Endogenous abscisic acid levels are linked to decreased growth of bush bean plants treated with NaCl

This paper studies the relative importance of endogenous ABA and ion toxicity in the leaf growth inhibition caused by NaCl in salt-adapted and unadapted bush beans. Adaptation to salt-stress was achieved by germination of seeds in 75 m M NaCl, while unadapted plants were germinated in tap water. The adaptation process caused a transitory increase in leaf ABA, Na⁺ and Cl⁻ concentrations, while leaf expansion was inhibited. However, when grown for 8 or 13 days in 75 m M NaCl-containing nutrient solution, primary and first trifoliolate leaves of salt-adapted plants had greater areas than those of unadapted plants. Concentrations of ABA, Na⁺ and Cl⁻ in these leaves were lower in adapted plants, and a strong negative correlation between leaf expansion growth and either leaf Na⁺, Cl⁻ or ABA concentrations could be established. However, in the second trifoliolate leaves only the ABA, but not the Na⁺ or Cl⁻, concentrations were significantly correlated with leaf expansion. Our results suggest that salt-induced inhibition of leaf expansion growth in bush beans is mediated by ABA rather than Na⁺ or Cl⁻ toxicity. Moreover, the increase of ABA, induced by the salt-pretreatment, seems to play an important role in limiting the accumulation of Na⁺ and Cl⁻ in the leaves, leading to adaptation of bush beans to salt-stress.     

Effect of Salt Stress on Callus Cultures of Oryza sativa L.

Kavi Kishor, P.B 1988. Effect of salt stress on callus culturesof Oryza sativa L.—. exp. Bot 39 235–240 Callus cultures of rice adapted to grow under increasing NaClstress were found to accumulate considerable amounts of freeproline, compared with unadapted cells. Salt-adapted cells grownfor 10 passages (25 d each) on NaCl-free medium accumulatedproline on re-exposure to salt as did cells which were growncontinuously on NaCl. On replacing NaCl (100 mol m^–3)with 100 mol m^–3 of KC1, fresh and dry weights as wellas free proline content of salt-adapted callus declined comparedto that attained on 100 mol m^–3 NaCl medium. However,equimolar concentrations of NaCl and KC1 (when added together)produced an increase in growth and free proline accumulationin salt adapted callus. Omission of Ca²⁺ from the growth mediuminhibited the growth of salt-adapted cells in the presence ofNaCl, while it had little effect on the growth of non-adaptedcells in the presence of NaCl. ABA increased the fresh and dryweights of the non-adapted callus only in the presence of 200mol m^–3 of NaCl but not in the absence of NaCl. ABA failedto evoke the same response in salt adapted cells in the presenceof the salt. Tissues exhibited good growth under inhibitorylevels of NaCl (500 mol m^–3) only when glycine betaine,choline and proline were added to the medium but showed no growthin the presence of sarcosine, glycine and dimethylglycine. Key words: Oryza saliva, callus cultures, NaCl stress     

Germination and seedling growth of cotton: salinity-calcium interactions

Abstract. The effects of NaCl salinity on germination and early seedling growth of cotton were studied. Germination was both delayed and reduced by 200 mol m⁻³ NaCl in the presence of a complete nutrient medium. Seedlings, 7–9 d old, were greatly reduced in fresh weight by salinity. The addition of supplemental Ca²⁺ (10 mol m⁻³ as SO₄²⁻ or Cl⁻) to the medium did not improve germination but, to a large degree, offset the reduction in root growth caused by NaCl. Roots growing in the high salt medium without supplemental Ca²⁺ appeared infected by microbes. The cation specificity of the beneficial Ca²⁺ effect on growth was ascertained by testing additions of MgSO₄ or KCl to the NaCl treatments. The contents of K⁴ and Ca²⁺ were reduced in both roots and shoots by the NaCl treatments. Supplemental Ca²⁺ partially offset this effect for K⁴ in the roots and for Ca²⁺ in both roots and shoots. Sodium contents were not affected by the supplemental Ca²⁺. It is concluded that the beneficial effect of high Ca²⁺ concentrations on root growth of cotton seedlings in a saline environment may be due to maintenance of K/Na-selectivity and adequate Ca status in the root.     

Changes in the proportion of endogenous osmotic solutes accumulated by Chlorella emersonii in the light and dark

Abstract. The type of endogenous osmotic solute accumulated by Chlorella emersonii grown at high external osmotic pressure (π_ext) depended on the light/dark conditions: proline accumulated to high concentrations in cells in the light, while sucrose accumulated to high concentrations in the dark. These findings were made during the alternating light dark cycles used to obtain synchronized cultures, i.e. cultures containing cells at only one stage of development at any one time. Similar decreases in proline and increases in sucrose in the dark were found for cells previously grown in continuous light to obtain non-synchronized cultures, i.e. cultures containing cells at all stages of development.
In cultures synchronized at 200 mol m ⁻³ NaCl (π_ext= 1.01 MPa), recently divided 'daughter cells' at the beginning of the light periods contained 60 mol m⁻³ proline and 100mol m⁻³ sucrose, while mature cells towards the end of light periods contained 130 mol m proline and 20 mol m⁻³ sucrose. The changes in proline and sucrose which occurred in synchronized cultures were due mainly to light/dark conditions and to a much lesser extent to different stages of cell development. The proportion of proline to sucrose in daughter cells collected from non-synchronized cultures in continuous light was not different from the proportion in heterogeneous populations of cells.
Results are discussed in relation to the accumulations of two, rather than one, endogenous osmotic solute and to growth reductions of C. emersonii exposed to high external osmotic pressures.     

Effects of sodium, potassium and calcium on salt-stressed barley. I. Growth analysis

Barley ( Hordeum vulgare L. cv. CM 72) was grown for a 28-day period and stressed with treatments of 125 mol m⁻³ NaCl or KC1 with low Ca²⁺ (0.4 mol m⁻³ Ca²⁺) or high Ca²⁺ (10 mol m⁻³ Ca²⁺). Plants were harvested periodically so that relative growth rate (RGR), net assimilation rate (NAR) and leaf area ratio (LAR) could be calculated using the functional approach to plant growth analysis. Relative growth rate declined with time for all treatments, including controls. Salinity inhibited RGR relative to control values by day 10. High Ca²⁺ improved the growth of salt-stressed plants in both NaCl-salinity and KCl-salinity. KC1 proved more toxic than NaCl, especially for KCI-salinity plants with low Ca²⁺, which died by day 28. Net assimilation rate, but not LAR, was highly correlated with RGR for all treatments. This indicates that the photosynthetic-assimilatory machinery was limiting RGR and not the leaf area of the plant.     

Effects of external NaCI, and of changes in turgor pressure and volume, on K+ concentrations in Chlorella emersonii

Summary. Soluble potassium concentrations were determined for the slightly vacuolated, unicellular, walled alga Chlorella emersonii. Sap of cells grown in 1 mol m⁻³ NaCI contained 140 mol m⁻³ K⁺ and sap of cells grown in 125, 200, and 335 mol m⁻³ NaCI contained 160-180 mol m⁻³ K ⁺.
The possible regulation of K ⁺ concentrations by a system of lurgor and volume maintenance was investigated by supplying 3-0-methylglucose. This solute accumulates to 85-230 mol m⁻³ in C. emersonii , but is not metabolized. Accumulation of 3-0-methylglucose increased the volume of cells grown at both low and high NaCI by about 10%. Furthermore, accumulation of 3-0-methylglucose also increased turgor pressures of cells grown in 1 and 125 mol m⁻³ NaCI by 0.3 and 0.2 MPa, respectively. (Similar measurements were not attempted for cells grown in 200 and 335 mol m⁻³ NaCI, because of the insensitivity of available methods to measure turgor pressure of cells exposed to high external osmotic pressures.)
At all NaCI concentrations, the K ⁺ concentrations of cells which had accumulated 3-0-methylglucose were only 10-20 mol m⁻³ lower than K⁺ in cells which had not been supplied with 3-0-methylglucose. In contrast, accumulation of 3-0-methylglucose greatly decreased concentrations of the endogenous osmotic solutes, proline and sucrose, which accumulated in cells grown in 125 mol m⁻³ and higher NaCI concentrations.
It is concluded that K⁺ concentrations in Chlorella emersonii are not controlled by a system of turgor and volume maintenance.     

Early effects of excess cadmium uptake in Phaseolus vulgaris

Abstract. Seedlings of Phaseolus vulgaris were exposed to solutions containing Cd²⁺ in the range 0 to 1 molm⁻³. Ethylene formation started following 3 h of exposure to 10⁻², 10⁻¹ and 1 mol m⁻³ Cd²⁺, peaked at 18 h and returned to a relatively low rate after 24 h. Cadmium-induced ethylene formation depended on the formation of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminoethoxyvinylglycine (AVG, 0.1 mol m⁻³) inhibited ACC accumulation and ethylene production during exposure to 0.2 mol m⁻³ Cd²⁺.
Activity of soluble and ionically-bound peroxidase increased after 18 h of exposure to Cd²⁺ concentrations above 10⁻³ mol m⁻³ due to an increase in activity of cathodic isoperoxidases. Stimulation of soluble and ionically-bound peroxidase by 0.2 mol m⁻³ Cd²⁺ was reduced in the presence of 0.1 mol m⁻³ AVG.
Accumulation of soluble and insoluble ('ligninlike') phenolics was found in plants exposed to Cd²⁺ (10⁻² mol m⁻³ or above) in the presence or absence of AVG. Deposition of insoluble (autofluorescing) material occurred in cell walls around vessels and was associated with reduced expansion and water content of leaves.     

Polyamine levels in relation to growth and NaCl concentration in normal and habituated sugarbeet callus cultures

Abstract. The concentrations of putrescine, spermidine and spermine, the only polyamines detectable in normal and habituated calli of Beta vulgaris L. ssp. altissima , were much higher in the habituated callus than the normal callus, irrespective of experimental conditions. These results suggest that, in normal (tolerant to NaCl) and habituated (sensitive to NaCl) calli, there exists a competition for the common precursor of ethylene and polyamine biosynthesis viz. S-adenosylmethionine. A disequilibrium favouring the synthesis of putrescine and spermidine in the habituated callus might be linked to structural deterioration of the cell membrane following extended culture or severe osmotic stress (68 mol m⁻¹ NaCl). The maintenance of membrane integrity by the normal callus coincides with ethylene production at the expense of polyamine synthesis. In contrast to the habituated callus, the salinity tolerance of the normal callus is accompanied by the accumulation of proline under hypersaline conditions (274mol m⁻³). The important osmoregulatory role played by quaternary ammonium compounds in the-aerial parts of Chenopodiaceae, especially the sugarbeet, is not observed in the calli, these compounds being found in very low concentrations in saline conditions.     

Changes in sodium and potassium in Nitellopsis cells treated with transient salt stress

Abstract. Nitellopsis cells grown in fresh water have a relatively low cytoplasmic Na⁺ (11 mol m⁻³) and high cytoplasmic K⁺ (90 mol m⁻³) content. A 30-min treatment with 100 mol m⁻³ external NaCl resulted in a high [Na⁺]_c (90 mol m⁻³) and a low [K⁺]_c (33 mol m⁻³), Subsequent addition of external Ca²⁺ (10 mol m⁻³) prevented Na⁺ influx and then [Na⁺]_c decreased slowly. Changes in [K⁺]_c were opposite to [Na⁺]_c. During the recovery time vacuolar Na⁺ increased, while vacuolar K⁺ decreased. Since all these processes proceeded also under ice-cold conditions, the restoration of original cytoplasmic ion compositions is suggested to be a passive nature. The notion that the passive movement of ions across the tonoplast can act as an effective and economic mechanism of salt tolerance under transient or under mild salt stress conditions is discussed.     

Response of Suaeda aegyptiaca to KCl, NaCl and Na2SO4 treatments

The response of Suaeda aegyptiaca (Hasselq.) Zoh. to various salinity treatments was tested in sand culture. Growth was promoted by NaCl and by Na₂SO₄ at all tested concentrations, but not by KCl. The effect of NaCl on growth was stronger than that of Na₂SO₄ and it increased gradually up to a 125 eq. m⁻³ optimum. Ion uptake was also affected by the different salts. Cl⁻ was taken up in similar quantities from KCl and from NaCl solutions and the content of the respective cations was also similar to one another. The presence of Na⁺ in the medium lowered the content of K⁺ in the plants and at the same time increased growth by as much as 900%. Transpiration was reduced and water use efficiency increased by Na⁺-salts. Highest water use efficiency was exhibited by plants which were treated with 125 eq. m⁻³ NaCl. It is concluded that Na⁺ at the macronutrient level has a specific promotive effect on the physiological processes of S. aegyptiaca. This effect is not due to replacement of K⁺ by Na⁺; neither can it be achieved by increasing the K⁺ concentration. Cl⁻ has an additional positive effect on growth of S. aegyptiaca. This effect is only expressed in the presence of Na⁺.     

Effects of abscisic acid and proline on adaptation of tobacco callus cultures to salinity and osmotic shock

Tobacco callus ( Nicotina tabacum cv. Badischer Geudertheimer) took up sorbitol rapidly and without a lag period from media with up to 0.7 M of the polyol. Accumulation of proline was greatly enhanced under these conditions and was proportional to the absorbed sorbitol, while the viability of the callus cultures was quite low after a few hours of incubation. Under moderate conditions (0.1 M sorbitol) as well as under severe osmotic shock (0.7 M sorbitol), the cells adapted by adjusting the sorbitol/proline ratio to ca 3. NaCl (0.1 M ) had the same effect as sorbitol (0.7 M ) on the survival rate, but only slightly affected proline synthesis in the first hours of incubation. Addition of 10⁷ or 10 ⁵ M abscisic acid (ABA) did not increase the proline content, but 10 ⁷ M ABA delayed the deleterious effect of NaCl and improved the state of the cells. No influence of abscisic acid during the incubation with sorbitol could be detected. Two different strategies for the adjustment of tobacco callus to salinity and sorbitol are suggested: Non-ionic stress is controlled by the accumulation of proline, whereas ABA could be involved in the adaptation to ionic stress.     

Effects of weak acids on proline accumulation in barley leaves: a comparison between abscisic acid and isobutyric acid

Abstract. When isobutyric acid (IBA) or abscisic acid (ABA) are supplied to leaf sections a similar rapid and marked decrease in the intracellular pH is observed. This acidification is accompanied by an increase in proline level which is about the same for both 3 mol m⁻³ IBA and 1 mol m⁻³ ABA treatments.
Fusicoccin (FC), known to act at the proton pump level, almost completely suppresses the ABA-induced acidification of the cell sap, whereas it only partially counteracts the acidifying effect of IBA, in particular during short periods of treatment. This effect of FC is paralleled by a similar inhibition of the induced proline accumulation: in fact, FC completely suppresses the ABA-induced increase in proline during short treatment periods, whereas it is only effective in inhibiting the IBA-induced proline accumulation after long treatment periods.
These data seem to suggest that the ABA- and IBA-induced changes in proline level might be mediated by changes in the intracellular pH.     

Effect of high external NaCl concentrations on ion transport within the shoot of Lupinus albus. I. Ions in xylem sap

Abstract. Xylem sap was collected from individual leaves of intact transpiring lupin plants exposed to increasing concentrations of NaCl by applying pneumatic pressure to the roots. Concentrations of Na⁺ and Cl⁻ in the xylem sap increased linearly with increases in the external NaCl concentration, averaging about 10% of the external concentration. Concentrations of K⁺ and NO₃⁻, the other major inorganic ions in the sap, were constant at about 2.5 and 1.5 mol m⁻³, respectively. There was no preferential direction of Na ⁺ or Cl⁻ to either young or old leaves: leaves of all ages received xylem sap having similar concentrations of Na⁺ and Cl⁻, and transpiration rates (per unit leaf area) were also similar for all leaves. Plants exposed to 120–160 mol m⁻³ NaCl rapidly developed injury of oldest leaves; when this occurred, the Na⁺ concentration in the leaflet midrib sap had increased to about 40 mol m⁻³ and the total solute concentration to 130 osmol m⁻³. This suggests that uptake of salts from the transpiration stream had fallen behind the rate of delivery to the leaf and that salts were building up in the apoplast.     

Relationship between salt accumulation and water content of dicotyledonous halophytes

Abstract. Growth rates and levels of minerals, Na⁺, K⁺, Mg⁺⁺, Ca⁺⁺, and water were measured in dicotyledonous halophytes grown along a salinity gradient from fresh water to 720 mol m⁻³ NaCl in a controlled environment greenhouse. Ten test species from the families Chenopodiaceae, Aizoaceae, and Batidaceae exhibited growth stimulation by 180 mol m⁻³ NaCl and were classified as euhalophytes. Ten others from the families Chenopodiaceae, Aizoaceae, Asteraceae, Brassicaceae, Polygonaceae, Boraginaceae, Malvaceae, and Plumbaginaceae showed their best growth on fresh water and were classified miohalophytes. Salt, and particularly sodium, accumulated in all halophytes but to a significantly greater extent among euhalophytes than miohalophytes. The water content of most species increased when grown on 180 mol m⁻³ NaCl compared to fresh water; but at higher salinities some of the species underwent dehydration. Dehydration of the succulent S. europaca was not coupled to a proportional decrease in growth. Water content and cation accumulation in euhalophytes appeared to be coordinated to produce a constant osmotic potential gradient within the shoot tissues relative to the external salinity. In contrast, miohalophytes did not appear to regulate osmotic potential as closely as euhalophytes.     

Supplemental manganese improves the relative growth, net assimilation and photosynthetic rates of salt-stressed barley

Previous results in our laboratory indicated that a reduced Mn concentration in the leaves of barley was highly correlated with the reduced relative growth and net assimilation rates of salt-stressed plants. If Mn deficiency limits the growth of salt-stressed barley, then increasing leaf Mn concentrations should increase growth. In the present study, the effect of supplemental Mn on the growth of salt-stressed barley ( Hordeum vulgare L. cv. CM 72) was tested to determine if a salinity-induced Mn deficiency was limiting growth. Plants were salinized with 125 mol m⁻³ NaCl and 9.6 mol m⁻³ CaCl₂. Supplemental Mn was applied in 2 ways: 1) by increasing the Mn concentration in the solution culture and 2) by spraying Mn solutions directly onto the leaves. Growth was markedly inhibited at this salinity level. Dry matter production was increased 100% in salt-stressed plants treated with supplemental Mn to about 32% of the level of nonsalinized controls. The optimum solution culture concentration was 2.0 mmol m⁻³, and the optimum concentration applied to the leaves was 5.0 mol m⁻³. Supplemental Mn did not affect the growth of control plants. Further experiments showed that supplemental Mn increased Mn concentrations and uptake to the shoot. Supplemental Mn increased the relative growth rate of salt-stressed plants and this increase was attributed to an increase in the net assimilation rate; there were no significant effects on the leaf area ratio. Supplemental Mn also increased the net photosynthetic rate of salt-stressed plants. The data support the hypothesis that salinity induced a Mn deficiency in the shoot, which partially reduced photosynthetic rates and growth.     

Abscisic acid induced stress-like polyamine pattern in wheat seedlings, and its reversal by potassium ions

In 3-day-old wheat ( Triticum aestivum L. cv. Marinat) seedlings, 100 μ M ABA blocked the growth and altered the level of K⁺ in both the shoot and root. The presence of ABA increased the putrescine titer during a 24-h treatment. Increasing the endogenous level of K⁺ by the addition of 10 m M KCl to the ABA-treated seedlings, inhibited the effect of ABA on growth and putrescine level. In both tissues, ABA increased putrescine content at low concentrations (1 μ M ), reaching the maximal effect at 100 μ M . Putrescine increase induced by ABA was inhibited by both α-difluoromethylarginine (DFMA) and α-difluoromethylornithine (DFMO) in shoots while only the inhibitor of arginine decarboxylase was effective in the root. The presence of ABA modulated, in opposite ways, ornithine and arginine decarboxylase activities. These results are discussed in relation to ion balance under stress.     

Stomatal limitation of photosynthesis and reduced growth of the halophyte, Plantago maritima L., at high salinity

Abstract. Plantago maritima L. was grown at three levels of salinity, 50, 200, 350 mol m⁻³ NaCl, and the effects on growth, ion content and photosynthetic capacity were studied. Shoot and root dry weight, leaf production and leaf length were all substantially reduced in plants grown at high salinity. Total leaf area of plants grown at 350 mol m⁻³ NaCl was only 20% of that in plants at low salinity. Both the Na⁺ and K⁺ content of leaves and roots increased with external salinity. There was no change in the Na⁺/K⁺ ratio of leaves or roots at different salinity levels. Despite the large reductions in growth and high accumulation of Na⁺ ions, leaf photosynthetic rate was only slightly reduced by salinity stress. The reduction in photosynthesis was not caused by reduced biochemical capacity as judged by photosynthetic response to intercellular CO₂ and by ribulose-1,5-bisphosphate carboxylase activity, but was due to reduced leaf conductance and low intercellular CO₂ concentration. The increased stomatal limitation of photosynthesis resulted in higher water-use efficiency of plants grown at high salinity.     

Effects of accumulation of 3-O-methylglucose on growth and osmotic regulation in Chlorella emersonii

Abstract. The effect of accumulation of 3- O -methylglucose (MG) on growth and steady-stale concentrations of the endogenous osmotic solutes proline and sucrose was studied in Chlorella emersonii grown at external osmotic pressure (II) of 0.08-1.64 MPa. NaCL was used as osmoticum. The total solute content of the cells was manipulated by supplying 2 mol m⁻³ MG for 4 and 48 h. MG accumulated to 50–230 mol m⁻³ within 4h and was not metabolized. Uptake of MG resulted in decreases in concentrations of proline and sucrose, the two solutes mainly responsible for osmotic adaptation of C. emersonii to high II. After 4 or 48 h growth in the presence of MG, the decreases in concentration of proline and sucrose were as predicted from the contribution of MG to the total solute content of the cell.     

A comparative study of the effects of NaCl salinity on respiration, photosynthesis, and leaf extension growth in Beta vulgaris L. (sugar beet)

Abstract. Three parameters influencing the capacity for carbon accumulation, i.e. photosynthesis, respiration, and leaf extension growth, were studied in Beta vulgaris L. (sugar beet) cultured in nutrient solution containing 0.5 to 500 mol m⁻³ NaCl. Leaf extension growth was the parameter most sensitive to salinity: the initial rate of leaf extension and final leaf length each declined linearly with increase in external NaCl concentration. Photosynthetic O₂ evolution of thin leaf slices did not decline until salinity levels reached 350 to 500 mol m⁻³ NaCl, while respiratory O₂ consumption was not affected by salinity throughout the range. The results suggest that the influence of salinity on the capacity for carbon accumulation in B. vulgaris occurs primarily through reduction in the area of photosynthetic surface.     

Effect of high external NaCl concentration on ion transport within the shoot of Lupinus albus. II. Ions in phloem sap

Abstract. Phloem sap was collected from petioles of growing and fully expanded leaves of lupins exposed to 0–150 mol m⁻³ [NaCl]_ext, for various periods of time. Sap bled from growing leaves only after the turgor of the shoot was raised by applying pneumatic pressure to the root. Increased pressure was also needed to obtain sap from fully expanded leaves of plants at high [NaCl]_ext. Exposure to NaCl caused a rapid rise in the Na⁺ concentration in phloem sap to high levels. The Na⁺ concentration reached 20 mol m⁻³ within a day of exposure and reached a plateau of about 60 mol m⁻³ in plants at 50–150 mol m⁻³ [NaCl]_ext, after a week. There was a slower, smaller increase in the Cl⁻ concentration. K⁺ concentrations in phloem sap were not affected by [NaCl]_ext. Cl⁻ concentrations in phloem sap collected from growing leaves were similar to those from old leaves while Na⁺ concentrations were somewhat increased, suggesting that there was no reduction in the salt content of the phloem sap while it flowed within the shoot to the apex. Calculations of ion fluxes in xylem and phloem sap indicated that Na⁺ and Cl⁻ fluxes in the phloem from leaves of plants at high NaCl could be equal to those in the xylem. This prediction was borne out by observations that Na⁺ and Cl⁻ concentrations in recently expanded leaves remained constant.