Involvement of Cell Wall-Bound Diferulic Acid in Light-Induced Decrease in Growth Rate and Cell Wall Extensibility of Oryza Coleoptiles

Irradiation of white fluorescent light (5 W m^–2) inhibitedthe growth of Oryza coleoptiles. Light irradiation increasedstress-relaxation parameters of coleoptile cell walls, minimumstressrelaxationtime and relaxation rate, and decreased cellwall extensibility (strain/load). Under light conditions, thecontents of ferulic and diferulic acids ester-linked to thehemicellulosic arabinose residue in cell walls increased andcorrelated with the modification of the cell wall mechanicalproperties. These results suggest that light irradiation enhancesthe formation of diferulic acid bridges in hemicelluloses, makingcell walls mechanically rigid and thus inhibits cell elongationin rice coleoptiles. Also, irrespective of coleoptile age orthe presence of light, the ratio of diferulic acid to ferulicacid was almost constant, suggesting that the rate limitingstep in the formation of diferulic acid bridges in Oryza cellwalls is in the step of feruloylation. (Received September 24, 1991; Accepted December 3, 1991)     

Using Viscoelastic Properties of the Woody Tissue from Tobacco Plants (Nicotiana tabacum) to Comment on the Molecular Structure of Cell Walls

Lignin in the cell walls of woody tissue has a much lower crosslinkdensity in tobacco wood than in tree wood. This causes tobaccowood to show very different viscoelastic behaviour. With theaid of genetically modified plants, it is shown that the ligninin tobacco plant cell walls behaves in much the same way asa polymer solution. It exhibits both rate stiffening and ratethinning behaviour due to the entangled nature of the ligninnetworks. The hydrophobic portions of lignin have a very lowpolymer chain crosslink density, hence entanglements make asignificant contribution.Copyright 1998 Annals of Botany Company Tobacco plants, lignin, polymer solution, shear thinning, stress relaxation.     

Ion-Exchange Properties of Isolated Cell Walls of Brown Algae: The Interstitial Solution

Kloareg, B., Demarty, M. and Mabeau, S. 1987. Ion-exchange propertiesof isolated cell walls of brown algae: the interstitial solution.—J.exp. Bot. 38: 1652–1662. Isolated cell walls of Pelvetia canaliculata (Dene) et Thur.,Laminaria digitata (L.) and other intertidal brown algae wereequilibrated in seawater and various mixtures of sodium andcalcium chlorides. After elution with distilled water, the interstitialsolution was assayed for ionic composition. With respect tothe external solution, it was less concentrated and containedlower proportions of divalent cations. Such modifications wererelated to the divalent ionic fraction and the total anion concentrationof the external solution. No significant differences were foundamong the brown algae investigated in spite of the differingcomposition of their walls. Results were analysed accordingto the polyelectrolyte condensation model. When the concentrationof the external solution was high (1000 meq dm³), experimentaldata were qualitatively in good agreement with the predictionsof the model. Key words: Cell walls, ion-exchange, Phaeophyta.     

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)     

Orientation of Microtubules during Regeneration of Cell Wall in Selected Giant Marine Algae

The microtubules in highly synchronized aplanospores of twogiant marine algae, Boergesenia forbesii and Valonia ventricosa,were examined by immunofluorescence microscopy throughout theregeneration of the cell wall. Microtubule orientation was alwaysrandom up to 20 h after wounding, although the orientation ofcellulose microfibrils changed from random to parallel withinthat time period. When the rhizoid cells were in the stage ofelongation at 7 to 10 days after wounding, highly ordered microtubuleswere always observed along the longitudinal cell axis exceptat the very tip of the cells where random ones were found. Incontrast, the microfibrils in the innermost lamellae of newlysynthesized cell walls showed three different orientations,that is, transverse, longitudinal and oblique to the longitudinalcell axis. These observations suggest that microtubules maycontrol cell shape, but not the orientation of microfibrils.The mechanism of cell wall construction in these algae is discussedin relation to the self-assembly mechanism thought to operatein the construction of helicoidal cell walls. ³ Present address: Polymer Research Laboratory, Mitsui ToatsuChemicals, Inc., Yokohama, Kanagawa 244, Japan. (Received November 18, 1987; Accepted April 11, 1988)     

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     

Food selection and growth of the planktonic ciliate Coleps hirtus isolated from a monomictic subtropical lake

A strain of Coleps hirtus (Ciliophora, Prorodontida) was isolatedfrom the epilimnion of monomictic Lake Kinneret. Growth of thisciliate was tested in response to 12 species of planktonic algaeand seven species of cultured bacteria from lake isolates whichwere offered as food. Eight species of algae (one Cryptophyceaeand seven Chlorophyceae) and four bacteria supported good toexcellent growth of C.hirtus. Growth rates (µ) and doublingtimes (DT) ranged from 0.008 to 0.029 h^–1 and from 23.9to 90.8 h respectively. C.hirtus was able to grow on bacteriaat concentration levels as low as 2–8 x 10⁵ cells ml^–1.No correlation was observed between growth rate of C.hirtusand cell volume of the prey. ^aPresent address: Istituto di Ecologia, Universita di Parma,43100 Parma, Italy     

Ammonium release by zooplankton in suspensions of heat-killed algae and an evaluation of the flow-cell method

The maximum excretion rate of NH₄ (39 nmol mg dry wt^–1h^–1) was directly measured for Daphnia pulex by measuringNH₄ accumulation in bottles containing D. pulex and dense, satiatingsuspensions of heat-killed algae. Ammonium release rates inthe algal suspensions were compared to those of individual animalsremoved from the suspension and placed in flow cells. Ammoniumrelease rate, R (nmol mg dry wt^–1 h^–1). in the flowcell decreased very rapidly with time, t (min), after removalaccording to the relation R = 26 + 25e^–0.16t. Ammoniumexcretion obtained by the flow cell method after extrapolationto time zero was not significantly different from that obtainedin the bottles. The considerable experiment-to-experiment variationin NH₄ excretion was in large part correlated (r² = 0.73) withthe feeding rate on the algae.     

Effect of temperature on growth and ingestion rates of Favella sp

This study describes the effect of temperature on the growthand ingestion rates of the tintinnid, Favella sp. cultured withthe dinoflagellate Heterocapsa triquetra. In vivo fluorescencewas used to monitor the change in density of the H. triquetrapopulation over 4- to 5-day periods in control tubes containingonly algae, and in experimental tubes containing algae and tintinnids.A ‘switchover point’ occurred in the temperaturedependency of the growth rate such that below 11.4°C, H.triquetra grew more quickly than Favella sp. and above thistemperature the situation was reversed. Ingestion rates of Favellaon H. triquetra were found to be temperature dependent in anonlinear fashion. The rate doubled (from 2.5 to 5.3 cells animal^–1h^–1) between 11.4 and 16.4°C whereas there was nochange in ingestion rates between 8.0 and 11.4°C, or between16.4 and 21.1°C.     

Physiological evaluation of temperature effect on the growth processes of the mysid, Neomysis intermedia Czerniawsky

Ingestion, respiration, and molting loss rates were measuredover the 3 – 29°C range in Neomysis intermedia. Weightspecific rates of these physiological processes ranged from2 to 140% body C day^–1 for ingestion, from 2 to 15% bodyC day^–1 for respiration, and from 0.1 to 5% body C day^–1for molting loss. All weight-specific rates showed a logarithmicdecrease with a logarithmic increase in body weight, and a logarithmicincrease with a linear increase in temperature below 20 or 25°C.The effect of temperature, however, was different between thephysiological rates, with a large temperature dependency foringestion (Q₁₀ = 2.6 –3.9) and molting loss (Q¹⁰ = 2.9– 3.6) and a moderate temperature dependency for respiration(Q₁₀ = 1.9 – 2.1). Calculated assimilation efficiencychanged with body size, but was constant over the temperaturerange examined. Allocation of assimilated materials varied witha change in temperature, reflecting the different temperaturedependence between physiological processes. It was deduced thatthe strong temperature dependency of the growth rate in N. intermediaobserved in the previous studies resulted from the large temperatureeffect on ingestion and assimilation rates, superimposed bythe different allocation of assimilated materials. ¹Present address: Department of Botany, University of Tokyo,Hongo, Tokyo 113, Japan     

In situ determinations of bacterial selectivity and filtration rates by five cladoceran zooplankters in a hypertrophic subtropical reservoir

Cladoceran in situ feeding rates on natural bacteria labelledwith [methyl-³H] were studied in parallel with feeding ratedeterminations on ¹⁴C-labelled Chlorella in a hypertrophic subtropicalreservoir (Lake Hartbeespoort) through spring and summer (1986/87).Community filtration rates (CFR5) on bacteria and algae weresimilar, but selection for Chlorella (relative to natural bacteria)increased from midsummer in association with declining bacterialdensity and increasing dominance of ‘inedible’ componentsof the natural phytoplankton. Species-specific filtration rates(SSFRs) were determined for Daphnia pulexllongispina, Ceriodaphniareticulata, Diaphanosoma excisum, Bosmina longirostris and Moinamicrura during their respective seasonal occurrence in the studyperiod. SSFRs on algae and bacteria increased with body length(L, mm) in all species apart from Bosmina. Species-specificdifferences in absolute feeding rate (FR, ml animal^–1day^–1), the slope of the FR-L relationship and bacteriaselectivity were evident. The feeding rate of all cladoceranson bacteria is described by the power equation FR 5.231L^1.42FR values on bacteria relative to FR values on algae averaged     

Thermal Analysis of Membrane Lipids from the Blue-Green Algae Anacystis nidulans and Anabaena variabilis

Differential scanning calorimetry was used on aqueous dispersionsof lipids extracted from the blue-green algae Anacystis nidulansand Anabaena variabilis. The reversible curves, exothermic oncooling and endothermic on heating, indicated that thermotrophicphase transition of the membrane lipids took place over a widerange of temperatures. These results suggest that the lipidsof the thylakoid membranes in these algae were in the liquidcrystalline state at the growth temperature, entered the phaseseparation state at about room temperature and went into thegel state below the freezing point. The temperature for theonset of phase separation was dependent on both the growth temperatureand the growth stage of the culture. ³Present address: Solar Energy Research Group, Institute ofPhysical and Chemical Research (RIKEN), Wako-shi, Saitama 351,Japan. (Received December 20, 1982; Accepted March 24, 1983)     

Lipid Metabolism in the Brown Marine Algae Fucus vesiculosus and Ascophyllum nodosum

Lipid metabolism and environmental effects on this process havebeen studied in the marine brown algae Fucus vesiculosus andAscophyllum nodosum. These algae showed very similar patternsof lipid metabolism during 24 h incubations. Labelling from[1-¹⁴C]acetate showed the major labelled lipids to be the ß-alanineether lipid and the neutral lipid fraction in both algae. Ofthe glycolipids, only sulphoquinovosyldiacylglycerol was welllabelled and the phosphoglycerides were all poorly labelled.The major labelled fatty acids were palmitate and oleate, againin both algae, although Fucus vesiculosus also showed significantlabelling of stearate and behenate. Although the amount of fattyacid labelling increased with time, the proportion of labelin palmitate and oleate remained approximately constant. Verylong chain fatty acids (arachidic, behenic) were increasinglylabelled with time. Lowered incubation temperatures decreased labelling of the saturatedfatty acids. Cu²⁺ increased the proportion of oleate labelledin both algae, and of linoleate in Fucus vesiculosus. This cationdecreased the percentage labelling of stearate and myristatein Ascophyllum nodosum. Lipid metabolism in Ascophyllum nodosumwas more sensitive to raised Cu²⁺ levels than in Fucus vesiculosus Key words: Acyl lipid metabolism, Fucus vesiculosus, temperature effects, Ascophyllum nodosum, copper pollution     

A salt-sensitive Dunaliella mutant I. Growth and osmoregulatory responses to increases in salt concentration

Dunaliella is a genus of green unicellular algae distributedin all the oceans and saline bodies of water throughout theworld and distinguished by unusual tolerance to salt. Sincethe cells of this genus do not possess a rigid cell-wall, theyrespond to changes in salt concentration by rapid alterationsin cell volume and then return to their original volume as aresult of adjustments in the amounts of intracellular ions andglycerol, this latter being the major organic osmoticum. Thepaper describes the behaviour of a mutant of D. parva 19/9 withreduced capabilities of growth above 0.5 kmol m^–3 NaCl.The mutant is unusual in that its abilities to synthesize glyceroland pump out Na⁺ and Cl^– do not appear to be impaired;volume changes in the hyperosmotic range also appear to be roughlythe same as in D. parva. The average cell volume of mutant cellsis reduced (206µm³ as opposed to 255 µm³ in D. parva)and their rate of change of cell volume after an increase insalt concentration is lower; it took about 10 min for mutantcells in the light to reach a new cell volume whereas D. parvacells reached their new volume in less than 1 min. Both factorsmay be dependent on components of the cytoskeleton. The mutantthrows light on adaptations necessary to allow Dunaliella cellsto grow at high salt concentrations and demonstrates that halotoleranceincludes, but is not equivalent to, osmoregulation. Key words: Dunaliella, salt tolerance, mutant     

A Cryomicroscopic Study of Cylindrocystis brebissonii De Bary and Two Species of Micrasterias Ralfs (Conjugatophyceae, Chiorophyta) during Freezing and Thawing

The changes in morphology of the unicellular algae Cylindrocystisbrebissonii and two species of Micrasterias during freezingand thawing were observed on a light microscope fitted witha temperature controlled stage. At slow rates of cooling extensiveshrinkage of the protoplast was observed. The response of thecell wall varied with cell-type. In C. brebissonii plasmolysiswas not observed and the cell wall and protoplast shrank together.In Micrasterias the cell wall did not contract and a distinctplasmolysis was observed. Following freezing to and thawingfrom –25?C cells of C. brebissonii were non-viable butremained osmotically responsive. Cooling at faster rates inducedintracellular ice formation in all cell-types. The criticalrate of cooling varied with cell-type and was determined bycell volume and suface area. Intracellular gas bubbles wereobserved during thawing following both rapid and slow cooling. Following cooling in dimethylsulphoxide cells of C. brebissoniiwere protected against freezing injury. The recovery on thawingfrom –196?C being determined by the rate of cooling, anoptimum rate of 1?C min^–1 was observed. During slow ratesof cooling (<2?C min^–1) cells remained unshrunken,at faster rates (10?C min^–1) the loss of cell viabilitywas related to osmotic shrinkage during cooling rather thanto nucleation of intracellular ice. Intracellular ice formationwas observed only following significantly faster rates of cooling(>20?C min^–1). Key words: Cylindrocystis, Micrasterias, cryomicroscopy, freezing injury     

Feeding of freshwater filter-feeders at very low food concentrations: Poor evidence for "threshold feeding" and "optimal foraging" in Daphnia longispina and Eudiaptomus gracilis

The feeding behaviour of two freshwater zooplankton species,the calanoid copepod Eudiaptomus gracilis and the cladoceranDaphnia longispina, at extremely low food concentrations hasbeen studied in order to test whether the animals react as predictedby the models of optimal foraging. Three different sized algae were offered in concentrations downto 1µg C/litre at 7°C during winter and 19°C duringsummer. Ingestion rates were measured by use of a flow-through¹⁴C-technique In contrast to the findings of several authors working withmarine copepods a threshold concentration where the animalsceased filtering could be detected only in one experiment byregression analysis (Daphnia, 19°C, food: Stichococcus).A depression of the filtering rate at low concentrations wasobserved only in Daphnia at 19°C with Stichococcus and Scenedesmusas food. It is supposed that in this case the reduction of thefiltering rate is not a direct response of the filtrator, butis caused by exhaustion due to the experimental conditions. As freshwater zooplankton is very rarely exposed to food abundancesas low as in the marine environment, there seems to be no selectionpressure favouring the evolution of an "optimal forager" behaviour. ⁽¹⁾Supported by Deutsche Forschungsgemeinschaft     

Feeding behaviour of the rotifer Ascomorpha ovalis: functional response, handling time and exploitation of individual Ceratium cells

The planktonic rotifer Ascomorpha ovalis feeds on large dinoflagellates(e.g. Ceratium sp., Peridinium sp.) and is able to extract theircell contents by means of its virgate mastax. This paper presentsthe results of experiments on the feeding behaviour of laboratory-culturedAscomorpha with Cerarium furcoides as food algae. Ascomorphaare three times larger than their prey Ceratium (by volume),but with regard to total length, their prey was even 20% larger.Ascomorpha showed a hyperbolic functional response curve witha plateau of the feeding rate at 8 Ceratium cells animal^–1dar^–1 when concentrations of Ceratium were >100 cellsml^–1. The mean handling time (time for capturing and extractingone Ceratium cell) was 3 min. The shape of the functional responsewas better described by a curvilinear model than by a rectilinearmodel. However, handling times cannot be responsible for this,since they were too short to set limits on ingestion rates.At low food concentrations, encounter rates with prey seemedto limit the feeding rates of Ascomorpha, whereas at mediumto high food concentrations, Satiation effects (lower attackrates) seemed to set limits on the feeding rates. Ascomorphashowed a significant decrease in the exploitation of singleCeratium cells at high prey concentrations. This decrease couldbe explained by a saturation effect in which the partly filledguts of Ascomorpha did not permit the total extraction of thecontents of a Ceratium cell.     

Light, temperature and nitrogen as interacting factors affecting diel vertical migrations of dinoflagellates in culture

Diel vertical migrations of the marine dinoflagellates Gonyaulaxpolyedra Stein and Ceratium furca (Ehr.) Clap, et Lachm. werefollowed in a laboratory tube (2.02 m x 0.25 m) under a 12:12hlight:dark cycle. The effects of temperature stratification,two levels of surface irradiance and nitrogen depletion on patternsof vertical migrations were examined. At temperatures between22–26°C with small temperature gradients, both speciesmigrated at a rate of 0.7 –1.0 m h^–1. Steeper thermoclines(ca. 0.8°C 0.1 m^–1) with temperatures below ca. 20°Ccaused a marked decrease in swimming speed which resulted inaccumulations of cells in these thermocline regions. Under conditionsof nutrient sufficiency both algae migrated into the surfacelayers at irradiance values of over 1000 µE m^–2s^–1. Increasing nitrogen depletion caused the downwardmigration of both algae to commence progressively earlier inthe day and before the end of the light period. The earlierdownward migrations enabled a more complete descent throughthe thermocline. Nitrogen depleted cells of Gonyaulax continuedto undertake vertical migrations but avoided high irradiancesthus forming subsurface maxima at irradiance levels close to150 µE m^–2 s^–1. Ceratium cells which exhaustedboth inorganic nitrogen and phosphorus ceased to migrate accompaniedby a large change in cellular fluorescence.     

Amino Acid-Sugar Linkages in Rice Coleoptile Cell Walls

The nature of amino acid-sugar linkages in cell walls was investigatedin a monocotyledonous tissue, rice coleoptiles. The molar ratiosof aspartic acid, threonine, and serine in cell walls were decreasedby hydrazinolysis in coleoptiles grown both on and under water.The molar ratios of threonine and serine were decreased alsoby a NaOHNaBH₄ treatment, while the alanine content was increased,and -aminobutyric acid was not formed. The cell walls were treated with NaOH in the presence of NaB³H₄,hydrolyzed, then divided into amino acid and sugar fractions.Two distinct radioactive peaks were detected in the thin-layerchromatography of the amino acid fractions. One was identifiedas alanine derived from glycosylated serine; the other was confirmedto be an oxidation product of glucosaminitol. There was justone ³H-labeled product in the sugar fractions, galactitol. Theseresults suggest the presence of serine-O-galactose and asparagine-N-N-acetylglucosamine linkages in rice coleoptile cell walls. The existence of glucosamine linked to amino acids was furthersupported by the incorporation of ¹⁴C-glucosamine into cellwalls. These linkages were also detected in the cell walls ofa dicotyledonous tissue, Vicia epicotyls. (Received April 2, 1981; Accepted June 24, 1981)     

Two Novel Endopeptidases Released into the Medium during Mating of Gametes of Chlamydomonas reinhardtii

During the mating reaction between mt⁺ and mt^- gametes of Chlamydomonasreinhardtii, two novel endopeptidases, each of which was ableto digest the B chain of insulin, were released into the culturemedium, together with a gamete lytic enzyme (GLE) which is responsiblefor digestion of the gametic cell walls. Both endopeptidasesand GLE were copurified from the mating medium by column chromatographyon DEAE-cellulose and concanavalin A. Gel filtration separatedthe peptidases, which were unable to digest gametic cell walls,into two fractions, endopeptidase-1 and endopeptidase-2. Theseenzymes were also separated from GLE, which was unable to digestthe B chain of insulin. Endopeptidase-1, with a molecular massof about 200 kDa, cleaved the B chain of insulin at the Ala¹⁴-Leu¹⁵peptide bond, and this activity was inhibited by EDTA. Endopeptidase-2,with a molecular mass of about 110 kDa, digested the peptideat the Leu¹⁵-Tyr¹⁶ peptide bond and was sensitive to iodoacetateand chymostatin. When the cell walls of gametes of either mating-typewere digested prior to mating with exogenously added GLE, thetwo endopeptidases were released into the medium, a result thatsuggests that they are stored, like GLE, outside the plasmalemma. (Received March 25, 1994; Accepted June 13, 1994)