Processes contributing to photoprotection of grapevine leaves illuminated at low temperature

Photoinactivation of photosystem II (PSII) and energy dissipation at low leaf temperatures were investigated in leaves of glasshouse-grown grapevine ( Vitis vinifera L. cv. Riesling). At low temperatures (< 15°C), photosynthetic rates of CO₂ assimilation were reduced. However, despite a significant increase in the amount of light excessive to that required by photosynthesis, grapevine leaves maintained high intrinsic quantum efficiencies of PSII ( F _v/ F _m) and were highly resistant to photoinactivation compared to other species. Non-photochemical energy dissipation involving xanthophylls and fast D1 repair were the main protective processes reducing the 'gross' rate of photoinactivation and the 'net' rate of photoinactivation, respectively. We developed an improved method of energy dissipation analysis that revealed up to 75% of absorbed light is dissipated thermally via pH- and xanthophyll-mediated non-photochemical quenching at low temperatures (5–15°C) and moderate (800 µmol quanta m⁻² s⁻¹) light. Up to 20% of the energy flux contributing to electron transport was dissipated via photorespiration when taking into account temperature-dependent mesophyll conductance; however, this flux used in photorespiration was only a relatively small amount of the total absorbed light energy. Photoreduction of O₂ at photosystem I (PSI) and subsequent superoxide detoxification (water-water cycle) was more sensitive to inhibition by low temperature than photorespiration. Therefore the water-water cycle represents a negligibly small energy sink below 15°C, irrespective of mesophyll conductance.     

Light-dependent inhibition of photosynthetic electron transport by zinc

The effects of zinc concentrations up to 400 μ M were examined on three photosynthetic electron transport reactions of thylakoids isolated from Pisum sativum L. cv. Meteor. Zinc (400 μ M ) had no effect on photosystem I mediated electron transport from reduced N,N,N',N'-tetramethyl- p -phenylenediamine to methyl viologen, but inhibited uncoupled electron flow from water to methyl viologen by ca 50% and to 2,6-dichlorophenol-indophenol (DCPIP) by ca 30% at saturating light levels. Zinc inhibition of DCPIP photoreduction was independent of the light intensity to which thylakoids were exposed. Decreasing the photon flux density below 400 μmol m⁻² s⁻¹ produced a logarithmic reduction in the zinc-induced inhibition of methyl viologen photoceduction; a stimulation of this reaction was observed below 80 μmol photons m⁻² s⁻¹. Increasing light intensity decreased the amount of zinc tightly bound to the thylakoid membranes, but increased the weakly associated zinc which could be removed by washing the membranes with buffer containing Mg². The results suggest that zinc acts on the photosynthetic electron transport system at two sites. Site 1 is on the oxidizing side of photosystem 2 and the inhibition by zinc is independent of the light intensity. Site 2 is between photosystems 1 and 2 and the electron flow can be positively or negatively affected by zinc depending on the light intensity.     

Apoplastic and membrane‐associated Ca2+ in leaves and roots as affected by boron deficiency

A combination of fluorescein‐isothiocyanate (FITC), coumarin‐benzothiazol (BTC), and chlorotetracycline (CTC) fluorescence was used to simultaneously monitor apoplastic pH, apoplastic free Ca²⁺, and plasma membrane‐bound Ca²⁺. As early boron deficiency reactions supposedly include alterations of plasma membrane‐bound transport processes besides rapid effects on cell wall physical properties, the corresponding changes were followed in leaves and roots of Vicia faba L. cv. Troy.
Boron deficiency did not alter the apoplastic pH, but it reduced plasma membrane‐bound Ca²⁺ in roots at 4 h and leaves at 3 days after starting the deficiency treatment. The decrease in plasma membrane‐bound Ca²⁺ coincided with an increase in apoplastic free Ca²⁺ and K⁺, and occurred before the first visible symptoms were noticed.
It is proposed that less Ca²⁺ is bound to the plasma membrane due to a reduction of specific Ca²⁺‐binding sites (borate esters with vic ‐diols or polyhydroxy‐carboxylates) before plasma membrane integrity deteriorates.     

Characterization of mitochondrial glutamate dehydrogenase from dark‐grown soybean seedlings

Purified preparations of NAD(H)‐glutamate dehydrogenase (GDH, EC 1.4.1.2.) were assayed to determine the effects of mono‐ and divalent cations, nucleotides and select carbon compounds on NAD(H)‐dependent GDH activity. The amination reaction was stimulated 2‐ to 17‐fold by divalent cations (Ca²⁺ > Cd²⁺ > Co²⁺ > Mg²⁺ > Mn²⁺ > Zn²⁺ between 1 and 1000 µ M ), but the reaction was unaffected by monovalent cations (Na ⁺ and K ⁺). The amination reaction was most responsive to changes in Ca²⁺ in a NADH‐dependent manner. The addition of EDTA or EGTA nullified the stimulatory effects of Ca²⁺. Calmodulin alone or in combination with calmodulin antagonists did not affect the amination reaction. Divalent cations (at 1 m M ) inhibited the rate of the deamination reaction by 15 to 25%, while monovalent cations had no effect. ATP inhibited the amination reaction by 10 to 60%, while ADP had little or no effect. ATP or ADP decreased the rate of the deamination reaction 23 to 60 or 20 to 38%, respectively. Many tricarboxylic acid cycle intermediates inhibited the amination reaction, 20 to 50% of the inhibition could be attributed to the chelating capacity of intermediates. Conversely, most of the carbon sources tested did not affect the deamination reaction, the only appreciable differences were increases in activity with sucrose (21%) and glucose (41%) and a decrease in activity with pyruvate (34%). Inhibitors of sulfhydryl groups were used to examine the importance of reduced thiol groups in the amination or deamination reactions. The amination was not dependent on reduced thiol groups, whereas the deamination reaction was dependent on reduced thiol groups.     

CO2, light and temperature influence senescence and protein degradation in wheat leaf segments

Effects of environmental conditions influencing photosynthesis and photorespiration on senescence and net protein degradation were investigated in segments from the first leaf of young wheat ( Triticum aestivum L. cv. Arina) plants. The segments were floated on H₂O at 25, 30 or 35°C in continuous light (PAR: 50 or 150 µmol m⁻² s⁻¹) in ambient air and in CO₂‐depleted air. Stromal enzymes, including phosphoglycolate phosphatase, glutamine synthetase, ferredoxin‐dependent glutamate synthase, phosphoribulokinase, and the peroxisomal enzyme, glycolate oxidase, were detected by SDS‐PAGE followed by immunoblotting with specific antibodies. In general, the net degradation of proteins and chlorophylls was delayed in CO₂‐depleted air. However, little effect of CO₂ on protein degradation was observed at 25°C under the lower level of irradiance. The senescence retardation by the removal of CO₂ was most pronounced at 30°C and at the higher irradiance. The stromal enzymes declined in a coordinated manner. Immunoreactive fragments from the degraded polypeptides were in most cases not detectable. However, an insolubilized fragment of glycolate oxidase accumulated in vivo, especially at 25°C in the presence of CO₂. Detection of this fragment was minimal after incubation at 30°C and completely absent on blots from segments kept at 35°C. In CO₂‐depleted air, the fragment was only weakly detectable after incubation at 25°C. The results from these investigations indicate that environmental conditions that influence photosynthesis may interfere with senescence and protein catabolism in wheat leaves.     

Direct and indirect effects of Cd2+ on photosynthesis in sugar beet (Beta vulgaris)

Sugar-beet plants ( Beta vulgaris L. cv. Monohill) were cultivated for 4 weeks in a complete nutrient solution. Indirect effects of cadmium were studied by adding 5, 10 or 20 μ M CdCl₂ to the culture medium while direct effects were determined by adding 1, 5, 20, 50 or 2 000 μ M CdCl₂ to the assay media. The photosynthetic properties were characterized by measurement of CO₂ fixation in intact plants, fluorescence emission by intact leaves and isolated chloroplasts, photosystem (PS) I and PSII mediated electron transport of isolated chloroplasts, and CO₂-dependent O₂ evolution by protoplasts. When directly applied to isolated leaves, protoplasts and chloroplasts. Cd²⁺ impeded CO₂ fixation without affecting the rates of electron transport of PSI or PSII or the rate of dark respiration. When Cd²⁺ was applied through the culture medium the capacity for, and the maximal quantum yield of CO₂ assimilation by intact plants both decreased. This was associated with: (1) decreased total as well as effective chlorophyll content (PSII antennae size), (2) decreased coupling of electron transport in isolated chloroplasts, (3) perturbed carbon reduction cycle as indicated by fluorescence measurements. Also, protoplasts isolated from leaves of Cd²⁺-cultivated plants showed an increased rate of dark respiration.     

Involvement of abscisic acid and ethylene in the responses of citrus seedlings to salt shock

The responses of salt‐sensitive citrus rootstocks to 200 m M NaCl were periodically determined on seedlings of citrange Carrizo ( Citrus sinensis [L.] Osbeck × Poncirus trifoliata [L.] Raf) during 30 days. The stressed seedlings adjusted osmotically, reduced stomatal conductance, increased proline content and ethylene production, and showed massive leaf abscission (92%). The salt shock also increased abscisic acid (ABA) and aminocyclopropane‐1‐carboxylic acid (ACC) in roots, xylem fluid and leaves, and in addition promoted Cl⁻ accumulation. The pattern of change of ABA, ACC and proline followed a two‐phase response: an initial transient increase (10‐12 days) overlapping with a gradual and continuous accumulation. This biphasic response appears to be compatible with the proposal that the transitory hormonal rises are induced by the osmotic component of salinity, whereas the Cl⁻ increase determines the subsequent accumulations. During the second phase, Cl⁻ levels correlated with abscission in leaves. Production of leaf ethylene was also concomitant with the increase in the abscission rate. Salt‐induced abscission was either reduced with CoCl₂ (52%) or inhibited with silver thiosulphate (14%). The results suggest that in salt‐stressed citrus, leaf abscission is induced by the chloride build‐up through a mechanism that stimulates leaf ACC synthesis and further conversion to ethylene.     

Effects of cadmium on growth of Helianthus annuus seedlings: physiological aspects

Sunflower seedlings ( Helianthus annuus hybrid Select) were grown in a complete nutrient solution in the absence or presence of Cd²⁺ (10 and 20 μM). Analyses were performed to establish whether there was a differential effect of Cd²⁺ on mature and young leaves. After 7 d the growth parameters as well as the leaf area had decreased in both mature and young leaves. Accumulation of Cd²⁺ in the roots exceeded that in the shoots. Seedlings treated with Cd²⁺ exhibited reduced contents of chlorophyll and CO₂ assimilation rate, with a greater decrease in young leaves. The photochemical efficiency of photosystem II (PSII) was not altered by Cd²⁺ treatment in either mature or young leaves, although during steady-state photosynthesis in young leaves there was a significant alteration in the following parameters: quantum yield of electron transport by PSII (Φ_PSII), photochemical quenching ( q _P), non-photochemical quenching ( q _NP), and excitation capture efficiency of PSII (Φ_exc).     

Loss of the precise control of photosynthesis and increased yield of non‐radiative dissipation of excitation energy after mild heat treatment of barley leaves

The after effects of a short exposure of intact barley leaves to moderately elevated temperature (40°C, 5 min) on the induction transients and the irradiance dependencies of photosynthesis and chlorophyll fluorescence are presented. This mild heat treatment strongly reduced the oscillations in the rate of photosynthesis and in the yield of chlorophyll fluorescence. However, only a 25% irreversible inhibition of maximum photosynthetic capacity of photosystem II (PSII) measured by oxygen evolution was produced and the intrinsic quantum yield of PSII measured by the chlorophyll fluorescence ratio (F_m‐ F_o)/F_m decreased by only 15%. In contrast, the above treatment increased radiationless dissipation processes in PSII by a factor of two. In heat‐treated leaves, photosynthesis was not saturated even by strong light. Both ΔpH‐dependent quenching of excitons in PSII (including formation of zeaxanthin) and state 1/state 2 transition were found to be stimulated. Heat exposure enhanced the control of PSII activity by PSI, as evidenced by a significant increase in the quenching effect of far‐red light on the maximum yield of chlorophyll fluorescence. It was deduced that after mild heat treatment, the photosynthetic apparatus in leaves lacks the precise coordinating control of electron transport and carbon metabolism owing to the inability of PSII to support electron transport at a level adequate for carbon metabolism. This effect was not related to the small irreversible thermal damage to PSII, but was rather due to a significant increase in non‐photochemical quenching of excitation energy.     

H+/PPi stoichiometry of a membrane‐bound pyrophosphatase of plant mitochondria

The H⁺/PP_i stoichiometry of the mitochondrial H⁺‐PP_iase from pea ( Pisum sativum L.) stem was determined by two kinetic approaches, and compared with the H⁺/substrate stoichiometries of the mitochondrial H⁺‐ATPase, and the vacuolar H⁺‐PP_iase and H⁺‐ATPase. Using sub‐mitochondrial particles or preparations enriched in vacuolar membranes, the rates of substrate‐dependent H⁺‐transport were evaluated: by a mathematical model, describing the time‐course of H⁺‐gradient (ΔpH) formation; or by determining the rate of H⁺‐leakage following H⁺‐pumping inhibition by EDTA at the steady‐state ΔpH. When the H⁺‐transport rates were divided by those of PP_i or ATP hydrolysis, measured under identical conditions, apparent stoichiometries of ca 2 were determined for the mitochondrial H⁺‐PP_iase and H⁺‐ATPase, and for the vacuolar H⁺‐ATPase. The stoichiometry of the vacuolar H⁺‐PP_iase was found to be ca 1. From these results, it is suggested that the mitochondrial H⁺‐PP_iase may, in theory, function as a primary H⁺‐pump poised towards synthesis of PP_i and, therefore, acting in parallel with the main H⁺‐ATPase.     

Relationship between photosystem II activity and CO2 fixation in leaves

There is now potential to estimate photosystem II (PSII) activity in vivo from chlorophyll fluorescence measurements and thus gauge PSII activity per CO₂ fixed. A measure of the quantum yield of photosystem II, Φ_II (electron/photon absorbed by PSII), can be obtained in leaves under steady-state conditions in the light using a modulated fluorescence system. The rate of electron transport from PSII equals Φ_II times incident light intensity times the fraction of incident light absorbed by PSII. In C₄ plants, there is a linear relationship between PSII activity and CO₂ fixation, since there are no other major sinks for electrons; thus measurements of quantum yield of PSII may be used to estimate rates of photosynthesis in C₄ species. In C₃ plants, both CO₂ fixation and photorespiration are major sinks for electrons from PSII (a minimum of 4 electrons are required per CO₂, or per O₂ reacting with RuBP). The rates of PSII activity associated with photosynthesis in C₃ plants, based on estimates of the rates of carboxylation (v_o) and oxygenation (v_o) at various levels of CO₂ and O₂, largely account for the PSII activity determined from fluorescence measurements. Thus, in C₃ plants, the partitioning of electron flow between photosynthesis and photorespiration can be evaluated from analysis of fluorescence and CO₂ fixation.     

Photosynthesis of two poplar clones under long‐term exposure to ozone

The photosynthetic response was studied in two clones ( Populus deltoides × maximowiczii Eridano and Populus × euramericana I‐214), known for their differential response to ozone (O₃) in terms of visible symptoms, when exposed to O₃ (60 nl l⁻¹ 5 h day⁻¹, 7 and 15 days). The photosynthetic ability was tested using gas exchange and chlorophyll fluorescence analysis. O₃ caused a decrease in the CO₂ assimilation rate at light saturation level in mature leaves of both clones. Alterations of Chl fluorescence parameters, in particular the F_v/F_m ratio and non‐photochemical quenching were also observed. The effects were similar for both clones and it could not be concluded that differential effects on electron transport capacity were responsible for the observed reduction in photosynthesis. The reduction of photosynthetic rate in Eridano was due mainly to a reduced mesophyll activity, as evidenced by the increase in intercellular CO₂ concentration and the minimal changes in stomatal conductance. In contrast, in I‐214, stomatal effects were primarily responsible, although effects on the mesophyll cannot be excluded. Data obtained indicate that the effects observed at the mesophyll level may be attributed to indirect effects caused by membrane disorders.     

Changes in photosystem II function during senescence of wheat leaves

Analyses of chlorophyll fluorescence were undertaken to investigate the alterations in photosystem II (PSII) function during senescence of wheat ( Triticum aestivum L. cv. Shannong 229) leaves. Senescence resulted in a decrease in the apparent quantum yield of photosynthesis and the maximal CO₂ assimilation capacity. Analyses of fluorescence quenching under steady‐state photosynthesis showed that senescence also resulted in a significant decrease in the efficiency of excitation energy capture by open PSII reaction centers (F'_v/F'_m) but only a slight decrease in the maximum efficiency of PSII photochemistry (F'_v/F'_m). At the same time, a significant increase in non‐photochemical quenching (q_N) and a considerable decrease in photochemical quenching (q_P) were observed in senescing leaves. Rapid fluorescence induction kinetics indicated a decrease in the rate of Q_A reduction and an increase in the proportion of Q_B‐non‐reducing PSII reaction during senescence. The decrease in both F'_v/F'_m and q_P explained the decrease in the actual quantum yield of PSII electron transport ((φ_PSII). We suggest that the modifications in PSII function, which led to the down‐regulation of photosynthetic electron transport, would be in concert with the lower demand for ATP and NADPH in the Calvin cycle which is often inhibited in senescing leaves.     

Light-dependent, chilling effects on phosphorylation of thylakoid proteins and consequences for associated photochemical activities in maize

When maize ( Zea mays L. cv. LG11) leaves are exposed to low temperatures and high light modifications to both photosystem 2 (PS2) and the light-harvesting chlorophyll a/b protein complex associated with photosystem 2 (LHC2) occur. This study examines the consequences of these modifications for phosphorylation of LHC2 and PS2 polypeptides and the associated changes in electron transport. Maize leaves were chilled at 5°C for 6 h under photon flux densities of 1 500 and 250 μmol m^-2 s^-1. Thylakoids were then isolated from the leaves and their abilities to phosphorylate LHC2 and PS2 polypeptides and modify electron transport activities were determined. Measurements of chlorophyll fluorescence induction in the thylakoids were also made. Thylakoids isolated from leaves chilled under high light and from leaves kept in the ambient growth environment had similar phosphoprotein profiles. However, polypeptide phosphorylation in thylakoids from the chilled leaves did not produce a decrease in PS2 electron transport. Chilling leaves under low light produced a decrease in the ability of isolated thylakoids to phosphorylate PS2, but not LHC2, polypeptides, which was not associated with any change in the phosphorylation-induced decrease in PS2 electron transport. Chilling under high, but not low, light appears to produce changes in membrane organisation that do not affect the ability of the thylakoids to phosphorylate PS2 and LHC2 polypeptides, but which do prevent the phosphorylation-induced decrease in excitation energy transfer from LHC2 to PS2.     

Photosystem I acceptor side limitation is a prerequisite for the reversible decrease in the maximum extent of P700 oxidation after short-term chilling in the light in four plant species with different chilling sensitivities

Changes in the extent of P700 oxidation (P700⁺) were investigated after chilling of barley, rice, pumpkin, and cucumber leaf segments at 4°C for 1 h under light with various photon flux densities. At 50 µmol photons m⁻² s⁻¹, the decrease in P700⁺ was observed only in cucumber, but at 150 µmol photons m⁻² s⁻¹, it was found in all plants except barley, revealing their expected chilling sensitivities. However, the decrease in P700⁺ by this short-term chilling was reversible in the presence of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or methyl viologen, and it did not show any causal relationship with the decrease in the electron transfer rate nor with the down-regulation of photosystem II through the accumulation of zeaxanthin and the development of non-photochemical quenching. These results led to the suggestion that photosystem I (PSI) acceptor side limitation is a prerequisite for the decrease of P700⁺. Furthermore, PSI acceptor side limitation could be mainly due to limitation of electron-sink pathways such as CO₂ assimilation and ascorbate–glutathione cycle, because treatment with glycolaldehyde which inhibits the former pathway, and with KCN which inhibits both pathways, decreased P700⁺ by 20–30% in barley leaves after chilling in the light.     

Superoxide‐producing NAD(P)H oxidases in plasma membrane vesicles from elicitor responsive bean plants

Higher plants produce active oxygen species (AOS) that regulate their defence responses against pathogenic elicitation. Etiolated bean seedlings ( Phaseolus vulgaris L. cv. Limburgse vroege) were used to measure the in vivo‐induced AOS production and to search for plasma membrane bound NAD(P)H‐dependent oxidases producing AOS. Immersed bean plants showed a substantial production of H₂O₂, as determined by the peroxidase (EC 1.11.1.7)‐dependent oxidation of 3,5‐dichloro‐2‐hydroxybenzenesulfonic acid (DHBS). Addition of the elicitor polygalacturonase (PGase, EC 3.2.1.15) from Aspergillus japonicus or the phosphatase inhibitor, cantharidin, resulted in a transient increase of AOS synthesis. Plasma membrane vesicles, purified from etiolated bean seedlings, showed an NAD(P)H‐dependent superoxide (O₂⁻) production that was highly stimulated with naphthoquinones. Protein solubilisation and anion exchange chromatography resolved a basal and three naphthoquinone‐stimulated NAD(P)H‐dependent O₂⁻ oxidase fractions. The natural phenol, apigenin, was also a strong inducer of the naphthoquinone‐dependent enzymes, when it was used in the presence of peroxidase. Although, the relation of these different in vitro‐determined plasma membrane NAD(P)H‐dependent O₂⁻ oxidases to the in vivo elicitation of H ₂O₂ has not been elucidated so far.     

Inhibition of electron flow around photosystem I in chloroplasts of Cd-treated maize plants is due to Cd-induced iron deficiency

Photosystem I activity of chloroplasts isolated from 21 days old maize seedlings ( Zea mays L. cv. Hidosil) cultivated in a nutrient solution containing different concentrations of Cd (10,20,30μM) was investigated. Cd markedly decreased ferredoxin(Fd)-dependent NADP⁺ photoreduction, while it had no effect on electron transport from 2. 6-dichlorophenolindophenol to methyl viologen, indicating that the metal interferred with electron transport on the reducing side of photosystem I. The decrease in electron transport correlated with a low Fd content, which in turn was correlated with a low Fe concentration, suggesting Cd-induced Fe deficiency. In in vitro experiments direct Cd inhibition of Fd-dependent NADP⁺ photoreduction required much higher Cd concentrations than those observed in Cd-treated plants.     

Mechanisms of cobalt uptake in plants: 60CO uptake and distribution in Chara

The mechanism of cobalt uptake was investigated using cells of the giant alga Chara corallina in which it is possible to resolve separately uptake by the cell wall and actual influx across the cell membrane. The absorption of ⁶⁰Co by Chara cells appeared to saturate within 2 h, but this was mainly due to rapid uptake into the cell wall which accounted for 87–92% of the total activity. Even after prolonged desorption most of the cell‐associated ⁶⁰Co was found on the cell wall. The intracellular distribution of absorbed ⁶⁰Co was investigated by fractionating the cell into cytoplasm and vacuole. It was shown that ⁶⁰Co influx to the vacuole occurs simultaneously with influx to the cytoplasm. The transported species appears to be Co²⁺ rather than the less charged Co(OH)⁺ or Co(OH)₂. ⁶⁰Co influx is pH dependent (optimum pH 7–9), and is sensitive to some other divalent metals. Influx from solutions containing 1 µ M ⁶⁰Co was inhibited by 5 µ M Cd²⁺, Cu²⁺, and Zn²⁺, but Mn²⁺ and Ni²⁺ had no significant effect. The sensitivity of Co uptake to N ‐ethyl maleimide (NEM) and cysteine suggests that the transport system involves direct binding of CO²⁺ to ‐SH groups.     

Responses of photosynthesis to NaCl in gametophytes of Acrostichum aureum

Gametophytes of Acrostichum aureum were cultured in 0.0 to 1.0% NaCl solutions or in NaCl‐free solution and then transferred to 1.0% NaCl solution. Photosynthetic light‐response curves, efficiency of the primary photochemical reaction, relative electron transport rate, and photochemical and non‐photochemical quenching at steady state were determined by photosynthetic O₂ evolution and in vivo chlorophyll fluorescence. Results obtained showed that the chlorophyll fluorescence parameters, F_v/F_m and F'_v/F'_m and αO₂ (the initial linear slope of the photosynthetic light‐response curve) increased in gametophytes grown in NaCl. Linear electron transport rate was stimulated by NaCl. Based on the chlorophyll content, light‐saturated photosynthesis in gametophytes grown in 0.2 to 0.7% NaCl increased slightly; it decreased in gametophytes grown in 1.0% NaCl. Photochemical quenching decreased in NaCl‐grown gametophytes at all photosynthetic photon flux density (PPFD) levels measured, but there was no increase in non‐photochemical quenching. The chlorophyll a/b ratio increased with increasing NaCl concentration in culture solutions. These results indicated that NaCl enhanced photochemical efficiency of photosystem II (PSII) and photosynthetic linear electron transport, thus resulting in the development of an excitation pressure in PSII. Such excitation pressure might act as a signal for photosynthetic acclimation to salt stress, thus allowing the gametophytes to grow in their natural habitats.