Effects of nitrogen nutrition on salt-stressed Nicotiana tabacum var. Samsun in vitro plantlets

Tobacco shoots were grown in vitro for 35 d, in MS culture mediummodified to include various sources (nitrate-N, ammonium-N ora mixture) and levels (0–120 mM) of N, and in the presenceof 0–180 mM NaCI or iso-osmotic concentrations of mannitol.Growth of control plantlets was significantly inhibited whenNH₄⁺-N was the sole N source, and at high (120 mM) NO^–₃-N supply. Under conditions of salt stress (90 and 180 mM NaCI)growth was repressed, with roots being more severely affectedthan shoots. Salinity also inhibited root emergence in vitro.The only alleviation of the salt stress by nitrate nutritionobserved in this study was on shoot growth parameters of plantletsgrown on 60 mM NO^–₃-N and 90 mM NaCI. Although both weresignificantly inhibited by NaCI, nitrate reduc-tase activitywas more severely affected than nitrate uptake. When mannitolreplaced NaCI in the culture medium, similar Inhibition of growth,nutrient uptake and enzyme activity were recorded. These observations,together with the relatively low recorded values for Na⁺ andCI^– uptake, indicate that under in vitro salt stress conditionsthe negative effects of NaCI are primarily osmotic. Key words: Growth, nitrogen metabolism, osmotic stress, salinity     

Inter-population Variations of Kochia indica During Germination Under Different Stresses

Germination of three populations of Kochia indica Wight fromcontrasting soil types were studied under various levels ofsalinity, alkalinity and osmotic stress with a view to evaluatetheir potential to establish through seeds in saline and alkalineenvironments of various magnitudes. The seeds were germinatedunder controlled conditions in Petri dishes using salinizedsolutions of NaCl, Na₂SO₄, MgCI₂ and CaSO₄ (electrical conductivity,EC, 4–16 ds m^–), sodium carbonate solutions (pH8.5–100), mannitol solutions ( – 2.5 to –100 bar) and de-ionised distilled water (control). Inter-populationdifferences were very marked, showing selective adaptation ofthe populations to a particular stress type. In general, allpopulations exhibited salt tolerance but were rather sensitiveto high osmotic stress. Kochia indica Wight (bui), germination, population, salinity, alkalinity, osmotic stress     

Further Characteristics of Salt-Dependent Bicarbonate Use by the Seagrass Zostera muelleri

Millhouse, J. and Strother, S. 1987. Further characteristicsof salt-dependent bicarbonate use by the seagrass Zostera muelleri.—J.exp. Bot. 38: 1055–1068. The contribution of HCO^–₃to photosynthetic O₂ evolutionin the seagrass Zostera muelleri Irmisch ex Aschers. increasedwith increasing salinity of the bathing seawater when the inorganiccarbon concentration was kept constant. K_1/2 (seawater salts)for HCO^–₃ -dependent photosynthesis was 66% of seawatersalinity. Both short- and long-term pretreatment at low salinitiesstimulated photosynthesis in full strength seawater. Twentyfour hours pre-incubation of seagrass plants in 3·0 molm^–3 NaHCO₃ resulted in increased photosynthesis at allsalinities, apparently due to stimulation of HCO^–₃ use(K_1/2 (seawater salts) = 26%). V_max (HCO^–₃) was not affectedby low salinity pretreatment. The kinetics of HCO^–₃ stimulationby the major seawater cations was investigated. Ca²⁺ was themost effective cation with the highest V_max (HCO^–₃) andwith K_1/2(Ca²⁺) = 14 mol m^–3. Mg²⁺ was also very effectiveat less than 50 mol m^–3 but higher concentrations wereinhibitory. This inhibition cannot be accounted for solely byprecipitation of MgCO₃. Na⁺ and K⁺ were both capable of stimulatingHCO^–₃ use. Stimulation was in two distinct parts. Up to500 mol m^–3, both citrate and chloride salts gave similarresults (K_1/2(Na⁺) 81 mol m^–3, V_max(HCO^–₃) 0·26µmol O₂ mg^–1 chl min^–1), but use of citratesalts above 500 mol m^–2 caused a second stimulation ofHCO^–₃ use (K_1/2(Na⁺) 830 mol m^–3, V_max(HCO^–₃)0·68 µmol O₂ mg^–1 chl min^–1). V_max(HCO^–₃)for the second-phase Na⁺ or K⁺ stimulation was of the same orderas for Ca²⁺-stimulated HCO^–₃ use. To further characterizesalt-dependent HCO^–₃ use, the sensitivity of photosynthesisto Tris and TES buffers was investigated. The effects of Trisappear to be due to the action of Tris⁺ causing stimulationof HCO^–₃ -dependent photosynthesis in the absence of salt,but inhibition of HCO^–₃ use in saline media. TES has noeffect on photosynthesis. External carbonic anhydrase, althoughimplicated in salt-dependent HCO^–₃ use in Z. muelleri,could not be detected in whole leaves. Key words: Zostera muelleri, HCO^–₃ use, salinity     

Role of K+ and Cl- in Osmotic Adjustment in Roots and Hypocotyls of Intact Mung Bean Seedlings

It was confirmed that osmotic adjustment occurred in young intactmung bean (Vigna mungo (L.) Hepper) seedlings exposed to highosmotic pressure stress. Root growth was not affected by osmoticpressure of less than 200 mOsra in the external solution, althoughhypocotyl growth was conspicuously reduced. Under this moderateosmotic stress, intracellular K⁺ concentration, [K⁺]_i, increaseddramatically during the osmotic adjustment in all the regionsof the root, but the intracellular Cl^– concentration,[Cl^–]_i, increased only in the aged mature region of theroot (28–33 mm from the root tip). About a half of theintracellular osmotic pressure in the aged mature region ofthe root could be ascribed to the contributions of [K⁺]_i and[Cl^–]_i, but in the hypocotyl, [K⁺]_i only contributed slightlyto the osmotic adjustment. (Received June 18, 1986; Accepted August 26, 1986)     

Induction of Increased Salt Tolerance in Sorghum bicolor by NaCl Pretreatment

Following 20 d of exposure to 75 or 150 mol m^–3 NaCl Sorghumbicolor (L.) Moench plants become capable of growing in mediumcontaining 300 mol m^–3 NaCl. Control plants, which havenot been pretreated, or plants pretreated for less than 20 ddie within 2 weeks when exposed to 300 mol m^–3 NaCl. Weconsider this induction of a capacity to survive in and toleratea high NaCl concentration as an adaptation to salinity. We suggestthat adaptation to salinity is more than osmotic adjustmentand that it takes longer to develop than osmotic adjustment.Concomitantly with the appearance of the ability to grow inhigh salinity, adaptation also comprises the development ofa capacity to regulate internal Na⁺ and Cl^– concentrations,even when external salinity is high. Shoot mean relative growthrates are similar for both control plants and for adapted plantsgrowing in 300 mol m^–3 NaCl, although their shoot Na⁺and Cl^– concentrations are quite different. Based on thesedata, we propose that adaptation of Sorghum to high salinityresults from a modulation of genome expression occurring duringextended exposure to non-lethal NaCl concentrations. Key words: Sorghum bicolor (L.) Moench, NaCl, salt tolerance, adaptation to salinity     

Lamprothamnium, a Euryhaline Charophyte: I. OSMOTIC RELATIONS AND MEMBRANE POTENTIAL AT STEADY STATE

The charophyte Lamprothamnium papulosum (Wallr.) J. Gr. is foundat salinities varying from nearly fresh water to twice thatof sea water. It can maintain its turgor constant at 302 mosmolkg^–1 (0.73 MPa) when exposed to external osmotic pressuresof 550 to 1350 mosmol kg^–1 (1.3–3.3 MPa). Turgorshows a tendency to rise slightly at lower osmotic pressure(388 mosmol kg^–1 of turgor at 150 mosmol kg^–1 externalosmolality). K⁺ and Cl^– are the main solutes in the vacuole,and are most important in controlling internal osmotic pressure.Mg²⁺, Ca²⁺, and SO^2–₄ are present in significant amountsbut their concentrations do not change with changes in externalsalinity. Na⁺ is present in lower concentration than K⁺, andplays a minor role in regulating turgor. Sucrose is presentin significant concentrations, but changes little with changesin salinity. Two enzymes involved in sucrose metabolism, sucrosephosphate synthetase (EC 2.4.1.14 [EC] ), and sucrose synthetase (EC2.4.1.13 [EC] ) are active in whole cell extracts of Lamprothamnium.As in the fresh water charophytes, Lamprothamnium membrane potentialmay be depolarized (close to E_K) or hyperpolarized, and presumablyof electrogenic origin. Both types of potential are found atall salinities tested.     

Salt Tolerance in the Succulent, Coastal Halophyte, Sarcocornia natalensis

The effects of 0, 50, 100, 200, 300, 400 and 500 mol m^–3NaCl on growth and ion accumulation in the succulent, coastalhalophyte Sarcocornia natalensis (Bunge ex Ung.-Sternb.) A.J. Scott were investigated. Increase in salinity from 0 to 300 mol m^–3 NaCl stimulatedproduction of fresh, dry, and organic dry mass, increased succulenceand shifted resource allocation from roots to shoots. Growthwas optimal at 300 mol m^–3 and decreased with furtherincrease in salinity. Water contributed to a large proportion of the increase in freshmass. Inorganic ions, especially Na⁺ and Cl– contributedsubstantially to the dry mass. At 300 mol m^–3 NaCl inorganicions contributed to 37% of total dry mass and NaCl concentrationin the shoots was 482 mol m^–3. Expressed sap osmotic potentialsdecreased from –2.10 to –3.95 MPa as salinity increasedfrom 0 to 300 mol m^–3 NaCl. Massive accumulation of inorganicions, especially Na⁺ and Cl^–, accounted for 86% of theosmotic adjustment at 300 mol m^–3 NaCl. Salinity treatments decreased the concentrations of K+ in shoots.Plant Na+ :K+ ratios increased steadily with salinity and reacheda maximum of 16.6 at 400 mol m3 NaCl. It is suggested that the exceptional salt tolerance of S. natalensisis achieved by massive inorganic ion accumulation which providessufficient solutes for osmoregulation, increased water fluxand turgor-induced growth. Key words: Sarcocornia natalensis, salt tolerance, halophyte     

Unequal distribution of potassium and anions within the Phaseolus pulvinus during circadian leaf movement

Ion and saccharide concentrations in the upper and lower partsof the laminar pulvinus of the primary leaf of Phaseolus vulgariswere measured in relation to the circadian movement. Concentrations of K⁺, Na⁺, Ca²⁺, Mg²⁺, Cl^–, organic acid,NO₃^–, H₂PO₄^–, fructose and fructose-yielding saccharidesin the pulvinus were 75–120, 0.3–0.7, 5–8,6–12, 40–60, 60–73, 19–35, 2–9and 1–5 mM, respectively, and the osmotic pressure ofthe pulvinus was considered to be due to these ions. The cell volume in the expanding part was larger than that inthe contracting part. The change of the cell volume alteredthe molar concentration in the cell sap and therefore the amountof solutes actually transported from the upper to the lowerpart and vice versa was estimated from the concentration expressedin moles per gram of dry weight. Results showed that K⁺, Cl^–, organic acid (or H⁺) andNO₃^– moved from the upper to lower parts or vice versain the pulvinus in relation to its deformation, keeping theelectroneutrality among those ions, whereas C_a2⁺ and Mg²⁺ didnot move. The difference in the K⁺ concentration between theupper and lower parts when the leaf was up or down amountedto 30% of the whole osmotic pressure. This lead to the conclusionthat the endogenous clock-controlled unequal distribution ofK⁺, Cl^–, organic acid (or H⁺) and NO₃^– in the pulvinuscould be the force for the circadian leaf movement. (Received August 7, 1979; )     

Effect of Divalent Cations on Na+ Permeability of Chara corallina and Freshwater Grown Chara buckellii

The effect of elevated Na⁺ concentration on Na⁺ permeability(P_Na) and Na⁺ influx in the presence of two levels of externaldivalent cations was determined in Chara corallina and freshwater-culturedChara buckellii. When Na⁺ in the medium was increased from 1.0to 70 mol m^–3, Na⁺ influx increased in both species ifCa²⁺ was low (0.1 mol m–3). If Ca²⁺ was increased to 7.0mol m^–3 when Na⁺ was increased, Na⁺ influx remained atthe low control level in C. corallina, and showed only a temporaryincrease in C. buckellii. Mg²⁺ was a better substitute for Ca²⁺in C. buckellii than in C. corallina. Na⁺ permeability data suggest that when the external Ca²⁺ concentrationis low, P_Na does not increase in the presence of elevated NaCl;the increase in Na⁺ influx appears to be due to the increasein external Na⁺ concentration alone. Ca^{2 +} supplementation appearsto decrease P_Na whereas supplemental Mg²⁺ has no effect. Na⁺ effluxes were computed from previously determined net fluxesand the influxes. It was found that for both species, fluxesin both directions were stimulated in response to all experimentaltreatments, but Na⁺ influx always exceeded efflux. This resultedin net Na⁺ accumulation in the vacuoles of both species. The results are discussed with reference to net flux and electrophysiologicaldata obtained previously under identical conditions, as wellas the comparative salinity tolerance of both species and theNa⁺/divalent cation ratio. Key words: Na⁺ influx, Na⁺ tolerance, membrane potential, permeability, Chara     

Dynamics of Salt Secretion by Sporobolus spicatus(Vahl) Kunth from Sites of Differing Salinity

Secretion of salts by bicellular salt glands and the water relationsof the grass Sporobolus spicatus were investigated at four sitesalong the coast of the Red Sea in Egypt that differed in theextremity of salinity and drought. Salt eliminated by the leaveswas similar in its composition at all sites. Na⁺and Cl^-werethe dominant ions in the soil, and together comprised about93% of the dry weight of secreted salt. The molar ratio of K⁺:Na⁺inthe plant leaves was more than ten-fold that in the interstitialsoil solution and thirteen-times that in the secreted salts,reflecting the high selectivity of the secretion mechanism forNa⁺. The concentration of Na⁺in the solution transported tothe leaves between 0900 and 1500 h was less than 0.1% of thatin the soil solution. Accumulation of salts by the plant shoots,which increased with increasing soil salinity and drought, wasmaximal during the day when the extent of secretion greatlyreduced. The ionic osmotic potential (     

Relative Importance of Ion Exclusion, Secretion and Accumulation in Spartina alterniflora Loisel.

The extent to which Spartina alterniflora Loisel. excluded,secreted or accumulated the major seawater ions (Cl^-, SO^2-₄,Na⁺, K⁺, Mg²⁺, and Ca²⁺) was investigated under varying salinitytreatments. From a quantitative viewpoint, ion exclusion wasmost prominent and accounted for 91–97% of the theoreticalmaximum ion uptake as a result of transpiration and growth.Of those ions taken up, approximately half was secreted fromthe shoots. Relative to K⁺, a disproportionate amount of Na⁺was excluded at the roots and secreted by the shoots. The concentrationwithin the tissues of S. alterniflora did not change with salinitytreatment for the majority of the ions examined, but Na⁺ wasmore than twice as concentrated at 40 g dm^-3 than at lOgdm^-3.Calculations of the flux of ions from salt marsh sediments tothe flood water via shoot secretion or stem/leaf turnover indicatethat these processes may be important to the ecology of S. alternifloraas mechanisms that limit the accumulation of salt within theroot zone.     

Effects of Na2SO4, K2SO4 and KCl on Growth and Ion Uptake of Callus Cultures of Vaccinium corymbosum L. cv. Blue Crop

Non-selected and Na₂SO-, K₂SO₄- or KCl-selected callus culturesof Vaccinium corymbosum L. cv. Blue Crop were grown on mediasupplemented with 0, 25 and 50 mM Na₂SO₄ (non-selected and Na₂SO(-selectedonly), 0, 25 and 50mMK₂SO₄ (non-selected and K₂SO₄-selectedonly) or 0, 50 and 100 mM KCl (non-selected and KCl-selectedonly). On all media, growth of selected callus (on a fresh-weightor dry-weight basis) was greater than that of non-selected callus,and selected callus grew optimally on the level and type ofsalt on which it was selected. Selected callus was friable andmaintained a higher f. wt:d. wt ratio. Tissue water potentialin selected callus was more negative than in non-selected callus. Flame photometry and chloridometry showed Na⁺, K⁺ and Cl^–accumulated in callus to concentrations equal to or greaterthan the initial concentration in the medium. Turbidometry showedthat tissue SO₄^2- concentration was lower than the concentrationin the medium. In most cases selected callus accumulated moreNa⁺, K^sup, SO₄^2– or Cl^– than non-selected callus.Vacuolar ion concentration was measured by electronprobe X-raymicroanalysis, and on most media selected callus had highervacuolar ion concentrations than non-selected callus. SO₄^2–and Cl^– were accumulated in the vacuoles at concentrationshigher than the external medium, but vacuolar Na⁺ concentrationdid not reach external concentration on Na₂SO₄ and on potassiumsalts was maintained between 12 and 17 mM. Vacuolar K⁺ concentration(approx. 142–191 mM on no salt) decreased on Na₂SO₄ andincreased on K₂SO₄ and KCl. There was no precise correlation between total or specific ionaccumulation (Na⁺, K⁺, SO₄^2– and Cl^– and fresh-weightyield. Results suggest that selection results in adaptationin response to decreased water potential of the medium. Vaccinium corymbosum, blueberry, electronprobe X-ray microanalysis, callus, in vitro selection, salt tolerance, KCl, K₂SO₄, Na₂SO₄     

Inter-population Variations of Kochia indica During Germination Under Different Stresses

Germination of three populations of Kochia indica Wight fromcontrasting soil types were studied under various levels ofsalinity, alkalinity and osmotic stress with a view to evaluatetheir potential to establish through seeds in saline and alkalineenvironments of various magnitudes. The seeds were germinatedunder controlled conditions in Petri dishes using salinizedsolutions of NaCl, Na₂SO₄, MgCl₂ and CaSO₄ (electrical conductivity,EC, 4–16 dsm¹), sodium carbonate solutions (pH 8.5ndash;10.0), mannitol solutions ( —2.5 to —10.0 bar) andde-ionised distilled water (control). Inter-population differenceswere very marked, showing selective adaptation of the populationsto a particular stress type. In general, all populations exhibitedsalt tolerance but were rather sensitive to high osmotic stress. Kochia indicaWight (bui), germination, populatio, salinity, alkalinity, osmotic stress     

Effect of NaCI Salinity on Flows and Partitioning of C, N, and Mineral Ions in Whole Plants of White Lupin, Lupinus albusL.

Plants of Lupinus albus L., cv. Ultra, were grown hydroponicallywith NO₃^–-nutrition for 51 d under control (0.05 mol m^–3Na⁺ and 10 mol m^–3 Cl^–) and saline (40 mol m^–3NaCI) conditions. Plants were harvested 41 and 51 d after germinationand analysed for content and net increment of C, N and the mineralcations K⁺, Na⁺, Mg²⁺, and Ca²⁺ and the anions Cl^–, NOJ,malate, phosphate, and SO₄^2–. Roots, stem interaodes,petioles and leaflets were analysed separately. During the studyperiod net photosynthesis, respiratory losses of CO₂ from shootand root and the composition of the spontaneously bleeding phloemsap and the root pressure xylem exudate were also determined.Using molar ratios of C over N in the transport fluids, incrementsof C and N, and photosynthetic gains as well as respiratorylosses of C, the net flows of C and N in the xylem and phloemwere then calculated as in earlier studies (Pate, Layzell andMcNeill, 1979a). Knowing the carbon flows, the ratios of ionto carbon in the phloem sap, and ion increments in individualorgans, net flows of K⁺, Na⁺, and Cl^– over the study periodwere also calculated. Salt stress led to a general decrease of all partial componentsof C and N partitioning indicating that inhibitions were notdue to specific effects of NaCI salinity on photosynthesis oron NO₃ uptake. However, there were differences between variouslyaged organs, and net phloem export of nitrogenous compoundsfrom ageing leaves was substantially enhanced under saline conditions.In addition, NO₃^–reduction in the roots was specificallyinhibited. Uptake and xylem transport of K⁺ was more severelyinhibited than photosynthetic carbon gain or NO₃^– uptakeby the root. K⁺ transport in the phloem was even more severelyrestricted under saline conditions. Na⁺ and Cl^– flowsand uptake, on the other hand, were substantially increasedin the presence of salt and, in particular, there were thenmassive flows of Na^– in the phloem. The results are discussedin relation to the causes of salt sensitivity of Lupinus albus.The data suggest that both a restriction of K⁺ supply and astrongly increased phloem translocation of Na⁺ contribute tothe adverse effects of salt in this species. Restriction ofK⁺ supply occurs by diminished K⁺ uptake and even more by reducedK⁺ cycling within the plant. Key words: Lupinus albus, salt stress, phloem transport, xylem transport, partitioning, carbon, nitrogen, K⁺, Na⁺, CI^–     

External Potassium Supply is not Required for Root Growth in Saline Conditions: Experiments with Ricinus communis L. Grown in a Reciprocal Split-Root System

Ricinus communis L. (castor bean) plants were grown in the absence(control) and in the presence of 100molm^–3NaCl with areciprocal split-root system, in which K⁺ was supplied to oneand NO₃^– to the other part of the root system. In theseplants shoot and, to a lesser extent, total root growth wereinhibited compared to plants with non-split roots. Without andwith NaCl, growth of roots receiving NO₃^– but noK⁺ (‘minusK/plus N-roots’) was substantially more vigorous thanunder the reverse conditions (‘plus K/minus N-roots¹).100mol m^–3 NaCl inhibited growth of minus K/plus N-roots¹to the same extent as that of non-split roots, indicating thatexternally supplied K⁺ was not required for root growth undersaline conditions. In growth media without added K⁺ the rootdepleted the external low K ⁺ levels resulting from chemicalsdown to a minimum value C_mln (1.0 to 1.4 mmol m^–3); inthe presence of 100 mol m^–3 NaCl, C_min, was higher (10–18mmol m^–3) and resulted from an initial net loss of K ⁺.C_min, was pH-dependent The distribution of K⁺, Na⁺ and Mg²⁺along the root was measured. In meristematic root tissues, K⁺ concentrations were scarcely affected by external K⁺ or byNaCl, where Na ⁺ concentrations were low, but somewhat elevatedat low external K+ and/or high NaCl. In differentiated, vacuolatedtissues K ⁺ concentrations were low and Na⁺ concentrations high,if K ⁺ was not supplied externally and/or NaCl was present.The longitudinal distribution of ions within the root was usedto estimate cytoplasmic and vacuolar ion concentrations. Thesedata showed a narrow homoeostasis of cytoplasmic K+ concentrations(100–140 mol m^–3) independent of external K ⁺ supplyeven in the presence of 100 mol m ^–3 NaCl. CytoplasmicNa ⁺ concentrations were maintained at remarkably low levels.Hence, external K⁺ concentrations above C_min, were not requiredfor maintaining K/Na selectivity, i.e. for controlling Na⁺ entry.The results are discussed with regard to mechanisms of K/Naselectivity and to the importance of phloem import of K⁺ forsalt tolerance of roots and for cytoplasmic K⁺ homoeostasis. Key words: Ricinus communis, nitrate, potassium, root (split-root), salt tolerance, phloem transport     

Lamprothamnium, a Euryhaline Charophyte: IV. MEMBRANE POTENTIAL, IONIC FLUXES AND METABOLIC ACTIVITY DURING TURGOR ADJUSTMENT

The early time-course of turgor adjustment following a hyper-or hypo-osomotic shock was examined in the brackish-water charophyteLamprothamnium papulosum. The response to a reduction in turgorwas a five to seven-fold stimulation of the influxes of Cl^–,K⁺ and Na⁺. The distribution of radioactive tracers in the cellsuggested that the ionic composition of the cytoplasm was strictlycontrolled during turgor adjustment. Metabolic activity wasrelatively unaffected by the loss of turgor. high fluxes throughthe cytoplasm, and a cytoplasmic K^– concentration possiblyas high as 280 mol m^–3. Osmotic adjustment to a lower salinity was achieved by largeincreases in the passive effluxes of K⁺ and Cl^– ratherthan by decreases in their influxes. The membrane remained hyperpolarized during hyperosmotic adjustmentbut depolarized after a hypo-osmotic change. This result isdiscussed in relation to changes in the driving forces for ionmovements during osmotic transitions. Key words: Lamprothamnium, Turgor, Osmotic stress     

The Effect of Salinity Changes Upon the Physiology of Eulittoral Green Macroalgae from Antarctica and Southern Chile: II INTRACELLULAR INORGANIC IONS AND ORGANIC COMPOUNDS

The effects of hypo- and hypersaline treatments ranging from7–68% on the intracellular inorganic ion and organic soluteconcentrations were determined in the eulittoral green macroalgaeUlothrix implexa, Ulothrix subflaccida, Enteromorpha bulbosa,Acrosiphonia arcta, and Ulva rigida from Antarctica and SouthernChile. The main inorganic cations were K⁺, Na⁺, and Mg²⁺ inall species. The major osmolyte in E. bulbosa, A. arcta, andU. rigida was K⁺ at increasing salinities. In both Ulothrixspecies, however, K⁺ levels declined during hypersaline stressand Na⁺ concentrations rose significantly. The main inorganicanions were Cl^-, SO²_4-, and PO³_4- in all algae, while E. bulbosaand U. rigida also contained NO⁺₃. A. arcta showed an extremelyhigh SO^2-₄ content. The organic solutes proline, sucrose, andß-dimethylsulphoniopropionate (DMSP) played an importantrole in osmotic acclimation. The occurrence of three organicosmolytes suggests an additional function of these solutes ascryoprotectants in the cold-water macroalgae investigated.     

Osmotic and Ionic Regulation in Chara L-cell Fragments

Ion absorption from rather complicated artificial pond water(APW) by cell fragments having a lower osmotic pressure thanthe intact internodal cell (L-cell fragments) was studied. L-cellfragments were prepared by taking advantage of trans cellularosmosis and ligating the cell with thread. The results wereas follows: (1) L-cell fragments absorbed more K⁺ than Na⁺ fromaKCL + NaCl mixture in the presence of Ca²⁺, Mg²⁺ and SO₂₄ inthe light; (2) the influx of KCI was larger than that of KNO₃;(3) the amount of positive charge carried by K⁺, Na⁺ and Mg²⁺across the cell membrane balanced well with the amount of negativecharge carried by Cl^– in Cl^–-containing and NO₃^–-free APW; (4) no conclusion could be made as to whether ornot the rule of electro neutrality held for the K⁺, Na⁺, Ca²⁺and NO₃^– fluxes across the cell membrane, because dilutedKNO₃ is unstable; (5) L-cell fragments in KCl-containing APWsurvived longer than those in KNO₃-containing and Cl^–-free APW; (6) after incubation in KNO₃-containing and Cl^–-freeAPW, L-cell fragments absorbed a great amount of KCI immediatelyafter being transferred to KCl-containing and NO₃^– -freeAPW; and (7) lowering the turgor pressure of the intact cellby raising the external osmotic pressure did not induce ionflux into the cell. Thus, we concluded that the L-cell fragmentsabsorbed ions from the external solution not because of theirlower turgor pressure, but because of the diluted ion concentrationof the cytoplasm and the vacuole. The electroneutrality ruleheld, at least, for K⁺, Na⁺, Mg²⁺ and Cl^– influxes acrossthe cell membrane inthe KCl-containing and NO₃^–-free APW.These results were analyzed on the basis of an extended poremodel which presumed the existence of ATP-dependent processesin the membrane, and suggested that K⁺, Na⁺ and Mg₂₊ inflowsinto an L-cell fragment are likely to be induced by active Cl^–inflow. (Received May 18, 1987; Accepted September 29, 1987)     

Salt Stress Causes Acceleration of Purine Catabolism and Inhibition of Pyrimidine Salvage in Zea mays Root Tips

Nucleotide metabolism was studied in apical 5.0 mm root tipsof corn plants (Zea mays L., cv. Pioneer 3906) hydroponicallycultured for 7 d and then salinized for 19 d at a rate calculatedto reduce the osmotic potential (_o) of the solutions by O.1MPad^–1 to a final _o = -0.4 MPa. Saline treatments withtwo different molar ratios of Ca₂₊/Na⁺ were employed, viz.,0–03 (2.5 mol m^–3 CaCl₂ + 86.5 mol m^–3 NaCl)for the NaCl treatment and 0.73 (31.5 mol m^–3 CaCl₂ +43.1 mol m^–3 NaCl) for the NaCl + CaCl₂ treatment. Bothsalt treatments reduced root growth by more than 30%. The capacityof roots to provide purine nucleotides either by de novo synthesisor by re-utilization of existing bases, e.g. salvage of hypoxanthineto adenine nucleotides, was not affected by either salt treatment.However, catabolism of hypoxanthine was accelerated more than3.5-fold by both salt treatments, demonstrating an increasedcapacity for purine catabolism which would shift the normal1: 1 ratio of synthesis: degradation of purine nucleotides observedfor the roots of healthy control plants to less than 0.2 duringsalt stress. The ratio of pyrimidine nucleotide synthesis: degradationwas also reduced. In this case, the unfavourable shift towardnucleotide degradation resulted because both salt treatmentsreduced salvage capacity by more than 25%, but had no compensatingeffect on de novo synthesis or catabolism of pyrimidines. Key words: Salinity, osmotic potential, nucleotide metabolism     

Na+-dependent HCO3- uptake into the rat choroid plexus epithelium is partially DIDS sensitive

Several studies suggest the involvement of Na⁺ and HCO₃^– transport in the formation of cerebrospinal fluid. Two Na⁺-dependent HCO₃^– transporters were recently localized to the epithelial cells of the rat choroid plexus (NBCn1 and NCBE), and the mRNA for a third protein was also detected (NBCe2) (Praetorius J, Nejsum LN, and Nielsen S. Am J Physiol Cell Physiol 286: C601–C610, 2004). Our goal was to immunolocalize the NBCe2 to the choroid plexus by immunohistochemistry and immunogold electronmicroscopy and to functionally characterize the bicarbonate transport in the isolated rat choroid plexus by measurements of intracellular pH (pH_i) using a dual-excitation wavelength pH-sensitive dye (BCECF). Both antisera derived from COOH-terminal and NH₂-terminal NBCe2 peptides localized NBCe2 to the brush-border membrane domain of choroid plexus epithelial cells. Steady-state pH_i in choroidal cells increased from 7.03 ± 0.02 to 7.38 ± 0.02 (n = 41) after addition of CO₂/HCO₃^– into the bath solution. This increase was Na⁺ dependent and inhibited by the Cl^– and HCO₃^– transport inhibitor DIDS (200 µM). This suggests the presence of Na⁺-dependent, partially DIDS-sensitive HCO₃^– uptake. The pH_i recovery after acid loading revealed an initial Na⁺ and HCO₃^–-dependent net base flux of 0.828 ± 0.116 mM/s (n = 8). The initial flux in the presence of CO₂/HCO₃^– was unaffected by DIDS. Our data support the existence of both DIDS-sensitive and -insensitive Na⁺- and HCO₃^–-dependent base loader uptake into the rat choroid plexus epithelial cells. This is consistent with the localization of the three base transporters NBCn1, Na⁺-driven Cl^– bicarbonate exchanger, and NBCe2 in this tissue. bicarbonate metabolism; BCECF; cerebrospinal fluid; acid/base transport; ammonium prepulse