Response of barley grains to the interactive e.ect of salinity and salicylic acid

Effect of grain soaking presowing in 1 mM salicylic acid (SA) and NaCl (0, 50, 100, 150 and 200 mM) on barley (Hordeum vulgare cv Gerbel) was studied. Increasing of NaCl level reduced the germination percentage, the growth parameters (fresh and dry weight), potassium, calcium, phosphorus and insoluble sugars content in both shoots and roots of 15-day old seedlings. Leaf relative water content (RWC) and the photosynthetic pigments (Chl a, b and carotenoids) contents also decreased with increasing NaCl concentration. On the other hand, Na, soluble sugars, soluble proteins, free amino acids including proline content and lipid peroxidation level and peroxidase activity were increased in the two plant organs with increasing of NaCl level. Electrolyte leakage from plant leaves was found to increase with salinity level. SA-pretreatment increased the RWC, fresh and dry weights, water, photosynthetic pigments, insolube saccharides, phosphorus content and peroxidase activity in the stressed seedlings. On the contrary, Na⁺, soluble proteins content, lipid peroxidation level, electrolyte leakage were markedly reduced under salt stress with SA than without. Under stress conditions, SA-pretreated plants exhibited less Ca²⁺ and more accumulation of K⁺, and soluble sugars in roots at the expense of these contents in the plant shoots. Exogenous application (Grain soaking presowing) of SA appeared to induce preadaptive response to salt stress leading to promoting protective reactions to the photosynthetic pigments and maintain the membranes integrity in barley plants, which reflected in improving the plant growth.     

Evaluation of salinity tolerance in seedlings of sugar beet (Beta vulgaris L.) cultivars using proline, soluble sugars and cation accumulation criteria

Salinity tolerance of sugar beet (Beta vulgaris L.) cultivars in terms of growth, proline and soluble sugars concentrations, and Na⁺/K⁺ and Na⁺/Ca²⁺ ratios were analyzed in this study. Three-week-old seedlings of three sugar beet cultivars, ‘Gantang7’, ‘SD13829’, and ‘ST21916’, differing in salinity tolerance, were treated with 0, 50, 100, and 200 mM NaCl. Plant shoots and roots were harvested at 7 days after treatment and subjected to analysis. Low concentration of NaCl (50 mM) enhanced fresh and dry weights of shoot and root in ‘Gantang7’, whereas high one (200 mM) reduced growth in all cultivars and the less reduction was observed in ‘ST21916’. Shoot proline was strongly induced by salinity stress in both ‘Gantang7’ and ‘SD13829’, while it remained unchanged in ‘ST21916’. The addition of 50 mM NaCl significantly increased shoot soluble sugars concentrations in ‘Gantang7’ while it had no significant effects in the other two cultivars. ‘Gantang7’ also showed a higher level of root soluble sugars concentration as compared to the other two cultivars. At 50 mM NaCl, the lower shoot Na⁺ concentration, and the higher shoot K⁺ and root Ca²⁺ concentration in ‘Gantang7’ resulted in the lower shoot Na⁺/K⁺ and root Na⁺/Ca²⁺ ratio. However, ‘SD13829’ maintained a lower Na⁺/K⁺ ratio in both shoot and root when subjected to 200 mM NaCl treatment. According to comprehensive evaluation on salinity tolerance, it is clear that ‘Gantang7’ is more tolerant to salinity than the other two cultivars. Therefore, it is suggested that ‘Gantang7’ should be more suitable for cultivating in the arid and semi-arid irrigated regions.     

等渗盐分、干旱胁迫下冬小麦叶片部分渗透调节物质的动态变化

分别以PEG-6000、NaCl模拟干旱胁迫及盐胁迫,采用水培方法研究了抗旱耐盐冬小麦沧-6001在干旱胁迫、盐胁迫条件下叶片可溶性糖、脯氨酸和可溶性蛋白质含量的动态变化以及Na⁺、K⁺在地上部和根系的分布。结果表明可溶性糖和可溶性蛋白变化趋势相似,其含量随干旱胁迫或盐胁迫时间延长而增加,但在胁迫处理后期下降,并且随胁迫强度增加,二者出现下降时间提前;脯氨酸在干旱胁迫条件下快速积累达到峰值后下降但在胁迫处理后期再次增加,在盐胁迫条件下,随胁迫强度的增加和胁迫时间的延长而增加;Na⁺在干旱胁迫下随胁迫程度增加而下降,盐胁迫条件下随胁迫程度的增加而增加,K⁺在干旱胁迫或盐胁迫下均随胁迫程度的增加而下降,且在根系中下降的速度大于地上部。     

Carbohydrate and Proline Contents in Leaves, Roots, and Apices of Salt-Tolerant and Salt-Sensitive Wheat Cultivars1

Intra-specific variations in nonstructural carbohydrates and free proline were determined in leaves, apices, roots, and maturing seeds of two salt-tolerant cultivars (CR and Kharchia-65) and one salt-sensitive cv. Ghods of spring wheat (Triticum aestivum L.) grown in sand culture at various levels of salinity (0, 100, 200, and 300 mM NaCl and CaCl₂ at 5 : 1 molar ratio) under controlled environmental conditions. The levels of leaf, apex, and root ethanol-soluble carbohydrates, fructans, starch, and proline increased in line with elevating level of salinity in all three cultivars under investigation. The contents of proline, soluble and insoluble carbohydrates in the apex increased to levels exceeding those in the leaves and roots. Soluble carbohydrate content of salt-sensitive cv. Ghods was higher in the leaves, apices, and roots and lower in the maturing seeds than in the other cultivars at all levels of salinity except at 300 mM. The results show considerable variation in the amount of soluble, insoluble sugars, and proline among plant tissues and wheat genotypes in response to salinity. Higher soluble carbohydrates and fructan in leaves, roots and maturing seeds of stressed plants indicate that their accumulation may help plant to tolerate salinity. Salt-sensitive cv. Ghods accumulated less soluble sugars in the maturing seeds and higher soluble sugars in the apices, which might be used as an indicator in screening wheat genotypes for salinity tolerance.     

Peroxidase activities of two rice cultivars differing in salinity tolerance as affected by proline and NaCl

Proline content, ion accumulation, cell wall and soluble peroxidase activities were determined in control and salt-treated calli (150 nM NaCl) and whole plants (30 mM NaCl) of two rice cultivars (salt sensitive cv. IKP and salt tolerant cv. Aiwu). Under salinity, the highest accumulation of Na⁺, Cl^? and proline occurred in calli, roots and younger leaves of cv. IKP, coupled with the highest decrease in K⁺ content; accumulations of Na⁺ and Cl^? were restricted to older leaves in cv. Aiwu. Relative growth rates of calli and roots or shoots from both cultivars were not linked to peroxidase activities. High concentrations (1 M) of exogenously applied glycerol did not inhibitin vitro activities of soluble peroxidase extracted from control and salt-treated calli or plants. Conversely, 35–55% (in cv. IKP) or 60–80% (in cv. Aiwu) of soluble peroxidase activities were found in presence of isosmotic proline concentration. There were no differences between proline and glycerol effects onin vitro cell wall peroxidase activities.     

Influence of Salt Stress on Growth, Ion Accumulation and Seed Oil Content in Sweet Fennel

A greenhouse experiment was conducted to assess the effect of 25, 50, 75, and 100 mM NaCl on growth, ion accumulation, seed yield, and seed oil content in 67-d-old plants of Foeniculum vulgare Mill. Increasing NaCl concentration caused a significant reduction in fresh and dry masses of both shoots and roots as well as seed yield. Na⁺ and Cl^– in both shoots and roots increased, whereas K⁺ and Ca²⁺ decreased consistently with the increase in NaCl concentration. Plants maintained markedly higher Ca²⁺/Na⁺ ratios in the shoots than those in the roots, whereas that of K⁺ /Na⁺ ratios remained almost uniform in both shoots and roots. Proline content in the shoots increased markedly at the highest NaCl concentration. Oil content in the seed decreased progressively with increase in salinity.     

Effect of NaCl,water stress or both on gas exchange and growth of wheat

Responses of wheat (Triticum aestivum L.) to various concentrations of NaCl and levels of drought were followed. With the rise of NaCl or drought, or NaCl and drought together, growth was retarded. The water content of shoots and roots was mostly unchanged. The chlorophyll and carotenoid contents were increased in plants subjected to salinity or drought or both. Only high salinity level induced a considerable decrease in net photosynthetic rate (P_N) and dark respiration rate (R_D). P_N and R_D were decreased with the decrease of soil moisture content. The content of Na⁺ in the shoots and roots of wheat plants increased with increasing salinity or decreasing soil moisture content or both treatments. Considerable variations in the content of K⁺, Ca²⁺ or Mg²⁺ were induced by the NaCl, drought or both treatments.     

Osmotic adjustment in triticales grown in presence of NaCl

Growth and Na⁺, K⁺, Cl^-, proteins, sugars and proline concentrations were measured in three triticale genotypes M2A, DF99 and Asseret grown on nutrient solution with or without 75 mM NaCl. In saline conditions, leaf area of the three triticales was reduced by 50 % and dry to fresh mass ratio increased. Total protein concentration was diminished by 10 %. K⁺ concentration decreased whereas Na⁺ and Cl^- accumulated in roots and shoots of salt-stressed plants. This ion accumulation was greater in roots of Asseret than in roots of the other triticales. Soluble sugar concentration increased in M2A and Asseret and decreased in DF99. Proline concentration increased in M2A and DF99 and decreased in Asseret. Osmotic adjustment was essentially realized by Na⁺ and Cl^- uptake. Non-reducing sugars and proline contributed too, but to a lesser extent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of salinity on osmotic adjustment characteristics of <Emphasis Type="Italic">Kandelia candel</Emphasis>

To elucidate the osmotic adjustment characteristics of mangrove plants, inorganic ion and organic solute contents of intermediate leaves were investigated in 3-month-old Kandelia candel (L.) Druce seedlings during 45 days of NaCl treatments (0, 200, and 500 mM NaCl). The contents of Na⁺, Cl⁻, total free amino acids, proline, total soluble sugars, pinitol and mannitol increased to different degree by salinity, whereas, K⁺ content decreased by salinity compared with control. NaCl treatment induced an increase of inorganic ion contribution while a decrease of organic solute contribution. It was concluded that accumulating a large amount of inorganic ions was used as the main osmotic adjustment mechanism under salinity treatment. However, accumulation of organic osmolytes might be considered to play much more important role in osmoregulation under severe salinity (500 mM NaCl) than under moderate salinity (200 mM NaCl), thus the damage caused by high toxic ions (Na⁺ and Cl⁻) concentration in K. candel leaves could be avoided.     

Mechanisms of salt tolerance and interactive effects of <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> inoculation on maize cultivars grown under salt stress conditions

The present work has been performed to study the growth and metabolic activities of two maize cultivars (cv. 323 and cv. 324) which are shown to have different tolerances to salt stress and to determine the effects of inoculation with Azospirillum spp. Along with identifying the mechanisms of maize salt tolerance and the role of Azospirillum (growth promoting rhizobacteria) in elevating salinity stress conditions is examined Maize cv. 323 was the most sensitive to salinity, while cultivar 324 was the most resistant of the 12 maize cultivars tested. Cultivars differences were apparent with certain growth criteria as well as related metabolic activities. The lack of a negative response to increasing NaCl concentration for water content, dry matter yield and leaf area of cv. 324 up to a concentration of – 0.6 MPa indicated salt tolerance. While for cv. 323 there was a marked inhibitory effect of salinity on growth. In the tolerant cv. 324, soluble and total saccharides, soluble protein in shoots and total protein in roots increased with salinity stress. The sensitivity of cv. 323 however was associated with depletion in saccharides and proteins. Proline accumulation was higher and detected earlier at a lower salinity concentration in the salt sensitive cv. 323 comapred to the salt tolerant cv. 324. When salt stressed maize was inoculated with Azospirillum, proline concentration declined significantly. The present study showed, in general, that the concentration of most amino acid increased on exposure to NaCl as well as when inoculated with Azospirillum. The relatively high salt tolerance of cv. 324, compared with cv. 323 was associated with a significantly high K⁺/Na⁺ ratio. Azospirillum inoculation markedly altered the selectivity of Na⁺, K⁺ and Ca⁺⁺ especially in the salt sensitive cultivar cv. 323. Azospirillum restricted Na⁺ uptake and enhanced the uptake of K⁺ and Ca⁺⁺ in cv. 323. A sharp reduction in the activity of nitrate reductase and nitrogenase in shoots and roots of both cultivars was induced by salinity stress. This reduction in NR and NA activity was highly significant at all salinity concentrations. Azospirillum inoculation stimulated NR and nitrogenase activity in both shoots and roots of both cultivars. The differential effect of Azospirillum inoculation on maize cv. 323 and cv. 324 illustrates the different sensitivity of these two cultivars to stress, but still does not provide any clues as to the key events leading to this difference.     

Effects of salinity and nitrogen on growth,ion relations and proline accumulation in Triglochin bulbosa

The effects of salinity and nitrogen on growth, ion relations and prolineaccumulation in the monocotyledonous halophyte, Triglochin bulbosa,was investigated in hydroponic culture over 5 months. The experimentaldesign was a 3 × 3 factorial with three salinity treatments (0, 150 and 300 mol m^-3 NaCl) and three levels of N (5, 10 and 20 gml^-1 N as NaNO₃). Total and root dry biomass accumulationwere significantly affected by salinity, but not by N or N × salinityinteraction. Increase in NaCl from 0 to 150 mol m^-3 had no effecton total or root dry biomass, while further increase in salinity to 300mol m^-3 significantly reduced biomass by 21% and 25%respectively. Shoot dry biomass, which was significantly affected by N andnot by salinity, increased with increase in N from 5 to 10 gml^-1. Ion concentrations in roots and shoots were significantlyaffected by salinity, but not by N or N × salinity interaction. Theconcentration of Na⁺ and Cl^- in roots and shoots increasedprogressively with an increase in salinity, while that of K⁺ decreased. Under non-saline conditions, Na⁺/K⁺ ratios were low (0.41to 0.44) and increased significantly with an increase in salinity in both rootsand shoots. Shoot sap osmotic potentials decreased progressively with anincrease in salinity. Increase in N in the hydroponic solution from 5 to20 g ml^-1 significantly increased root and shoot N by 66%and 41% respectively. Tissue concentrations of proline were significantlyaffected by salinity and substrate N but not by N × salinity interaction. Theconcentration of proline in roots and shoots increased significantly by334% and 48%, respectively, with an increase in salinity from 0 to 300mol m^-3 NaCl. Increase in substrate N from 5 to 20 g ml^-1 significantly increased proline in roots and shoots by 66% and41% respectively. The significance of substrate N on the accumulationof proline is discussed in relation to salt tolerance.     

Superoxide dismutase isozyme activity and antioxidant responses of hydroponically cultured Lepidium sativum L. to NaCl stress

The present study was focused to assess the physiological behavior and antioxidant responses of the medicinal plant Lepidium sativum L. (commonly called Garden cress) subjected hydroponically to NaCl stress during its vegetative growth stage. The results showed that the addition of NaCl to growth medium significantly reduced plant growth. The magnitude of the response was also linked to the plant organ considered and NaCl concentration supplemented to the medium. Tissue hydration seemed unaffected by salinity. Reduction in dry weight (DW) production was associated with a high accumulation of Na⁺ and Cl^? and a significant reduction of K⁺ content in shoots. The accumulation of osmoregulatory compounds (proline and total sugars) in shoots and roots was greatly increased by NaCl. Activity staining of antioxidants after a native polyacrylamide gel electrophores (PAGE) showed four superoxide dismutase (SOD) isozymes in the extract of leaf-soluble proteins (one Mn-SOD, two Fe-SODs, and one CuZn-SOD), and three isoforms in roots (Mn-SOD, Fe-SOD, and CuZn-SOD). Four peroxidase (POD) isozymes in the roots and only one isozyme in the leaves were detected. The work demonstrated that activities of antioxidant defense enzymes changed in parallel with the increased salinity. In summary, these findings proved that L. sativum can be classified as a moderately tolerant plant to salinity.     

Effects of NaCl and Na2SO4 on red-osier dogwood (Cornus stolonifera Michx) seedlings

Sodium chloride and sodium sulfate are commonly present in extraction tailings waters produced as a result of surface mining and affect plants on reclaimed areas. Red-osier dogwood (Cornus stolonifera Michx) seedlings were demonstrated to be relatively resistant to these high salinity oil sands tailings waters. The objectives of this study were to compare the effects of Na₂SO₄ and NaCl, on growth, tissue ion content, water relations and gas exchange in red-osier dogwood (Cornus stolonifera Michx) seedlings. In the present study, red-osier dogwood seedlings were grown in aerated half-strength modified Hoagland's mineral solution containing 0, 25, 50 or 100 mM of NaCl or Na₂SO₄. After four weeks of treatment, plant dry weights decreased and the amount of Na⁺ in plant tissues increased with increasing salt concentration. Na⁺ tissue content was higher in plants treated with NaCl than Na₂SO₄ and it was greater in roots than shoots. However, Cl^– concentration in the NaCl treated plants was higher in shoots than in roots. The decrease in stomatal conductance and photosynthetic rates observed in presence of salts is likely to contribute to the growth reduction. Our results suggest that red-osier dogwood is able to control the transport of Na⁺ from roots to shoots when external concentrations are 50 mM or less.     

Changes in water relations,photosynthetic activity and proline accumulation in one-year-old olive trees (<Emphasis Type="Italic">Olea europaea</Emphasis> L. cv. Chemlali) in response to NaCl salinity

The comparative responses of young olive trees (Olea europaea L. cv “Chemlali”) to different NaCl salinity levels were investigated over 11 months. One-year-old own rooted plants were grown in 10-L pots containing sand and perlite mixture (1:3 v/v). Trees were subjected to three irrigation treatments: CP (control plants that were irrigated with fresh water); SS1 (salt stressed plants irrigated with water containing 100 mM NaCl) and SS2 plants (salt stressed plants irrigated with water containing 200 mM NaCl). Shoot elongation rate, relative water content, leaf water potential and net carbon dioxide exchange rates decreased significantly with increased NaCl salinity level. Under stressed conditions, the increase of Na⁺ and Cl⁻ ions in both leaves and roots was accompanied with that of proline and soluble sugars. The above results show that the accumulation of proline and sugars under stressed conditions could play a role in salt tolerance. The absence of toxicity symptoms under both stress treatments and the superior photosynthetic activity recorded in SS1-treated plants suggest that cv Chemlali is better able to acclimatize to 100 mM NaCl than at 200 mM NaCl. Our findings indicate that saline water containing 100 mM NaCl, the most available water in arid region in Tunisia, can be recommended for the irrigation of cv Chemlali in the arid south of Tunisia.     

Salinity-induced changes in two cultivars of Vigna radiata: responses of antioxidative and proline metabolism

The influence of increasing salinity stress on plant growth, antioxidant enzymes and proline metabolism in two cultivars of Vigna radiata L. (cv. Pusa Bold and cv. CO 4) was investigated. Salt stress was imposed on 30-days-old cultivars with four different concentrations of NaCl (0, 100, 200 and 300 mM). The roots and shoots of CO 4 showed greater reduction in fresh weight, dry weight and water content when compared to Pusa Bold with increasing salt stress. Under salinity stress, the roots and shoots of CO 4 exhibited higher Na⁺: K⁺ ratio than Pusa Bold. The activities of reactive oxygen species (ROS) scavenging enzymes and reduced glutathione (GSH) concentration were found to be higher in the leaves of Pusa Bold than in CO 4, whereas oxidized glutathione (GSSG) concentration was found to be higher in the leaves of CO 4 compared to those in Pusa Bold. Our studies on oxidative damage in two Vigna cultivars showed lower levels of lipid peroxidation and H₂O₂ concentration in Pusa Bold than in CO 4 under salt stress conditions. High accumulation of proline and glycine betaine under salt stress was also observed in Pusa Bold when compared to CO 4. The activities of proline biosynthetic enzymes were significantly high in Pusa Bold. However, under salinity stress, Pusa Bold showed a greater decline in proline dehydrogenase (ProDH) activity compared to CO 4. Our data in this investigation demonstrate that oxidative stress plays a major role in salt-stressed Vigna cultivars and Pusa Bold has efficient antioxidative characteristics which could provide better protection against oxidative damage in leaves under salt-stressed conditions.     

Inducible and constitutive mechanisms of salt stress resistance in <Emphasis Type="Italic">Geum urbanum</Emphasis> L.

The avens (Geum urbanum L.) seedlings were grown for 6 weeks until the expansion of five to six leaves and then exposed to salinity shock (300 mM NaCl in the nutrient medium) or to a gradual (within 4 days) increase in NaCl concentration from 100 to 400 mM. The dynamics of stress-dependent accumulation of Na⁺, Cl^?, proline, and polyamines in leaves and roots was measured, together with activities of antioxidant enzymes, namely, superoxide dismutase (SOD) and guaiacol-dependent peroxidase occurring in soluble, ionically bound, and covalently bound forms. It is shown that avens plants can adapt to gradual salinization by mobilizing stressinducible protective mechanisms (accumulation of proline and spermine) and by activating constitutive enzyme systems (SOD and peroxidase).     

The role of xylopodium in Na+ exclusion and osmolyte accumulation in faveleira [Cnidoscolus phyllacanthus (d. arg.) Pax et K. Hoffm] under salt stress

The aim of this work was to evaluate physiological and biochemical responses of faveleira under salinity. Plants were grown in nutrient solution containing 0, 50, 100 or 150 mM NaCl. After 8 days of stress, plants were harvested and separated into roots, xylopodium, stem + petiole (SP), and basal, median and apical leaves. Salinity reduced the dry weight of all plant parts, although the indicators of water status were not changed. Salt stress increased the content of Na⁺ in the different plant parts, especially in xylopodium, in which it increased approximately eightfold while the content of K⁺ decreased by approximately 40 % under 150 mM NaCl. As a consequence, the K⁺/Na⁺ ratio decreased in all plant organs. In stressed plants, the content of soluble sugars was increased in the roots, SP and leaf strata and the content of soluble proteins increased in all organs. The content of total free amino acids increased in the roots, SP and apical leaves, while the proline content increased in all organs except in xylopodium. It is suggested that the xylopodium may be involved in a mechanism of exclusion and/or compartmentalization of Na⁺ in faveleira under salinity to avoid ionic toxicity in the leaves.     

Effect of NaCl on Biomass and Contents of Sugars, Proline and Proteins in Seedlings and Leaf Explants of Nicotiana tabacum Grown in vitro

Effects of NaCl on growth in vitro and contents of sugars, free proline and proteins in the seedlings and leaf explants of Nicotiana tabacum cv. Virginia were investigated. The fresh and dry mass of the seedlings decreased under salinity. These growth parameters in leaf explants decreased at 50 mM NaCl and increased up to 150 mM NaCl and then decreased at higher level of salinity. Free proline content in both seedlings and leaf explants increased and polysaccharide content decreased continuously with increasing of NaCl concentration. Reducing sugars, oligosaccharides, soluble sugars and total sugars contents in both seedlings and leaf explants decreased up to 150 mM NaCl and then increased at higher concentrations of NaCl.     

Contractile roots are the most sensitive organ in <Emphasis Type="Italic">Crocus sativus</Emphasis> to salt stress

Crocus sativus corms were grown in Perlite and watered by half-strength modified Hoagland nutrient solution containing 0, 50, 100, 150, 200 mM NaCl. Growth parameters and contents of proteins, proline, polyphenols, minerals and saccharides were studied in fibrous roots, contractile roots, corms and leaves. All plants remained alive and did not display any sign of foliar damage even at 200 mM NaCl. However, the salinity decreased growth, relative water content and increased contents of proline and Na⁺ in all organs. Total protein content was increased in corms and contractile roots but decreased in fibrous roots. Changes in protein pattern were also observed. Polyphenol content was increased by salinity in all organs except the leaves. As salinity increased, content of soluble saccharides decreased except in the contractile roots.     

Sodium,potassium, chloride and proline concentrations of chloroplasts isolated from a halophyte,Mesembryanthemum crystallinum L.

Concentrations of four major solutes (Na⁺, K⁺, Cl^-, proline) were determined in isolated, intact chloroplasts from the halophyte Mesembryanthemum crystallinum L. following long-term exposure of plants to three levels of NaCl salinity in the rooting medium. Chloroplasts were obtained by gentle rupture of leaf protoplasts. There was either no or only small leakage of inorganic ions from the chloroplasts to the medium during three rapidly performed washing steps involving precipitation and re-suspension of chloroplast pellets. Increasing NaCl salinity of the rooting medium resulted in a rise of Na⁺ und Cl^- in the total leaf sap, up to approximately 500 and 400 mM, respectively, for plants grown at 400 mM NaCl. However, chloroplast levels of Na⁺ und Cl^- did not exceed 160–230 and 40–60 mM, respectively, based upon a chloroplast osmotic volume of 20–30 l per mg chlorophyll. At 20 mM NaCl in the rooting medium, the Na⁺/K⁺ ratio of the chloroplasts was about 1; at 400 mM NaCl the ratio was about 5. Growth at 400 mM NaCl led to markedly increased concentrations of proline in the leaf sap (8 mM) compared with the leaf sap of plants grown in culture solution without added NaCl (proline 0.25 mM). Although proline was fivefold more concentrated in the chloroplasts than in the total leaf sap of plants treated with 400 mM NaCl, the overall contribution of proline to the osmotic adjustment of chloroplasts was small. The capacity to limit chloroplast Cl^- concentrations under conditions of high external salinity was in contrast to an apparent affinity of chloroplasts for Cl^- under conditions of low Cl^- availability.Abbreviation Chl chlorophyll