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PubMed2005

Isolation and characterization of a carotenoid oxygenase gene from Chlorella zofingiensis (Chlorophyta).

The green alga Chlorella zofingiensis produces large amounts of the valuable ketocarotenoid astaxanthin under dark, heterotrophic growth conditions, making it potentially employable for commercial production of astaxanthin as feed additives, colorants, and health products. Here, we report the identification and characterization of a beta-carotene oxygenase (CRTO) gene that is directly involved in the biosynthesis of ketocarotenoids in C. zofingiensis. The open reading frame of the crtO gene, which is interrupted by three introns of 243, 318, and 351 bp, respectively, encodes a polypeptide of 312 amino acid residues. Only one crtO gene was detected in the genome of C. zofingiensis. Furthermore, the expression of the crtO gene was transiently up-regulated upon glucose treatment. Functional complementation in Escherichia coli showed that the coding protein of the crtO gene not only exhibits normal CRTO activity by converting beta-carotene to canthaxanthin via echinenone, but also displays a high enzymatic activity of converting zeaxanthin to astaxanthin via adonixanthin. Based on the bifunctional CRTO, a predicted pathway for astaxanthin biosynthesis in C. zofingiensis is described, and the CRTO is termed as carotenoid 4,4'-beta-ionone ring oxygenase.

Blotting, Southern↗
PubMed2004

Regeneration and maturation of daughter cell walls in the autospore-forming green alga Chlorella vulgaris (Chlorophyta, Trebouxiophyceae).

Cell-wall synthesis in Chlorella vulgaris, an autospore-forming alga, was observed using the cell wall-specific fluorescent dye Fluostain I. The observation suggested two clearly distinguishable stages in cell-wall synthesis: moderate synthesis during the cell-growth process and rapid synthesis at the cell-division stage. We used electron microscopy to examine the structural changes that occurred with growth in the premature daughter cell wall during the cell-growth and cell-division phases. The cell began to synthesize a new daughter cell wall shortly after its release from the autosporangium. A very thin daughter cell wall, with a thickness of about 2 nm, was formed inside the mother cell wall and completely enveloped the outer surface of the plasma membrane of the cell. The daughter cell wall gradually increased in thickness from 2 to 3.8 nm. During the protoplast-division phase in the cell-division stage, the daughter cell wall expanded on the surface of the invaginating plasma membrane of the cleavage furrow, accompanied by active synthesis of the cell wall, which increased in thickness from 3.8 to 6.1 nm. The daughter cell matured into an autospore while completely enclosed by its own thickening (from 6.1 to 17 nm) wall. Finally, the released daughter cell was enclosed by its own cell wall after the mother cell wall burst. The daughter cell with mature wall thickness (17-21 nm) emerged as a small, but complete, autospore.

Cell Cycle↗
PubMed1990

Characterization of nuclear DNA in 12 species of Chlorella (Chlorococcales, Chlorophyta) by DNA reassociation.

Strains of 12 different species of the genus Chlorella were analyzed for amount, reiteration frequency and kinetic complexity of chromosomal DNA components by C0t analysis. The resulting C0t curves reveal at least two different DNA components consisting of single copy DNA (up to 95%) and of repetitive DNA with complexities of 4.1 x 10(3) base pairs (bp) to approximately 11.7 x 10(3) bp and a reiteration frequency of 100-760. The total amount of repetitive DNA is less than 9% of the nuclear genome and similar in all strains studied. In contrast, the total kinetic complexity varies in a wide range from 1.26 x 10(7) bp to 8.08 x 10(7) bp which is mainly due to differences in the size of single copy DNA. The genome sizes in Chlorella seem not to be correlated with biochemical and physiological characteristics and therefore are unlikely to be useful as a taxonomical marker. A comparison of thermal denaturation profiles showed that the melting points of repetitive and single copy DNA differ by approximately 7 degrees C which may result from base mismatch and/or from a distinct base composition of the repetitive DNA.

Cell Nucleus↗
PubMed2005

A novel marine algal toxicity bioassay based on sporulation inhibition in the green macroalga Ulva pertusa (Chlorophyta).

A 5-day aquatic toxicity test based on sporulation inhibition of the green macroalga Ulva pertusa Kjellman has been developed. Optimal test conditions determined for photon irradiance, salinity and temperature were 60-200 micromol photons m(-2)s(-1), 25-35 per thousand and 15-20 degrees C, respectively. Tests were conducted by exposing U. pertusa thallus disks to a reference toxicant (sodium dodecyl sulfate; SDS), metals (Cd2+, Cu2+, Zn2+, Pb2+) and elutriates of sludge collected from nine different locations. The EC(50) values for SDS was 5.35 mg L(-1). When four heavy metals were assayed, the NOECs were highest for lead (0.625 mg L(-1)) and lowest for copper (0.031 mg L(-1)). The EC50 values showed the following toxicity rankings: Cu(2+) (0.061 mg L(-1))>Cd2+ (0.326 mg L(-1))>Zn2+ (0.738 mg L(-1))>Pb2+ (0.877 mg L(-1)). The bioassay indicated also that the sporulation endpoint could be a sensitive indicator of toxicity effects of elutriates of sludge as reflected from the NOEC values equal to or lower than the lowest concentration employed (6.25%). Sporulation was significantly inhibitied in all elutriates with the greatest and least effects observed in elutriates of sludge from industrial waste (EC50 6.78%) and filtration bed (EC50 15.0%), respectively. The results of the Spearman rank correlation analysis for EC50 data versus the concentrations of toxicants in the sludge presented a significant correlation between toxicity and four heavy metals (Cd2+, Cu2+, Pb2+, Zn2+). Introduction of the concept of toxicity unit (TU) showed that these metals were the main cause of toxicity in elutriates of at least four out of nine sludge samples. Members of the order Ulvales show a wide geographic distribution and have similar reproductive characteristics, thus making it possible to apply the present test method to other algae of this taxa, elsewhere. This novel method will be a useful tool for assessing the aquatic toxicity of a wide range of toxicants, once the respective sensitivities are demonstrated.

Analysis of Variance↗
PubMed2005

Tolerance to cadmium in Chlamydomonas sp. (Chlorophyta) strains isolated from an extreme acidic environment, the Tinto River (SW, Spain).

The effects of selected concentrations of Cd on the growth and ultrastructure of three strains of Chlamydomonas sp. isolated from a highly acidic river, Río Tinto (SW Spain) were examined. The river is characterized by its extreme physico-chemical conditions in terms of low pH, mean 2.2 and high concentrations of heavy metals. Growth, Cd accumulation, chlorophyll a, influence of Fe in Cd toxicity and ultrastructural localization were determined. The strains were cultured in both, artificial chemically defined media as well as in natural water from the river. Since iron is the main component of the river water, the effect of different concentrations of this element in relation with Cd toxicity was also analysed. The three strains analysed showed comparable growth and ultrastructural changes. Cd concentration corresponding to 50% growth inhibition (EC50) was 0.2 mM when cells were grown in artificial media. When cells were grown in natural water, no significant differences were found between the controls and the Cd supplemented media even at the highest concentration of 0.8 mM. At an inhibitory level of 0.1 mM of Cd, increasing the concentration of iron up to 90 or 180 mM resulted in a dramatic recovery in algal growth rates in artificial media, reaching normal growth curves. The accumulation of Cd depended on dose and time in the artificial media. The maximal accumulation of Cd was reached after 3 days for all Cd doses, and remained almost unchanged in the subsequent period of time. Chlorophyll a amount depended on dose but not on time in the artificial growth media. At the ultrastructural level, an increase in the periplasmalemmal space was observed due to the presence of a large number of vacuoles, together with a decrease in the relative volume of the nucleus when the cells were incubated in the presence of Cd. Pyrenoid and starch granules were observed and accumulation of spherical electron-dense bodies were also detected. X-ray spectra of these bodies for cells growing in artificial acid media showed intense C, O and Cd signals. In addition to these peaks, a strong Fe signal was also observed when cells were grown in natural acidic water.

Analysis of Variance↗
PubMed2006

In vitro antioxidant activity of acetylated and benzoylated derivatives of polysaccharide extracted from Ulva pertusa (Chlorophyta).

The antioxidant activity of natural ulvan and its derivatives (acetylated and benzoylated ulvans) in vitro was determined, including scavenging activity against superoxide and hydroxyl radicals, reducing power, and chelating ability. Obvious differences in antioxidant activity between natural ulvan and its derivatives were observed, moreover, the antioxidant activity of acetylated and benzoylated ulvans was stronger than that of natural ulvan.

Acetylation↗
PubMed2005

First assessment of the Caulerpa racemosa (Caulerpales, Chlorophyta) invasion along the French Mediterranean coast.

The introduced green alga Caulerpa racemosa var. cylindracea has been rapidly spreading in the Mediterranean Sea since 1990. It was first observed in France in 1997 (Marseilles). In early 2004, the stretch of the French Mediterranean coastline and the surface area affected by the invasion were estimated at about 83 km and 4014 ha, respectively. The depth range of colonized areas was usually 10-35 m depth. Shallow (0-10 m) and deep (down to 40 m) dense meadows were rarely observed. In contrast to the dead matte of Posidonia oceanica, which constituted the most widely colonized substratum, dense P. oceanica meadows and fine sand with large ripple-marks were not invaded. Few rocky areas were colonized and coarse sand bottoms were usually colonized below 20 m depth. All the colonized areas were exposed to human activities and more than 40% were fishing areas. Mild climate, suitable substrata, presence of vectors of dispersal and absence of efficient biological control make the French Mediterranean coast particularly vulnerable to the further spread of the alga.

Caulerpa↗
PubMed2003

Phylogeny of the non-photosynthetic green micro-algal genus Prototheca (Trebouxiophyceae, Chlorophyta) and related taxa inferred from SSU and LSU ribosomal DNA partial sequence data.

All five species in the heterotrophic micro-algal genus Prototheca and their relatives were compared for the extent of nucleotide divergence in the nuclear small-subunit (SSU) and in the 5' end of large-subunit (LSU) ribosomal RNA genes (rDNAs). Phylogenetic analysis based on combined SSU and LSU rDNA sequence alignment was implemented with the neighbor-joining, the maximum-parsimony, and the maximum-likelihood methods. The relationships among the species of Prototheca based on this data set were largely concordant with those inferred from SSU or LSU rDNA sequences alone. The obtained phylogenetic trees indicated that P. stagnora and P. ulmea should be regarded as different species and that both of the species as well as P. moriformis were placed in a cluster represented by P. zopfii, whereas P. wickerhamii was not directly grouped together with the other members of Prototheca and was more closely related to the autotrophic alga Auxenochlorella protothecoides. Therefore, the genus Prototheca is paraphyletic in its present circumscription; and these conclusions lead us to propose the transfer of P. wickerhamii to Auxenochlorella or to a new genus. On the basis of nucleotide sequence similarities, unlike SSU rDNA, the LSU rDNA region examined in this study appeared to be variable in recognizing a heterogeneity within a single species P. zopfii, which had been shown earlier in a chemotaxonomic study.

DNA, Ribosomal↗
PubMed2003

Antihyperlipidemic effects of different molecular weight sulfated polysaccharides from Ulva pertusa (Chlorophyta).

Ulvan, a sulfated polysaccharide from Ulva pertusa, was degraded to yield two low molecular weight fractions U1 and U2. The molecular weights of ulvan and its fractions were determined and varied from 151.6 to 28.2 kDa. They were fed to rats on a hypercholesterolemic diet for 21 days to evaluate and compare the antihyperlipidemic actions. Ulvan-based diet significantly lowered the levels of serum total cholesterol (-45.2%, P<0.05) and low density lipoprotein cholesterol (LDL-cholesterol, -54.1%, P<0.05). While U1- and U2-based diets significantly elevated the levels of serum high density lipoprotein cholesterol (HDL-cholesterol, +22.0% for U1, not significant; +61.0% for U2; P<0.05) and reduced triglyceride (TG, -82.4% for U1, -77.7% for U2; P<0.05) in rats as compared to control diet. In addition, consumptions of various ulvans significantly increased fecal bile acid excrement. The results indicated that ulvans with different molecular weights exhibited diverse effects on lipid metabolism. The high molecular weight ulvan was effective in serum total and LDL-cholesterol, whereas low molecular weight fractions were in TG and HDL-cholesterol. The fractions were considered to be more beneficial to hyperlipidemia associated with diabetes over ulvan.

Adipose Tissue↗
PubMed2001

Molecular phylogeny and taxonomic revision of Chlamydomonas (Chlorophyta). I. Emendation of Chlamydomonas Ehrenberg and Chloromonas Gobi, and description of Oogamochlamys gen. nov. and Lobochlamys gen. nov.

The genus Chlamydomonas (including Chloromonas) is one of the largest green algal genera comprising more than 600 species. To initiate a comprehensive analysis of the phylogeny and systematics of the genus, we determined nuclear-encoded SSU rRNA sequences from 32 strains of Chlamydomonas, Chloromonas and Chlorogonium with emphasis on oogamous taxa and related strains, and incorporated these into global molecular phylogenetic analyses of 132 strains of Chlorophyceae. In addition, we studied the morphology and reproduction of oogamous and related strains by light microscopy. We recognize and designate 18 monophyletic lineages (clades) within the Chlorophyceae, 11 of which are confined to the CW (basal bodies displaced clockwise) subgroup. The majority of clades recognized within the Chlorophyceae do not correspond to any of the traditional classification systems, which are still largely based on the organization level. Strains assigned to Chlamydomonas and Chloromonas were found in seven different clades confirming the polyphyly of the two genera as presently conceived. To initiate the taxonomic revision of Chlamydomonas, C. reinhardtii is proposed as the conserved type of the genus. In consequence, species in clades other than the clade containing C. reinhardtii must be transferred to other genera, a process initiated in this contribution. The oogamous strains studied represent a monophyletic lineage, which is described as Oogamochlamys gen. nov. comprising three species (O. gigantea, O. zimbabwiensis and O. ettlii spec. nov.). The sister clade to Oogamochlamys consists of isogamous strains characterized by chloroplasts with incisions and is described as Lobochlamys gen. nov. with two species (L. culleus and L. segnis). Another clade is characterized by asteroid or perforated, parietal chloroplasts and contains the type species of Chloromonas (C. reticulata). Thus, the polyphyletic Chloromonas (traditionally defined as "Chlamydomonas without pyrenoids") can be legitimized as a monophyletic genus by restriction to this clade and is here emended on the basis of chloroplast characters (the clade contains strains with or without pyrenoids thus rejecting the character "absence of pyrenoids").

Animals↗
PubMed2004

Isolation of a lactic acid bacterium and yeast consortium from a fermented material of Ulva spp. (Chlorophyta).

AIMS: Microbiota in a fermented culture of Ulva spp. was examined with the objective to characterize the type of fermentation and to obtain starter microbes for performing seaweed fermentation. METHOD AND RESULTS: Fermented Ulva spp. cultures which were obtained and transferred in a laboratory were examined for their microbiota. With phenotypic characterization and phylogenetic analysis based on rRNA gene nucleotide sequences, the predominant micro-organisms were identified as Lactobacillus brevis, Debaryomyces hanseni var. hansenii, and a Candida zeylanoides-related specimen, suggesting that the observed fermentation can be categorized to lactic acid and ethanol fermentation. Inoculating the individually cultured cell suspensions of the three kinds of micro-organisms with cellulase induced the fermentation in various kinds of seaweed. CONCLUSIONS: A microbial consortium composed of a lactic acid bacterium, L. brevis, and yeasts, D. hansenii and a C. zeylanoides-related specimen, were predominant in a fermented culture of Ulva spp. Lactic acid and ethanol fermentation could be induced in various kinds of seaweed by adding this microbial consortium along with cellulase. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of lactic acid and ethanol fermentation in seaweed, which is expected to provide a new material for food and dietary applications.

Candida↗
PubMed2004

Phylogeographical structure, distribution and genetic variation of the green algae Ulva intestinalis and U. compressa (Chlorophyta) in the Baltic Sea area.

The marine algae Ulva intestinalis and U. compressa are morphologically plastic with many overlapping characters and are therefore difficult to distinguish from each other. The present distribution of U. intestinalis and U. compressa is investigated along the salinity gradient in the Baltic Sea area through analyses of internal transcribed spacer (ITS) sequence data. Also, the amount and distribution of intraspecific genetic polymorphism in the ITS region is studied allowing inferences on the phylogeographical pattern and postglacial recolonization of the Baltic Sea area. The data show that of the two species only U. intestinalis occurs in the Baltic Sea. The distribution of U. compressa is more restricted than previously reported, and it was not found in salinities lower than 15 ppt. All of Scandinavia and the Baltic Sea were covered with ice during the last ice age and the organisms in the Baltic Sea must have colonized the area after the ice had started to melt. The genetic diversity of U. intestinalis and U. compressa in the Baltic Sea and the neighbouring area was found to be reduced compared to that in the British Isles. This reduction may be the result of either a historical reduction of diversity or an adaptation of specific clones to the northern environmental conditions.

Base Sequence↗
PubMed2003

Karyotypic analysis of two algae species Scenedesmus incrassatulus Bohl and Scenedesmus antennatus Bréb (Chlorophyta, Chlorococcales).

The karyotypes (number, morphology and size of the chromosomes) of two algae species of Scenedesmus genus, S. incrassatulus and S. antennatus, were studied. The karyotype of S. incrassatulus (n=4) was asymmetric, characterized by two large metacentric, one large submetacentric and one small metacentric chromosomes. The karyotype assembly of S. antennatus (n=6) reveals two metacentrics and four submetacentrics. This karyotype was symmetric. The general chromosomal formulae of both species, as well as the total average metaphase length of their haploid set are presented. The results of chromosomal studies of other related species are compared and discussed. Data from the karyotypic analysis showed that S. incrassatulus, S. antennatus and S. obliquus are separate biological species from taxonomical point of view.

Chromosome Banding↗
PubMed2026

Genome-scale insights into metabolic streamlining and photosynthetic energy balance in the extremophile green alga Picocystis salinarum (Picocystophyceae, Chlorophyta).

Picocystis salinarum is an early-diverging chlorophyte and the sole described member of the Picocystophyceae, frequently dominating hypersaline and alkaline lakes despite extreme physicochemical constraints. To elucidate the genomic foundations of its ecological success, we generated a fully annotated, chromosome-scale nuclear genome assembly of the type strain originally isolated from a saline pond in San Francisco Bay. The 18.5-Mb genome comprises 30 chromosomal assemblies, exhibits clear diploidy, and contains multiple copies of intact Ty3/Gypsy and Ty1/Copia long terminal repeat retrotransposons encoding polyproteins with atypical accessory domains. Phylogenomic analyses reveal strong affinity with the Nephroselmidophyceae. Comparative analyses reveal extensive metabolic streamlining, including the absence of a queuosine salvage pathway, the 2-methylcitrate cycle, &#x3b2;-oxidation of propionate, and branched-chain amino acid catabolism, traits retained in several marine prasinophyte lineages. In contrast, the genome preserves multiple ancestral bacterial derived systems. Notably, P. salinarum features a complete chloroplast NADH dehydrogenase-like complex, including all membrane, electron binding, and assembly components, a configuration not previously reported in sequenced chlorophyte algae. This retention implies substantial capacity for cyclic electron flow and chlororespiration, processes expected to be critical in chronically low-light and chemically extreme environments. The genome further reveals a distinctive biochemical CO2-concentrating mechanism centered on plastid-targeted phosphoenolpyruvate carboxykinase, complete plastid peptidoglycan biosynthetic and remodeling pathways, and partial retention of lipid-A-related machinery. Conversely, P. salinarum lacks canonical non-photochemical quenching proteins while retaining xanthophyll-cycle enzymes that support slower photoprotective responses. Together, these features define a coordinated genomic architecture that underpins the specialization of P. salinarum to hypersaline, alkaline, and persistently low-light ecosystems.

3&#x2010;deoxy&#x2010;D&#x2010;manno&#x2010;octulo↗
PubMed1995

Behavior of mitochondria in synchronized cells of Chlamydomonas reinhardtii (Chlorophyta).

Cells of Chlamydomonas reinhardtii Dangeard were synchronized under a 12 hour:12 hour light:dark regimen. Behavior of mitochondria in these cells was studied by fluorescence microscopy using a mitochondrial membrane-binding fluorescent dye, dimethylaminostyrylmethyl-pyridiniumiodine (DASMPI), as well as by electron microscopy. Following time courses of change in frequency of occurrence of five typical morphologies of mitochondria in synchronized cells, strikingly dynamic behavior of mitochondria was demonstrated. The five types are (A) a giant global mitochondrion with large matrix and peripherally localized cristae, a part of which is in close contact with the nucleus, (B) a mitochondrion composed of thick-corded bodies connected to each other, a part of which is in contact with the nucleus, (C) thin-corded forms with a few branches, (D) small lump forms scattered in the cytoplasm, and (E) stringy forms with intricate branchings extended throughout the cytoplasm. During the early half of the light period, changes of C-->B-->C-->D occur, while the inversely sequential changes of D-->C-->B-->C proceed during the later half of the light period. The appearance of the B-type mitochondrion is accompanied by a transient decrease of O2-uptake activity of cells. The early appearing B-type mitochondrion is temporarily turned into a giant A-type mitochondrion, concomitant with discharge of membranes into the cytoplasm and their retake by the A-type form in the process of reversion to B-type. In the reversion process, partitioning membranes are also formed in the large matrix of A-type mitochondrion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Isolation, growth, ultrastructure, and metal tolerance of the green alga, Chlamydomonas acidophila (Chlorophyta).

An acidophilic volvocine flagellate, Chlamydomonas acidophila (Volvocales) that was isolated from an acid lake, Katanuma, in Miyagi prefecture, Japan was studied for growth, ultrastructural characterization, and metal tolerance. Chlamydomonas acidophila is obligately photoautotrophic, and did not grow in the cultures containing acetate or citrate even in the light. The optimum pH for growth was 3.5-4.5. To characterize metal tolerance, the toxic effects of Cd, Co, Cu, and Zn on this alga were also studied. Effective metal concentrations, which limited the growth by 50%, EC50 were measured, after 72 h of static exposure. EC50s were 14.4 microM Cd2+, 81.3 microM Co2+, 141 microM Cu2+, and 1.16 mM Zn2+ for 72 h of exposure. Thus, this alga had stronger tolerance to these metals than other species in the genus Chlamydomonas.

Adaptation, Physiological↗
PubMed2004

Functional sulfur amino acid production and seawater remediation system by sterile Ulva sp. (Chlorophyta).

Sterile Ulva, which is a macroalga, has the potential to grow stably; therefore, this seaweed is expected to be an efficient resource of functional food containing various nutrients such as sulfur amino acids, proteins, carbohydrates, and minerals. Ulva latuca was selected from the "Marine Park" in Tokyo Bay, and its growth rate (g-dry/[m2.d]) was measured using model reactors located on the land or on the surface of the sea at Yokohama. The growth rate of U. lactuca was recorded to be approx 20 g-dry/(m2.d), which is estimated to be 10 times greater than that in a natural field in the Marine Park. In addition, this growth rate was higher than that of conventional crops such as corn and rice on a farm or paddy. These data led us to newly design and propose a floating type of labor-efficient U. lactuca production system. d-Cysteinolic acid, which is included in U. lactuca as a major sulfur amino acid, inhibited the Fenton reaction, resulting in suppression of hydroxyl radical production and singlet oxygen. Addition of the sulfur amino acid (1 microM) to HepG2 cells markedly decreased the intracellular triglyceride level. Hence, this proposed facility also has the potential for industrial production of a valuable resource for the primary prevention of lifestyle-related diseases using enriched or eutrophied seawater.

Amino Acids, Sulfur↗