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PubMed2000

Light-harvesting complex II pigments and proteins in association with Cbr, a homolog of higher-plant early light-inducible proteins in the unicellular green alga Dunaliella.

Like higher plants, unicellular green algae of the genus Dunaliella respond to light stress by enhanced de-epoxidation of violaxanthin and accumulation of Cbr, a protein homologous to early light-inducible proteins (Elips) in plants. Earlier studies indicated that Cbr was associated with the light-harvesting complex of photosystem II (LHCII) and suggested it acted as a zeaxanthin-binding protein and fulfilled a photo-protective function (Levy et al. 1993, J. Biol. Chem. 268: 20892-20896). To characterize the protein-pigment subcomplexes containing Cbr in greater detail than attained so far, thylakoid membranes from Dunaliella salina grown in high light or normal light were solubilized with dodecyl maltoside and fractionated by isoelectric-focusing. Analysis of the resolved LHCII subcomplexes indicated preferred associations among the four LHCIIb polypeptides and between them and Cbr: subcomplexes including Cbr contained one or two of the more acidic of the four LHCIIb polypeptides as well as large amounts of lutein and zeaxanthin relative to chlorophyll a/b. After sucrose gradient centrifugation, Cbr free of LHCIIb polypeptides was detected together with released pigments; this Cbr possibly originated in subcomplexes dissociated in the course of the analysis. These results agree with the conclusion that Cbr is part of the network of LHCIIb protein-pigment complexes and suggest that the role played by Cbr involves the organization and/or stabilization of assemblies highly enriched in zeaxanthin and lutein. Such assemblies may function to protect PSII from photodamage due to overexcitation.

Centrifugation, Density Gradient↗
PubMed2002

The effect of physical and chemical treatment on the mechanical properties of the cell wall of the alga Chara corallina.

Single large internode cells of the charophyte (giant alga) Chara corallina were dissected to give sheets of cell wall, which were then notched and their mechanical properties in tension determined. The cells were subjected to a thermal treatment in excess water (cf. cooking), which had little effect on strength but increased the stiffness, contrasting with the behaviour of higher-plant tissues. Extraction in CDTA (cyclohexane-trans-1,2-diamine-N,N,N',N'-tetraacetate) or 4 M KOH reduced the strength from 17 MPa to 10 MPa, although sequential extraction in CDTA and 4 M KOH reduced the strength further to 4 MPa. The stiffness decreased from 500 MPa to 300 MPa on extraction in CDTA or 4 M KOH, while falling to 70 MPa after extraction in CDTA followed by 4 M KOH. Conventional sequential extraction in CDTA, Na2CO3 at 1 degrees C and 20 degrees C, and KOH at 0.5 M, 1 M, 2 M and 4 M caused a gradual decrease in stiffness and strength after the CDTA treatment to the same lower values. This result is in keeping with mechanical properties for plant tissues, but in contrast to the removal of pectic polysaccharides from model cell wall systems, which does not reduce the stiffness.

Algorithms↗
PubMed2001

Potassium drives daily reversible thallus enlargement in the marine red alga Porphyra leucosticta (Rhodophyta).

Growth rate in terms of area expansion per 30 min was measured in the marine red algae Porphyra leucosticta under light/dark cycles of 6:6 h. Thalli grown in artificial seawater (ASW), under controlled ionic concentrations, showed a rapid thallus expansion just after light-on (morning-peak). Dark phase began with a significant thallus contraction (dark-peak) but no growth was observed until the next light phase. The removal of K+ from the medium inhibited the reversible growth peak that this species shows after light-on. On the other hand, the removal of Na+ did not have an apparent effect on growth pattern. Addition of Rb+ to K+-free ASW restored the morning-peak to 60% of its value in ASW, but addition of Li+ failed to restore the morning-peak. Intracellular ion analyses revealed that after light-on, the internal K+ content of the cells of this species increased 4-fold in 90 min, reaching an intracellular concentration of up to 300 micromol K+ per gram fresh weight, and that this value remained fairly constant over the light phase. Addition of 100 mM of tetraethylammonium, a specific K+-channel blocker, inhibited the morning-peak by 40%. The Na+ and Cl- contents in the cells increased rapidly in the first 45 min of the light phase, but then the internal concentrations of both ions decreased to their minimum values in the light phase. The carbonic anhydrase inhibitors acetazolamide and 6-ethoxyzolamide did not affect thallus expansion during the light phase. In contrast, thallus expansion in the light phase was completely inhibited by diethylstilbestrol, an inhibitor of the putative primary pump, and by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU), an inhibitor of photosynthesis, but not by sodium vanadate, a specific inhibitor of the plasma-membrane H+-ATPase. We suggest that the periodic oscillation of the short-term growth rate of P. leucosticta occurs in response to potassium fluxes, which control the osmotic pressure and eventually the relative cell volume. The possible effects of the loosening of the cell wall and the internal K+ concentration on the growth of this species are also discussed.

Acetazolamide↗
PubMed2002

The genes encoding subunits of ATP synthase are conserved in the reduced plastid genome of the heterotrophic alga Prototheca wickerhamii.

The heterotrophic unicellular alga Prototheca wickerhamii is closely related to the photoautotrophic Chlorella vulgaris but has a 54,100-bp plastid DNA (ptDNA) that is much smaller than the chloroplast DNA of C. vulgaris (150,613 bp). The nucleotide sequence of 28,093 bp of the Prototheca ptDNA has been determined. No genes for photosynthetic functions have been found, except for sequences encoding six subunits of the ATP synthase ( atpA, atpB, atpE, atpF, atpH, and atpI). Transcripts of these atp genes have also been detected. Whether the leucoplasts of Prototheca contain a functional ATP synthase has still to be elucidated. Identified genes further include tufA, minD, cysT, and genes coding for three rRNAs, 22 tRNAs, and 12 ribosomal proteins. The results support the idea that, in the reduced plastid genome of Prototheca, genes coding for components of the plastid translational apparatus have been preferentially retained, and might be needed for the expression of the atp genes and some unassigned ORFs.

Chloroplast Proton-Translocating ATPases↗
PubMed2006

Distinctive architecture of the chloroplast genome in the chlorophycean green alga Stigeoclonium helveticum.

The chloroplast genome has experienced many architectural changes during the evolution of chlorophyte green algae, with the class Chlorophyceae displaying the lowest degree of ancestral traits. We have previously shown that the completely sequenced chloroplast DNAs (cpDNAs) of Chamydomonas reinhardtii (Chlamydomonadales) and Scenedesmus obliquus (Sphaeropleales) are highly scrambled in gene order relative to one another. Here, we report the complete cpDNA sequence of Stigeoclonium helveticum (Chaetophorales), a member of a third chlorophycean lineage. This genome, which encodes 97 genes and contains 21 introns (including four putatively trans-spliced group II introns inserted at novel sites), is remarkably rich in derived features and extremely rearranged relative to its chlorophycean counterparts. At 223,902 bp, Stigeoclonium cpDNA is the largest chloroplast genome sequenced thus far, and in contrast to those of Chlamydomonas and Scenedesmus, features no large inverted repeat. Interestingly, the pattern of gene distribution between the DNA strands and the bias in base composition along each strand suggest that the Stigeoclonium genome replicates bidirectionally from a single origin. Unlike most known trans-spliced group II introns, those of Stigeoclonium exhibit breaks in domains I and II. By placing our comparative genome analyses in a phylogenetic framework, we inferred an evolutionary scenario of the mutational events that led to changes in genome architecture in the Chlorophyceae.

Base Sequence↗
PubMed2006

The family of DOF transcription factors: from green unicellular algae to vascular plants.

This article deals with the origin and evolution of the DOF transcription factor family through a phylogenetic analysis of those DOF sequences identified from a variety of representative organisms from different taxonomic groups: the green unicellular alga Chlamydomonas reinhardtii, the moss Physcomitrella patens, the fern Selaginella moellendorffii, the gymnosperm Pinus taeda, the dicotyledoneous Arabidopsis thaliana and the monocotyledoneous angiosperms Oryza sativa and Hordeum vulgare. In barley, we have identified 26 different DOF genes by sequence analyses of clones isolated from the screening of genomic libraries and of ESTs, whereas a single DOF gene was identified by bioinformatics searches in the Chlamydomonas genome. The phylogenetic analysis groups all these genes into six major clusters of orthologs originated from a primary basal grade. Our results suggest that duplications of an ancestral DOF, probably formed in the photosynthetic eukaryotic ancestor, followed by subsequent neo-, sub-functionalization and pseudogenization processes would have triggered the expansion of the DOF family. Loss, acquisition and shuffling of conserved motifs among the new DOFs likely underlie the mechanism of formation of the distinct subfamilies.

Algal Proteins↗
PubMed2000

A putative mitochondrial ftsZ gene is present in the unicellular primitive red alga Cyanidioschyzon merolae.

Two ftsZ homologues were isolated from the unicellular primitive red alga Cyanidioschyzon merolae (CmftsZ1 and CmftsZ2). Phylogenetic analysis revealed that CmftsZ1 is most closely related to the ftsZ genes of alpha-Proteobacteria, suggesting that it is a mitochondrial-type ftsZ gene, whereas CmftsZ2 is most closely related to the ftsZ genes of cyanobacteria, suggesting that it is a plastid-type ftsZ gene. Southern analysis indicates that CmftsZ1 and CmftsZ2 are both single-copy genes located on chromosome XIV in the C. merolae genome. Northern analysis revealed that both CmftsZ1 and CmftsZ2 are transcribed, and accumulate specifically before cell and organelle division. The results of Western analysis suggest that CmFtsZ1 is localized in mitochondria.

Amino Acid Sequence↗
PubMed1997

A novel homologue of the prokaryotic htrA gene is differentially expressed in the alga Haematococcus pluvialis following stress.

The alga Haematococcus pluvialis is able to respond to environmental stress by changing from the motile, vegetative green cell form to red stationary aplanospores. This differentiation program is accompanied by dramatic morphological changes that must result, at least in part, from differential gene expression. To begin to identify genes that are differentially expressed as a response to stress, we applied the differential display technique to identify and isolate differentially expressed RNA sequences. Here we report on the isolation and characterization of one such RNA sequence that codes for Haematococcus htrA, a member of a heat shock serine protease family previously described only in prokaryotes. Interestingly, database searches of mouse and human cDNA sequences showed that a previously unreported homologue is also found in a number of tissues. htrA mRNA was not detectable in vegetative cells, but was found at high levels in developing aplanospores. Evidence presented suggests that RNA transcripts encoding this protein are differentially spliced, and that the different splice products are differentially expressed during the developmental process. These experiments provide the groundwork upon which the developmental program of H. pluvialis can be investigated. In addition, they indicate that the htrA family of heat shock serine proteases may play an important role in stress response in higher organisms as well as in bacteria.

Amino Acid Sequence↗
PubMed2001

The flanking regions of PsaD drive efficient gene expression in the nucleus of the green alga Chlamydomonas reinhardtii.

The nuclear gene PsaD encodes an abundant chloroplast protein located on the stromal side of the Photosystem I complex. We have cloned and sequenced a genomic fragment containing the PsaD gene from the green alga Chlamydomonas reinhardtii. Sequence comparison with its cDNA revealed that the PsaD ORF contains no introns. Thus, the regulatory sequences required for high-level expression of PsaD must lie in the flanking promoter and untranslated regions. We used this genomic fragment to construct a vector that allows for high-level expression of endogenous and exogenous genes, as well as cDNAs that could not be expressed from existing vectors. It is also possible to use the PsaD transit sequence to target the expressed protein to the chloroplast compartment.

Animals↗
PubMed2001

ATP citrate lyase in the glaucocystophyte alga Cyanophora paradoxa is a cytosolic enzyme: characterisation of the gene for the large subunit at the cDNA and genomic levels.

The single-copy gene for the large subunit of ATP citrate lyase (ACL) from the alga Cyanophora paradoxa was characterized at the cDNA and genome levels. The gene product showed high sequence similarity to its mammalian counterparts, but is smaller in size, as is also found for the fungal subunit Acl1 and, most probably, for the corresponding subunit from Arabidopsis thaliana. The C. paradoxa gene is interrupted by at least 12 introns of 53-65 bp with conserved border and branchpoint sequences, and the product lacks a stroma-targeting peptide. Enzyme activity was found in the cytosol of C. paradoxa but not in the plastid (cyanelle) fraction. This is in contrast to the subcellular distribution of ATP citrate lyases in higher plants, where both chloroplast and cytosolic enzymes have been reported.

ATP Citrate (pro-S)-Lyase↗
PubMed2004

Spatial associations among algae affect host use in a herbivorous marine amphipod.

Host plant choice by herbivores may be constrained by the spatial arrangement of plants with selection of poor quality hosts being more likely if preferred hosts are locally rare. I tested whether the presence of a herbivorous marine amphipod, Peramphithoe parmerong, on a poor quality host could be explained by the relative abundance and spatial arrangement of alternative brown algal hosts. Amphipods strongly preferred the high quality Sargassum linearifolium over the poor quality Padina crassa in laboratory assays with a higher tendency to select Padina crassa when it was relatively more abundant. In the field, however, adults colonised both hosts to equal densities. Rates of colonisation were independent of host quality but strongly affected by the identity of neighbouring hosts with those algae in close proximity to S. linearifolium receiving far greater densities of amphipods than those surrounded by Padina crassa. There was no evidence that the presence of adult Peramphithoe parmerong on Padina crassa was due to local scarcity of S. linearifolium. In contrast, the distribution and behaviour of juveniles were predictable from differences in food quality, with most inhabiting the high quality S. linearifolium and few emigrating from this host. The success of a simple adaptive model based on food quality to predict juvenile, but not adult, behaviour indicates that the relative importance of food quality varies with age, and that other factors are important determinants of adult movements among hosts.

Adaptation, Physiological↗
PubMed2006

Dopamine functions as an antiherbivore defense in the temperate green alga Ulvaria obscura.

On northeastern Pacific coasts, Ulvaria obscura is a dominant component of subtidal "green tide" blooms, which can be harmful to marine communities, fisheries, and aquaculture facilities. U. obscura is avoided by herbivores relative to many other locally common macrophytes, which may contribute to its ability to form persistent blooms. We used a bioassay-guided fractionation method to experimentally determine the cause of reduced feeding on Ulvaria by echinoderms, molluscs, and arthropods. Our results indicated that dopamine, which constituted an average of 4.4% of the alga's dry mass, was responsible for decreased feeding by sea urchins (Strongylocentrotus droebachiensis). Subsequent experiments demonstrated that dopamine also reduced the feeding rates of snails (Littorina sitkana) and isopods (Idotea wosnesenskii). Dopamine is a catecholamine that is a common neurotransmitter in animals. The catecholamines dopamine, epinephrine (adrenaline), and norepinephrine also occur in at least 44 families of higher plants. The functions of catecholamines in plants are less well known than in animals but are likely to be diverse and include both physiological and ecological roles. Our results are the first experimental demonstration of a plant or algal catecholamine functioning as a feeding deterrent. This novel use of dopamine by Ulvaria may contribute to the formation and persistence of harmful Ulvaria blooms in northeastern Pacific coastal waters.

Animals↗
PubMed2005

Nonlinear control for algae growth models in the chemostat.

This paper deals with output feedback control of phytoplanktonic algae growth models in the chemostat. The considered class of model is of variable yield type, meaning that the ratio between the environmental nutrient absorption rate and the cells' growth rate varies, which is different from classical bioprocesses assumptions. On the basis of weak qualitative hypotheses on the analytical expressions of the involved biological phenomena (which guarantee robustness of the procedure toward modeling uncertainties) we propose a nonlinear controller and prove its ability to globally stabilize such processes. Finally, we illustrate our approach with numerical simulations and show its benefits for biological laboratory experiments, especially for ensuring persistence of the culture facing classical experimental problems.

Algorithms↗
PubMed2002

Localization of p210-related proteins in green flagellates and analysis of flagellar assembly in the green alga Dunaliella bioculatawith monoclonal anti-p210.

Recently, p210 was identified as a component of the flagellar basal apparatus in the green flagellate Spermatozopsis similis. In a search for potential homologues to p210, isolated cytoskeletons of several green flagellates were probed with a monoclonal antibody, BAS4.13, against p210. In Western blots, cross-reacting bands in the molecular-mass range of 210 kDa were detected only in the quadriflagellate Spermatozopsis exsultans. As described earlier for S. similis, the flagellar transition region was decorated in Chlamydomonas reinhardtii and several other green flagellates, whereas in the marine alga Dunaliella bioculata the antigen was present in the proximal part of the axoneme. Double immunofluorescence of D. bioculata with an antitubulin antibody further revealed dotlike signals at sites where the probasal bodies are located. Since most of the antigen in D. bioculata was located in the axoneme, deflagellation offered a possibility to study the kinetics of its incorporation during flagellar regeneration. The antigen was only detected after a flagellum reached a length of 3-4 microm and its integration into the growing flagellar proceeded from proximal to distal. A similar delay in the incorporation of the antigen was also observed during flagellar assembly on new basal bodies during cell division. Thus, the antigen of BAS4.13 was incorporated late and from proximal to distal into the growing flagellum. We conclude that the pace and site by which individual proteins are integrated into the flagellum differ greatly.

Algal Proteins↗
PubMed2004

Cytokinesis in brown algae: studies of asymmetric division in fucoid zygotes.

The mechanism of cytokinesis was investigated during the first asymmetric division in fucoid zygotes. A plate of actin assembled midway between daughter nuclei where microtubules interdigitated and defined the cytokinetic plane. A membrane was then deposited in islands throughout the cytokinetic plane; the islands eventually fused into a continuous partition membrane and cell plate material was deposited in the intermembrane space. All of these structures matured from the center of the cell outward (centrifugal maturation). Pharmacological agents were used to investigate the roles of microtubules, actin, and secretion in cytokinesis. The findings indicate a mechanism of cytokinesis that may be unique to the brown algae.

Bridged Bicyclo Compounds, Heterocyclic↗
PubMed2006

Rhizoids and protonemata of characean algae: model cells for research on polarized growth and plant gravity sensing.

Gravitropically tip-growing rhizoids and protonemata of characean algae are well-established unicellular plant model systems for research on gravitropism. In recent years, considerable progress has been made in the understanding of the cellular and molecular mechanisms underlying gravity sensing and gravity-oriented growth. While in higher-plant statocytes the role of cytoskeletal elements, especially the actin cytoskeleton, in the mechanisms of gravity sensing is still enigmatic, there is clear evidence that in the characean cells actin is intimately involved in polarized growth, gravity sensing, and the gravitropic response mechanisms. The multiple functions of actin are orchestrated by a variety of actin-binding proteins which control actin polymerisation, regulate the dynamic remodelling of the actin filament architecture, and mediate the transport of vesicles and organelles. Actin and a steep gradient of cytoplasmic free calcium are crucial components of a feedback mechanism that controls polarized growth. Experiments performed in microgravity provided evidence that actomyosin is a key player for gravity sensing: it coordinates the position of statoliths and, upon a change in the cell's orientation, directs sedimenting statoliths to specific areas of the plasma membrane, where contact with membrane-bound gravisensor molecules elicits short gravitropic pathways. In rhizoids, gravitropic signalling leads to a local reduction of cytoplasmic free calcium and results in differential growth of the opposite subapical cell flanks. The negative gravitropic response of protonemata involves actin-dependent relocation of the calcium gradient and displacement of the centre of maximal growth towards the upper flank. On the basis of the results obtained from the gravitropic model cells, a similar fine-tuning function of the actomyosin system is discussed for the early steps of gravity sensing in higher-plant statocytes.

Actomyosin↗
PubMed2001

Purification and Characterization of Mannitol-l-Phosphatase in the Red Alga Caloglossa continua (Ceramiales, Rhodophyta).

Purification of mannitol-l-phosphatase, an enzyme catalyzing the final step of mannitol biosynthesis, was first achieved in the mannitol-accumulating red alga Caloglossa continua (Okamura) King et Puttock. The enzyme was shown to be a monomer, since gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave close values of apparent molecular weights of 28,500 and 30,200, respectively. The protein exhibited an isoelectric point of 4.8. The substrate specificity for mannitol-l-phosphate (MIP) was very high, and that for K(m)(MIP) was 0.41 mM. The catalytic activity was optimal at pH 7.4. The enzyme was activated by Mg(2+), but was strongly inhibited by Ca(2+), NaF, N-ethylmaleimide, and p-hydroxymercuribenzoic acid. Seawater levels of NaCl and physiological levels of mannitol also inhibited the activity by 50% or more. Changes in the concentrations of those ions and metabolites may regulate the biosynthesis of mannitol as an osmoregulant in vivo.

Journal Article↗
PubMed2002

Screening of red algae filaments as a potential alternative source of eicosapentaenoic acid.

Lipids were extracted by a supercritical fluid extraction method from 10 species of filamentous red algae obtained from culture collections and their fatty acid compositions were determined. The fatty acid profiles of the 10 species were similar. The major fatty acids were 16:0, 20:4 omega 6 and 20:5 omega 3 (eicosapentaenoic acid, EPA), which amounted to over 70% of the total fatty acids. The highest EPA content (29.8 mg/L), as a percentage of total fatty acids, was produced by Liagora boergesenii filaments, which has good potential for EPA mass production in pilot plants.

Journal Article↗