The influence of salinity on phosphate uptake and distribution in lupin roots

Treeby, M. T. and van Steveninck, R. F. M. 1988. The influence of salinity on phosphate uptake and distribution in lupin roots. - Physiol. Plant. 72: 617–622.
The uptake and distribution of phosphate in lupin ( Lupinus luteus L. cv. Weiko III) roots under moderate salt (NaCl) stress was studied. Vacuolar inorganic phosphate (PJ concentrations in high phosphate plants were decreased by salt, although whole root P| was unaffected. In low phosphate plants, vacuolar P_i was unaffected by salt while whole root P_i was increased. Phosphate uptake was not altered by salt in high phosphate plants, but was depressed in low phosphate plants. These observations lead to the conclusion that in high phosphate plants P_i accumulates in cytoplasm and/or stele, ultimately giving rise to phosphate toxicity in shoots. Increasing phosphate supply had no effect on Na⁺ accumulation in root cell vacuoles in the epidermis or cortex, but the concentration of Cl⁻ in endodermal vacuoles was lowered.     

Role of sodium in the ABA‐mediated long‐term growth response of bean to salt stress

Long‐term salt effects on plant growth have often been related to direct ion toxicity due to the accumulation of high ion concentrations in plant tissue. This work examines the relative importance of endogenous ABA, as well as Na⁺ and Cl⁻ toxicity, in the inhibition of leaf growth and photosynthesis, in bean plants grown at 1, 25, 50 and 75 m M NaCl until the fruit‐bearing stage. All salt‐treated plants showed very high leaf Cl⁻ concentrations, with little difference between plants exposed to 50 or 75 m M NaCl. The 25 and 50 mM salt‐treated plants were able to successfully exclude Na⁺ from their leaves, and only suffered an initial decline in the rate of leaf growth. Plants exposed to 75 m M NaCl showed an increase in Na⁺ leaf concentrations with an accompanying decrease in growth and photosynthesis as salt exposure progressed. A high correlation was found between leaf Na⁺ and leaf growth. Leaf ABA significantly increased with salt supply, and was highly correlated with both leaf Na⁺ and leaf growth. Our results suggest that in bean plants under long‐term salt stress, leaf ABA may participate in the regulation of leaf growth, and leaf Na⁺ would be at least partly responsible for increased ABA levels.     

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.     

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.     

Seasonal changes in ionoregulatory variables of the glass eel Anguilla anguilla following estuarine entry: comparison with resident elvers

In the present study, glass eels Anguilla anguilla in the Minho River estuary (41·5° N, 8·5° W) decreased in size (standard length, L _S and mass, M ) from the beginning (autumn) to the end of the sampling season (summer). On the other hand elvers increased in L _S and M from spring to summer and were significantly larger than glass eels in paired comparisons. Branchial Na⁺/K⁺-ATPase and vacuolar (V-type) proton ATPase ( in vitro activities), two important ion transporting pumps, did not show significant seasonal changes in either glass eels or elvers although in glass eels Na⁺/K⁺-ATPase (activity) expression was significantly higher than in elvers. In a single month comparison Na⁺/K⁺-ATPase branchial mRNA expression was also higher in glass eels as was the protein level expression of both Na⁺/K⁺-ATPase and NKCC (Na⁺:K⁺:2Cl⁻ co-transporter). Immunofluorescence microscopy indicated apical CFTR Cl⁻ channel labelling in Na⁺/K⁺-ATPase positive chloride cell in glass eels which was absent in elvers. Whole body sodium concentration and percentage water did not show significant seasonal differences in either glass eels or elvers although there were significant differences between these two groups during some months.     

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⁺.     

Physiological function of water channels as affected by salinity in roots of paprika pepper

The sap flow (Jv) and the osmotic pressure-dependent hydraulic conductance (L₀) of detached exuding root systems from paprika pepper plants (cv. Albar) were measured. Plants stressed with NaCl (30 m M ) and with six times the macronutrients of the Hoagland nutrient solution (6×HNS) were compared with controls grown in complete Hoagland nutrient solution. Jv of +NaCl and +6×HNS plants decreased markedly, but recovered to values similar to those of controls after removal of the treatments. Hydraulic conductance L₀ was always less in NaCl plants than in controls and 6×HNS. A total increase in the ion concentration of the xylem (except Na⁺ and Cl⁻) was observed with both treatments. In control and 6×HNS plants, HgCl₂ treatment (50 μ M ) caused a sharp decline in L₀ to values similar to those of NaCl-stressed roots, but were restored by treating with 5 m M dithiothreitol (DTT). However, in NaCl roots only a slight effect of Hg²⁺ and DTT was observed. In each treatment, there was no difference in the flux of K⁺ into the xylem after HgCl₂ and DTT application. The results suggest that NaCl decreased L₀ of the roots by reducing either the activity or abundance of Hg-sensitive water channels. The putative reduction in water-channel function of NaCl-treated plants did not seem to be due to the osmotic effect.     

Differences in osmoacclimation between sporophytes and gametophytes of the brown alga Ectocarpus siliculosus

The osmoacclimation of Ectocarpus siliculosus isolates known to have different salt tolerances was investigated. Included were isolates originating from 5 different locations in the northern hemisphere, and sporophyte and gametophyte phases of different ploidy from two of the locations were compared. The effect of salinity treatment (8–64%₀) on inorganic ions (K⁺, Na⁺, Mg²⁺, Cl⁻, SO²⁻₄, phosphate) and the low molecular weight carbohydrate mannitol was measured, together with complimentary measurements of cell viability. Very different responses between isolates were obtained, both between isolates of different geographic origin and between sporophytes and gametophytes from the same parent material. A similarity in response between haploid and diploid gametophytes, and diploid and triploid sporophytes indicates that physiological differences between gametophyte and sporophyte generations are not necessarily based on ploidy changes alone. There were no identifiable differences in the responses of male and female gametophytes. K⁺ is the major osmolyte within the species, and differences in the regulation of K⁺ largely account for the observed variation in osmoacclimation, both between life history phases and between isolates from different localities. Isolates with broader salt tolerances had the higher concentrations of mannitol. There were differences between isolates in the amounts and regulation of Cl⁻ and phosphate, the latter being present in unusually high concentrations. There were also isolate differences in the concentrations of Mg²⁺ and SO²⁻₄, although these divalent ions were present only in low concentrations.     

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⁺.     

Growth, contents of K+ and kinetics of K+(86Rb) uptake in barley cultured at different low supply rates of potassium

Plants of barley ( Hordeum vulgare L. cv. Salve) were grown with 6.5–35% relative increase of K⁺ supply per day (RKR) using a special computer-controlled culture unit. After a few days on the culture solution the plants adapted their relative growth rate (RGR) to the rate of nutrient supply. The roots of the plants remained in a low salt status irrespective of the rate of nutrient supply, whereas the concentration of K⁺ in shoots increased with RKR. Both V_max and K_m for K⁺(⁸⁶Rb) influx increased with RKR. It is concluded that with a continuous and stable K⁺ stress, the K⁺ uptake system is adjusted to provide an effective K⁺ uptake at each given RKR. Allosteric regulation of K⁺ influx does not occur and efflux of K⁺ is very small.     

Effects of sulfate and nitrate on K+ uptake and growth of wheat and cucumber

The uptake of K⁺ ion was studied in the roots of wheat ( Triuicum aestivum L. cv. GK Szeged) and cucumber ( Cucumis sativus L. cv. Budai csemege) seedlings grown in nutrient solution under nitrogen and sulfate stress conditions. Seedlings pretreated with 1 or 10 m M NaNO₃, absorbed more K⁺ than those treated with 0.1 m M NaNO₃. However, the posteffect of NaNO₃ was considerably influenced by the Na₂SO₄, treatment. The results suggest that, at least partly, a feed-back regulation of K⁺ uptake may occur. However, due to the high Na⁺ contents of the roots, a Na⁺ effect in this process cannot be excluded. The growth and dry matter yields of the roots and shoots were strongly influenced by the SO²⁻/₄ and NO⁻/₃ supply of the plants. Appreciable differences were experienced between wheat and cucumber seedlings. The optimum SO²⁻/₄ concentration of the growth solution for maximal growth varied considerably between the species, and was also different for the roots and the shoots in a given species.     

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.     

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.     

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.     

Physiological effects of Na2SO4 and NaCl on callus cultures of Brassica campestris (Chinese cabbage)

Hypocotyl-derived callus cultures of Brassica campestris L. ssp. pekinensis cv. Kim-jung (Chinese cabbage) were grown on Murashige and Skoog medium containing no additional salt, NaCl or Na₂SO₄. Na₂SO₄ was more than twice as inhibitory in comparison to the same concentration of NaCl when growth and fresh:dry weight ratios of established callus were measured. Levels of protein, starch, sucrose and α-amino nitrogen were not significantly altered in salt-grown callus. Concentrations of reducing sugars and chlorophyll were 2–3 times greater in callus grown on either salt. Proline concentration increased 15–20 fold on the highest levels of salt. Final concentrations (reached in 20–24 days) were closely correlated to the initial Na⁺ concentration of the medium, regardless of salt type. The osmotic potential in callus transferred to NaCl or Na₂SO₄ reached a maximum negative value after 16 days. For both salts, subsequent increases were correlated to increases in fresh:dry weight and growth. On both salts, turgor remained relatively constant (0. 6–0.75 MPa). Changes in Na⁺, K⁺, Mg²⁺ and Ca²⁺ content were correlated to initial Na⁺ concentration in the medium, not salt type. Accumulation of Na⁺ was accompanied by loss of K⁺ and Mg²⁺. Six to seven times less sulfate was measured in callus grown on Na₂SO₄ than chloride in callus grown on similar concentrations of NaCl.     

K+ status and ABA affect both exudation rate and hydraulic conductivity in sunflower roots

Twenty‐day‐old sunflower plants ( Helianthus annuus L. cv. Sun‐Gro 380) grown in nutrient solutions with different KCl levels were used to study the effects of K⁺ status of the root and of abcisic acid (ABA) on the exudation rate (J_v), the hydraulic conductivity of the root (L_p), the fluxes of exuded K⁺ and Na⁺ (J_K and J_Na), and the gradient of osmotic pressure between the xylem and the external medium. J_v and L_p increased in direct proportion to the K⁺ starvation of the root. Also addition of ABA (4 µ M ) at the onset of exudation in the external medium made J_v and L_p rise, and this effect also increased with the degree of K⁺ starvation. Similarly, K⁺ starvation and ABA promoted both the flux of exuded Na⁺ and the accumulation of Na⁺ in the root. We suggest that ABA acts as a regulating signal for the radial transport of water across the root, and that potassium may be an effector of this mechanism.     

Impairment of Na+,K+-ATPase activity following enterotoxigenic Campylobacter jejuni infection: changes in Na+, Cl− and 3-O-methyl-D-glucose transport in vitro, in rat ileum

Abstract Unidirectional fluxes of Na⁺, Cl⁻ and 3-O-methyl-D-glucose (3-MG) were measured in vitro across Campylobacter jejuni live culture-infected and control rat ileal short-circuited tissues by the Using Chamber technique. Net secretion of Na⁺ and enhanced secretion of Cl⁻ ions was observed in the infected animals ( P < 0.001, n =6) as compared to the net absorption of Na⁺ and marginal secretion of Cl⁻ ions in the control animals. There was a significant decrease in the mucosal-to-serosal fluxes of 3-MG in C. jejuni -infected rat ileum. The specific Na⁺,K⁺-ATPase activity when measured biochemically in the membrane-rich fraction of enterocytes was found to be significantly lower (58%) in the infected group as compared to the control group ( P < 0.001). Our results therefore suggest that infection with an enterotoxigenic C. jejuni inhibits the Na⁺,K⁺-ATPase activity in rat enterocytes. The impairment of Na⁺,K⁺-ATPase activity thus appears to induce a secondary change in Na⁺,Cl⁻ and 3-MG transport in vitro in rat ileum.     

Heterogeneous Na+ Sensitivity of Na+,K+-ATPase Isoenzymes in Whole Brain Membranes

Abstract: The Na⁺ sensitivity of whole brain membrane Na⁺,K⁺-ATPase isoenzymes was studied using the differential inhibitory effect of ouabain (α1, low affinity for ouabain; α2, high affinity; and α3, very high affinity). At 100 m M Na⁺, we found that the proportion of isoforms with low, high, and very high ouabain affinity was 21, 38, and 41%, respectively. Using two ouabain concentrations (10⁻⁵ and 10⁻⁷ M ), we were able to discriminate Na⁺ sensitivity of Na⁺, K⁺-ATPase isoenzymes using nonlinear regression. The ouabain low-affinity isoform, α1, exhibited high Na⁺ sensitivity [ K _a of 3.88 ± 0.25 m M Na⁺ and a Hill coefficient ( n ) of 1.98 ± 0.13]; the ouabain high-affinity isoform, α2, had two Na⁺ sensitivities, a high ( K _a of 4.98 ± 0.2 m M Na⁺ and n of 1.34 ± 0.10) and a low ( K _a of 28 ± 0.5 m M Na⁺ and an n of 1.92 ± 0.18) Na⁺ sensitivity activated above a thresh old (22 ± 0.3 m M Na⁺); and the ouabain very-high-affinity isoform, α3, was resolved by two processes and appears to have two Na⁺ sensitivities (apparent K _a values of 3.5 and 20 m M Na⁺). We show that Na⁺ dependence in the absence of ouabain is the result of at least of five Na⁺ reactivities. This molecular functional characteristic of isoenzymes in membranes could explain the diversity of physiological roles attributed to isoenzymes.     

Excretion of ions (Cd2+, Li+, Na+ and Cl−) by Tamarix aphylla

Excretion of minerals by the NaCl-resistant and comparatively cadmium-resistant tree Tamarix aphylla (L.) Karst, was investigated. Cd²⁺ was excreted by plants exposed for 1–10 days to 9 or 45 μ M Cd²⁺ solutions. Excretion of this toxic ion increased considerably with time but was less than 5% of the quantities that had been accumulated in the shoots. Excretion of Na⁺ and Cl⁻ was positively correlated with NaCl concentration (1.5, 10, 50 m M ) of the medium. The Na⁺/Cl⁻ ratios of the excrete were positively correlated with the concentration of the treatment solution. Ca²⁺ excretion decreased with increasing NaCl concentrations of the solution. Excretion of K⁺ and Mg²⁺ was only little affected by NaCl. Excretion of Li⁺ occurred whenever this element was supplied in the uptake solution; daily excretion rates of Li⁺ increased with time. The ecological significance of excretion is discussed in relation to the low selectivity of the mechanism in T. aphylla .