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Structural similarities between psbA genes from red and brown algae.

The single copy psbA genes from the multicellular red alga Antithamnion spec. and the brown alga Ectocarpus siliculosus have been cloned and sequenced and monocistronic transcripts have been detected. Both genes contain an insertion of 21 bp at the 3' end which was also found in cyanobacteria and which is absent in chloroplasts and the chlorophyll b-containing prochlorophyte Prochlorothrix hollandica. These findings are in agreement with the hypothesis of a polyphyletic origin of plastids. Plastids of red and brown algae appear to be closely related.

Amino Acid Sequence↗

The structural analysis of the mitochondrial SSUrRNA implies a close phylogenetic relationship between mitochondria from plants and from the heterotrophic alga Prototheca wickerhamii.

The gene for the mitochondrial small subunit rRNA (SSUrRNA) from the heterotrophic alga Prototheca wickerhamii has been isolated from a gene library of extranuclear DNA. Sequence and structural analyses allow the determination of a secondary structure model for this rRNA. In addition, several sequence motifs are present which are typically found in SSUrRNAs of various mitochondrial origins. Unexpectedly, the Prototheca RNA sequence has more features in common with mitochondrial SSUrRNAs from plants than with that from the green alga Chlamydomonas reinhardtii. The phylogenetic relationship between mitochondria from plants and algae is discussed.

Base Sequence↗

Chloroplast genome characterization in the red alga Griffithsia pacifica.

It has been suggested that cyanobacteria served as the ancestors for rhodophytic algae whose chloroplasts contain chlorophyll a and phycobilins, and that a rodophyte served as the plastid source for chromophytic plants that contain chlorophylls a and c. Although organellar DNA has been used to assess phylogenetic relatedness among terrestrial plants and green algae whose chloroplasts contain chlorophylls a and b, few data are presently available on the molecular profile of plastid DNA in chromophytes or rhodophytes. In this study the chloroplast genome of the rhodophytic, filamentous alga Griffithsia pacifica has been characterized. DNA was purified from isolated chloroplasts using protease k treatment and sodium dodecyl sulfate lysis followed by density centrifugation in Hoechst-33258 dye-CsCl gradients. Single and double restriction enzyme digests demonstrate that the DNA prepared from purified chloroplasts has a genome size of about 178 kilobase pairs (kb). A restriction map of this chloroplast genome demonstrates that it is circular and, unlike the chloroplast DNA (cpDNA) in most other plants, contains only a single ribosomal DNA operon. DNA was also purified from the mitochondria that co-isolated with chloroplasts. Mitochondrial DNA consists of molecules that range in size from 27 to 350 kb based on restriction endonuclease digestion and electron microscopic analysis.

Chloroplasts↗

Structure, expression, and phylogenetic relationships of a family of ypt genes encoding small G-proteins in the green alga Volvox carteri.

In addition to the previously described gene yptV1 encoding a small G-protein we have now identified and sequenced four more ras-related ypt genes (yptV2-yptV5) from the green alga Volvox carteri. The four new genes encode polypeptides consisting of 203 to 217 amino-acid residues that contain the typical sequence elements (GTP-binding domains, effector domain) of the ypt/rab subgroup of the Ras superfamily. Comparison of the derived amino-acid sequences from the V. carteri ypt gene products and their Ypt homologs from other species revealed similarity values ranging from 60% to 85%, whereas intraspecies similarities were found to approach only 55%. The coding sequences are interrupted by 5-7 introns of variable size (70-1000 nucleotides) occupying different positions in the genes. Reverse-transcribed samples of stage-specific RNAs were PCR-amplified with primers specific to yptV1, yptV3, yptV4, and yptV5 to determine if yptV transcription might be restricted to either cell type or to a specific stage of the life cycle. These experiments demonstrated that each of these genes is expressed throughout the entire Volvox life cycle and in both the somatic and the reproductive cells of the alga. The transcription start sites of yptV1 and yptV5 were mapped by primer extension. Expression of recombinant yptV cDNA in E. coli yielded recombinant proteins that bound GTP specifically, demonstrating a property which is typical for small G-proteins. The derived YptV polypeptide sequences were used to group them into four distinct classes of Ras-like proteins. These are the first proteins of the Ras superfamily to be identified in a green alga. We discuss the possible role of the YptV-proteins in the intracellular vesicle transport of Volvox.

Amino Acid Sequence↗

The role of vanadium in gree plants. II. Vanadium in green algae--two sites of action.

Cells of Chlorella pyrenoidosa, derived from vanadium free agar slants, respond with great sensitivity to microamounts of vanadium, added as NH4VO3 to autotrophic liquid cultures. Between 0.01 and 1 microgram V per litre nutrient medium (2-10(-10)-2-10(-8) g-at/1), the algae respond with a continuous incrase in dry weight. At higher V-concentrations, further enhancement in biomass is accompanied by a additional increase in chlorophyll content. Maximum V-effect on both parameters was found to be at 500 microgram V/1 (10(-5) G-AT/1). Dry weight as well as chlorophyll content of Chlorella are decreased by concentrations above 25 mg V/1; 100 mg V/1 (2-10(-3) g-at/1) stop growth and cause death of the cells. The toxic threshold for the V-content in the algae was determined to be at 150-200 microgram V/g (3-4-10(-6) g-at/g) dry weight. Two different pH-optima for a positive vanadium action on dry weight and chlorophyll biosynthesis were established, the first at pH 7, the other in the range pH 7.5--8. Two sites of vanadium action in green algae are discussed.

Chlorella↗

Requirement of low oxidation-reduction potential for photosynthesis in a blue-green alga (Phormidium sp.).

Photosynthesis in a Phormidium species which forms dense conical-shaped structures in thermal springs is strongly inhibited by aeration but is stimulated by sulfide and other agents (cysteine, thioglycolate, sulfite) which lower the oxidation-reduction potential. The compact structures which this alga forms in nature may restrict oxygen penetration from the enviroment so that the anaerobic or microaerophilic conditions necessary ofr photosynthesis can develop. The alga may be defective in a regulatory mechanism that controls the reoxidation of reduced pyridine nucleotides formed during photosynthesis. It is suggested that other mat-forming and benthic blue-green algae may also prefer anaerobib conditions for growth and photosynthesis.

Anaerobiosis↗

Structure of the Rubisco operon from the unicellular red alga Cyanidium caldarium: evidence for a polyphyletic origin of the plastids.

The genes for both subunits of ribulose-1,5-bisphosphate-carboxylase/oxygenase (Rubisco) were located on the plastid DNA (ptDNA) of the unicellular red alga Cyanidium caldarium. Both genes are organized together in an operon. The sequence homology of both genes to the corresponding genes from the unicellular red alga Porphyridium aerugineum is remarkably high, whereas homology to Rubisco genes from chloroplasts and two recent cyanobacteria is significantly lower. These data provide strong evidence for a polyphyletic origin of chloroplasts and rhodoplasts. In addition the genes for the small subunit of Rubisco (rbcS) from red algae show about 60% homology to rbcS genes from cryptophytes and chromophytes. Thus, homologies in the rbcS gene indicate a close phylogenetic relationship between rhodoplasts and the plastids of Chromophyta.

Amino Acid Sequence↗

Chemical composition, in vitro protein digestibility and in vitro available iron of blue green alga, Nostoc commune.

Blue-green alga, Nostoc commune, contained moderate amounts of protein and iron. Its in vitro protein digestibility was 43.50%. The soluble and ionic iron from the alga was extractable to some extent at pH 1.5 but was not detectable at pH 8.0. The digestion by protease did not affect the iron detection. Heat processing at 100 and 120 degrees C failed to increase the digestibility and the content of available iron. The dietary fiber in the alga may be responsible for low protein digestion and low iron availability.

Biological Availability↗

Toxicity of carbofuran to blue-green alga Nostoc muscorum.

Effect of commercial grade pesticide furadan (3% a.i. as carbofuran) was studied on the survival, growth and nitrogen fixation of blue-green alga Nostoc muscorum. The lower concentration of furadan i.e. 25 microgram/ml enhanced survival, growth and nitrogen fixation in the alga whereas these were gradually inhibited in higher concentrations (50--1000 microgram/ml) and the presence of more than 1200 microgram/ml was algicidal. The preliminary observations revelaed that pesticide is biodegraded by the alga.

Carbofuran↗

Detection of green algae (Chlorophyceae) for the diagnosis of drowning.

The plankton test (generally, diatom test) is one of the methods available to diagnose the cause of death of submerged bodies. The solubilization method using tissue solubilizer Soluene-350 was used in this study to detect not only diatoms but also green algae, based on the fact that the solubilizer does not digest the cell walls of green algae which are made from cellulose. Detection of green algae from organs of submerged cadavers is very informative to determine drowning in fresh water, and also in cases where only few diatoms are detected in the organs.

Acetone↗

Antimutagenic properties of fresh-water blue-green algae.

The antimutagenic properties of whole fresh-water blue-green algae Aphanisomenon flos-aquae, marketed under the commercial name "Alpha Sun" were tested using the Ames test. Simultaneous addition of both algae and Nitrovin (a mutagen) to the test medium did not reduce the mutagenic activity. On the other hand, addition of freeze-dried blue-green algae to the test medium 2-24 h before the application of mutagen reduced its mutagenic activity.

Antimutagenic Agents↗

Complete large subunit ribosomal RNA sequences from the heterokont algae Ochromonas danica, Nannochloropsis salina, and Tribonema aequale, and phylogenetic analysis.

The large subunit ribosomal RNA sequences from the heterokont algae Ochromonas danica, Nannochloropsis salina, and Tribonema aequale were determined. These sequences were combined with small subunit ribosomal RNA sequences in order to carry out a phylogenetic analysis based on neighbor-joining, maximum parsimony, and maximum likelihood methods. Our results indicate that heterokont fungi and heterokont algae each are monophyletic, and confirm that they together form a monophyletic group called "stramenopiles." Within the heterokont algae, the eustigmatophyte Nannochloropsis salina either clusters with the chrysophyte Ochromonas danica or forms a sister group to a cluster comprising the phaeophyte Scytosiphon lomentaria and the xanthophyte Tribonema aequale. The alveolates were identified as the closest relatives of the stramenopiles, but the exact order of divergence between the eukaryotic crown taxa could not be established with confidence.

Animals↗

Are red algae plants? A critical evaluation of three key molecular data sets.

Whether red algae are related to green plants has been debated for over a century. Features present due to their shared photosynthetic habit have been interpreted as support for an evolutionary sisterhood of the two groups but, until very recently, characters endogenous to the host cell have provided no reliable indication of such a relationship. In this investigation, we examine three molecular data sets that have provided key evidence of a possible relationship between green plants and red algae. Analyses of an expanded alignment of DNA-dependent RNA polymerase II largest subunit sequences indicate that their support for independent origins of rhodophytes and chlorophytes is not the result of long-branch attraction, as has been proposed elsewhere. Differences in the pol II C-terminal domain, an essential component of plant mRNA transcription, also suggest different host cell ancestors for the two groups. In contrast, concatenated sequences of two groups of mitochondrial genes, those encoding subunits of NADH-dehydrogenase as well as cytochrome c oxidase subunits plus apocytochrome B, appear to cluster red algal and green plant sequences together because both groups have evolved relatively slowly and share a super-abundance of ancestral positions. Finally, analyses of elongation factor 2 sequences demonstrate a strong phylogenetic signal favoring a rhodophyte/chlorophyte sister relationship, but that signal is restricted to a contiguous segment comprising approximately half of the EF2 gene. These results argue for great caution in the interpretation of phylogenetic analyses of ancient evolutionary events but, in combination, indicate that there is no emerging consensus from molecular data supporting a sister relationship between red algae and green plants.

Apoproteins↗

The development of marine Toxicity Identification Evaluation (TIE) procedures using the unicellular alga Nitzschia closterium.

Unicellular algae are highly sensitive to a wide range of toxicants and have been used extensively in ecotoxicological testing. This, along with their ability to grow in very small test volumes over short test durations, make them ideal test organisms for use in Toxicity Identification Evaluations (TIEs). Despite this, microalgae have not previously been used in marine TIE studies. In this study, the marine diatom Nitzschia closterium was shown to be a highly suitable test organism after modification of the standard test protocol to reduce test volumes to 6 mL and test duration to 48 h. The alga was tolerant to the chemicals used in phase I of the standard USEPA TIE protocol, and physical TIE manipulations had no effect on algal growth. The cation exchange procedure, however, inhibited algal growth, while the anion exchange procedure stimulated growth, making these two procedures unsuitable for use with this species. Of the buffers trialed for the graduated pH procedure, 0.01 M PIPES buffer was found to be suitable for buffering at pH 7 because it maintained the required pH over the duration of the test and did not affect the growth or sensitivity of the algae to one reference toxicant (copper). A trial TIE on a secondary-treated sewage effluent for discharge into coastal waters showed that the developed protocols could successfully be used to identify ammonia as the major toxicant in the effluent.

Ammonia↗

The effect of anthracene and phenanthrene on the growth, photosynthesis, and SOD activity of the green alga Scenedesmus armatus depends on the PAR irradiance and CO2 level.

Short-term (24-h) experiments were performed to examine the effect of phenanthrene (PHE) and anthracene (ANT) on algae grown in a batch culture system at irradiances of 12, 33, 48, and 64 W m(-2) of the PAR range. Cultures were aerated (0.1 or 2% CO(2)) or nonaerated. As a result of aeration the concentration of ANT dropped from 0.45 mg L(-1) at the beginning of the experiment (t(0)) to an undetectable value after 10 h. The PHE concentration dropped from 9.36 mg L(-1) at t0 to 0.17 mg L(-1) after 24 h. ANT at nominal concentrations exceeding 0.05 mg L(-1) inhibited the growth of the algae in a concentration- and irradiance-dependent manner. The algistatic effect of ANT observed at 64 and 48 W m(-2) was independent of the CO(2) level, whereas the growth inhibition at 33 and 12 W m(-2) was much greater in cultures aerated with 2% than with 0.1% CO(2). PHE inhibited the growth only at a concentration of 10 mg L(-1) (about 50% of the control) regardless of the irradiance or the CO(2) concentration. The toxicity of both PHE and ANT was similar in aerated (0.1% CO(2)) and nonaerated cultures. ANT and PHE had a distinct effect on the photosynthesis. At 0.1% CO(2), the irradiance-dependent inhibition (ANT) and stimulation (PHE) of the photosynthesis was observed. The inhibitory effect of ANT was enhanced when algae were grown at 2% CO(2), but at the same CO(2) concentration PHE did not affect the photosynthesis. ANT stimulated the total superoxide dismutase (SOD) activity in the cells, this effect being generally more pronounced at elevated CO(2) levels and increasing with the irradiance. The total SOD activity was 250-300% higher in cells treated with PHE aerated with 0.1% CO(2). At 2% CO(2) no effect of PHE on the enzyme's activity was noted. The results obtained indicate that ANT acts as a photosensitizer causing an oxidative damage of cells, while PHE seems to affect the macromolecular synthesis.

Anthracenes↗

Growth stimulation and inhibition effects of 4-hydroxybenzoic acid and some related compounds on the freshwater green alga Pseudokirchneriella subcapitata.

4-hydroxybenzoic acid (4-HBA) exhibited low algal toxicity with the 72-h median inhibition concentration (IC50) of 9.9 mmol/L in the standard growth inhibition test using the freshwater green alga Pseudokirchneriella subcapitata. In contrast, it stimulated the algal growth at lower concentrations ranging from 0.1 to 1.0 mmol/L. Comparative studies with benzoic acid and 2- and 3-hydroxybenzoic acids (2-HBA and 3-HBA) indicated that 2-HBA was the most toxic, giving a 72-h IC50 of 0.172 mmol/L, and 4-HBA was the least toxic and that only 4-HBA had the pronounced growth stimulation activity. In a semicontinuous exposure to 4-HBA (0.15 and 0.3 mmol/L), algae maintained increased cell growth compared with controls during up to 10 times consecutive batch cultures, without any indication of adaptive responses to the growth enhancing effect of 4-HBA. Return to the clean standard medium of the exposed cells resulted in the quick recovery from the stimulant effect. Furthermore, 4-HBA (0.3 mmol/L) was found to diminish the toxicity of 2-HBA (<1.0 mmol/L). The presence of such compounds in the complex mixture might interact with other toxicants to alter the whole toxicity when assessed by the algal growth inhibition test. The effects of 4-HBA on P. subcapitata growth observed in the present study are not expected for planktonic algae in the aquatic environments, because known environmental concentrations are far below the effective concentration range.

Chlorophyta↗

The cell wall as a barrier to uptake of metal ions in the unicellular green alga Chlamydomonas reinhardtii (Chlorophyceae).

The cell walls of plants, including those of algae, have the capacity to bind metal ions in negatively charged sites. The authors had already shown that the wild type (walled) strain of the unicellular green alga Chlamydomonas reinhardtii Dangeard was more tolerant to Cd, Co, Cu, and Ni than a wall-less mutant of the same species. The objective of the present study was to determine if the tolerance to metals was associated with an increased adsorption of the same metals to the cell wall. Adsorbed metal was defined as that fraction that could be removed with a solution containing Na(2)EDTA and CaCl(2). The fraction that remained after the EDTA/CaCl(2) wash was considered to be strongly bound in the cell. When exposed to metals, singly, in solution for 24 h, cells of both strains accumulated the metals. The original hypothesis was supported by the results for Cd, Co, and Ni insofar as significantly higher concentrations of these metals were in the loosely bound fraction of the walled strain in comparison with the wall-less strain. However, there are three reasons why the potentially protective effect of the cell wall did not explain differential tolerance of the two strains. After 24 h of exposure (1) less Cd was accumulated internally by the wall-less strain than by the walled strain, (2) very little of the accumulated Cu was in the loosely bound fraction of the walled strain, and (3) the two strains accumulated comparable and relatively high amounts of internal Cu. Unexpectedly, significant amounts of Cd and Cu were also removable from the surface of the wall-less cells. One possible explanation for these apparently externally bound metals in the wall-less strain is that the cells exuded metal-chelating molecules that decreased the ability of metal ions to penetrate the plasma membrane. It was concluded that metal tolerance in this alga must involve a complex of mechanisms involving both internal and external detoxification of metal ions.

Animals↗

Toxin-Producing Anabaena flos-aquae Induces Settling of Chlamydomonas reinhardtii, a Competing Motile Alga.

Toxin production is an adaptation that allows cyanobacteria in resource-limiting environments to ameliorate the effects of herbivory and competition with other phototrophs. We demonstrate that the cyanobacterial toxins anatoxin-a and microcystin-LR paralyze the motile green alga Chlamydomonas reinhardtii. In addition, both purified toxins and cyanobacterial extracellular products containing these toxins cause the alga to settle faster than in nontoxic media. In microcosm experiments, the presence of either the cyanobacterium or its extracellular products induce settling in the alga, similar to the response observed with the addition of both anatoxin-a and microcystin-LR. The cyanobacterial production of paralyzing toxins represents a novel mechanism for phytoplankton settling. This prokaryotic/eukaryotic chemical interaction may create a competitor-free zone for cyanobacteria in lake environments, predicating optimal conditions for a toxic cyanobacterial bloom.

Journal Article↗