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PubMed1994

Novel touch-induced, Ca(2+)-dependent phobic response in a flagellate green alga.

The biflagellate green alga Spermatozopsis similis exhibits a remarkable avoidance reaction in addition to the photophobic or stop response characteristic of such algae. S. similis normally swims forward with its anteriorly attached flagella directed posteriorly and propagating sine-like waves base to tip. Upon contact with surfaces or other cells, S. similis responds with rapid backward swimming, covering distances of up to 50 microns in 140 to 220 msec. This reaction, which we term the mechanoshock response, also can be triggered by vigorous mechanical stimulation, but not by physiological light intensities. It consists of 3 phases: (1) a rapid acceleration phase with average duration of 31 msec; (2) a phase of about 66 msec with constant high speed (maximal velocities of > 600 microns.sec-1) or slow deceleration; and (3) a deceleration phase of approximately 83 msec, followed by a stop or short period of circling. The cells then resume forward swimming in a random direction. Prior to the mechanoshock response the flagella rapidly are brought together into a close parallel configuration extending anteriorly of the cell body. They then appear to propel the cell by undulatory beating, while the cell describes a pronounced helical path. Small decreases in the extracellular Ca2+ concentration, as well as low concentrations of Ba2+, strongly suppress the probability of this phobic reaction. We conclude that this mechanoshock response involves large Ca2+ influxes, probably mediated by mechanosensitive and/or stretch-activated ion-channel(s).

Barium↗
PubMed1988

Isoenzyme analysis of lichen algae in immobilized pH gradients.

A base for a modern species' concept of chlorococcal algae can be obtained not by morphological analysis, but by biochemical characters, e.g. isoenzyme banding patterns. From isolated lichen algae of the genus Trebouxia de Puymaly a set of five such enzymes has been studied by isoelectric focusing in immobilized pH gradients (IPG): phosphoglucomutase, phosphoglucose isomerase, malate dehydrogenase, mannitol dehydrogenase and leucine aminopeptidase. The first four are resolved into isoforms in a pH 4-7 IPG interval, while the last one is analyzed in an IPG pH 3.5-5 span. The patterns are specific for distinct populations, inter- and intraspecifically varying in dependence from their geographical distribution or the lichen species from which they have been isolated. Their limited heterogeneity (one to four isoforms) suggests that they are the products of specific genes rather than artefacts of the extraction procedure or the IPG analysis. Sharp isozyme patterns can only be obtained in a mixed-bed, carrier-ampholyte (CA)-IPG gel and by anodic application, suggesting that the recently proposed mechanism of hydrophobic protein-IPG matrix interaction (Electrophoresis, 1987, 8, 62-70) is fully operative here. As an additional mechanism, it is proposed that, in some cases, CA might simply act, when added to an IPG gel, by buffering, in the transient state, the sample zone before the protein migrates from the liquid phase into the IPG matrix.

Ampholyte Mixtures↗
PubMed1995

Nutritive value of marine algae Laminaria japonica and Undaria pinnatifida.

The chemical composition, that is crude proteins, fats, carbohydrates, cellulose, ashes, minerals and nucleic acids were determined in two commercially available marine algae: Laminaria japonica and Undaria pinnatifida. The energetic value and energetic share of proteins, fats and carbohydrates were calculated. The results were discussed in terms of importance of marine algae in human diet.

Dietary Carbohydrates↗
PubMed1985

Major light-harvesting polypeptides encoded in polycistronic transcripts in a eukaryotic alga.

By sequence analysis of previously identified fragments and low stringency hybridization of an identified gene for a phycobiliprotein subunit to total plastid DNA, we have identified four phycobiliprotein subunit genes in a eukaryotic alga, Cyanophora paradoxa. The four phycobiliprotein subunits, alpha and beta of phycocyanin (PC) and allophycocyanin (APC), comprise the bulk of the light-harvesting complex in this alga. The alpha and beta subunit genes encoding each phycobiliprotein (the products of which are required in a 1:1 ratio in the light-harvesting complex) are contiguous; however, the genes for different phycobiliproteins, PC and APC, are located in different regions of the genome. The two PC subunit genes are in the small single copy region of the plastid genome whereas the APC subunit genes are in the large single copy region and the two sets of phycobiliprotein genes are transcribed from opposite strands. The alpha and beta subunits of both PC and APC are encoded in dicistronic transcripts and this arrangement may provide a mechanism by which the two subunits can be synthesized in equimolar amounts. Levels of the PC transcript are approximately five times that of the APC transcript which may reflect the relative abundance of their gene products in the phycobilisome. The 5' ends of the transcripts for PC and APC were mapped and the regulatory regions identified. Several features of the promoter regions for these highly transcribed genes are described.

Amino Acid Sequence↗
PubMedDate not supplied

Improved erythrocyte lysis assay in microtitre plates for sensitive detection and efficient measurement of haemolytic compounds from ichthyotoxic algae.

Haemolytic substances produced by ichthyotoxic algae often are unknown in molecular structure or specific mechanism of toxicity. Detection and quantification of such substances are dependent on bioassays, using markers that are sensitive for haemolytic impairment and generation of a recordable response. The erythrocyte lysis assay (ELA) represents an advantageous bioassay in this respect, because the lytic response can be measured photometrically by the amount of released haemoglobin. The aim of the present study was to establish an improved assay based on the ELA principle, for sensitive determination of haemolytic substances of microalgae and for high sample throughput. For this purpose we adapted the ELA to a 96-well microtitre plate format, which significantly reduced the sample volumes and allowed rapid processing of samples. Further improvement was achieved by measuring absorption of lysed erythrocytes at 414 nm, which significantly increased the sensitivity of the ELA compared to the measurements at 540 nm that are usually applied in this type of assay. Using carp (Cyprinus carpio) erythrocytes it was possible to detect haemolysis induced by 4 microg ml(-1) of saponin and as little as two haemolytic Alexandrium tamarense cells. It is suggested that this improved ELA in microtitre plates be used as a low-cost monitoring tool for detection and analysis of potential harmful algae. Furthermore, this ELA can be useful as a sensitive screening system for substances of pharmacological interest, e.g. selectively acting cytolytic antibiotics.

Animals↗
PubMed1991

Scale formation in algae.

Scale biogenesis in algae represents a unique model system to study the transport of secretory macromolecules through the Golgi apparatus (GA) and their exocytosis. The larger scales can be visualized in the light microscope, and thus the kinetics of scale assembly, transport, and secretion can be studied in vivo. In addition, scales are osmiophilic and readily visible in conventional transmission electron microscopy; thus, details of scale assembly and sorting can be studied without invoking immunolabeling techniques. The following are distinctive features of scale biogenesis in algae: 1) transport of scales through the GA-stack occurs by cisternal progression; 2) scale secretion may be very rapid (in some cases a single GA-cisterna leaves the stack every 15-20 s); 3) sorting of different scale types does not occur in the GA, but in a post-GA-compartment. Recent progress in the analysis of scale formation in the green flagellates Tetraselmis and Scherffelia is reviewed.

Animals↗
PubMed1986

Impact of symbiotic algae on sea anemone metabolism: analysis by in vivo 31P nuclear magnetic resonance spectroscopy.

High-field pulsed Fourier-transform nuclear magnetic resonance spectroscopy (NMR) was used to quantify the adenylate levels of sea anemones (Aiptasia pulchella) with and without symbiotic dinoflagellates (Symbiodinium sp.). Animals were fed to repletion, then starved in darkness for up to six days before collection of in vivo NMR spectra. The host adenylate ratio of ATP: (ATP + ADP) declined significantly with increasing periods of starvation in both symbiotic and aposymbiotic hosts (P less than 0.05). However, the decline in the animal adenylate ratio was significantly more rapid in animals bearing symbiotic algae (P less than 0.05). This suggests that symbiotic algae in darkness cause more rapid depletion of host energy reserves, possibly by drawing on host pools of organic substrates. In vivo NMR spectroscopy was also used to evaluate the effect on A. pulchella of photosynthesis by zooxanthellae. Symbiotic anemones were fed to repletion, then starved under high irradiance (300 to 320 mu Ein m-2 s-1) or low irradiance (70 to 80 mu Ein m-2 s-1) conditions for up to five days. The host adenylate ratio declined significantly (P less than 0.01) with starvation under both treatments, but no significant difference was detected between treatments (P greater than 0.35). Blotted wet weight of anemones under high and low irradiance declined by 50% over eight days of starvation, but there was no significant difference in the rate of weight loss by anemones in the two treatments. There results suggest that translocation of photosynthate from symbiotic zooxanthellae does not significantly affect host adenylate ratio or have a sparing effect on host biomass during starvation in this symbiotic sea anemone.

Adenine Nucleotides↗
PubMed1978

Pesticide (hexachlorocyclohexane) inhibition of growth and nitrogen fixation in blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides.

The effects of the pesticide hexachlorocyclohexane (HCH) on the nitrogen fixing blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides commonly found as blooms in fish ponds were studied. These algae were very sensitive to HCH, and a distinct decrease in growth rate was observed on prolonged incubation. Lower concentrations (10 microgram/ml) were algistatic and higher concentrations (60 microgram/ml) were algicidal. The inhibition of nitrogen fixation indicated that the presence of HCH might affect overall nitrogen economy of inland waters.

Cyanobacteria↗
PubMed1981

Heterotrophic growth and nitrogen fixation in the filamentous blue-green alga Anabaena sp.

Experiments were conducted to study the effect of five different organic substrates viz. lactose, fructose, mannose, xylose and sodium acetate on the heterotrophic growth characteristics with emphasis on growth, pigment composition, heterocyst frequency and nitrogen fixation of Anabaena sp. under light and dark conditions. Lactose was found to be the best carbon source to support the heterotrophic growth of the alga. Phycoerythrin pigments were synthesized in photoheterotrophic cultures but pigment production was not observed in dark-grown cultures. The alga fixed nitrogen to a limited extent in the dark in pure inorganic medium. But when suitable organic substrates were supplemented, appreciable nitrogen fixation was observed in dark heterotrophic as well as in photoheterotrophic cultures.

Acetates↗
PubMed1983

Cardiac inhibitory action of constituents of the marine green alga Ulva pertusa.

As part of a search for substances from marine organisms, which exhibit inotropic effects, the chemical constituents of the marine green alga, Ulva pertusa were investigated. The fractionated extract was tested for inotropic effects on the isolated guinea pig atria. The aqueous layer obtained from the acetone extract of fresh algae was concentrated, and the residue was extracted with methanol. The methanolic extract was fractionated by chromatography using mixtures of aqueous methanol. Elution with 50% aqueous methanol afforded material that had a significant negative inotropic effect. Further purification of this material by high-performance liquid chromatography (HPLC) using methanol-water (2:5) afforded crystalline adenosine, which was shown to be the active substance of U. pertusa, causing a negative inotropic action.

Adenosine↗
PubMed1995

Interactions between bacteria and harmful algae: a review.

The ways in which bacteria interact with eukaryotic, unicellular algae are extremely diverse. Such relationships vary widely according to a number of criteria, including spatial and temporal scales, the degree of specificity, and if the relationship can be characterized as beneficial or detrimental to any of the organisms involved. These criteria can be applied to our assessment of how microbes interact with those species involved in the formation of harmful algal blooms (HABs). The aim of this paper is to assess the current state of our knowledge of bacterial/HAB interactions as they pertain to the influence of bacteria on HAB population dynamics, the role of bacteria in the production of toxins normally attributed to the algae, and the suggestion that HABs may act as vectors for pathogenic bacteria. Given that viruses are now considered to play a potentially important role in structuring phytoplankton communities, the possible effects of viruses on the population dynamics of harmful algal species are also addressed.

Bacterial Physiological Phenomena↗
PubMed2003

Acute toxicity of fatty acids to the freshwater green alga Selenastrum capricornutum.

The acute toxicity of fatty acids (C14 to C18) commonly found in wood was determined by the standard algal growth inhibition test using the freshwater green alga Selenastrum capricornutum. Toxicity, quantified as IC50 values, varied depending on the number of total carbons and double bonds. Of the tested acids, oleic (cis-9-octadecenoic) acid showed the highest toxicity (72-h IC50 = 0.47 mg/L) to the alga, and triolein, a triglyceride of oleic acid, showed no apparent toxicity. Further examination of a series of C18:1 acids with a double bond at the 6, 11, or 12 position revealed that both double-bond position and cis or trans configuration affected toxicity. The 72-h IC50 data for these fatty acids and related compounds seemed to correlate well with the melting point (mp), showing two separate linear relationships: at mp < 35 degrees C toxicity increased with increasing melting point, and at mp > 40 degrees C toxicity decreased with melting point.

Chlorophyta↗
PubMed2004

Acute toxicity investigations of ester-based lubricants by using biotests with algae and bacteria.

Although ester-based lubricants are ecologically acceptable due to their good biodegradability, there are still some environmental ecotoxicological impacts that have to be considered. Information on the acute ecotoxicological behavior of lubricants is obtained in this work using several single species bioassays. In previous studies it was observed that lubricating fluids containing additives for the enhancement of their technical performance were more problematic than base fluids especially with respect to algae growth inhibition. In order to clarify the influence of additives, the anti-wear additive tri-n-butyl phosphate was tested. It was very toxic to algae though not to bacteria. Additionally, a mixture of this additive with a base fluid is characterized. Despite the high toxicity of the single additive, the water extract of the mixture of tri-n-butyl phosphate with hydraulic base fluid caused almost no toxicity. Therefore, tri-n-butyl phosphate cannot explain the effect observed for the toxicity of water extracts of the commercially available lubricants.

Biological Assay↗
PubMed1998

Two unusual amino acid substitutions in cytochrome b of the colorless alga Polytomella spp.: correlation with the atypical spectral properties of the bH heme.

The dithionite-reduced spectra of the purified bc1 complexes from the colorless alga Polytomella spp. and the closely related green alga Chlamydomonas reinhardtii were compared. The spectrum of the bc1 complex from C. reinhardtii showed a profile similar to those of the bc1 complexes from other species. In contrast, the bc1 complex from Polytomella spp. exhibits a double-peak spectrum in the alpha-band region, where the absorption bands of cytochrome c1 and cytochrome b are completely resolved. To further understand the molecular basis of these spectroscopic differences, the mitochondrial gene encoding cytochrome b of Polytomella spp. was cloned, sequenced, and compared with that of C. reinhardtii. The Polytomella spp. cytochrome b gene is 1113 bp long and does not contain introns. The deduced protein sequence exhibits 56% identity and 68% similarity with the cytochrome b of C. reinhardtii, and in a phylogenetic analysis it clearly affiliated with the b-type cytochromes of C. reinhardtii and C. smithii. A comparison of the primary sequences of the Polytomella spp. cytochrome b with other b-type cytochromes, and its analysis based on the structure featuring eight transmembrane stretches, allowed the identification of a tyrosine in position 114, which substitutes for a tryptophan present in all mitochondrial b-type cytochromes sequenced to date. In addition, the primary sequence of the cytochrome b from Polytomella spp. has a serine at position 36, instead of a nonpolar residue (alanine or leucine) found in all other species. In the proposed model for cytochrome b, both residues Tyr114 and Ser36 are in close proximity to the high-potential bH heme. The above data suggest that the polar residues Y114 and S36, each one by itself or in combination, may interact with heme bH of Polytomella spp. and, thus, may be responsible for the unique spectroscopic characteristics of cytochrome b.

Amino Acid Sequence↗
PubMed1999

Spectrin-like proteins in green algae (Desmidiaceae).

Immunochemical detection of actin as well as spectrin-like proteins have been carried out in the green algae Micrasterias denticulata, Closterium lunula, and Euastrum oblongum. In these algae, actin is detected on Western blots at 43 kDa with antibodies to actin from higher plant and animal origin. By use of antibodies to human and chicken erythrocyte spectrin a cross-reactivity with desmid proteins is found at about the molecular mass of 220 kDa, where also human erythrocyte spectrin is detected. Additional bands are present at 120 kDa and 70 kDa, which are probably breakdown products. An antibody against chicken alpha-actinin, a small protein of the spectrin superfamily, recognizes bands at 90 kDa, where it is expected, and 70 kDa, probably the same breakdown product as mentioned for spectrin. Isoelectric focusing provides staining at pI 4.6 with antibodies against spectrin. Immunogold labelling of spectrin and alpha-actinin antigens on high-pressure frozen, freeze-substituted Micrasterias denticulata cells with the same antibodies exhibits staining, especially at membranes of different populations of secretory vesicles, at dictyosomes, and the plasma membrane. However, no clear correlation to the growth pattern of the cell could be observed. Taken together, our results demonstrate the presence of spectrin-like proteins in desmid cells which are probably functional in exocytosis.

Actinin↗
PubMed1999

Influence of centriole behavior on the first spindle formation in zygotes of the brown alga Fucus distichus (Fucales, Phaeophyceae).

The influence of centrioles, derived from the sperm flagellar basal bodies, and the centrosomal material (MTOCs) on spindle formation in the brown alga Fucus distichus (oogamous) was studied by immunofluorescence microscopy using anti-centrin and anti-beta-tubulin antibodies. In contrast to a bipolar spindle, which is formed after normal fertilization, a multipolar spindle was formed in polyspermic zygote. The number of mitotic poles in polyspermic zygotes was double the number of sperm involved in fertilization. As an anti-centrin staining spot (centrioles) was located at these poles, the multipolar spindles in polyspermic zygotes were produced by the supplementary centrioles. When anucleate egg fragments were fertilized, chromosome condensation and mitosis did not occur in the sperm nucleus. Two anti-centrin staining spots could be detected, microtubules (MTs) radiated from nearby, but the mitotic spindle was never produced. When a single sperm fertilized multinucleate eggs (polygyny), abnormal spindles were also observed. In addition to two mitotic poles containing anti-centrin staining spots, extra mitotic poles without anti-centrin staining spots were also formed, and as a result multipolar spindles were formed. When karyogamy was blocked with colchicine, it became clear that the egg nucleus proceeded independently into mitosis accompanying chromosome condensation. A monoastral spindle could be frequently observed, and in rare cases a barrel-shaped spindle was formed. However, when a sperm nucleus was located near an egg nucleus, the two anti-centrin staining spots shifted to the egg nucleus from the sperm nucleus. In this case, a normal spindle was formed, the egg chromosomes arranged at the equator, and the associated MTs elongated from one pole of the egg spindle toward the sperm chromosomes which were scattered. From these results, it became clear that paternal centrioles derived from the sperm have a crucial role in spindle formation in the brown algae, such as they do during animal fertilization. However, paternal centrioles were not adequate for the functional centrosome during spindle formation. We speculated that centrosomal materials from the egg cytoplasm aggregate around the sperm centrioles and are needed for centrosomal activation.

Antibodies↗
PubMed1994

Biotests using unicellular algae and ciliates for predicting long-term effects of toxicants.

Test systems for predicting long-term effects with the freshwater algae Chlamydomonas reinhardi and Scenedesmus subspicatus and the ciliate Tetrahymena pyriformis were evaluated with respect to the following reference chemicals: atrazine, bromacil, diuron, methyl parathion, lindane, 3,4-dichloroaniline, pentachlorophenol, cadmium, copper, and the volatile 1,2-dichloropropane. In growth-inhibition tests under static conditions the algae revealed a higher sensitivity to the toxicants than the ciliate except for lindane and methyl parathion. Comparison of the impairment of photosynthetic efficiency (EPR, NOEC 24 hr) with the inhibition of growth (NOEC 72 hr) of S. subspicatus revealed a higher sensitivity of the EPR parameter for inhibitors of the photosynthesis. A flowthrough system was developed for long-term tests and testing of volatile and instable substances. Under flowthrough conditions C. reinhardi was more susceptible to the chemicals than under static test conditions, except for pentachlorophenol. Due to the high volatility, 1,2-dichloropropane was only tested in the flowthrough system. The data obtained from these toxicity tests provide information about effects on organisms representing different levels of the aquatic food web, possessing differences in sensitivity against toxicants. The presented flowthrough system allows the testing of volatile and instable chemicals, problematic in static test systems, and the EPR parameter is suitable for the early characterization of chemicals acting as specific inhibitors of the photosynthetic electron transport chain.

Animals↗
PubMed1995

Polarography as a tool in peptide and protein analysis: studies on metal-chelating substances induced by cadmium in the algae Pheodactylum tricornutum and the graminae Agrostis capillaris.

Methods to determine characteristics and quantities of metal-binding substances are needed in order to identify phytochemical systems for use in agriculture and waste treatment. A metal-binding polypeptide (phytochelatin) was prepared by liquid chromatography from the algae Pheodactylum tricornutum and roots of the graminae Agrostis capillaris after exposure to cadmium. Polarographic (voltammetric) analysis indicated binding of cadmium by the polypeptide at different stabilities. No catalytic hydrogen peak at -1.7 V appeared in the polarogram of plant and algae metal-binding substance, but did appear in the polarogram of the metal-chelating protein metallothionein. In all polarograms, two significant peaks were detected. One at approximately -520 mV probably represents the reduction of single cadmium-sulfur bonds, and one at -930 mV the reduction of cadmium-thiolate complexes. A dose-response relationship between cadmium concentration and peak current at -930 mV was observed after exposure of roots of Agrostis at 5 and 10 microM.

Cadmium↗