Reactivation of Cytoplasmic Streaming in a Tonoplast-Permeabilized Cell Model of Acetabularia

To find whether cytoplasmic streaming in Acetabularia is controlledby Ca²⁺, a tonoplast-permeabilized cell model was prepared usinga vacuolar perfusion technique. The cytoplasmic streaming remainedalmost normal after perfusion with EGTA medium (10 mM EGTA,40 mM PIPES, 5mM MgCl₂ and 800 mM sorbitol, pH 6.9), but stoppedwithin 10 min when saponin medium (EGTA medium plus 50 µg/mlsaponin, 50 µg/ml hexokinase and 5 mM glucose) was perfused.This model system was reactivated with a solution containing0.5 mM ATP and different concentrations of Ca²⁺ (reactivationmedium). With the reactivation medium at pCa 6–5, theresumed streaming lasted for about 10 min before the cytoplasmaggregated. At pCa 4–3, the streaming was observed onlyfor a few minutes because the cytoplasm aggregated quickly.At pCa 7, no reactivated movement was observed. Reactivationwas not induced in an ATP- or Mg²⁺-deficient medium even inthe presence of an adequate concentration of Ca²⁺, and was inhibitedby 50 µg/ml cytochalasin B or 1 mM N-ethylmaleimide. We concluded from these observations that the cytoplasmic streamingin Acetabularia is very likely to be driven by the actomyosinsystem in the presence of Mg-ATP and Ca²⁺ at pCa 6–5. (Received October 31, 1984; Accepted April 1, 1985)     

Effect of GA3 on the Molecular Mass of Polyuronides in the Cell Walls of Alaska Pea Roots

The role of cell wall matrix polysaccharides in gibberellin-regulatedroot growth is unknown. We examined pectic polysaccharides frompea roots treated with or without gibberellin A₃ (GA₃) in thepresence of ancymidol, an inhibitor of gibberellin biosynthesis.Pectic polymers solubilized by CDTA (trans-l,2-cyclohexanediamine-N,N,N',N'-tetraaceticacid) at 23°C and subjected to gel permeation analysis exhibitedhigh polydispersity with a molecular mass in excess of 500 kDa.Subsequent extraction of cell walls with CDTA at 100°C solubilizedpolymers with an average mol mass of 10 to 40 kDa. Subjectingthe high molecular mass pectic polymers extracted at 23°Cto 70–100°C for 2h generated 10 to 40 kDa fragments,similar in size distribution to those solubilized directly fromcell walls by CDTA solutions at 100°C. Pectic polymers from(GA₃+Anc)-treated roots were of higher average mol mass thanthose from Anc-treated roots in both the elongation zone andin the basal maturation zone. Since (GA₃+Anc)-treated rootselongate more quickly than Anc-treated roots [Tanimoto (1994)Plant Cell Physiol. 35:1019], the slender, GA₃-treated rootsmay produce and deposit highly integrated pectins more rapidlythan the thicker, Anc-treated roots in the elongating or elongatedcell walls. ²Present address: Horticultural Sciences Department, POB 110690IFAS, University of Florida, Gainesville, FL 32611-0690 U.S.A.     

Effects of lanthanum on rhythmic and nyctinastic leaflet movements in Albizzia lophantha

The involvement of extracellular calcium in rhythmic and nyctinasticmovement oi Albizzia lophantha Benth. leaflets has been studiedby testing the effect of LaCl₃ and its interaction with thephytochrome control of these movements. A 2h pulse of LaCl₃(10–50 mM) promotes a loss of rhythmicity, leaving leafletsin an open position, and also overrides the phase shift causedby phytochrome. A 2 h pulse of LaCl₃ (1 mM) decreases the amplitudeof rhythmic oscillations but does not promote arhythmicity normodify the phase shift caused by red light. The red light pulseabolishes the damping effect of 1 mM La³⁺. LaCl₃ inhibits nyctinasticclosure and decreases the phytochrome control of nyctinasticclosure. A subsequent supply of CaCl₂ (10 to 100 mM) does notreverse La³⁺ (10 mM) inhibition of closure. Light-induced openingis independent of LaCl, but rhythmic opening in darkness showsdifferent responses to La³⁺ depending on the time at which La³⁺is applied. Data suggest that extracellular calcium is requiredfor the closure mechanism and for the expression of rhythmicmovement. It could also be involved in the phytochrome transductionpathway and/or in the linking steps between phytochrome andthe circadian clock. Key words: Albizzia lophantha, calcium, circadian rhythm, lanthanum, phytochrome     

Effects of Chilling, Light and Nitrogen-containing Compounds on Germination, Rate of Germination and Seed Imbibition of Clematis vitalba L.

Effects of chilling (5 °C) period, light and applied nitrogen(N) on germination (%), rate of germination (d to 50% of totalgermination; T_50%) and seed imbibition were examined inClematisvitalba L. In the absence of chilling, light and N, germinationwas minimal (3%). When applied alone, both chilling and N increasedgermination. Chilling for 12 weeks increased germination to64%, and 2.5 mM NO^-₃or NH⁺₄increased germination to 10–12%.Light did not increase germination when applied alone, but didwhen applied in combination with chilling and/or N. Half theseed germinated when light was combined with 2.5 mM NO^-₃or NH⁺₄.The influence of chilling, light and/or N on germination wasgreater when combined, than when either factor was applied alone.Both oxidized (NO^-₃) and reduced (NH⁺₄) forms of N increasedgermination, but non-N-containing compounds did not, suggestingthe response was due to N and not ionic or osmotic effects. Without additional N, T_50%decreased from 16–20 d at zerochilling, to around 5 d at 8 and 12 weeks chilling. AlthoughT_50%was not influenced by an increase in NO^-₃or NH⁺₄from 0.5to 5.0 mM , it did increase with additional applied N thereafter.However, the magnitude of the N effect was small compared tothat of chilling. Like germination, seed imbibition increasedwith a longer chilling period, but in contrast imbibition decreasedslightly with increased applied NO^-₃or NH⁺₄. It is argued thatincreased imbibition is not directly related to an increasein total germination, but that it may be related to the rateof germination. Possible mechanisms involved in the reductionin dormancy ofC. vitalba seed are discussed. Clematis vitalba L.; germination; dormancy; imbibition; rate of germination; chilling; light; nitrate; ammonium; nitrogen; phytochrome     

Effects of ABA on Synthesis of Cell-Wall Polysaccharides in Segments of Etiolated Squash Hypocotyl I. Changes in Incorporation of Glucose and myo-Inositol into Cell-Wall Components

Externally supplied [³H]myo-inositol and [¹⁴C]glucose were incorporatedin cell-wall fractions of segments of etiolated squash hypocotyl.The extent of incorporation of [¹⁴C]glucose into cell-wall fractionswas very much greater than that of [³H]myo-inositol. Radioactivityfrom [¹⁴C]-glucose was effectively incorporated into hemicelluloseB and cellulose fractions and was incorporated uniformly intohexose, pentose and uronic acid residues, but radioactivityfrom [³H]myo-inositol was incorporated predominantly into uronicacid and pentose residues in the pectin and hemicellulose Bfractions. Exogenously applied ABA significantly suppressed the elongationof segments of squash hypocotyl and the incorporation of radioactivityfrom [^l4C]glucose and [³H]myo-inositol into the segments. Furthermore,ABA significantly inhibited the distribution of incorporatedradioactivity from [¹⁴C]glucose into the cellulose fraction,but did not affect distribution into the pectic fraction. Bycontrast, ABA only slightly inhibited the distribution of theincorporated radioactivity from [³H]myo-inositol into the pecticfraction. These results suggest that most of the cell-wall polysaccharidesin segments of squash hypocotyl are synthesized via the UDP-sugarpathway, and that ABA significantly inhibits the synthesis ofcellulose but not the synthesis of pectic polysaccharides whenABA suppresses the elongation of the segments. (Received March 25, 1988; Accepted November 15, 1988)     

Mutagenesis of the N-glycosylation site of hNaSi-1 reduces transport activity

The human Na⁺-sulfate cotransporter (hNaSi-1) belongs to the SLC13 gene family, which also includes the high-affinity Na⁺-sulfate cotransporter (hSUT-1) and the Na⁺-dicarboxylate cotransporters (NaDC). In this study, the location and functional role of the N-glycosylation site of hNaSi-1 were studied using antifusion protein antibodies. Polyclonal antibodies against a glutathione S-transferase fusion protein containing a 65-amino acid peptide of hNaSi-1 (GST-Si65) were raised in rabbits, purified, and then used in Western blotting and immunofluorescence experiments. The antibodies recognized native NaSi-1 proteins in pig and rat brush-border membrane vesicles as well as the recombinant proteins expressed in Xenopus oocytes. Wild-type hNaSi-1 and two N-glycosylation site mutant proteins, N591Y and N591A, were functionally expressed and studied in Xenopus oocytes. The apparent mass of N591Y was not affected by treatment with peptide-N-glycosylase F, in contrast to the mass of wild-type hNaSi-1, which was reduced by up to 15 kDa, indicating that Asn⁵⁹¹ is the N-glycosylation site. Although the cell surface abundance of the two glycosylation site mutants, N591Y and N591A, was greater than that of wild-type hNaSi-1, both mutants had greatly reduced V_max, with no change in K_m. These results suggest that Asn⁵⁹¹ and/or N-glycosylation is critical for transport activity in NaSi-1. antifusion protein antibodies; Xenopus oocytes; sulfate; immunofluorescence     

Effects of Osmotic Stress, Ionic Stress and IAA on the Cell-Membrane Resistance of Vigna Hypocotyls

The cell-membrane resistance (R_m) of Vigna hypocotyls was examined,and the effects of osmotic stress, ionic stress and IAA on R_mwere investigated. R_m decreased by 64 to 77% under osmotic stressin the presence of absorbable solutes (40 mM sorbitol, 15 mMKC1, 30 mM sucrose; or 40 mM sorbitol, 15 mM KC1, 30 mM sucroseplus 10^–4 M IAA) or under ionic stress (50 mM NaCl or50 mM KC1). R_m was not changed by perfusion with 10^–4M IAA. Therefore, the hyper-polarizations of the membrane potentialobserved in both cases should be ascribed totally to the activationof the electrogenic proton pump. Although R_m showed an increaseof 1.6 fold when the hypocotyls were subjected to osmotic stress(100 mM sorbitol or 100 mM sorbitol plus 10^–4 M IAA),83.6% or 92.4% of the hyperpolarization of the membrane potential(V_px was also the result of the activation of the pump. Theresults, calculated on the basis of the current source model,support the viewpoint that the hyperpolarization of the cellmembrane potential of Vigna hypocotyls under osmotic stress,ionic stress or in the presence of IAA is an expression of theactivation of the proton pump, and is not caused by an increasein R_m. ¹ Present address: Researchers and Planners of Natural Environment, Yotsugi Bldg. (2F), 1-5-4 Horinouchi, Suginami-Ku, Tokyo,166 Japan ² Present address: Graduate School of Integrated Science, YokohamaCity University, 22-2 Seto, Kanazawa-Ku, Yokohama, 236 Japan (Received February 14, 1991; Accepted July 24, 1991)     

Changes of Cell Wall Composition and Polymer Size in Primary Roots of Cotton Seedlings Under High Salinity

The aim of this study was to investigate changes in cell wallchemical composition and polymer size in the root tip of intactcotton seedlings (Gossypium hirsutum L. cv. Acala SJ-2) grownin saline environments, in order to relate the interaction betweenhigh salinity and root growth to possible changes in cell wallmetabolism. Cotton seedlings were grown in modified Hoagland nutrient solutionwith various combinations of NaCl and CaCl₂. Cell walls werefractionated into four fractions (pectin, hemicellulose 1 and2, cellulose), and analysed for their total sugar content, neutralsugar composition and size of polysaccharides. At 1 mol m^–3Ca, 150 mol m^–3 NaCl resulted in a significant increasein the cell wall uronic acid content, but a reduction in cellulosecontent on a per unit dry weight basis. Supplemental Ca overcamethe inhibitory effect of high Na on cellulose content. The neutralsugar composition of the cell wall fractions showed no majorchanges caused by varied Na/Ca ratios. Determinations of polysaccharidepolymer size showed that high Na at 1 mol m^–3 Ca led toan increase in the amount of polysaccharides of intermediatemolecular size and a decrease in that of small size in the hemicellulose1 fraction, indicating a possible inhibition of polysaccharidedegradation by high Na. This change was not observed in the10 mol m^–3 Ca treatments. The results reveal a relationshipbetween the effects of high salinity on root growth and cellwall metabolism, particularly in regard to cellulose biosynthesis Key words: Gossypium hirsutum, salinity, root, cell wall     

Direct Transfer of Plasmid DNA from Intact Yeast Spheroplasts into Plant Protoplasts

We developed a polyethylene glycol (PEG)-mediated direct DNAtransfer method from intact Saccharomyces cerevisiae spheroplastsinto Arabidopsis thaliana protoplasts. To monitor the DNA transferfrom yeast to plant cells, ß-glucuronidase (GUS) reportergene in which a plant intron was inserted was used as a reporter.This intron-GUS reporter gene on a 2µm-based plasmid vectorwas not expressed in yeast transformants, while it expressedGUS activity when the plasmid DNA was introduced into plantcells. When a mixture of 1 x 10⁸ of S. cerevisiae spheroplastsharboring the plasmid and 2 x 10⁶ of A. thaliana protoplastswas treated with PEG and high pH-high Ca²⁺ solution (0.4 M mannitol,50 mM CaCl₂, 50 mM glycine-NaOH pH 10.5), GUS activity was detectedin the extract of the plant cells after a three-day culture.The GUS activity was higher than that of a reconstitution experimentin which the mixture of 1 x 10⁸ of S. cerevisiae spheroplastswhich did not carry the reporter gene, 2 x 10⁶ of A. thalianaprotoplasts and the same amount of the reporter plasmid DNAas that contained in 1 x 10⁸ of S. cerevisiae spheroplasts,was treated with PEG and high pH-high Ca²⁺ solution. Moreover,the GUS gene expression was resistant to micrococcal nucleasetreatment before and during PEG treatment. From these results,we concluded that plasmid DNA can be directly transferred fromintact yeast spheroplasts to plant protoplasts by a nuclease-resistantprocess, possibly by the cell fusion. ²Deceased on September 15, 1992.     

23Na and 7Li NMR study of Nitella cell walls before and after an ion-induced loss of the cationic exchange capacity

²³Na- and ⁷Li-NMR studies were conducted on isolated cell wallsof Nitella to monitor the changes induced in the pectin matrixafter the cation exchange capacity was reduced to about 60%of its initial value by a pretreatment with a 500 mM LiCl solution.Walls in Ca²⁺ form were exchanged with NaCl and LiCl solutionsbefore and after this pretreatment. In the whole cell wall equilibratedwith 10, 35 and 100 mM NaCl solutions or mixtures at the sametotal concentrations of Na⁺ and Li⁺ in the same proportions,the ²³Na- and ⁷Li-NMR spectra were all characterized by onlyone signal. The Na⁺ and Li⁺ relaxation times were drasticallyshorter than those recorded in aqueous solution but the twoNa⁺ relaxation rates increased linearly with the Na⁺ concentrationin the external solutions. In contrast, in the cell wall wherethe CEC had been reduced, the Na⁺ relaxation times were longerand no correlation with external concentrations was obtained.The ⁷Li spectra in walls equilibrated with 500 mM LiCl displayedtwo distinct lines suggesting different Li⁺ bonding affinitiesfor the wall polyuronides. These data are interpreted in thescheme proposed by Gillet et al. (1994) to explain the competitiveeffects of alkaline ions on the wall pectin disorganizationvia disruption of divalent cation crosslinks, but also of solvation-likebonds between cell wall polymers. Key words: ²³Na, ⁷Li, NMR, Nitella, pectic polysaccharides, cell wall     

Glycanases Associated with Cell Walls of Cicer arietinum L: Arabinogalactan Degradation

The proteins extracted from Cicer arietinum cell walls showedendo-glycanase activity against the water-soluble hemicellulosicpolysaccharides extracted with 4% KOH. Such endo-glycanase activitywas able to shift the molecular mass distribution of the arabinogalactancausing a decrease in its average molecular mass, as well asto release small oligosaccharides constituted exclusively ofarabinose. Exo-glycanase activities such as glucosidase andgalactosidase were also present. Key words: Arabinogalactan, cell wall, Cicer arietinum, glycanase, hemicellulose     

Involvement of Water Channels in the Hydration-Induced Calcium Release in Nitella flexilis

An internodal cell of Nitella flexilis treated with 10 mM KC1was vacuole-perfused with an isotonic solution containing ethyleneglycol-bis-(rß-aminoethylether)N,N,N,N-tetraaceticacid (EGTA) and its content including cytoplasm was squeezedout into a vessel and covered with silicone oil. When the hypotonicsolution was added into the cytoplasmic drop which had beenmixed with aequorin, a significant increase in the light emissionfrom aequorin was detected with the photomultiplier, indicatinga release of Ca²⁺ from some cell organdies storing Ca²⁺. Thisincrease in the light emission was strongly inhibited by treatingcells with 0.1 mM HgCl₂ which is known to inhibit water channelsin the plasma membrane. The inhibition was completely recoveredby washing HgCl₂ with 2-mer-captoethanol. This suggests thatwater channels may exist in the membrane of Ca²⁺ stores andplay an essential role in the hydration-induced Ca²⁺ release. (Received February 12, 1998; Accepted May 21, 1998)     

Distribution of Calanus species in Kongsfjorden, a glacial fjord in Svalbard

The distribution of Calanus species was investigated in Kongsfjordenin summer of 1996 and 1997. In both years Calanus finmarchicusand Calanus glacialis dominated, although the boreal C. finmarchicuswas more abundant than the Arctic C. glacialis in 1997. Thiscoincided with a 2°C higher water temperature at 50 m in1997, indicating stronger influence of Atlantic origin waterthat year. Advected Calanus finmarchicus occurred in deep andsubsurface layers of the outer fjord in 1996 (200 ind. m^-3,mainly CIII). A less abundant local population aggregated insurface layers of the inner fjord (100 ind. m^-3). Similarly,advected C. finmarchicus occurred in subsurface layers in 1997(446 ind. m^-3, mainly CIII and CIV) and a local population insurface layers (183 ind. m^-3, mainly CI). Calanus glacialisin 1996 aggregated as CII and CIII in the deep layers of theouter fjord (272 ind. m^-3), whereas CIII–CV were abundant(216 ind. m^-3) in cold surface waters of the inner fjord. In1997 C. glacialis (mostly CIII–CV) was more abundant inthe outer than in the inner part of the fjord (40 and 192 ind.m^-3, respectively). Within Kongsfjorden, Calanus finmarchicusneeds one year to complete its life cycle, whereas Calanus glacialisneeds two. Calanus hyperboreus seems to be an expatriate inthe fjord system.     

Effect of Brefeldin A on the synthesis and transport of cell wall polysaccharides and proteins in pea root seedlings

The in vivo effect of Brefeldin A (BFA) on the synthesis andtransport of cell wall polysaccharides and proteins in the rootsof pea seedlings (Pisum sativum L. cv. Alaska) was investigated.BFA (10µgml^—1 inhibited the synthesis of cell wallmatrix polysaccharides by approximately 43%. Under the sameconditions, cellulose synthesis was inhibited by approximately77%. The percentage of incorporation of L—[U—¹⁴C]leucineand L-[U-14C]proline into cytosolic, membrane and cell wallproteins was only slightly changed in the presence of BFA. Inaddition, the drug did not change the pattern of newly synthesizedproteins in the three fractions as judged by SDS—PAGEfluorography. Double labelling of proteins and cell wall polysaccharidesconfirmed the above reported data. All these results showedthat the synthesis and transport of proteins to the cell wallwas only slightly affected by BFA under similar conditions tothose which brought about a strong inhibition of the synthesisof matrix and cellulosic polysaccharides. BFA had no effecton the activity of membrane-bound and digitonin-solubilizedmannan and glucomannan synthase isolated from the third internodeof pea seedlings. This would exclude an effect of BFA at thelevel of the catalytic site of the synthases. The inhibitionof polysaccharide synthesis by the drug was rapidly eliminatedafter its removal. It is concluded that the effect of BFA onthe biosynthesis of cell wall polysaccharides could be causedby an interaction of the drug with the topological organizationof the synthase complexes in the membranes. This effect wouldprecede the action of the drug at the level of vesicle transportto the walls. Key words: Brefeldin A, cell wall polysaccharides (synthesis and transport), Pisum sativum L, polysaccharide synthases, proteins (synthesis and transport)     

Effect of EGTA on Nyctinastic Closure Mediated by Phytochrome in Albizzia lophantha

The role of extracellular calcium on nyctinastic closure ofAlbizzia lophantha leaflets has been studied by testing theeffect of ethyleneglycol-bis-(ß-aminoethylether)-N,N,N',N'-tetraaceticacid (EGTA) and its reversibility by calcium. EGTA (1 and 5mM) causes an inhibition of nyctinastic closure and at a concentrationof 1 mM EGTA it decreases the difference between the effectof red light (R) and far-red light (FR) irradiation on leafletclosure. A simultaneous or subsequent supply of CaCl₂ (5 or10 mM) reverses EGTA (5 mM) inhibition on closure as well ascausing an additional promotion of closure. We suggest that external calcium could play a dual role in nyctinasticclosure. Phytochrome control of leaflet closure probably needsexternal Ca²⁺ and, in addition, Ca²⁺ could regulate the closuremechanism by controlling ionic fluxes through the plasma membranein pulvinular motor cells. (Received June 9, 1989; Accepted November 27, 1989)     

Specific Labeling of Cell Wall Polysaccharides with myo-[2-3H] Inositol during Germination and Growth of Phaseolus vulgaris L.

myo-[2-³H]Inositol was fed to bean seeds by imbibition and itsmetabolic fate was studied during germination and seedling growth.The largest amount of myo-inositol was taken up from a 500 HIMsupply (8 mg/seed) and the highest percentage was from 1 HIM(29%). myo-Inositol was incorporated to new cell wall polysaccharidesof hypocotyl and roots, mostly as uronic acid and pentose residues.In the 80% ethanolinsoluble cell walls of hypocotyls at 3, 4and 5 days after imbibition, 47 to 52% of ³H was detected asuronic acids, 20 to 24% as arabinose and 11 to 19% as xylose.Glucogenesis from myo-inositol was low: less than 6% was recoveredas hexoses. The ³H in uronic acid and arabinose residues decreasedwith increasing age (i.e. 0 to 6 cm from cotyledons) and increasedin older segments (further than 6 cm from cotyledons). In theoldest segment of 5-day-old hypocotyl (> 10 cm), ³H in thesugar residues was more than that in the youngest part (0–2cm). On the other hand, ³H in xylose residues increased steadilyin the older part, but did not exceed that in arabinose. The results show that the myo-inositol oxidation pathway functionsin growing hypocotyls and roots of bean seedlings to provideexclusively uronic acid and pentose units for cell wall synthesis.Results also show that incorporation of arabinose and uronicacids derived from myo-[2-³H]inositol to cell wall polysaccharidesis active in two regions of the hypocotyl; first, for the constructionof the primary walls in the young, growing region of the hypocotyl,and second, for thickening of the walls after completion ofelongation growth. ¹Supported by NSERC of Canada. (Received April 10, 1984; Accepted June 12, 1984)     

Effect of Ammonia on Cellular pH of Anabaena cylindrica Determined with 31P NMR Spectroscopy

The pH changes in the blue-green alga (cyanobacterium) Anabaenacylindrica caused by addition of ammonia were investigated using³¹P NMR spectroscopy. A pH shift of 0.9 or more was observedwhen 30 nM NH₄OH was added to the cell suspension, but no significantcellular pH change was observed with 50 mM NH₄CI, a concentrationhigh enough to stimulate dark CO₂ fixation of this alga. Thechange in cellular pH does not seem to cause ammonia-inducedstimulation of dark CO₂ fixation. (Received June 22, 1985; Accepted January 10, 1986)     

Role of extracellular [Ca2+] in fatigue of isolated mammalian skeletal muscle

The possiblerole of altered extracellular Ca²⁺concentration([Ca²⁺]_o)in skeletal muscle fatigue was tested on isolated slow-twitch soleusand fast-twitch extensor digitorum longus muscles of the mouse. Thefollowing findings were made. 1) Achange from the control solution (1.3 mM[Ca²⁺]_o)to 10 mM[Ca²⁺]_o,or to nominally Ca²⁺-freesolutions, had little effect on tetanic force in nonfatigued muscle.2) Almost complete restoration oftetanic force was induced by 10 mM[Ca²⁺]_oin severely K⁺-depressed muscle(extracellular K⁺ concentration of10-12 mM). This effect was attributed to a 5-mV reversal of theK⁺-induced depolarization andsubsequent restoration of ability to generate action potentials(inferred by using the twitch force-stimulation strength relationship).3) Tetanic force depressed bylowered extracellular Na⁺concentration (40 mM) was further reduced with 10 mM[Ca²⁺]_o.4) Tetanic force loss at elevatedextracellular K⁺ concentration (8 mM) and lowered extracellular Na⁺concentration (100 mM) was partially reversed with 10 mM[Ca²⁺]_oor markedly exacerbated with low[Ca²⁺]_o.5) Fatigue induced by using repeatedtetani in soleus was attenuated at 10 mM[Ca²⁺]_o(due to increased resting and evoked forces) and exacerbated at low[Ca²⁺]_o.These combined results suggest, first, that raised[Ca²⁺]_oprotects against fatigue rather than inducing it and, second, that aconsiderable depletion of[Ca²⁺]_oin the transverse tubules may contribute to fatigue.

     

Ionic Relations of Garden Orache, A triplex hortensis L.: Growth and Ion Distribution at Moderate Salinity and the Function of Bladder Hairs

The growth of garden orache, A triplex hortensis was studiedunder conditions of mild NaCl or Na₂SO₄ salinity. Growth, drymatter production and leaf size were substantially stimulatedat 10 mM and 50 mM Na⁺ salts. Increased growth, however, appearedto be due to a K⁺-sparing effect of Na⁺ rather than to salinityper se. The distribution of K⁺ and Na⁺ in the plant revealeda remarkable preference for K+ in the roots and the hypocotyl.In the shoot the K/Na ratio decreased strongly with leaf age.However, the inverse changes in K⁺ and Na⁺ content with leafage were dependent on the presence of bladder hairs, which removedalmost all of the Na⁺ from the young leaf lamina. Measurementsof net fluxes of K⁺ and Na⁺ into roots and shoots of growingAtriplex plants showed a higher K/Na selectivity of the netion flux to the root compared to the shoot. With increasingsalinity the selectivity ratio S_{K, Na}^* of net ion fluxes tothe roots and to the shoots was increased. The data suggestthat recirculation of K⁺ from leaves to roots is an importantlink in establishing the K/Na selectivity in A. hortensis plants.The importance of K⁺ recirculation and phloem transport forsalt tolerance is discussed. Key words: Atriplex hortensis, Salinity, Potassium, Sodium, K⁺ retranslocation, Bladder hairs, Growth stimulation