Effects of Light and Dark on the Ultrastructure of Lichen Algae

Quantitative ultrastructural observations have been made onthe algal cells (Trebouxia) in two lichens, Parmelia sulcataand P. laevigata, stored for 48 h in the dark or under a light/darkregime. The response of the alga was found to differ in theselichens. In P. sulcata the dark treatment caused a decreasein starch grains, lipid-rich pyrenoglobuli and peripheral cytoplasmicstorage bodies and an increase in pyrenoid and chloroplast proteinbodies. The algae in P. laevigata contained little starch andno chloroplast protein bodies. However, after dark treatment,starch, cytoplasmic storage bodies and pyrenoid dimensions sometimesdeclined, while pyrenoglobuli numbers increased. Some of theseapparent changes depended upon the units used for calculatingthe cross-sectional areas of structures, e.g. absolute units,percentage of cell wall, protoplast or chloroplast cross-sectionalarea. Chloroplast area increased in the dark (as a % of cellwall area) in both species while mitochondria were larger inthe dark in P. sulcata but not in P. laevigata. Ultrastructuralchanges were not clearly correlated with changes in photosyntheticand respiratory rates. These results directly support the suggestionthat some intra-cellular structures are energy-generating reservesthe dimensions of which can rapidly change. Parmelia sulcata, Parmelia laevigata, lichen algae, light and dark storage, starvation, reserve substances, organdie dimensions     

Immunochemical Studies on Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase in the Chloroplasts of the Marine Alga Bryopsis maxima

The localization of ribulose 1,5-bisphosphate carboxylase/oxygenase(RuBisCO) in chloroplasts of the green alga Bryopsis maximawas examined by immunological techniques. Three strains of hybridomaswere established between myeloma cells and the spleen cellsfrom mouse immunized against B. maxima RuBisCO. The antibodiesreacted with the large subunit of B. maxima RuBisCO but notwith spinach RuBisCO. Immunofluorescence and immunoenzymaticstudies showed that the large subunit of B. maxima RuBisCO wasconcentrated in pyrenoids and on the surface of starch grainssurrounding the pyrenoids. (Received September 22, 1987; Accepted March 2, 1988)     

An analysis of seed development in Pisum sativum XIX. Effect of mutant alleles at the r and rb loci on starch grain size and on the content and composition of starch in developing pea seeds

The effects of mutant alleles at the r and rb loci on starchgrain size and the levels of starch and amylose in developingpea (Pisum sativum L.) seeds have been examined. Four lines,near-isogenic except for genes at these loci, have been usedto show that both mutations reduce levels of starch throughoutembryo development and reduce levels still further when combinedin the ‘double mutant’. The reduction in starchcontent was due, at least in part, to a reduction in starchgranule size. Although the proportion of starch in mature embryoswas similar in the rrRbRb and RRrbrb lines, the starch contentdiffered between these two lines during development, as a percentageof embryo dry weight. This difference was due to a reductionin the absolute growth of the embryo caused by the rb mutation.Lines homozygous for the mutant r allele with either wild-type(RbRb) or mutant (rbrb) alleles at the rb locus contained increasedproportions of amylose in their starch throughout development,due to a reduced production of amylopectin. The presence ofthe rb mutation, however, also reduced the amount of amylosein relation to the reduction in total starch levels. Mutantalleles at both loci also reduced starch levels in the testaduring development, the reduction due to rb being more extreme.Reciprocal crosses showed a maternal effect of the rb mutationon final seed size and on the absolute amount of starch in theembryo. Key words: Pisum sativum L., seed, starch, development, mutant     

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.     

Changes of Starch Localization within the Chloroplast Induced by Changes in CO2 Concentration during Growth of Chlamydomonas reinhardtii: Independent Regulation of Pyrenoid Starch and Stroma Starch

To understand the physiological function of the pyrenoid, aprotein complex in algal chloroplast stroma with surroundingstarch sheaths, the effects of environmental conditions on thepyrenoid and pyrenoid starch were investigated in the unicellulargreen alga Chlamydomonas reinhardtii. Pyrenoid starch was rapidlyaccumulated within 5 hours when the extracellular CO₂ concentrationwas lowered from 4% to ordinary air level (0.04%). Startingwith high-CO₂ grown cells containing well-developed stroma starchgranules, degradation of stroma starch and accumulation of pyrenoidstarch were observed in parallel during the adaptation to lowCO₂ condition. This pyrenoid-starch accumulation was light dependentand completely inhibited by DCMU. The starch relocalizationprocess was reversible, but the breakdown of pyrenoid starchwas slower than its accumulation. The time courses of accumulation(or degradation) of starch around the pyrenoid paralleled increases(or decreases) in carbonic anhydrase (EC 4.2.1.1 [EC] ) activity,and the pyrenoid starch accumulation is thought to be one ofthe adaptation phenomena to CO₂ concentration. When nitrogenassimilation was inhibited, stroma starch and total starch contentincreased, while that of pyrenoid starch decreased. These resultsindicate that the synthesis and degradation of the two formsof starch were regulated independently by the environmentalconditions. ADP-glucose starch synthase (EC 2.4.1.21 [EC] ) activitywas detected at a physiological level, but the change of starchlocalization could not be explained by total starch synthaseactivity nor by starch-degrading enzyme activities. We assumethat starch metabolism around the pyrenoid is regulated independentlyfrom that in other stromal spaces. (Received May 16, 1988; Accepted July 17, 1988)     

The Ultrastructure of Tetrasporogenesis in the Marine Red Alga Chondria tenuissima (Good, et Woodw.) (Ceramiales, Rhodomelaceae)

Tetraspore development has been studied in Chondria tenuissimausing light and electron microscopy. The transformation of tetrasporangialmother cells into mature tetrasporangia involves a series ofstructural changes, especially of dictyosomes and of the nucleus.The youngest stage of tetrasporogenesis consists of a uninucleatetetraspore mother cell with synaptonemal complexes present duringearly prophase of meiosis I. Mitochondria are aggregated aroundthe nucleus, dictyosome activity is low, and proplastids occurin the peripheral cytoplasm. The cleavage furrows are initiatedalmost concomitantly with commencement of meiosis. When thecleavage furrows are initiated, spherical bodies bounded bytwo membranes are found within the cytoplasm; they develop intovacuoles with fibrillar contents (fv₁), which increase in sizeduring tetraspore development by fusing with each other andwith Golgi vesicles. The Golgi vesicles and the vacuoles withfibrillar contents (fv₁) contribute material to the developingtetraspore wall. During the middle stage of tetraspore formationthe vacuoles with fibrillar contents (fv₁) are dominant, dictyosomeactivity increases, as well as the number of plastids and mitochondria;starch formation also increases. Stacked cisternae of the endoplasmicreticulum are found within the peripheral part of the nucleus.The same nuclear structures are also observed in tetrasporangiaof the marine red alga Gastroclonium clavalum. The final stageis characterized by the disappearance of vacuoles with fibrillarcontents (fv₁) and of the stacked ER within the nucleus, presenceof straight, large dictyosomes which produce cored vesicles,an abundance of starch grains and by the formation of fullydeveloped chlorqplasts. The cored vesicles contain Thiéry-positivematerial and contribute to the formation of vacuoles with fibrouscontents (fv₂) as they are dominant in the tetraspores beforetheir liberation. Rhodophlyla, Chondria, tetrasporogenesis, ultrastructure, Golgi apparatus     

Lipid Biosynthesis in the Blue-Green Alga (Cyanobacterium), Anabaena variabilis III. UDPglucose:Diacylglycerol Glucosyltransferase Activity in vitro

The synthesis of glyceroglycolipids was studied in membraneand soluble fractions of Anabaena variabilis. The membrane fractionexhibited a high activity of UDPglucose: diacylglycerol glucosyltransferase,but practically no activity of UDPgalactose: diacylglycerolgalactosyltransferase. The glucosyltransferase activity wasmaximal at about pH 7.0 and dependent on Mg²⁺ The Michaelisconstant (K_m) for UDPglucose was 45?10^–6 M. The solublefraction catalyzed the incorporation of galactose from UDP galactoseinto digalactosyl diacylglycerol. These in vitro results werecompatible with the biosynthetic pathway of glyceroglycolipidsin this alga that we previously elucidated on the basis of tracerexperiments in vivo. ¹ Present address: Department of Biology, Faculty of Science,University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan. (Received June 1, 1982; Accepted July 1, 1982)     

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)     

Microfibrillar Structure, Cortical Microtubule Arrangement and the Effect of Amiprophos-methyl on Microfibril Orientation in the Thallus Cells of the Filamentous Green Alga, Chamaedoris orientalis

Microfibrillar structure, cortical microtubule orientation andthe effect of amiprophos-methyl (APM) on the arrangement ofthe most recently deposited cellulose microfibrils were investigatedin the marine filamentous green alga, Chamaedoris orientalis.The thallus cells of Chamaedoris showed typical tip growth.The orientation of microfibrils in the thick cell wall showedorderly change in longitudinal, transverse and oblique directionsin a polar dependent manner. Microtubules run parallel to thelongitudinally arranged microfibrils in the innermost layerof the wall but they are never parallel to either transverseor obliquely arranged microfibrils. The ordered change in microfibrilorientation is altered by the disruption of the microtubuleswith APM. The walls, deposited in the absence of the microtubules,showed typical helicoidal pattern. However, the original crossedpolylamellate pattern was restored by the removal of APM. Thissuggests that cortical microtubules in this alga do not controlthe direction of microfibril orientation but control the orderedchange of microfibril orientation. Amiprophos-methyl, Chamaedoris orientalis, coenocytic green alga, cortical microtubule, microfibrillar structure, tip growth     

The Effects of Salinity upon Cellular Volume of the Marine Red Alga Porphyra purpurea (Roth) C.Ag.

Changes in cell volume of the marine red alga Porphyra purpureahave been investigated using photomicroscopic and radioisotopictechniques. There is an inverse relationship between cell volumeand external salt content. The alga responds to changes in thewater potential of its bathing medium by rapid swelling in hyposalinemedia and shrinkage in hypersaline conditions. Cells P. purpureabehave as osmometers in concentrated sea-waters, obeying theBoyle-Van't Hoff law. A non-osmotic volume, 20–25% ofthe total cell volume in sea-water, can be predicted from thelinear plot of volume versus reciprocal pressure in concentratedsea-water media. In dilute sea-waters the presence of non-rigidcell walls serves to limit any increases in cell volume. Theprimary response to dilution stress is thus an increase in turgor.Cell volume is not returned to its original value followingprolonged immersion in either hyposaline or hypersaline media,showing that the alga does not ‘osmoregulate’ sensustricto.     

The Significance of Root Starch in Post-fire Shoot Recovery of the Resprouter Stirlingia latifolia R. Br. (Proteaceae)

Stirlingia latifolia, a common shrub of Banksia woodlands ofSW Australia, is a highly successful resprouter species recoveringfrom fire by multiple sprouting of new shoots from its upperroot stock. in comparison with the congeneric fire-sensitive(obligate seeder) species Stirlingia tenuifolia it exhibitsa low shoot:root dry weight ratio and high concentrations ofstored starch in the cortical tissue of its roots. The relationshipbetween root reserves of starch and development of newly sproutingshoot material following fire is examined in S. latifolia afterspring and summer burns. During the initial 2-5 month periodafter fire, levels of stored starch in the roots fall by 50-75%,followed by a slow increase as plants reproduce and the attainmentof pre-fire starch levels by 1·5-2 years after the fire.Starch reserves of roots can be further reduced by shading theregenerating shoots to limit their input of photosynthates andalmost totally eliminated by monthly removal of successive flushesof new shoots over a 10-12 month period. New shoots continueto sprout until all the starch is eliminated. The data are discussedin relation to the fire-induced reproduction of S. latifoliaand its ability to thrive in very frequently burnt habitats.Copyright1993, 1999 Academic Press Fire response, Proteaceae, resprouter, shoot:root ratio, starch storage, Stirlingia latifolia     

Cellular and Extracellular Polysaccharides of the Blue green Alga Nostoc

The carbohydrate content of various cellular fractions of theblue-green alga Nostoc was studied as a function of age of theculture. The production of extracellular and intracellular polysaccharideswas higher in actively growing cultures. Mannose and glucosewere the main components of cell wall polysaccharides. Glucosamineand diaminopimelic acid were also detected in the cell walls.The kinetics of incorporation of radioactivity from sodium [¹⁴C]bicarbonate showed that the extracellular polysaccharides werelabelled within 10 min of incubation suggesting the active exudationof polysaccharides by this alga. The selective excretion ofpolysaccharides by the alga Nostoc is also suggested.     

Further Observations on the Structure of Plant Cilia, by a Combination of Visual and Electron Microscopy

By combining visual microscopy of stained material with electronmicroscopy it has been possible to demonstrate fibrillar disintegrationin the cilia of Saprolegnia (front and back), Chlorosaccus (frontand back), Allomyces, and Olpidium. These represent three fungiand a yellow-green alga. All flagella showed eleven componentstrands, in most cases clearly differentiated into nine strandsand a central pair. In addition, stained and unstained materialof Saprolegnia and Chlorosaccus have been closely compared inthe electron microscope and the comparison has included observationson one and the same cell with die electron microscope and thelight microscope. The nature of the artifacts produced by staininghas been investigated and the value of the combination of thesevarious technical processes demonstrated. The phyletic importanceof the hairs on the front flagellum in the yellow-green algae,brown algae, and water moulds is discussed.     

Change in Levels of Starch and Sugars in Epidermis of Commelina benghalensis during Fusicoccin Stimulated Stomatal Opening

The concentrations of malate, starch and sugars were determinedin presonicated epidermal strips of Commelina benghalensis.During opening, the starch content of epidermis decreased whilethe level of sugars or malate increased. Fusicoccin (FC) stimulatedstomatal opening and elevated the levels of malate and sugars.However, the contribution from sugars was nearly 50% of theosmotic effect of malate and it increased to more than 60% inthe presence of FC. We conclude that FC stimulates stomatalopening by enhancing not only potassium influx into guard cellsbut also hydrolysis of starch into sugars (and malate). Significantcorrelations were noticed between the width of stomatal apertureand epidermal starch (negative), malate and sugars (both positive).The negative relationship between starch and malate or sugarswithin epidermis indicated that starch hydrolysis lead to formationof sugars as well as malate. Starch—sugar interconversioncan therefore play a significant role in modulating the solutepotential of guard cells. Key words: Commelina benghalensis, Stomatal opening, Fusicoccin, Epidermal starch and sugars     

Cytokinins from the tRNA of the Red Alga Porphyra perforata J. Ag.

Isopentenyladenosine and cis-zeatin riboside have been identifiedby GC-MS from the tRNA of a red alga Porphyra perforata. (Received March 12, 1991; Accepted December 7, 1991)     

The in vivo cleavage of carotenoids into retinoids in Chlamydomonas reinhardtii

A variety of carotenoids have been incorporated in vivo intoa carotenoid mutant of the unicellular alga, Chlamydomonas reinhardtii.The mutant can synthesize retinal from exogenous carotenoids.In this way the first step in biosynthesis of retinoids, namelyhow ß-carotene is converted into retinal, has beenstudied. Since retinal forms with opsin the photoreceptor forphototaxis in Chlamydomonas, the action-spectral peaks of thephototaxis restored by carotenoid incorporation were used topredict the products formed by the enzyme that cleaves thesecarotenoids. The data suggest that the physiologically relevantcleavage of ß-carotene into retinal is central ratherthan excentric. When apocarotenoids are substrates, the enzymetargets the double bond located a constant distance away fromthe carbonyl group on the acyclic end and, consequently, retinalis not produced. Interestingly, dehydro-analogues containinga triple bond are preferentially cleaved and oxidized at thetriple bond relative to the corresponding analogue with onlydouble bonds. Key words: Chlamydomonas reinhardtii, Chlorophyceae, retinoid biosynthesis, carotenoid cleavage, action spectrum     

The Effect of Osmotic and Ionic Stress on the Primary Processes of Photosynthesis in Dunaliella tertiolecta

Cells of the euryhaline unicellular green alga Dunaliella tertiolectawere exposed to a range of osmotic potentials imposed by theaddition of NaCl, KCl, sucrose or ethylene glycol to the algalsuspending medium. The alga synthesized glycerol in responseto increased concentrations of NaCl, KCl or sucrose, but notto increased concentrations of ethylene glycol. The overallprocess of photosynthesis was inhibited by all four treatmentsduring the first 100 min, but especially by NaCl and KCl. Theinduction of chlorophyll fluorescence indicated that photosystemIl was inhibited by such treatment although to a lesser extentthan the overall process. The size and half-time of decay ofthe flash-induced field-indicating absorption change at 519nm were decreased by NaCl and KCl but the effect of sucroseand ethylene glycol on the absorption change was much less.Therefore, the photosynthetic processes in intact Dunaliellacells are more markedly inhibited by the combined osmotic andionic stresses due to NaCl and KCl than by osmotic stress alone.In most cases some recovery of the inhibition of photosynthesiswas seen concomitant with the synthesis of intracellular glycerol,suggesting that the alga has the ability to overcome osmoticand ionic stresses. Key words: Dunaliella, Photosynthesis, Osmotic stress     

Localization of Starch in the Base of Narcissus Leaves

The localization of starch in the leaves and flower stalk ofNarcissus tazetta is related to stages of development includingleaf abscission. Starch begins to appear in the newly vacuolatedparenchyma of the leaf primordia when primordia are about 0.5mm in length, and becomes localized in the leaf bases at theonset of differentiation into sheath and blade. The part playedby the intercalary meristems as a physiological and anatomicalbarrier to the distribution of starch in these organs is stressed,and the possible mechanism is discussed.     

On the Chemical Composition and Developmental Role of the Mesoglea of Hydra

Two aspects of the chemistry and function of the mesoglea ofhydra were studied, (i) Chemical composition: Its componentneutral sugars and amino acids were analyzed to determine ifthis structure contains collagen, (ii) Role in morphogenesis:Hydra were exposed to the lathyrogen 3-aminopropionitrile, aninhibitor of collagen crosslinking, to discover if new cross-linkedmesoglea is required for normal regeneration to occur. Large amounts of the neutral sugars glucose and galactose andsmall amounts of fucose and rhamnose were found, as was theamino sugar glucosamine. Evidence that the major protein componentof the mesoglea is collagen was revealed by the detection ofhydroxylysine, hydroxyproline, and proline, and of large amountsof glycine. The glucose and galactose are joined as a dimerby an alkali-stable bond to the polypeptide backbone of thecollagen. The lathyrogen 3-aminopropionitrile, known to block the assemblyof newly synthesized collagen into fibers, causes abnormal headregeneration in hydra. The drug has its maximum effect between24 and 48 hr after the previous head is removed.     

Isolation and characterization of the 337 m{micro} UV-absorbing substance in red alga, Porphyra yezoensis UEDA

Using a variety of Sephadex gel filtrations, starch block zoneelectrophoresis, Avicel SF preparative TLC and DEAE cellulosecolumn chromatography, the characteristic 337 mµ UV-absorbingsubstance from marine red alga, Porphyra yezoensis UEDA wasisolated. The molecular weight of this substance was about 1,000,and its chemical composition was 43.11% C, 5.85% H, 7.23% N,34.73% O and 9.08% Na. In vivo it is located in the chloroplast.On irradiation at 378 mµ, it has a fluorescence actionspectrum peak at 470 mµ. Structural studies on this substanceare still underway, but it could be a kind of aminosugar judgingfrom NMR, IR spectra and other chemical properties. ¹This work was partially supported by the grant of the Ministryof Education in 1969 ²Present address: Department of Biochemistry and Biophysics,University of Hawaii, Honolulu, Hawaii 96822, U.S.A. (Received December 21, 1969; )