The Effects of Salinity upon Galactosyl-Glycerol Content and Concentration of the Marine Red Alga Porphyra purpurea (Roth) C.Ag.

Changes in the major alcohol-soluble, low molecular weight carbohydratesof P. purpurea, O--D-galactopyranosyl-(1-2)-glycerol (‘floridoside’)and O--D-galactopyranosyl-(1-1)-glycerol (‘isofloridoside’),have been examined in response to salinity variation. ‘Floridoside’is shown to vary in absolute amount, increasing in hypersalineand decreasing in hyposaline media. ‘Isofloridoside’content per cell does not change in a similar manner. Responsesare almost identical under light or dark conditions, ‘floridoside’changes being complete within 24 h. Decreasing the externalwater potential using ionic and non-ionic solutes has the sameeffect upon galactosyl-glycerol content. The amount of ‘floridoside’synthesized, and degraded under hypersaline and hyposaline conditionsrespectively is shown to be insufficient to restore cell volumeto its original value. It is therefore suggested that the primaryfunction of ‘floridoside’ increases in concentratedsea-waters is that of a compatible solute, serving to protectthe cell during periods when the external salt content is increaseddramatically.     

The role of hypersalinity in the persistence of the Texas 'brown tide' in the Laguna Madre

A brown tide bloom of the alga Aureoumbra lagunensis was presentwithout interruption in the Laguna Madre of Texas from January1990 through October 1997. This is the longest continual phytoplanktonbloom of which we are aware. Although the factors leading tothe initiation of this bloom have been well documented, thefactors contributing to its persistence are still being investigated.Two physical characteristics of the Laguna Madre may play animportant role: the long turnover time for waters in this coastallagoon (–1 year) and the hypersaline conditions that usuallyexist (40–60 PSU) due to evaporation exceeding precipitation.In this study, we examined the effects of salinity on the growthrates of the brown tide alga and on the growth of one of itsprotozoan grazers. Historical data from before the onset ofthe brown tide provide evidence for the suppression of microzooplanktonpopulations and mesozooplankton growth caused by hypersalinity.The brown tide alga will grow in a remarkably wide range ofsalinities ranging from 10 to 90 PSU. Maximum growth rates areachieved at salinities ranging from 20 to 60 PSU. One commongrazer on the brown tide alga, the heterotrophic dinoftagellateOxyrrhis marina, was found to grow more slowly under hypersalineconditions. The normally hypersaline conditions of the LagunaMadre may, therefore, favor the brown tide alga over other phytoplanktonspecies that do not grow well under hypersaline conditions,and also suppress the growth and feeding rates of potentialgrazers.     

Ecotypic Variation in the Osmotic Responses of Enteromorpha intestinalis (L.) Link.

Young, A. J., Collins, J. C. and Russell, G. 1987. Ecotypicvariation in the osmotic responses of Enteromorpha intestinalis(L.) Link.—J. exp. Bot. 38: 1309–1324. The physiological basis for salt tolerance has been studiedin the euryhaline marine alga Enteromorpha intestinalis. Adaptationto dilute and concentrated seawaters has been investigated inthree separate populations of this alga: marine, rock pool andestuarine. Internal K⁺, Na⁺ and Cl^– levels have been determined usingtracer efflux analyses. K⁺ has been shown to be the major osmoticsolute within this alga. Cellular levels of Cl^– and, inparticular, Na⁺ are low although levels in the cell wall arehigh. Levels of these ions varied considerably between the separateplants; K⁺ levels within marine plants of E. intestinalis aretwo to four times those found in the other populations. Thetertiary sulphonium compound ß-dimethylsulphonio-propionateis maintained at relatively high levels, although it remainsfairly insensitive to change in the external salinity. Changes in the tissue water content and cell volume are large,particularly within the estuarine plants. The thin cell wallsof these plants allow large changes in volume in the diluteconditions experienced in an estuary, while low turgor preventscell rupture. Thicker cell walls and small cells of the marineand rock pool plants assist in tolerating high and low externalosmotic potential—the estuarine plants respond poorlyto concentrated seawater. Key words: Enteromorpha, osmoregulation, ecotypes     

Solute Regulation in the Euryhaline Marine Alga Enteromorpha prolifera (O. F. Mull) J. Ag.

Young, A. J., Collins, J. C. and Russell, G. 1987. Solute regulationin the euryhaline marine alga Enteromorpha prolifera (O. F.Mll) J. Ag.—J. exp. Bot. 38: 1298–1308. The physiological basis for salt tolerance has been studiedin the euryhaline alga Enteromorpha prolifera. Levels of inorganicions and organic (compatible) solutes have been measured. K⁺makes the major contribution towards the internal osmotic potentialof the cell, while Cl^– and, in particular, Na⁺ contentsare low. Levels of the organic solute ß-dimethylsulphonio-propionate(DMSP) are high but are fairly insensitive to changes in theexternal salinity. Levels of amino-acids, calcium, phosphateand sulphate contribute relatively little towards the internalosmotic potential of the alga. As salinity is altered there are marked changes in the tissuewater content and volume. These changes directly affect theconcentration of the osmotic solutes within the cell. In diluteseawaters there is an increase in turgor as there is littlechange in the internal solute content of the cell compared tovalues in normal sea water. Inorganic ions, in particular K⁺,and organic solutes are accumulated in concentrated seawaters,although concentrations greater than 2·00 x seawaterresult in a reduction in the internal osmotic potential of thecell, mainly through loss of K⁺. Key words: Enteromorpha, salinity, osmoregulation     

Isolation of Protoplasts from Kappaphycus alvarezii var. tambalang (Rhodophyta) and Secretion of i-Carrageenan Fragments by Cultured Cells

A method for generating protoplasts from the carrageenan-producingred alga Kappaphycus alvarezii was developed. Digestions withcellulase and _k-carrageenase produced only a few cortical cellprotoplasts, while digestions with cellulase and i-carrageenaseonly produced epidermal cell protoplasts. When both carrageenaseswere used in the digestion media with cellulase, protoplastswere released from all cell types and yields ranged from 1·0to 1·2x10⁷ cells g^–1 with sizes from 5 to 200 µmdiameter. Protoplasts were subsequently cultured to study cellwall regeneration. Calcofluor-positive material (probably cellulose)was detected within 6 h after removal of protoplasts from thewall digestion media, whereas, i-carrageenan fragments weredetected in all regenerating protoplast cultures 24 h afterremoval from the digestion media. Protoplasts continued to produceCalcofluorpositive material and secrete carrageenan fragmentsinto culture media for several days. However, cells culturedin media augmented with K⁺ ions stopped secreting carrageenanfragments after 24 h. Cells cultured for 48 h in seawater labelledweakly with an i-carrageenan hybridization probe, but not atall with a corresponding _k-probe. Cells cultured for 48 h, blottedto nylon membranes and probed with anti-carrageenan monoclonalantibodies, showed the presence of gelling carrageenan subunitsin the cell walls. Key words: -Carrageenan, Kappaphycus, protoplasts, Rhodophyta     

Dunaliella salina: A model System for Studying the Response of Plant Cells to Stress

This review focuses on the biochemical and physiological responseof the halotolerant green alga, Dunaliella salina, to conditionsof stress. It is now well established that in response to stress,cells of Dunaliella salina var. bardawil show increased glycerolproduction, massive ßcarotene accumulation and enhancedabscisic acid metabolism. In this respect, cellular responsesare regulatory and seem to depend on a diversity of mechanismswhich may be linked to a modification of the abscisic acid balance.Dunaliella lacks a rigid cell wall and the cellular contentsare enclosed by an elastic plasma membrane that permits rapidcell volume changes in response to extracellular changes inosmolarity. Based on the ‘stretch activated ion channelsmodel’ reviewed recently by Kirst (1990) we propose thefollowing cascade of responses: volume change/distortion ofplasmalemma     

Effects of Metabolic Inhibitors on the Calcification of a Freshwater Green Alga Gloeotaenium loitlesbergarianum Hansgirg. 2. Effects of Some Inhibitors of Protein Synthesis

The effects of five inhibitors of protein synthesis, viz. streptomycin,aurin tricarboxylic acid, tetracycline, chloramphenicol, andcycloheximide, on the calcification of Gloeotaenium loitlesbergarianumHansgirg, a freshwater green alga were studied. Streptomycinhad no effect while aurin tricarboxylic acid at 50 µgml^–1 and tetracyline, chloramphenicol and cycloheximideat 20 µg ml^–1 completely inhibited calcificationin the alga. High concentrations of chloramphenicol and cycloheximidewere not completely inhibitory when added 26 h and 32 h respectivelyafter the material was incubated in the induction medium. Itis concluded that the effects by these substrates are the resultsof inhibition of protein synthesis, which is directly or indirectlylinked to calcification. calcification, Gloeotaenium loitlesbergarianum Hansgirg, green alga, chlorophyceae, protein synthesis inhibitors     

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     

Determination of Volume of Dunaliella Cells by Lithium Dilution Measurements and Derivation of Internal Solute Concentrations

A method has been developed to measure the cell volume of theunicellular green alga Dunaliella parva 19/9 using Li⁺ measurementsonly. Concentrations of internal solutes can also be calculatedif they are assayed in the same samples as Li⁺. We found thatD. parva cells grown in 0.4 kmol m^–3 NaCl have an averageaqueous cell volume of 65.1 ?2.9 µm³, a K⁺ concentrationof 126?6 mol m^–3, a Na⁺ concentration of 11?11 mol m^–3and a glycerol concentration of 615?27 mol m–3 (n= 12).Algae grown in 1.5 kmol m^–3 NaCl have an average aqueouscell volume of 131 ?7.5 µm³, a K⁺ concentration of 109?4mol m^–3, a Na⁺ concentration of 10?39 mol m^–3 anda glycerol concentration of 1 425?59 mol m^–3 (n = 12).These results indicate that D. parva cells adapted to high salinitieshave larger cell volumes than those adapted to lower salinities.However, there is no evidence for a significant difference ininternal Na⁺ concentration, despite the almost 4-fold differencein the concentration of external NaCl. The intracellular glycerolconcentration alone accounts for 65% and 54%, respectively,of the osmotic balance in low and high salt grown cells. Key words: Dunaliella, cell volume, intracellular solutes     

Regulation of Free Calcium Concentration in the Pitchers of the Carnivorous Plant Sarracenia purpurea: A Model for Calcium in the Higher Plant Apoplast?

The pitcher of the carnivorous plant Sarracenia purpurea L.contains an entrapped body of liquid within which its prey isdigested. Free calcium in the pitcher is derived from eitherthe pitcher walls or from prey falling into the pitcher; inthe absence of exogenous (prey-derived) calcium it will dependon the active and passive calcium regulatory properties of thepitcher walls and may to some extent therefore mimic calciumin the apoplast of plant cells. Using a calcium-specific electrode,the free calcium concentration of the pitchers of Sarraceniaplants was investigated and the effect of adding a variety ofconcentrations of calcium in water determined. The mean pitcherfree calcium concentration in vivo was 2.3 x 10^–5 M±2.5x 10^–5 M; when pitchers were washed and filled with watercontaining lower calcium concentrations, the concentration inthe pitcher water rose to 1–5 x 10^–5 M. When highercalcium concentrations (up to 1 x 10^–4 M) were added,the pitcher calcium concentration declined to 1–7 x 10^–5M. Concentrations of calcium above 1 x 10^–4 M were alsoreduced, but to a lesser extent. Metabolic inhibition of activeion transport, while inhibiting pitcher acidification, did notinhibit regulation of pitcher free calcium, suggested that itoccurs as a result of calcium exchange sites in the pitcherwalls. The data are discussed in relation to the physiologyof Sarracenia pitchers and to the usefulness of the pitcheras a model for free calcium in the higher plant apoplast. Sarracenia purpurea L., carnivorous plant, pitcher, free calcium, plant apoplast     

BIOTIN LIBERATION BY THE LICHEN ALGA COCCOMYXA SP. AND BY CHLORELLA PYRENOIDOSA

The unicellular green alga Coccomyxa, a component of the lichenPeltigera aphthosa, liberated about 7.2mµg biotin permg dry weight of cells into the culture medium during a growthperiod of 15–20 days. The corresponding figure for thefree-living alga Chlorella pyrenoidosa was 0.45mµg ofbiotin. Chromatographic analysis indicated that this was freebiotin and not a bound form of the vitamin. The biotin concentrationof rinsed Coccomyxa cells was 1.88mµg per mg dry weightof cells, of which less than 0.01mµg was extractable byhot water. Cells of Chlorella contained 0.16mµg of biotinper mg dry weight, of which 0.11mµg was extractable byhot water. The biotin content of Coccomyxa, which was about12 times that of Chlorella, is thus almost entirely in the boundform. The importance of biotin in the symbiotic interactionsbetween the alga and the fungus in Peltigera is discussed. ¹Present address: University Department of Agriculture, Oxford,England. ²Present address: Institute of Marine Resources, Universityof California, La Jolla, California, U.S.A.     

Cation Exchange Properties of the Cell Walls of Enteromorpha intestinalis (L.) Link. (Ulvales, Chlorophyta

The cell wall of Enteromorpha intestinalis (a marine alga) hasbeen found to behave as a weakly cross-linked cation exchangerin NaCl solutions from 0.1–1020 mMolal (0.1–1000mMolar). Anion adsorption could be described by Freundlich isothermsover this concentration range. The large anion, inulin carboxylate,was found to be a tracer of the anion free space of plant tissuesonly in salt solutions above 10 mMolal. The cell wall of Enteromorphahas a cation exchange capacity of about 2500 µ mol g^–1dry wt. (Na+ form). The cell wallvolume is a complex functionof p^H and the NaCl concentration. As a result, the cation exchangecapacity is only predictable on a dry weight basis. The fixednegative charges of the cell wall have a pK_a of2 in situ and1.75 in vitro, and seem to be a mixture of sulphate and carboxylsugar esters. The applicability of the Donnan equation to plant cell wallsis discussed. Interpretation of the cell wall as a single thermodynamicphase is shown to be inappropriate. A large proportion of thecell wall solution is unaffected by the fixed anions.     

The importance of crustacean zooplankton in structuring rotifer and phytoplankton communities; an enclosure study

During the spring clear-water phase of 1993, an enclosure experimentwas performed in the mesotrophic Schöhsee (Plön, FRG)in order to assess the impact of crustacean zooplankton on therotifer and phytoplankton community. Among the crustacean plankton,calanoid and cyclopoid cope-pods were abundant, but Daphnia‘longispina’ reached the highest densities in thisexperiment. The colonial rotifer Conochilus unicomis was notaffected by crustacean plankton. The two most abundant species,Synchaeta peclinata and Keratella cochlearis, increased exponentiallywhen macrozooplankton had been excluded from the enclosures,but did not increase when crustaceans were present. Birth anddeath rates of K.cochlearis could be reliably determined inthis field experiment, suggesting that this rotifer specieswas mainly controlled by exploitative competition rather thanby mechanical interference or predation. Daphnia ‘longispina’generally grazed selectively on the smaller ciliates and algae,thus depriving the rotifers of their phytoplankton resources.The dominant alga, the chrysophycean Dinobryon, increased, whethercrustaceans were present or not, but appeared to be grazed uponto a certain extent despite its considerable cell size and colonialorganization.     

Physiological Responses of Phytoflagellates to Dissolved Organic Substrate Additions. 2. Dominant Role of Autotrophic Nutrition in Pyrenomonas salina (Cryptophyceae)

The enhancement of algal growth by organic substrate assimilationis a common laboratory observation, yet few studies have addressedthe interaction of dissolved organic compounds and environmentalfactors for controlling the relative contribution of heterotrophyand autotrophy to the nutrition of these algae. The effectsof light intensity and glycerol addition on the growth, cellvolume, pigmentation, and carbon uptake of the facultative heterotroph,Pyrenomonas salina Santore, were examined. Glycerol additionto cultures growing at a limiting light intensity increasedthe growth rate, increased the average cell volume and cellularstarch content, decreased the cellular phycoerythrin to chlorophyll ratio, and had no effect on the CO₂ fixation rate cell^–1.Glycerol addition to cultures growing at a moderate light intensitythat was saturating for photo-autotrophic growth increased theaverage cell volume and cellular starch content but had no effecton the CO₂ fixation rate cell^–1. The results indicatethat autotrophy was the major process for carbon acquisitionduring the growth of P. salina, but that carbon acquisitionfrom glycerol catabolism also was used to partially supportgrowth of the alga at the limiting light intensity. In addition,glycerol presumably was used to fulfill the energy and/or reductantrequirements of the alga, and to increase the reserve carbohydrate(starch). ¹ Current address and address for correspondences: Horn PointEnvironmental Laboratories, University of Maryland, PO Box 775,Cambridge, Maryland 21613, U.S.A. (Received October 29, 1990; Accepted May 31, 1991)     

Purification of Ribulose 5-phosphate Kinase and Minor Polypeptides of Pyrenoid from the Green Alga Bryopsis maxima

Ribulose 5-phosphate (Ru5P) kinase (ATP:D-ribulose 5-phosphate1-phosphotrans- ferase; EC 2.7.1.19 [EC] ), an enzyme in the reductivepentose phosphate cycle, was purified from the green alga Bryopsismaxima and its activity and peptide composition were studied.The specific activity of purified Ru5P kinase was 20 µmoleRuBP formed (mg protein)^–1 min^–1 corresponding toa 490-fold purification from the supernatant of chloroplasts.The K_m values of Ru5P kinase for ATP and Ru5P were 69 µMand 330 µM, respectively. The molecular size of Ru5P kinase was estimated as 90 kDa bygel filtration and that of its polypeptide as 41 kDa by SDS-polyacrylamidegel electrophoresis. A small portion of the Ru5P kinase wasfound in a large molecular state (500 kDa) which was consideredto be an inactive form of the enzyme. Ru5P kinase activity has been reported in the pyrenoid of Eremosphaeraviridis as well as ribulose 1,5-bisphosphate carboxylase-oxygenase(RuBisCO) and ribose 5-phosphate isomerase activity (Holdsworth1971). In Bryopsis maxima, among the pyrenoid polypeptides otherthan that of RuBisCO, we found a polypeptide of 42 kDa, similarto that of Ru5P kinase in molecular size and ratio to RuBisCO.A peptide map of the 42 kDa pyrenoid polypeptide, however, showedthat it differed from that of Ru5P kinase. In conclusion, Ru5Pkinase may be not involved in the pyrenoid of this alga. (Received January 19, 1985; Accepted May 15, 1985)     

Changes in Nuclear DNA Content, Cell and Nuclear Size, and Frequency of Cell Division in the Cotyledons of Brassica napus L. during Embryogenesis

Cellular behaviour was examined during embryogenesis in Brassicanapus to test whether or not polyploidy occurs in the cotyledonsduring the phase of oil deposition. Nuclear DNA content, nuclearand cell size, and the mitotic index were measured in the cotyledonson various days post anthesis (dpa). In squashed monolayersfrom 15 dpa cotyledons, a polyploid (>5C) population wasdetected together with a substantial number of cells in G2 (4C).Nuclear volume was measured on sectioned tissues and, at 15dpa, the range of values from the cotyledons (40–500 *m³)contrasted with that in the vestigial suspensor and endosperm(50–> 600 µm³). At 15 dpa the nuclear volumedata suggest that whilst cells in the cotyledons were in Gland G2 many endosperm and suspensor cells were polyploid. Thus,polyploidy observed in the squashed monolayers was probablydue to contaminating endosperm/suspensor cells. At 25 and 35dpa, polyploidy was not detected; all cells were in Gl (2C)and cell area increased. The mitotic index peaked at 20 dpabefore declining and given the narrower distribution of nuclearvolumes at 25 and 35 dpa (50–300 µm³), these dataare consistent with cell arrest in Gl. Thus, polyploidy wasnot detected in the cotyledons of B. napus which differs fromwhat is known about cellular development in legume cotyledons. Key words: Brassica napus L., DNA, nuclear volume     

Studies on the Growth in Culture of Plant Cells: XI. THE INFLUENCE OF SHAKING RATE ON THE GROWTH OF SUSPENSION CULTUEES

The influence of shaking rates (expressed as revolutions permin) on orbital shaking platforms (1 in (2.54 cm) diam. rotarymotion) on the growth of cell suspension cultures of Acer pseudoplatanusL. and Atropa belladonna cultivar lutea Döll are described.By following cell growth and respiration and the levels of oxygenand carbon dioxide in the media during the progress of incubationit is concluded that the reduction of growth at sub-optimalshaking rates is not due to oxygen deficiency or toxic accumulationof carbon dioxide. The growth of the Atropa cell suspensionin ‘closed systems’ has been studied by the developmentof modified culture vessels and evidence obtained that the reducedgrowth in the systems is due to the formation by the culturesof an unidentified volatile growth inhibitor and not to eitheroxygen depletion or toxic accumulation of either carbon dioxideor ethylene. It is suggested that the reduced growth in ‘opensystems’ cultures at sub-optimal shaking speeds is eitherdue to retention of this volatile inhibitor or to restrictionof nutrient uptake by the existence of a stationary liquid-phaseboundary to the cells.     

Effects of xylopine on physiological activities of Scenedesmus obliquus

Xylopine, an aporphine alkaloid contained in the bark of Xylopiadiscreta strongly inhibits growth of Scenedesmus obliquus; 50%inhibition was caused by 2x10^–5 M of xylopine. Photosynthesis was also strongly inhibited by xylopine at concentrationsabove 10^–4 M. Effects on photosynthesis, however, differeddepending on the ratio of alkaloid concentration to cell concentrationin the reaction mixture, so that under certain experimentalconditions with concentrations of xylopine below 10^–4M, there was some acceleration of photosynthesis. Xylopine strongly inhibited the HILL reaction with benzoquinoneas the HILL oxidant. Respiration was not affected at concentrationscompletely inhibiting photosynthesisd growth of the alga. A probable role of the aporphine alkaloid in controlling lichengrowth in nature is discussed. (Received March 25, 1970; )     

The Transport and Metabolism of Urea in Chara australis: III. TWO SPECIFIC TRANSPORT SYSTEMS

Reciprocal competitive inhibition studies were used to showthat N-methyl-urea (NMU), acetamide and urea all compete forbinding to a common transport system, designated system I andthat this system is one of two specific mechanisms transportingurea in Chara. System I binds urea with a Km of about 0–3mmol m-3 and is strongly influenced by metabolic controls. SystemI is active and electrogenic and may be energized by the couplingof urea uptake to an influx of protons. This is the first reportof an electrogenic urea transport system in an alga. The secondspecific mechanism for urea transport, designated system II,binds urea with a relatively low affinity (K_m c. 7–0 mmolm-3) and does not transport NMU to a significant extent. SystemII is less subject to metabolic control than system I and, thoughit may be active, is not electrogenic. Key words: Urea, methylurea, proton cotransport, metabolic control