the cleavage plane of quench frozen tissue, which was maintained at liquid nitrogen temperatures. The accuracy of quantitative data obtained with this technique is greatly improved by the adoption of a peak to background ratio method and use of carbon slurry standards, which mimic the quench frozen tissue and its X-ray fluorescence. Chloroplasts were incapable of maintaining relatively high levels of Cl^? under conditions of low Cl^? availability (zero Cl^? or 20 μM Cl^? in nutrient solution), and under conditions of Cl^? stress (100 or 200 mM Cl^?) chloroplasts had only a limited capacity to maintain a Cl^? concentration at a level below that of the cytoplasm and vacuole. However, under conditions of Cl^? stress the concentration of Cl^? in cytoplasm immediately adjacent to chloroplasts was substantially higher than in the chloroplasts or more distant cytoplasm. Thus, Cl^? levels in chloroplasts are apparently not as tightly regulated as was suggested by estimates of Cl^? concentration based on aqueous isolation of chloroplasts. Levels of S in chloroplasts were relatively high (equivalent to 40–60 mM SO₄^2? in S standards) and constant for all treatments, with the possible exception of lower S levels in chloroplasts of leaves approaching premature senescence as a result of salt stress. It is implied that the stability of the S-content results largely from its presence in macromolecular components of chloroplasts (sulfolipids and proteins).     

S‐deficiency responsive accumulation of amino acids is mainly due to hydrolysis of the previously synthesized proteins – not to de novo synthesis in Brassica napus

To characterize the mechanisms of amino acid accumulation under sulphur (S)‐deficiency and its physiological significance in Brassica napus, stable isotopes ¹⁵N and ³⁴S were employed. The plants were exposed for 9 days to S‐deficient conditions (0.05 mM vs 1.5 mM sulphate). After 9 days of S‐deficiency, leaf‐osmotic potential and total chlorophyll content significantly decreased. S uptake decreased by 94%, whereas N uptake and biomass were not significantly changed. Using ¹⁵N and ³⁴S labelling, de novo synthesis of amino acids and proteins derived from newly absorbed NO₃^? and SO₄^2? and the content of N and S in the previously synthesized amino acids and proteins were quantified. At the whole plant level, S‐deficiency increased the pool of amino acids but resulted in strong decrease of incorporation of newly absorbed NO₃^? and SO₄^2? into amino acids by 22.2 and 76.6%, respectively, compared to the controls. Total amount of N and S incorporated into proteins also decreased by 28.8 and 62.1%, respectively. The levels of ¹⁴N‐ and ³²S‐proteins (previously synthesized proteins) strongly decreased, mainly in mature leaves. The data thus indicate that amino acid accumulation under short‐term S‐deficiency results from the degradation of previously synthesized proteins rather than from de novo synthesis.     

Enhanced tolerance of photosynthesis against high temperature damage in salt-adapted halophyte Atriplex centralasiatica plants

Thermotolerance of photosynthesis in salt‐adapted Atriplex centralasiatica plants (100–400 mm NaCl) was evaluated in this study after detached leaves and whole plants were exposed to high temperature stress (30–48 °C) either in the dark or under high light (1200 mol m^?2 s^?1). In parallel with the decrease in stomatal conductance, intercellular CO₂ concentration and CO₂ assimilation rate decreased significantly with increasing salt concentration. There was no change in the maximal efficiency of PSII photochemistry (F_v/F_m) with increasing salt concentration, suggesting that there was no damage to PSII in salt‐adapted plants. On the other hand, there was a striking difference in the response of PSII and CO₂ assimilation capacity to heat stress in non‐salt‐adapted and salt‐adapted leaves. Leaves from salt‐adapted plants maintained significantly higher F_v/F_m values than those from non‐salt‐adapted leaves at temperatures higher than 42 °C. The F_v/F_m differences between non‐salt‐adapted and salt‐adapted plants persisted for at least 24 h following heat stress. Leaves from salt‐adapted plants also maintained a higher net CO₂ assimilation rate than those in non‐salt‐adapted plants at temperatures higher than 42 °C. This increased thermotolerance was independent of the degree of salinity since no significant changes in F_v/F_m and net CO₂ assimilation rate were observed among the plants treated with different concentrations of NaCl. The increased thermotolerance of PSII induced by salinity was still evident when heat treatments were carried out under high light. Given that photosynthesis is considered to be the physiological process most sensitive to high temperature damage, increased thermotolerance of photosynthesis may be of significance since A. centralasiatica, a typical halophyte, grows in the high salinity regions in the north of China, where the temperature in the summer is often as high as 45 °C.     

Antioxidant enzyme activities and hormonal status in response to Cd stress in the wetland halophyte Kosteletzkya virginica under saline conditions

Salt marshes constitute major sinks for heavy metal accumulation but the precise impact of salinity on heavy metal toxicity for halophyte plant species remains largely unknown. Young seedlings of Kosteletzkya virginica were exposed during 3 weeks in nutrient solution to Cd 5 µM in the presence or absence of 50 mM NaCl. Cadmium (Cd) reduced growth and shoot water content and had major detrimental effect on maximum quantum efficiency (F_v/F_m), effective quantum yield of photosystem II (Y(II)) and electron transport rates (ETRs). Cd induced an oxidative stress in relation to an increase in O₂^?? and H₂O₂ concentration and lead to a decrease in endogenous glutathione (GSH) and α‐tocopherol in the leaves. Cd not only increased leaf zeatin and zeatin riboside concentration but also increased the senescing compounds 1‐aminocyclopropane‐1‐carboxylic acid (ACC) and abscisic acid (ABA). Salinity reduced Cd accumulation already after 1 week of stress but was unable to restore shoot growth and thus did not induce any dilution effect. Salinity delayed the Cd‐induced leaf senescence: NaCl reduced the deleterious impact of Cd on photosynthesis apparatus through an improvement of F_v/F_m, Y(II) and ETR. Salt reduced oxidative stress in Cd‐treated plants through an increase in GSH, α‐tocopherol and ascorbic acid synthesis and an increase in glutathione reductase (EC 1.6.4.2) activity. Additional salt reduced ACC and ABA accumulation in Cd+NaCl‐treated leaves comparing to Cd alone. It is concluded that salinity affords efficient protection against Cd to the halophyte species K. virginica, in relation to an improved management of oxidative stress and hormonal status.     

Photosynthetic limitations in olive cultivars with different sensitivity to salt stress

Olive (Olea europea L) is one of the most valuable and widespread fruit trees in the Mediterranean area. To breed olive for resistance to salinity, an environmental constraint typical of the Mediterranean, is an important goal. The photosynthetic limitations associated with salt stress caused by irrigation with saline (200 mm ) water were assessed with simultaneous gas‐exchange and fluorescence field measurements in six olive cultivars. Cultivars were found to possess inherently different photosynthesis when non‐stressed. When exposed to salt stress, cultivars with inherently high photosynthesis showed the highest photosynthetic reductions. There was no relationship between salt accumulation and photosynthesis reduction in either young or old leaves. Thus photosynthetic sensitivity to salt did not depend on salt exclusion or compartmentalization in the old leaves of the olive cultivars investigated. Salt reduced the photochemical efficiency, but this reduction was also not associated with photosynthesis reduction. Salt caused a reduction of stomatal and mesophyll conductance, especially in cultivars with inherently high photosynthesis. Mesophyll conductance was generally strongly associated with photosynthesis, but not in salt‐stressed leaves with a mesophyll conductance higher than 50 mmol m^?2 s^?1. The combined reduction of stomatal and mesophyll conductances in salt‐stressed leaves increased the CO₂ draw‐down between ambient air and the chloroplasts. The CO₂ draw‐down was strongly associated with photosynthesis reduction of salt‐stressed leaves but also with the variable photosynthesis of controls. The relationship between photosynthesis and CO₂ draw‐down remained unchanged in most of the cultivars, suggesting no or small changes in Rubisco activity of salt‐stressed leaves. The present results indicate that the low chloroplast CO₂ concentration set by both low stomatal and mesophyll conductances were the main limitations of photosynthesis in salt‐stressed olive as well as in cultivars with inherently low photosynthesis. It is consequently suggested that, independently of the apparent sensitivity of photosynthesis to salt, this effect may be relieved if conductances to CO₂ diffusion are restored.     

Photosynthesis,carbon allocation,and growth of sulfur dioxide ecotypes ofGeranium carolinianum L.

Summary This study investigated ways in which genetically determined differences in SO₂ susceptibility resulting from ecotypic differentiation inGeranium carolinianum were expressed physiologically. The SO₂-resistant and SO₂-sensitive ecotypes were exposed to a combination of short- and long-term SO₂ exposures to evaluate the responses of photosynthesis, H₂S efflux from foliage (sulfur detoxification), photoassimilate retention, leaf-diffusive resistance to CO₂, and growth. When exposed to SO₂, both ecotypes re-emit sulfur in a volatile, reduced form, presumably as H₂S. Because H₂S efflux rates at various SO₂ concentrations were comparable between ecotypes, genetic differences inG. carolinianum could not be attributed to a re-emission of excess sulfur as H₂S. Incipient SO₂ effects on photosynthesis were observed as cumulative SO₂ flux into the leaf interior excecded 0.40 nmol·m^–2 in the resistant ecotype and 0.26 nmol·m^–2 in the sensitive ecotype. Although initial SO₂-induced changes in photosynthesis in both ecotypes were mediated through an increase in stomatal resistance to CO₂, the ecotype-specific patterns as a function of pollutant concentration and exposure time were associated with marked increases in residual resistance to CO₂. Patterns in photosynthesis, photoassimilate retention, and growth following long-term SO₂ exposures were also ecotype-specific. Although physiological accommodation of SO₂ stress was observed in both ecotypes, it was more pronounced in the resistant ecotype. The physiological mechanisms underlying genetic differences inG. carolinianum in response to SO₂ stress were concluded to be (1) dissimilar threshold levels of response to SO₂ and/or its toxic derivatives and (2) differences in homeostatic processes governing the rate of repair or compensation for physiological injury.Research sponsored by the Office of Health and Environmental Research, U.S. Department of Energy, under contract No. DEAC05-840R21400 with Martin Marietta, Energy Systems, Inc. and the U.S. Environmental Protection AgencyPublication No. 2610, Environmental Sciences Division, Oak Ridge National Laboratory     

Salicylic acid differently impacts ethylene and polyamine synthesis in the glycophyte Solanum lycopersicum and the wild‐related halophyte Solanum chilense exposed to mild salt stress

This study aimed to determine the effects of exogenous application of salicylic acid (SA) on the toxic effects of salt in relation to ethylene and polyamine synthesis, and to correlate these traits with the expression of genes involved in ethylene and polyamine metabolism in two tomato species differing in their sensitivity to salt stress, Solanum lycopersicum cv Ailsa Craig and its wild salt‐resistant relative Solanum chilense. In S. chilense, treatment with 125 mM NaCl improved plant growth, increased production of ethylene, endogenous salicylic acid and spermine. The production was related to a modification of expression of genes involved in ethylene and polyamine metabolism. In contrast, salinity decreased plant growth in S. lycopersicum without affecting endogenous ethylene, salicylic or polyamine concentrations. Exogenous application of salicylic acid at 0.01 mM enhanced shoot growth in both species and affected ethylene and polyamine production in S. chilense. Concomitant application of NaCl and salicylic acid improved osmotic adjustment, thus suggesting that salt and SA may act in synergy on osmolyte synthesis. However, the beneficial impact of exogenous application of salicylic acid was mitigated by salt stress since NaCl impaired endogenous SA accumulation in the shoot and salicylic acid did not improve plant growth in salt‐treated plants. Our results thus revealed that both species respond differently to salinity and that salicylic acid, ethylene and polyamine metabolisms are involved in salt resistance in S. chilense.     

Interaction between CO2 enrichment and salinity stress in the C4 non-halophyte Andropogon glomeratus (Walter) BSP

Abstract Increasing atmospheric CO₂ may result in alleviation of salinity stress in salt-sensitive plants. In order to assess the effect of enriched CO₂ on salinity stress in Andropogon glomeratus, a C₄ non-halophyte found in the higher regions of salt marshes, plants were grown at 350, 500, and 650 cm³ m^?3 CO₂ with 0 or 100 mol m^?3 NaCl watering treatments. Increases in leaf area and biomass with increasing CO₂ were measured in salt-stressed plants, while decreases in these same parameters were measured in non-salt-stressed plants. Tillering increased substantially with increasing CO₂ in salt-stressed plants, resulting in the increased biomass. Six weeks following initiation of treatments, there was no difference in photosynthesis on a leaf area basis with increasing CO₂ in salt-stressed plants, although short-term increases probably occurred. Stomatal conductance decreased with increasing CO₂ in salt-stressed plants, resulting in higher water-use efficiency, and may have improved the diurnal water status of the plants. Concentrations of Na⁺ and Cl^? were higher in salt stressed-plants while the converse was found for K ⁺. There were no differences in leaf ion content between CO₂ treatments in the salt-stressed plants. Decreases in photosynthesis in salt-stressed plants occurred primarily as a result of decreased internal (non-stomatal) conductance.     

Water relations and leaf gas exchange properties in some elite canola (Brassica napus) lines under salt stress

In order to examine whether growth of eight genetically diverse canola (Brassica napus) lines under salt stress is positively associated with their rate of photosynthesis and other gas exchange related attributes, 20‐day old plants of all eight lines were subjected to salinised soil containing 2.4 dS m^?1 NaCl (control), 4 dS m^?1 NaCl, 8 dS m^?1 NaCl or 12 dS m‐¹ NaCl. The lines DGL (non canola) and Dunkeld were found to be salt tolerant and Rainbow and Cyclon salt sensitive with regard to shoot dry matter production and seed yield under saline conditions. In most of the lines there was a negative relationship between growth and net CO₂ assimilation rate. For example, the salt sensitive line Cyclon was the lowest and Con‐III the highest, and the salt tolerant line Dunkeld intermediate in net CO₂ assimilation rate under salt stress. Stomatal conductance was found to be lower in the salt sensitive line Cyclon, followed by the salt tolerant line Dunkeld along with Oscar. Water use efficiency estimated as Pn/E was moderate in the salt sensitive line Cyclon and the salt tolerant line Dunkeld. In conclusion, high salt tolerance of Dunkeld and DGL (non‐canola) was not positively associated with net CO₂ assimilation rate or Pn/E.     

Multiple effects of salinity on photosynthesis of the protist Euglena gracilis

The effect of NaCl in the culture medium on growth, photosynthesis and cell content of chlorophyll, K⁺, Na⁺, Ca²⁺ and Mg²⁺ in Euglena gracilis was studied. O₂ production, quantum yield of photosystem II (PSII), the non-photochemical quenching of chlorophyll fluorescence (q_N) and the chlorophyll alb ratio all diminished by 0.2 M NaCl. Respiration and chlorophyll a and b increased, whereas the photochemical quenching (q_p) of chlorophyll fluorescence was not affected by 0.2 M NaCl. Salt stress also induced an increase in cell volume and in K⁺ and Na⁺ concentrations, but decreased the concentrations of Ca²⁺ and Mg²⁺. Except for a protective effect on O₂ production, additional Ca²⁺ in the culture medium did not attenuate the salt effect on the parameters measured. The addition of HCO₃^? restored the PSII quantum yield of O₂ production in cells grown in high salt. Salt stress promoted a decrease in the apparent rate of quinone A (Q_A) reduction and an apparent obstruction of Q_B reduction, which were not prevented by excess HCO₃^?; the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) did not increase chlorophyll fluorescence in salt-grown cells. These results indicate that photosynthesis in Euglena grown under salt stress exhibits: (1) diminution of the HCO₃^? dependent water-splitting activity of PSII; (2) inhibition of the electron transfer at the quinone pool level; (3) probable increase in thylakoid stacking (as indicated by the effect on the chlorophyll alb ratio); and (4) dissipation of the H⁺ gradient across the thylakoid membranes (as indicated by the decrease of q_N).     

ACCLIMATION OF EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) TO PHOTON FLUX DENSITY1

Growth rate, pigment composition, and noninvasive chl a fluorescence parameters were assessed for a noncalcifying strain of the prymnesiophyte Emiliania huxleyi Lohman grown at 50, 100, 200, and 800 μmol photons·m^?2·s^?1. Emiliania huxleyi grown at high photon flux density (PFD) was characterized by increased specific growth rates, 0.82 d^?1 for high PFD grown cells compared with 0.38 d^?1 for low PFD grown cells, and higher in vivo chl a specific attenuation coefficients that were most likely due to a decreased pigment package, consistent with the observed decrease in cellular photosynthetic pigment content. High PFD growth conditions also induced a 2.5‐fold increase in the pool of the xanthophyll cycle pigments diadinoxanthin and diatoxanthin responsible for dissipation of excess energy. Dark‐adapted maximal photochemical efficiency (F_v/F_m) remained constant at around 0.58 for cells grown over the range of PFDs, and therefore the observed decline, from 0.57 to 0.33, in the PSII maximum efficiency in the light‐adapted state, (F_v′/F_m′), with increasing growth PFD was due to increased dissipation of excess energy, most likely via the xanthophyll cycle and not due to photoinhibition. The PSII operating efficiency (F_q′/F_m′) decreased from 0.48 to 0.21 with increasing growth PFD due to both saturation of photochemistry and an increase in nonphotochemical quenching. The changes in the physiological parameters with growth PFD enable E. huxleyi to maximize rates of photosynthesis under subsaturating conditions and protect the photosynthetic apparatus from excess energy while supporting higher saturating rates of photosynthesis under saturating PFDs.     

Increasing NaCl and CaCl2 concentrations in the growth medium of quince leaves: II. Effects on shoot regeneration

Summary The effects of NaCl and CaCl₂ on shoot regeneration from quince (Cydonia oblonga BA L29 clone) leaves were investigated. Caulogenesis was induced on in vitro-grown leaves treated for 2d in liquid Murashige and Skoog (MS) medium with 11.3 μM 2,4-dichlorophenoxyacetic acid and cultured on MS gelled medium supplemented with 4.5 μM thidiazuron and 0.5 μM naphthaleneacetic acid. Three experiments were performed: in the first, we compared the effects of NaCl at 0, 25, 50, 100, and 200 mM in factorial combination with 3, 9, and 27 mM CaCl₂. In the second, NaCl was tested at 0, 5, 10, 20, 40, and 80 mM with CaCl₂ at 0.3, 1.0, and 3.0 mM. The third experiment was carried out with the same experimental design as the second one but replacing NaCl with Na₂SO₄. Shoot regeneration was evaluated after 50 d of culturing: 25 in darkness and 25 in white light. In the first experiment, shoot regeneration was very poor and was observed only at the lower salt concentrations. In the second experiment, the percentages of caulogenic leaves were much higher, but decreased with increasing NaCl concentration. The more pronounced negative effect of the highest NaCl concentrations appeared to be partly mitigated by CaCl₂ at 1 and 3 mM. The presence of 3 mM CaCl₂, in the experiment with Na₂SO₄, appeared to be even more effective in reducing the adverse effect of sodium stress on caulogenesis. This result was attributed to the lower Cl⁻ concentration in the growth medium, which resulted from replacing NaCl with Na₂SO₄. NaCl applied at low concentrations (5 and 10 mM) in combination with 3 mM CaCl₂ exerted a favorable effect on adventitious shoot regeneration. As regards the Na⁺ and Ca²⁺ interaction, when the Na⁺/Ca²⁺ ratio was below roughly 35 and 20, with NaCl and Na₂SO₄, respectively, at least 60% of leaves showed regenerating capacity, but optimal values of this ratio were not derived.     

The use of low [CO2] to estimate diffusional and non-diffusional limitations of photosynthetic capacity of salt-stressed olive saplings

In this study it has been shown that increased diffusional resistances caused by salt stress may be fully overcome by exposing attached leaves to very low [CO₂] (～ 50 µmol mol^?1), and, thus a non‐destructive‐in vivo method to correctly estimate photosynthetic capacity in stressed plants is reported. Diffusional (i.e. stomatal conductance, g_s, and mesophyll conductance to CO₂, g_m) and biochemical limitations to photosynthesis (A) were measured in two 1‐year‐old Greek olive cultivars (Chalkidikis and Kerkiras) subjected to salt stress by adding 200 mm NaCl to the irrigation water. Two sets of A–C_i curves were measured. A first set of standard A–C_i curves (i.e. without pre‐conditioning plants at low [CO₂]), were generated for salt‐stressed plants. A second set of A–C_i curves were measured, on both control and salt‐stressed plants, after pre‐conditioning leaves at [CO₂] of ～ 50 µmol mol^?1 for about 1.5 h to force stomatal opening. This forced stomata to be wide open, and g_s increased to similar values in control and salt‐stressed plants of both cultivars. After g_s had approached the maximum value, the A–C_i response was again measured. The analysis of the photosynthetic capacity of the salt‐stressed plants based on the standard A–C_i curves, showed low values of the J_max (maximum rate of electron transport) to V_cmax (RuBP‐saturated rate of Rubisco) ratio (1.06), that would implicate a reduced rate of RuBP regeneration, and, thus, a metabolic impairment. However, the analysis of the A–C_i curves made on pre‐conditioned leaves, showed that the estimates of the photosynthetic capacity parameters were much higher than in the standard A–C_i responses. Moreover, these values were similar in magnitude to the average values reported by Wullschleger (Journal of Experimental Botany 44, 907–920, 1993) in a survey of 109 C₃ species. These findings clearly indicates that: (1) salt stress did affect g_s and g_m but not the biochemical capacity to assimilate CO₂ and therefore, in these conditions, the sum of the diffusional resistances set the limit to photosynthesis rates; (2) there was a linear relationship (r² = 0.68) between g_m and g_s, and, thus, changes of g_m can be as fast as those of g_s; (3) the estimates of photosynthetic capacity based on A–C_i curves made without removing diffusional limitations are artificially low and lead to incorrect interpretations of the actual limitations of photosynthesis; and (4) the analysis of the photosynthetic properties in terms of stomatal and non‐stomatal limitations should be replaced by the analysis of diffusional and non‐diffusional limitations of photosynthesis. Finally, the C₃ photosynthesis model parameterization using in vitro‐measured and in vivo‐measured kinetics parameters was compared. Applying the in vivo‐measured Rubisco kinetics parameters resulted in a better parameterization of the photosynthesis model.     

Constitutive and inducible aspects of nitrate-nitrogen uptake by Chlamydomonas reinhardtii

Similarly to higher plant root systems, Chlamydomonas reinhardtii Dangeard (UTEX 90) cells exhibited biphasic NO₃^? uptake kinetics. The uptake pattern was similar in cells cultured in 10 mM NO₃^? (NO₃^?-grown), 0.25 mM NO₃^? (N-limited) or 10 mM NO₃^? followed by an 18-h period of N-deprivation (N-starved). In all cell types there was an apparent phase transition in uptake at 1.1 mM NO₃^?, although there were variations in the uptake V_max of both isotherms. The rate of uptake via isotherm 0 ([NO₃^?]<1.1 mM) in N-limited cells was higher than that of either NO₃^?-grown or N-starved cells. In contrast, NO₃^?-grown and N-limited cells exhibited comparable V_max values when supplied with 1.1 to 1.8 mM NO₃^? (isotherm 1). When supplied with 1.6 mM NO₃^?, both N-limited and N-starved cells exhibited enhanced linear uptake after 60 min of incubation. We ascribed this to an induction phenomenon. This trend was not observed when NO₃^?-grown cells were supplied with 1.6 mM NO₃^?, or when N-limited and N-starved cells were supplied with 0.6 mM NO₃^?. The ‘inducible’ aspect of uptake by N-limited cells was blocked by cycloheximide (10 mg l^?1), but not by actinomycin D (5 mg l^?1), thus indicating the involvement of a translational or post-translational event. To investigate this phenomenon further, we analysed the cell proteins of N-limited cells supplied with either 0.6 or 1.6 mM NO₃^? for 90 min, using two-dimensional gel electrophoresis. Comparison of protein profiles enabled the identification of a single cell membrane-associated polypeptide (21 kDa, pI ca 5.5) and ten soluble fraction polypeptides (17–73 kDa, pI ca 5.0 to 7.1) unique to the high NO₃^? treatment. We propose that the ‘inducible’ portion of NO₃^? uptake may provide the means by which C. reinhardtii cells regulate uptake in accordance with assimilatory capacity.     

Whole Plant Regulation of Sulfur Nutrition of Deciduous Trees-Influences of the Environment

Abstract: The current view of sulfur nutrition is based on the source‐to‐sink relationship of carbohydrates. SO₄^2‐ reduction is thought to occur mainly in leaves. Surplus reduced sulfur must be transported out of the leaves, loaded into the phloem and transported to other tissues, in particular tissues assumed to be sink organs. However, it has not been proved that tissues which are sinks for carbohydrates are also sink organs for reduced sulfur. It is evident that sinks must communicate with sources, and vice versa, to signal demand and to transport the surplus of reduced sulfur that is produced. The demand‐driven control model of sulfur nutrition proposes that the tripeptide glutathione is the signal which regulates S nutrition of the whole plant at the level of SO₄^2‐ uptake. Acclimatization to environmental changes has been shown to result in several changes in S nutrition of deciduous trees: (i) Drought stress diminished SO₄^2‐ transport into the xylem, although the GSH content in lateral roots remained unaffected, possibly due to an overall reduction in water status. (ii) Flooding decreased APS reductase activity in the anoxic roots. This may be due to enhanced GSH transport to the roots, but it is more likely to be the result of a change in metabolism leading to diminished energy gain in the roots. (iii) Mycorrhization enhanced the GSH content in the phloem, while SO₄^2‐ uptake was not affected. This clearly goes against the demand‐driven control model. (iv) Under both short‐ and long‐term exposure to elevated pCO₂, the APS reductase activity in leaves and lateral roots did not correlate with the GSH contents therein. Therefore, it must be assumed that, under these conditions, regulation of S nutrition goes beyond the demand‐driven control model, and occurs within the network of other nutrient metabolism. (v) Atmospheric S in the form of H₂S enhanced the reduced sulfur content of the phloem and lateral roots. Under these conditions, the SO₄^2‐ loaded into the xylem decreased. It would appear that the demand‐driven control model of sulfur nutrition is not always valid in the case of deciduous trees.     

Diffusional conductance to CO2 is the key limitation to photosynthesis in salt‐stressed leaves of rice (Oryza sativa)

Salinity significantly limits leaf photosynthesis but the factors causing the limitation in salt‐stressed leaves remain unclear. In the present work, photosynthetic and biochemical traits were investigated in four rice genotypes under two NaCl concentration (0 and 150 mM) to assess the stomatal, mesophyll and biochemical contributions to reduced photosynthetic rate (A) in salt‐stressed leaves. Our results indicated that salinity led to a decrease in A, leaf osmotic potential, electron transport rate and CO₂ concentrations in the chloroplasts (C_c) of rice leaves. Decreased A in salt‐stressed leaves was mainly attributable to low C_c, which was determined by stomatal and mesophyll conductance. The increased stomatal limitation was mainly related to the low leaf osmotic potential caused by soil salinity. However, the increased mesophyll limitation in salt‐stressed leaves was related to both osmotic stress and ion stress. These findings highlight the importance of considering mesophyll conductance when developing salinity‐tolerant rice cultivars.     

Nitric Oxide Alleviates Oxidative Damage Induced by Enhanced Ultraviolet-B Radiation in Cyanobacterium

To study the role of nitric oxide (NO) on enhanced ultraviolet-B (UV-B) radiation (280–320 nm)-induced damage of Cyanobacterium, the growth, pigment content, and antioxidative activity of Spirulina platensis-794 cells were investigated under enhanced UV-B radiation and under different chemical treatments with or without UV-B radiation for 6 h. The changes in chlorophyll-a, malondialdehyde content, and biomass confirmed that 0.5 mM sodium nitroprusside (SNP), a donor of nitric oxide (NO), could markedly alleviate the damage caused by enhanced UV-B. Specifically, the biomass and the chlorophyll-a content in S. platensis-794 cells decreased 40% and 42%, respectively under enhanced UV-B stress alone, but they only decreased 10% and 18% in the cells treated with UV-B irradiation and 0.5 mM SNP. Further experiments suggested that NO treatment significantly increased the activities of superoxide dismutase (SOD) and catalase (CAT), and decreased the accumulation of O ₂ ⁻ in enhanced UV-B-irradiated cells. SOD and CAT activity increased 0.95- and 6.73-fold, respectively. The accumulation of reduced glutathione (GSH) increased during treatment with 0.5 mM SNP in normal S. platensis cells, but SNP treatment could inhibit the increase of GSH in enhanced UV-B-stressed S. platensis cells. Thus, these results suggest that NO can strongly alleviate oxidative damage caused by UV-B stress by increasing the activities of SOD, peroxidase, CAT, and the accumulation of GSH, and by eliminating O ₂ ⁻ in S. platensis-794 cells. In addition, the difference of NO origin between plants and cyanobacteria are discussed.