CELL‐WALL POLYMER MAPPING IN THE COENOCYTIC MACROALGA CODIUM VERMILARA (BRYOPSIDALES,CHLOROPHYTA)1

Cell walls in the coenocytic green seaweed Codium vermilara (Olivi) Chiaje (Bryopsidales, Chlorophyta) are composed of ～32% (w/w) β‐(1→4)‐d‐mannans, ～12% sulfated polysaccharides (SPs), and small amounts of hydroxyproline‐rich glycoprotein‐like (HRGP‐L) compounds of the arabinogalactan proteins (AGPs) and arabinosides (extensins). Similar quantities of mannans and SPs were reported previously in the related seaweed C. fragile (Suringar) Hariot. Overall, both seaweed cell walls comprise ～40%–44% of their dry weights. Within the SP group, a variety of polysaccharide structures from pyruvylated arabinogalactan sulfate and pyruvylated galactan sulfate to pyranosic arabinan sulfate are present in Codium cell walls. In this paper, the in situ distribution of the main cell‐wall polymers in the green seaweed C. vermilara was studied, comparing their arrangements with those observed in cell walls from C. fragile. The utricle cell wall in C. vermilara showed by TEM a sandwich structure of two fibrillar‐like layers of similar width delimiting a middle amorphous‐like zone. By immuno‐ and chemical imaging, the in situ distribution of β‐(1→4)‐d‐mannans and HRGP‐like epitopes was shown to consist of two distinct cell‐wall layers, whereas SPs are distributed in the middle area of the wall. The overall cell‐wall polymer arrangement of the SPs, HRGP‐like epitopes, and mannans in the utricles of C. vermilara is different from the ubiquitous green algae C. fragile, in spite of both being phylogenetically very close. In addition, a preliminary cell‐wall model of the utricle moiety is proposed for both seaweeds, C. fragile and C. vermilara.     

CHARACTERIZATION OF CELL WALL POLYSACCHARIDES OF THE COENCOCYTIC GREEN SEAWEED BRYOPSIS PLUMOSA (BRYOPSIDACEAE,CHLOROPHYTA) FROM THE ARGENTINE COAST1

Bryopsis sp. from a restricted area of the rocky shore of Mar del Plata (Argentina) on the Atlantic coast was identified as Bryopsis plumosa (Hudson) C. Agardh (Bryopsidales, Chlorophyta) based on morphological characters and rbcL and tufA DNA barcodes. To analyze the cell wall polysaccharides of this seaweed, the major room temperature (B1) and 90°C (X1) water extracts were studied. By linkage analysis and NMR spectroscopy, the structure of a sulfated galactan was determined, and putative sulfated rhamnan structures and furanosidic nonsulfated arabinan structures were also found. By anion exchange chromatography of X1, a fraction (F4), comprising a sulfated galactan as major structure was isolated. Structural analysis showed a linear backbone constituted of 3‐linked β‐d ‐galactose units, partially sulfated on C‐6 and partially substituted with pyruvic acid forming an acetal linked to O‐4 and O‐6. This galactan has common structural features with those of green seaweeds of the genus Codium (Bryopsidales, Chlorophyta), but some important differences were also found. This is the first report about the structure of the water‐soluble polysaccharides biosynthesized by seaweeds of the genus Bryopsis. These sulfated galactans and rhamnans were in situ localized mostly in two layers, one close to the plasma membrane and the other close to the apoplast, leaving a middle amorphous, unstained cell wall zone. In addition, fibrillar polysaccharides, comprising (1→3)‐β‐d ‐xylans and cellulose, were obtained by treatment of the residue from the water extractions with an LiCl/DMSO solution at high temperature. These polymers were also localized in a bilayer arrangement.     

DISINTEGRATION OF THE CELLS OF SIPHONOUS GREEN ALGA CODIUM EDULE (BRYOPSIDALES,CHLOROPHYTA) UNDER MILD HEAT STRESS1

The siphonous green alga Codium edule P. C. Silva (Bryopsidales, Chlorophyta) has the highest covering ratio among the macroalgae on the coral reef of Nanwan Bay in southern Taiwan, but its population in the subtidal region drastically decreases from July to September each year. The objective of this study was to determine whether the high temperature of summer could be the basis for this population decrease. Chlorophyll fluorescence measurements revealed that when the algae were incubated at 35°C (a temperature that can be reached in southern Taiwan during the summer), their photosynthetic activities were almost completely inhibited after about 8 h. The circadian rhythm of photosynthesis was disrupted at a temperature as low as 32°C. TEM studies showed that 4 h incubation at 35°C induced a decrease in turgidity accompanied by vacuole shrinkage and plasmolysis. The marked disintegrative changes, including damage to organelles, such as chloroplasts and nuclei, occurred after about 8 h, at which time central vacuoles collapsed and the cell interior was then filled with numerous small vesicles. Our results suggested that the rise in seawater temperature during the summer could be one of the major causes of the massive death of C. edule in the field.     

CELL WALL CARBOHYDRATE EPITOPES IN THE GREEN ALGA OEDOGONIUM BHARUCHAE F. MINOR (OEDOGONIALES,CHLOROPHYTA)1

Cell wall changes in vegetative and suffultory cells (SCs) and in oogonial structures from Oedogonium bharuchae N. D. Kamat f. minor Vélez were characterized using monoclonal antibodies against several carbohydrate epitopes. Vegetative cells and SCs develop only a primary cell wall (PCW), whereas mature oogonial cells secrete a second wall, the oogonium cell wall (OCW). Based on histochemical and immunolabeling results, (1→4)‐β‐glucans in the form of crystalline cellulose together with a variable degree of Me‐esterified homogalacturonans (HGs) and hydroxyproline‐rich glycoprotein (HRGP) epitopes were detected in the PCW. The OCW showed arabinosides of the extensin type and low levels of arabinogalactan‐protein (AGP) glycans but lacked cellulose, at least in its crystalline form. Surprisingly, strong colabeling in the cytoplasm of mature oogonia cells with three different antibodies (LM‐5, LM‐6, and CCRC‐M2) was found, suggesting the presence of rhamnogalacturonan I (RG‐I)–like structures. Our results are discussed relating the possible functions of these cell wall epitopes with polysaccharides and O‐glycoproteins during oogonium differentiation. This study represents the first attempt to characterize these two types of cell walls in O. bharuchae, comparing their similarities and differences with those from other green algae and land plants. This work represents a contribution to the understanding of how cell walls have evolved from simple few‐celled to complex multicelled organisms.     

THE CELL WALL (THECA) OF TETRASELMIS STRIATA (CHLOROPHYTA): MACROMOLECULAR COMPOSITION AND STRUCTURAL ELEMENTS OF THE COMPLEX POLYSACCHARIDES

Isolated cell walls (thecae) from the scaly flagellate green alga Tetraselmis striata Butcher contain the unusual 2-keto-sugar acids 3-deoxy-manno-2-octulosonic acid (Kdo), 3-deoxy-5-O-methyl-manno-2-octulosonic acid (5OMeKdo), and 3-deoxy-lyxo-2-heptulosaric acid (Dha). In addition, galacturonic acid, galactose, gulose, and arabinose are present. EDTA-extraction yielded an insoluble fraction that retains the shape of the cell walls and contains no 2-keto-sugar acids. Methylation analysis demonstrated the presence of terminal hexose, GalA, Dha, and Kdo as well as 2-substituted hexose, 4-or 8-substituted Kdo, and 4,8-disubstituted Kdo. However, most of the carbohydrate material (about 60%) was not methylated. Periodate oxidation of the cell wall preparation showed the presence of 2-substituted Gul, 4- or/and 5-substituted and 7- or/and 8-substituted Kdo, which is in agreement with the methylation analysis. Again, a significant amount of carbohydrate material was not degraded, indicating complex substitution patterns. Oligosaccharides were generated by partial hydrolysis and fractionated using gel permeation chromatography and high-pH anion-exchange chromatography. Oligosaccharides contained either GalA and Kdo, or Gal, Kdo, Dha, and Gul, respectively. The structure of a GalA and Kdo containing disaccharide was established using ¹ H NMR spectroscopy.     

CELL DIVISION AND THE ROLE OF THE PRIMARY WALL IN THE FILAMENTOUS DESMID ONYCHONEMA LAEVE (CHLOROPHYTA)1

Cell division and the role of the primary wall in filament formation in the desmid Onychonema laeve Nordst. were investigated by transmission and scanning electron microscopy. In addition, sequential chemical extractions and enzyme treatments were performed, on cell walls of intact filaments. Interphase cells are deeply constricted, consisting of two semicells, each elliptical in front view and circular in side view. In addition to two short lateral spines, each semicell has two apical processes that originate on opposite sides at an angle of about 15° from the central axis and overlap the adjacent cell. Division is initiated as in other desmids by a slight separation of the semicells and development of a girdle of primary wall material at the isthmus. In O. laeve the girdle of primary wall expands to form a spherical vesicle (termed a division vesicle) between the separating semicells. Nuclear division and septum formation occur in this vesicle when it is nearly the full diameter of the filament. Morphogenesis of the apical processes begins with completion of the septum, before the secondary wall appears. At maturity each apical process is surrounded by a thick layer of both secondary and primary wall, except that its capitate tip protrudes through the shroud of primary wall. Sequential treatment with hot ammonium oxalate, 4% NaOH, 17.5% NaOH and 10% chromic acid or various enzyme solutions did not cause filament breakage. SEM and TEM views of O. laeve after these treatments show intact secondary walls and intact primary wall material covering and connecting the apical processes of adjacent cells. It is the persistence of the primary wall between cells and around the apical processes that maintains the long, unbranched filamentous morphology of Onychonema laeve.     

CELL WALL STRUCTURE AND IRON DISTRIBUTION IN THE GREEN ALGA STAURASTRUM LUETKEMUELLERI (DESMIDIACEAE)1

The cell wall of Staurastrum luetkemuelleri Donnat & Ruttner was examined with scanning electron microscope (SEM) using whole cells, in thin sections with transmission electron microscope (TEM), and in air dried whole cells and unstained thin sections with X-ray microanalysis in the scanning-transmission electron microscope (STEM). The cell wall was ornamented with spines and wartlike structures. Spines were solid structures, consisting of deposits of cell wall material between two main cell wall layers. The wart-like structures were pore organs extending through the cell wall and the mucilaginous layer outside the cell wall. The pore cylinder was surrounded by deposits of cell wall material similar to the ones in the spines. X-ray microanalysis of selected areas on whole cells from a natural population showed iron accumulation in discrete locations on the cell extensions of S. luetkemuelleri. In the unstained thin sections iron was found only in the cell wall deposits in the spines. Cells grown in laboratory cultures failed to show iron accumulation regardless of readdition of iron-EDTA (Fe-EDTA) to the culture medium.     

THE CRYSTALLINE CELL WALL OF TETRASELMIS CONVOLUTAE (CHLOROPHYTA): A FREEZE FRACTURE ANALYSIS1

A freeze fracture analysis of the cell wall of Tetraselmis convolutae (Parke et Manton) revealed the existence of a crystalline median layer consisting of regular repeating subunits of 27 nm. These circular subunits lie in curved, interlocking, longitudinal rows with some irregular discontinuities appearing in the subunit pattern of the crystalline lattice. A comparison with the cell walls of other green algal flagellates is presented, revealing similarities and suggesting evolutionary patterns.     

SEASONAL VARIABILITY OF PHYSICOCHEMICAL AND RHEOLOGICAL PROPERTIES OF ULVAN IN TWO ULVA SPECIES (CHLOROPHYTA) FROM THE BRITTANY COAST1

The seasonal variability in the extraction yield, physicochemical characteristics, and rheological properties of ulvan from two Ulva species contributing to Brittany “green tides” has been studied. These seaweeds were collected in the water column for Ulva armoricana Dion, de Reviers et Coat and on hard substrata for Ulva rotundata Bliding. The maximum ulvan extraction efficiency was not related to the maximum ulvan content in the seaweeds, but with the active growth period of the seaweeds. Ulvan chemical structure, macromolecular characteristics, and rheological properties were affected by both species and seasons. The proportion of high‐molecular‐weight ulvan was the major factor positively correlated with the gelling properties. Characteristics of ulvan from U. rotundata subjected to tides were more affected by seasons than ulvan from U. armoricana living in a more constant environment. These results point to several useful recommendations concerning Ulva sp. biomass collected with regard to ulvan characteristics and uses.     

A STUDY OF CELL WALL AND FLAGELLA FORMATION DURING CELL DIVISION IN THE SCALY GREEN ALGA,SCHERFFELIA DUBIA (CHLOROPHYTA)1

The thecate green flagellate Scherffelia dubia (Perty) Pascher divides within the parental cell wall into two progeny cells. It sheds all four flagella before cell division, and the maturing progeny cells regenerate new walls and flagella. By synchronizing cell division, we observed mitosis, cytokinesis, cell maturation, flagella extension, and cell wall formation via differential interference contrast microscopy of live cells and serial thin‐section EM. Synthesis of thecal and flagellar scales is spatially and temporally strictly separated. Flagellar scales are collected in a pool during late interphase. Before prophase, Golgi stacks divide, flagella are shed, the parental theca separates from the plasma membrane, and flagellar scales are deposited on the plasma membrane near the flagellar bases. At prophase, Golgi bodies start to synthesize thecal scales, continuing into interphase after cytokinesis. During cytokinesis, vesicles containing thecal scales coalesce near the cell posterior, forming a cleavage furrow that is initially oriented slightly diagonal to the longitudinal cell axis but later becomes transverse. After the progeny nuclei have moved into opposite directions, resulting in a “head to tail” orientation of the progeny cells, theca biogenesis is completed and flagellar scale synthesis resumes. Progeny cells emerge through a hole near the posterior end of the parental theca with four flagella of about 8 μm long. The precise timing of flagellar and thecal scale synthesis appears to be an evolutionary adaptation in a scaly green flagellate for the thecal condition, necessary for the evolution of the phycoplast and thus multicellularity in the Chlorophyta.     

A ZYGOTE-SPECIFIC PROTEIN WITH HYDROXYPROLINE-RICH GLYCOPROTEIN DOMAINS AND LECTIN-LIKE DOMAINS INVOLVED IN THE ASSEMBLY OF THE CELL WALL OF CHLAMYDOMONAS REINHARDTII (CHLOROPHYTA)

The cell wall of Chlamydomonas reinhardtii zygotes, which forms rapidly after the fusion of wall-free gametes, provides a tractable system for studying the properties and assembly of hydroxyproline-rich glycoproteins, the major proteinaceous components of green algal and plant cell walls. We report the cloning of the zsp2 gene and the analysis of its ZSP-2 product, a 58.9 kDa polypeptide that is synthesized exclusively by zygotes. The protein contains two (SP)_x repeats, establishing it as a member of the cell wall hydroxyproline-rich glycoproteins family. It also contains a 4-fold iteration of an amino acid sequence centered around cysteine residues, a configuration found in both plant and animal lectins. Furthermore, we report four observations on pellicle composition and production. First, cell-free preparations of the pellicle matrix are rich in hydroxyproline, arabinose, and galactose and contain bundles of very long fibrils. Second, glutathione blocks pellicle formation and results in the accumulation of long fibrils in the growth medium. Third, antibody to ZSP-2 also blocks pellicle formation. Fourth, ZSP-2 immunolocalizes to the boundary between the outer layers of the wall proper and the pellicle matrix. These observations are consistent with the possibility that the Cys-rich (glutathione-sensitive) lectin-like domains of ZSP-2 may bind to sugar residues on the long fibrils and anchor them to the cell wall, thereby initiating and maintaining pellicle formation.     

FLORIDOSIDE AS A CARBON PRECURSOR FOR THE SYNTHESIS OF CELL‐WALL POLYSACCHARIDE IN THE RED MICROALGA PORPHYRIDIUM SP. (RHODOPHYTA)1

Although red algae are known to be obligatory photoautotrophs, the red microalga Porphyridium sp. was shown to assimilate and metabolize floridoside. A pulse‐chase experiment with [¹⁴C]floridoside showed that at the end of a 240‐min pulse, 70% of total ¹⁴C‐uptake by the cells remained in the floridoside fraction. To evaluate the assimilation of floridoside by Porphyridium sp. cells, we exposed Porphyridium sp. not only to [¹⁴C]floridoside but also to its constituents, [¹⁴C]glycerol and [¹⁴C]galactose, as compared with [¹⁴C]bicarbonate. The extent of incorporation of [¹⁴C] galactose by the Porphyridium sp. cells was insignificant (50–80 dpm·mL^?1), whereas uptake of ¹⁴C from [¹⁴C]glycerol into the algal cells was evident (2.4 × 10³ dpm·mL^?1) after 60 min of the pulse. The pattern of ¹⁴C distribution among the major constituent sugars, xylose, glucose and galactose, of the labeled soluble polysaccharide was dependent on the ¹⁴C source. The relative content of [¹⁴C]galactose in the soluble polysaccharide was highest (28.8%) for [¹⁴C]floridoside‐labeled culture and lowest (19.8%) for the [¹⁴C]glycerol‐labeled culture. Upon incubation of [¹⁴C]floridoside with a crude extract of a cell‐free system prepared from nonlabeled cells of Porphyridium sp., the label was indeed found to be incorporated into the sulfated polysaccharide. Our results suggested that the carbon metabolic pathway in Porphyridium sp. passes through the low molecular weight photoassimilatory product—floridoside—toward sulfated cell‐wall polysaccharide production.     

ANALYSIS OF EXPRESSED SEQUENCE TAGS FROM THE GREEN ALGA ULVA LINZA (CHLOROPHYTA)1

There is a general lack of genomic information available for chlorophyte seaweed genera such as Ulva, and in particular there is no information concerning the genes that contribute to adhesion and cell wall biosynthesis for this organism. Partial sequencing of cDNA libraries to generate expressed sequence tags (ESTs) is an effective means of gene discovery and characterization of expression patterns. In this study, a cDNA library was created from sporulating tissue of Ulva linza L. Initially, 650 ESTs were randomly selected from a cDNA library and sequenced from their 5′ ends to obtain an indication of the level of redundancy of the library (21%). The library was normalized to enrich for rarer sequences, and a further 1920 ESTs were sequenced. These sequences were subjected to contig assembly that resulted in a unigene set of approximately 1104 ESTs. Forty‐eight percent of these sequences exhibited significant similarity to sequences in the databases. Phylogenetic comparisons are made between selected sequences with similarity in the databases to proteins involved in aspects of extracellular matrix/cell wall assembly and adhesion.     

EFFECT OF LOW TEMPERATURE ON THE STEREOISOMER COMPOSITION OF β-CAROTENE IN THE HALOTOLERANT ALGA DUNALIELLA BARDAWIL (CHLOROPHYTA)1

Dunaliella bardawil Ben-Amotz & Avron, a β-carotene-accumulating halotolerant alga, was analyzed for the effect of growth temperatures on its pigment content and on the stereoisomeric composition of β-carotene by the use of advanced liquid chromatography and photodiode array detection. Decreasing culture temperature from 30° to 10°C increased the β-carotene content twofold and the ratio of 9-cis to all-trans β-carotene fourfold, with no significant changes in the other cell pigments. The variation of total β-carotene content by temperature was correlated with the integral irradiance received by the algal culture during a cell division cycle, whereas the 9-cis stereoisomer increased over the amount expected by that integration. The massive accumulation of 9-cisβ-carotene within the β-carotene globules is interpreted as indicating that the oily 9-cis stereoisomer protects against the crystallization of all-trans β-carotene at low temperatures.     

OCCURRENCE AND CHARACTERIZATION OF ARABINOGALACTAN‐LIKE PROTEINS AND HEMICELLULOSES IN MICRASTERIAS (STREPTOPHYTA)1

The cell wall of the green alga Micrasterias denticulata Bréb. ex Ralfs (Desmidiaceae, Zygnematophyceae, Streptophyta) was investigated to obtain information on the composition of component polysaccharides and proteoglycans to allow comparison with higher plants and to understand cell wall functions during development. Various epitopes currently assigned to arabinogalactan‐proteins (AGPs) of higher plants could be detected in Micrasterias by immuno TEM and immunofluorescence methods, but the walls did not bind the β‐d ‐glycosyl‐Yariv (β‐GlcY) reagent. Secretory vesicles and the primary wall were labeled by antibodies against AGPs (JIM8, JIM13, JIM14). Dot and Western blot experiments indicated a proteoglycan nature of the epitopes recognized, which consisted of galactose and xylose as major sugars by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC‐PAD). Epitopes of alkali‐soluble polysaccharides assigned to noncellulosic polysaccharides in higher plants could be detected and located in the wall during its formation. The polyclonal anti‐xyloglucan (anti‐XG) antibody labeled primary and secondary wall of Micrasterias, whereas the monoclonal antibody CCRC‐M1, directed against the fucose/galactose side chain of xyloglucan (XyG), did not recognize any structures. Labeling by anti‐XG antibody at the trans‐sites of the dictyosomes and at wall material containing vesicles indicated that secretion of the epitopes occurred similar to higher plants. The presence of (1→3, 1→4)‐β‐glucan (mixed linked glucan) in the secondary cell wall but not in the primary cell wall of Micrasterias could be demonstrated by an antibody recognizing this glucan type, whereas (1→3)‐β‐glucan (callose) could not be detected. The analytical results revealed that alkali‐soluble polysaccharides in the secondary wall of Micrasterias consist mostly of (1→3, 1→4)‐β‐d ‐glucan.     

THE FIBRILLAR POLYSACCHARIDES AND THEIR LINKAGE TO ALGAENAN IN THE TRILAMINAR LAYER OF THE CELL WALL OF COELASTRUM SPHAERICUM (CHLOROPHYCEAE)

Methylation and spectroscopical analyses of DMSO-LiCl-solubilized fractions of the fibrillar cell wall of Coelastrum sphaericum Näg. established the presence of cellulose and β-1,4-linked mannan. A small proportion (2–7%) of (1→2) linkages in β-mannan that introduced an interruption in the regular ribbon chain conformation was interpreted as a component that modulated the mechanical strength of the cell wall. The trilaminar layer fractions consisted mostly of algaenan and cellulose. Evidence for ether linkages between glucose C-6 in the β-1,4-glucan and algaenan was obtained.     

GROWTH-INHIBITORS AND GROWTH-PROMOTERS IN ENTEROMORPHA COMPRESS A (CHLOROPHYTA)1

Methanol extracts from the alga Enteromorpha compressa (L.) Grev. contain substances which inhibit the elongation of Lepidium roots. Chromatographic separation of the inhibiting substances revealed that one of the inhibitory zones of the chromatograms had properties of the so-called inhibitor β. Neither abscisic acid (ABA) nor lunularic acid proved to be responsible for the growth-inhibiting property of this zone. Moreover, the extracts contain substances which promote the elongation of Avena coleoptile segments. One of these substances could be tentatively identified as indole-3-acetic acid by thin-layer and gas-liquid chromatography. (In addition to indole-3-acetic acid a second growth-promoting factor with the properties of the so-called accelerator α could be detected.)     

RELATIONSHIP BETWEEN PRESENCE OF A MOTHER CELL WALL AND SPECIATION IN THE UNICELLULAR MICROALGA NANNOCHLORIS (CHLOROPHYTA)1

The cell division mechanisms of seven strains from six species of Nannochloris Naumann were analyzed and compared with those of three species of Chlorella Beijerinck and Trebouxia erici Ahmadjian using differential interference microscopy and fluorescence microscopy. Nannochloris bacillaris Naumann divides by binary fission and N. coccoides Naumann divides by budding. Distinct triangular spaces or mother cell walls were found in the dividing autosporangia of the other five strains from four species of Nannochloris, three species of Chlorella, and T. erici. In an attempt to infer an evolutionary relationship between nonautosporic and autosporic species of Nannochloris, we constructed a phylogenetic tree of the actin genes using seven strains from six species of Nannochloris, three species of Chlorella, and T. erici. Nannochloris species were polyphyletic in the Trebouxiophyceae group. Two nonautosporic species of N. bacillaris and N. coccoides were monophyletic and positioned distally. Moreover, to determine their phylogenetic position within the Trebouxiophyceae, we constructed phylogenetic tree of 18S rRNA genes adding other species of Trebouxiophyceae. Nannochloris species were polyphyletic in the Trebouxiophyceae and appeared in two different lineages, a Chlorella–Nannochloris group and a Trebouxia–Choricystis group. The nonautosporic species, N. bacillaris and N. coccoides, and three autosporic species of Nannochloris belonged to the Chlorella–Nannochloris group. Nannochloris bacillaris and N. coccoides were also monophyletic and positioned distally in the phylogenetic tree of 18S rRNA genes. These results suggest that autosporulation is the ancestral mode of cell division in Nannochloris and that nonautosporulative mechanisms, such as binary fission and budding, evolved secondarily.     

THE WAVELENGTH DEPENDENCE OF MASSIVE CAROTENE SYNTHESIS IN DUNALIELLA BARDAWIL (CHLOROPHYCEAE)1

Dunaliella bardawil Ben-Amotz & Avron accumulates high concentrations of β-carotene when grown under high light intensity. The β-carotene is composed mainly of 9-cis and all-trans β-carotene. Accumulation of β-carotene and an increase in the ratio of the 9-cis to the all-trans isomer are strongly dependent on the light intensity under which the algae are cultivated but are independent of light quality within the photosynthetically active radiation range. Cells grown under continuous red (>645 nm) or white light of 500 W·m^?2 reach a value of about 32 pg β-carotene·cell^?1 and a ratio of 9-cis to all-trans β-carotene of around 2, whereas cells grown under low red or white light intensity of 25 W·m^?2 contain about 3 pg·cell^?1 and a ratio of isomers of around 0.3.