Salinity effects on oxygen consumption, gill Na+, K+-ATPase and ion regulation in juvenile coho salmon

The metabolic response of juvenile coho salmon Oncorhynchus kisutch to different salinities was examined, using whole-animal oxygen consumption rates and gill Na⁺, K⁺-ATPase activities as indicators of osmoregulatory energetics. Coho salmon smolts were acclimated to fresh water (FW), isosmotic salinity (ISO, 10‰) and sea water (SW, 28‰) and were sampled for up to 6 weeks for plasma levels of cortisol, glucose and ions (Na⁺, K⁺, Cl⁻), gill Na⁺, K⁺-ATPase activity and oxygen consumption rates. Following an initial adjustment period, plasma constituents in SW fish returned to near-FW values, indicating that the fish were acclimated to SW by day 21. Gill Na⁺, K⁺-ATPase activities on days 21 and 42 were lowest in ISO, higher in FW and highest in SW. This result is consistent with the idea that less energy would be required to maintain ion balance in an isosmotic environment, where the ionic gradients between extracellular fluid and water would be minimal. Oxygen consumption rates of swimming fish (1 body length s⁻¹), however, did not differ significantly between the three test salinities after 6 weeks. The results of this study suggest that the metabolic response of juvenile salmonids to changes in salinity is dependent on life-history stage (e.g. fry v . smolt), and that oxygen consumption rates do not necessarily reflect osmoregulatory costs.     

Salinity adaptation in the snakehead, Ophiocephalus maculatus Lacépède: changes in oxygen consumption, branchial Na+-K+-ATPase and body composition

Snakeheads were adapted to fresh water (1 mOsm kg⁻¹), 25% sea water (230 mOsm kg⁻¹), 33% sea water (320 mOsm kg⁻¹) and 40% sea water (380 mOsm kg⁻¹) for 20 days. Exposure to salt water resulted in tissue dehydration, elevations of plasma osmolality, Na⁺, Mg²⁺, Cl⁻and protein concentrations and stimulation of branchial Na⁺-K⁺-ATPase activity. These changes were accompanied by concomitant decline of the hepatosomatic index and liver glycogen concentration. The routine rate of oxygen consumption was increased in snake-heads adapted to 33% sea water. These data were taken to indicate a stressful effect of salinity to the snakehead despite documentation of its ability to penetrate into brackish waters.     

Limiting cytosolic Na+ confers salt tolerance to rice cells in culture: a two-photon microscopy study of SBFI-loaded cells

Rice ( Oryza sativa L.), a staple food in Asia, is very sensitive to soil salinity. However, intraspecific variations exist, with the coastal cultivar Pokkali tolerating even brackish water. This study explores cellular mechanisms that contribute to salt tolerance in rice. It is widely accepted that limiting cytosolic Na⁺ should improve the survival of plants subjected to saline stress. However, an understanding of the mechanisms by which Na⁺ levels are controlled in relatively tolerant cultivars requires monitoring cytosolic Na⁺ non-invasively and in real time, which is technically challenging. We have used two-photon excitation for the ratiometric estimation of cytosolic Na⁺ in cultured cells using sodium-binding benzofuran isophthalate. Pokkali cells maintained low cytosolic Na⁺ (approximately 25 m M ), and a viability of over 85% under high salinity , while Jaya cells were unable to maintain low cytosolic Na⁺ and suffered decreased viability even at moderate saline stress. Here we show that the permeability of the Pokkali plasma membrane to Na⁺ is significantly lower than that of Jaya, to the extent that it is comparable with permeabilities reported for halophytes. Pokkali effectively sequesters Na⁺ in intracellular compartments utilizing a Ca²⁺-regulated transport system(s). Together these cellular mechanisms allow Pokkali to maintain low cytosolic Na⁺ up to a stress of 250 m M NaCl. The findings demonstrate that differences in survival between these contrasting varieties of rice are mainly because of differences in membrane transport mechanisms and thus have significance in crop improvement.     

Salinity tolerance of two tropical fishes, Oreochromis aureus and O. niloticus. I. Biochemical and morphological changes in the gill epithelium

Cichlids of the genus Oreochromis are fish of economic importance in African countries. They tolerate brackish water, however, with great variations between species. In this work, two species, both from the Ivory Coast but of different origins, O. niloticus (field and laboratory strains) and O. aureus (field strain) were compared during osmotic challenges (10, 20 and 30%o salinity) in order to provide physiological support for their specific behaviour when confronted with natural hypertonic environments. Tolerance to salinity was assessed by correlated observations on gill structure, plasma sodium levels and gill Na⁺/K⁺ ATPase activity. In fresh water (FW), all fish presented a gill epithelium structure characteristic of FW stenohaline fish: no chloride cells (CC) on the lamellae and few CC on the filaments. An increase in external salinity induced the proliferation of CC on filaments, a feature typical of seawater teleosts. This change in gill structure was accompanied by an increase of gill Na⁺/K⁺ ATPase activity. In the most tolerant strains, plasma Na⁺ did not change, indicating successful ion regulation in the hypertonic media. With regard to potential interest of field strains in fish culture, O. aureus acclimated more easily to brackish water than O. niloticus . Interestingly, O. niloticus , kept for several generations in the laboratory, performed best in our challenge studies. Plasma Na⁺ levels and gill CC proliferation upon transfer to an isotonic medium may be the parameters of choice when testing these fish for their response to a salinity change.     

Salt tolerance of the reed plant Phragmites communis

Reed plants ( Phragmites communis Trinius) were grown at NaCl concentrations up to 500 m M and their growth, mineral contents and leaf blade osmotic potential were determined. Addition of NaCl up to 300 m M did not affect growth significantly. Sucrose, Cl^-and Na⁺ concentrations in the shoots increased with the salinity of the medium and the shoot water content decreased. K⁺ always contributed most to the leaf osmotic potential. Even in the presence of 250 m M NaCl in the rooting medium, the leaf blade contained only 50 mM Na⁺, suggesting that the plants have an efficient mechanism for Na⁺ exclusion. ²²Na⁺ uptake experiments suggested that the retranslo-cation of absorbed Na⁺ from shoots to the rooting medium lowered the uptake of Na⁺.     

Quantifying the three main components of salinity tolerance in cereals

Salinity stress is a major factor inhibiting cereal yield throughout the world. Tolerance to salinity stress can be considered to contain three main components: Na⁺ exclusion, tolerance to Na⁺ in the tissues and osmotic tolerance. To date, most experimental work on salinity tolerance in cereals has focused on Na⁺ exclusion due in part to its ease of measurement. It has become apparent, however, that Na⁺ exclusion is not the sole mechanism for salinity tolerance in cereals, and research needs to expand to study osmotic tolerance and tissue tolerance. Here, we develop assays for high throughput quantification of Na⁺ exclusion, Na⁺ tissue tolerance and osmotic tolerance in 12 Triticum monococcum accessions, mainly using commercially available image capture and analysis equipment. We show that different lines use different combinations of the three tolerance mechanisms to increase their total salinity tolerance, with a positive correlation observed between a plant's total salinity tolerance and the sum of its proficiency in Na⁺ exclusion, osmotic tolerance and tissue tolerance. The assays developed in this study can be easily adapted for other cereals and used in high throughput, forward genetic experiments to elucidate the molecular basis of these components of salinity tolerance.     

Effects of chloride and sodium on gas exchange parameters and water relations of Citrus plants

Gas exchange parameters, water relations and Na⁺/Cl^- content were measured on leaves of one-year-old sweet orange ( Citrus sinensis [L.] Osbeck cv. Hamlin) seedlings grown at increasing levels of salinity. Different salts (NaCl, KCl and NaNO₃) were used to separate the effects of Cl⁻ and Na⁺ on the investigated parameters. The chloride salts reduced plant dry weight and increased defoliation. Accumulation of Cl⁻ in the leaf tissue caused a sharp reduction in photosynthesis and stomatal conductance. By contrast, these parameters were not affected by leaf Na⁺ concentrations of up to 478 m M in the tissue water. Leaf water potentials reached values near −1.8 MPa at high NaCl and KCl supplies. This reduction was offset by a decrease in the osmotic potential so that turgor was maintained at or above control values. The changes in osmotic potential were closely correlated with changes in leaf proline concentrations. Addition of Ca²⁺ (as calcium acetate) increased growth and halved defoliation of salt stressed plants. Furthermore, calcium acetate decreased the concentration of Cl⁻ and Na⁺ in the leaves, and increased photosynthesis and stomatal conductance. Calcium acetate also counteracted the reductions in leaf water and osmotic potentials induced by salinity. In addition, calcium acetate inhibited the accumulation of proline in the leaves which affected the reduction in osmotic potential. These results indicate that adverse effects of salinity in Citrus leaves are caused by accumulation of chloride.     

Changes in free polyamine levels induced by salt stress in leaves of cultivated and wild tomato species

The effects of N_aCl on endogenous free levels of the poluamines putrescine, spermi dine and spermine, and the relationships between polyamines, K⁺ levels and Na⁺ accumulation were determined in leaves of the cultivated tomato ( Lycopersicon esculentum Mill.) and its wild, salt-tolerant relative L. pennellii (Correll) D' Arcy at different exposure times during a 32-day period. Both stress treatments (100 and 200 m M NaCl) decreased the levels of putrescine and spermidine, although to a different degree for the cultivated and wild tomato species. The spermine levels did not decrease with salinity in L. pennellii over the salinization period, whereas they decreased in L. esculentum , except at the first application of the 100m M NaCl treatment. In both species, the changes induced by salinity in total polyamines and K⁺ were very similar, with the accumulation of Na⁺ in the leaf being concomitant with a decrease in both total polyamines and K⁺. This suggests that the main role of the polyamines in the leaf tissues. In this sense, a direct relationship between total polyamines and K⁺, and inverse relationship between polyamines and Na⁺ and between K⁺ and Na⁺ were found for both species. In the short term (up to 4 days) a peculiar physiological behavior was found in L. pennellii , as the total polyamine and K⁺ levels decreased at 100 m M but not at 200 m M NaCl, while after this time the latter plants had values lower than those of the 100 m M NaCl-treated plants at day 11.     

Salinity tolerance of Kosteletzkya virginica. I. Shoot growth, ion and water relations

Abstract. Kosteletzkya virginica (L.) Presl., a dicotyledonous halophyte native to brackish tidal marshes, was grown on nutrient solution containing 0. 85, 170 or 255 mol m^-3 NaCl, and the effects of external salinity on shoot growth and ion content of individual leaves were studied in successive harvests. Growth was stimulated by 85 mol m^-3 NaCl and was progressively reduced at the two higher salinities. Growth suppression at high salinity resulted principally from decreased leaf production and area, not from accelerated leaf death. As is characteristic of halophytic dicots. K. virginica accumulated inorganic ions in its leaves, particularly Na⁺ and K⁺. However, the Na⁺ concentration of individual leaves did not increase with time, but remained constant or even declined, seeming to be well-coordinated with changes in water content. A striking feature of the ion composition of salinized plants was the development of a dramatic gradient in sodium content, with Na⁺ partitioned away from the most actively growing leaves. Salt-treated plants exhibited a strong potassium affinity, with foliar K⁺ levels higher in salinized plants than unsalinized plants after an initial decrease. These results suggest that selective uptake and transport, foliar compartmentation of Na⁺ and K⁺ in opposite directions along the shoot axis, and the regulation of leaf salt loads over time to prevent build-up of toxic concentrations are whole-plant features which enable K. virginica to establish favourable K⁺-Na⁺ relations under saline conditions.     

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 salinity on the ionic balance and growth of juvenile turbot

The effects of salinity changes (27, 19 and 10‰) on seawater-adapted juvenile turbot were studied on their plasma osmolarity and ion concentrations, on oxygen consumption, on gill Na⁺,K⁺-ATPase activity after 3 months and on growth parameters. All plasma concentrations (except chloride) were unchanged, suggesting that fish were well adapted to their environment. Oxygen consumption was significantly decreased in the 19 and 10‰ groups, where fish weighed significantly more 105 days after transfer than fish maintained in sea water. These results, and the fact that apparent food conversion rates were lower in a diluted environment, suggest that on a long term schedule growth conditions could be improved by adaptation to brackish waters (salinities between 10 and 19‰). The effects of transfer from sea water to 27, 19, 10 and 5‰ were also followed during the first 3 weeks. With salinity 10‰ a steady state was reached on day 21 with all plasma values within the same range. The significant differences observed in osmolarity, plasma ion concentrations and Na⁺,K⁺-ATPase activity 3 weeks after transfer of juveniles to 5‰ salinity, compared with transfers in higher salinities, suggest that there is a threshold of acclimation of turbot to a hypotonic environment.     

Extracellular ionic and acid-base adjustments of Atlantic salmon presmolts and smolts in fresh water and after transfer to sea water: the effects of ovine growth hormone on the acquisition of euryhalinity

In order to better understand the basis for the acquisition of euryhalinity by juvenile salmon and the role of endogeneous stimuli, experiments have been carried out to examine the dynamics of ionic and acid-base adjustments in fresh water (FW) and after direct transfer to full salinity (32 g l⁻¹) sea water (SW) (1) on Atlantic salmon smolt during the natural period of smoltification in spring, (2) on presmolt salmon in autumn, after intraperitoneal implantation of pellets containing ovine growth hormone (oGH). During parr-smolt transformation in FW, gill Na⁺/K⁺ ATPase activity gradually rises, the plasma osmolality (Posm) is unaffected and the total CO₂ of the plasma decreases significantly while whole blood pH fluctuates slightly. Direct transfer of smolt from FW to SW provokes only a slight increase in Posm and emphasizes the acid-base balance disruptions shown in FW. An oGHimplant in a presmolt stimulates gill Na⁺/K⁺ ATPase activity in FW, and affects the acid-base balance. After SW transfer (12 days after implantation), oGH treatment prevents the increase of osmotic pressure and the restoration of the acid-base, ionic equilibrium was faster for oGH-implanted fish than for sham-operated fish. These observations show that in FW smelting salmon develop most of the systems they need for migration and growth in SW and that oGH implants induce the development of physiological characteristics of smolts in a non-natural period of smolting.     

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.     

Changes in ion content and transport during development of embryonic rainbow trout

Wet mass and water content of four lots of whole eggs did not change throughout embryonic development of rainbow trout Oncorhynchus mykiss. Eggs in all four lots accumulated Na⁺. Eggs in lots 2 and 4 also accumulated Ca²⁺ and Cl^-, whereas eggs in lot 1 showed no significant change in Ca²⁺ or Cl^- and eggs in lot 3 showed no change in Cl^-and a small loss of Ca²⁺. Although the Na⁺ content of embryonic tissues increases in the later stages of development, the yolk sac content remained constant, indicating uptake of Na⁺ from the environment. Na+ uptake by whole eggs was non-saturable, consistent with diffusion of Na⁺ across the chorion into the perivitelline fluid. Na⁺ uptake in dechorionated embryos was saturable, as was Ca²⁺ uptake by both whole eggs and dechorionated embryos, consistent with active uptake or facilitated diffusion mechanisms at the surface of embryos. Very low Ca²⁺ uptake rates in dechorionated embryos suggest that the Ca²⁺ uptake mechanism is not fully developed until after hatching.     

Plasma ionic regulation and gill Na+/K+-ATPase changes during rapid transfer to sea water of yearling rainbow trout, Salmo gairdneri: time course and seasonal variation

Danish rainbow trout, Salmo gairdneri Richardson, (40–65 g) were transferred to 28%o sea water at intervals throughout the early spring and summer. Gill Na⁺/K⁺-ATPase of fish kept in fresh water surged distinctly during May. Simultaneously, a body silvering occurred and plasma concentrations of Cl⁻, Na⁺ and total thyroxine (T4) decreased. The seawater transfer-induced adaptive response in plasma electrolytes comprised a biphasic change, i.e., an adjustive peak phase and a regulatory phase lasting for 2 days and 1 week after transfer, respectively. Further, gill Na⁺/K⁺-ATPase activity increased to a new level after an initial lag phase of 2–3 days, but electrolyte regulation was mostly initiated prior to the adaptive change in ATPase activity. In spite of increasing pre-transfer freshwater Na⁺/K⁺-ATPase activity during the spring, the electrolyte peak level, the degree of muscle dehydration and the mortality of fish transferred to sea water increased from April to July. The apparent uncoupling of freshwater Na⁺/K⁺-ATPase activity and plasma electrolyte regulation in sea water is discussed in relation to smelting and prediction of hypo-osmoregulatory performance.     

Effects of salinity and replacement of K+ by Na+ on lipid composition in two sugar beet inbred lines

The effects of NaCl and replacement of K⁺ by Na⁺ on the lipid composition of the two sugar beet inbred lines FIA and ADA were studied (a) with increasing additions of NaCl to the basal medium, and (b) with increasing replacement of K⁺ by Na⁺ at the same total concentration as in the basal medium. Direct relations were noted between NaCl concentration of the nutrient solution and the phospholipid concentration in the roots of FIA, the genotype characterized by a low K⁺/Na⁺ ratio, as well as between NaCl in the medium and the phospholipid concentration in the shoots of ADA, the genotype with a high K ⁺/Na ⁺ ratio. The sulfolipid level in the roots of FIA was maintained at higher NaCl concentrations, while it was decreased in ADA. The glycolipid concentration in the shoots of ADA and the degree of unsaturation of the fatty acids of the total lipid fraction were decreased by salinity, indicating reduced biosynthesis of chloroplast glycolipids and/or accelerated oxidation of these lipids in the presence of NaCl.
In the Na⁺ for K⁺ replacement experiment a low content of K⁺ in the medium resulted in decreased levels of total lipids, phospholipids and sulfolipid in the roots of both genotypes, which did not relate to root growth. K⁺-leakage from the roots at low K⁺-level in the medium may be reduced by the increase in saturation of the lipids. In the shoots of ADA increased levels of total lipids, phospholipids and Sulfolipid were noted at a low K⁺-concentration of the nutrient solution.     

Environmental induction of Na+ transporter affinity in Atlantic salmon embryos

Atlantic salmon ( Salmo salar L.) alevins hatched from eggs transferred from high- to low-Na water at 250° days, before the onset of the phase of increasing whole egg sodium content (at ∼380°days), showed a significantly reduced K _m for Na⁺ transport, whereas transfer at 400° days did not produce any change in K _m . Alevins hatched from eggs given acid shocks of 1, 3, 7 or 14 days duration initiated at 250 or 400° days showed no significant changes in Na⁺ transporter K _m . Extended acid exposure (38 days) from 250°days to hatching resulted in a slight lowering of K _m (P<0.05). A 24-day acid exposure from 400°days to hatching had no effect on Na⁺ transporter K _m . Alevins hatched from eggs incubated throughout in acidified water had a significantly reduced K _m compared to controls (P<0.01).
The timing and duration of periods of Na depletion of eggs is considered with respect to environmental induction of increased Na transporter affinity in teleost embryos as a mechanism of long-term physiological adaptation to the gradual acidification of natural waters.     

Salinity tolerance of Kosteletzkya virginica. II. Root growth, lipid content, ion and water relations

Abstract. Kosteletzkya virginica (L.) Presl., a dicot halophyte native to brackish tidal marshes, was grown on nutrient solution containing 0. 85, 170 or 255 mol m ³ NaCl, and the effects of external salinity on root growth, ion and water levels, and lipid content were examined in successive harvests. Root growth paralleled shoot growth trends, with some enhancement observed at 85 mol m ³ NaCl and a reduction noted at the higher salinities. Root Na⁺ content increased with increasing external NaCl, but remained constant with time for each treatment. K⁺ content, although lower in salt-grown plants after 14 d salinization, subsequently increased to levels comparable to unsalinized plants. A strong K⁺ affinity was reflected in the increased K⁺/Na⁺ selectivity of salt-grown plants and by their low Na⁺/K⁺ ratios. Cl levels rose in salinized plants and values were double or more those for Na⁺, indicating the possibility of a sodium-excluding mechanism in roots. Root phospholipids and sterols, principal membrane constituents, were maintained or elevated and the free sterol/phospholipids ratio increased in salinized K. virginica plants, suggesting retention of overall membrane structure and decreased permeability. This response, considered in light of root calcium maintenance and high potassium levels, suggests that salinity-induced changes in membrane lipid composition may be important in preventing K⁺ leakage from cells.     

Characterization of Na+ exclusion mechanisms of salt-tolerant reed plants in comparison with salt-sensitive rice plants

Salt-tolerant reed plants ( Phragmites communis Trinius) and salt-sensitive rice plants ( Oryza sativa L. cv. Kinmaze) were grown in salinized nutrient solutions up to 50 m M NaCl, and growth, Na⁺ contents and kinetics of ²²Na⁺ uptake and translocation were compared between the species to characterize the salt tolerance mechanisms operating in reed plants. When both plants were grown under the same salinity, Na⁺ contents of the shoots were lower in reed plants, although those of the roots were quite similar. The shoot base region of both species accumulated Na⁺ more than the leaf blades did. Sodium-22 uptake and pulse-chase experiments suggested that the lower Na⁺ transport rate from root to shoot could limit excessive Na⁺ accumulation in the reed shoot. There was a possibility that the apparently lower ²²Na⁺ transport rate to the shoot of reed plants was due to net downward Na⁺ transport from shoot base to root.     

Control of Na+ and K+ transport in Spergularia marina I. Transpiration effects

In this paper we begin our study of factors controlling Na⁺ and K⁺ uptake in the halophyte Spergularia marina (L.) Griseb., with emphasis on plants growing at moderate salinity (0.2x sea water). The involvement of transpiration was considered first because of its potential to account for much or all of the transport of ions, and particularly of Na⁺, to the shoot under these growth conditions. Transpiration was constant with time through most of the light period, quickly dropping to 6% of the day time rate at night. ²²Na⁺ uptake, on the other hand, showed much less day/night variation, and relative transport to the shoot was constant. After establishing that transpiration was linearly related to leaf weight, possible transpiration effects were further considered as correlations between leaf weight and transport to the shoot. Under constant, day-time conditions, with linear effects of time and plant size removed, total transport of ²²Na⁺ to the shoot (per plant) was not correlated to leaf weight. A similar result was found when transport was expressed per gram of root, and when partitioning of total label to the shoot was considered. Finally, the correlation was considered between leaf weight and a Na⁺/K⁺ enrichment factor defined as the Na⁺/K⁺ ratio in the leaves divided by that in the roots. This correlation was also insignificant. The results indicate that analysis of control of Na⁺ and K⁺ uptake and transport in this experimental system need not consider effects of transpiration.