Fine Structure of the Cell Surface and Golgi Apparatus of Ochromonas*

SYNOPSIS. Ochromonas danica has an unusually flexible cell surface capable of producing projections of varying sizes and shapes: large projections, 340–360 nm long, and small projections, 50–110 nm long. These projections have been demonstrated by transmission and scanning electron microscopy; some of them may break off into the medium and be the source of extracellular membranes and vesicles reported in the cell-free O. danica growth medium. Ruthenium red stained the acid mucopolysaccharide layer just outside the cell surface as well as small blebs at the cell surface. The Golgi complex of O. danica, Ochromonas malhamensis, Ochromonas sociabilis and Ochromonas sp. produced small coated vesicles which may move toward and fuse with the plasma membrane. The role of the several vesicles is unknown but possible functions are discussed.     

Conservation of Glycerolated Cultures of Ochromonas danica and O. malhamensis at −10 C

SYNOPSIS. Ochromonas danica in a complex natural growth medium dies at 6–10 C in 4 days; O. malhamensis in ∼2 days. O. danica grown in the medium supplemented with 4.0% glycerol survived at −10±2 C for 35 days, and with 8% glycerol 29 days. O. malhamensis lasted only to 5 days in these media supplemented with 4% glycerol. Ethylene glycol and dimethylsulfoxide were too toxic to be effective. Difficulties in freeze-preservation of certain other phagocytic cells, notably blood granulocytes having comparatively simple flexuous outer membranes, add interest to use of O. danica and O. malhamensis as test organisms for preservation methods, especially in the convenient, inexpensive -10 to -20 C range. Biphasic media with an overlay of distilled water serve for conservation at room temperature. Problems of mutational erosion of these photosynthetic phagotrophs are discussed.     

Comparative study on hydrolases in five species of Ochromonas (chrysomonadina).

Acid phosphatase and beta-glucosidase were shown to be present in five species of Ochromonas grown in organic media (O. danica, O. malhanesis, O. munuta, O. sociabilis and Ochromonas sp. 933/4). Acid phosphatase was found to have a pH optimum at 4.0 in O. danica, and at 5.1 in the four other species. No alkaline phosphatase was found in any of the above mentioned species. Beta-glucosidase in the species studied has a pH optimum at 4.6. Low alpha-glucosidase activity was found only in O. danica. Acid phosphatase in all the five species shows an increase in activity during the logarithmic phase of growth and a decrease during the early stationary phase. Beta-glucosidase shows a similar behavior only in O. danica.     

Element content of Ochromonas danica: a replicated chemostat study controlling the growth rate and temperature

Ecological stoichiometry focuses on the balance between multiple nutrient elements in resources and in consumers of those resources. The major consumers of bacteria in aquatic food webs are heterotrophic and mixotrophic nanoflagellates. Despite the importance of this consumer-resource interaction to understanding nutrient dynamics in the aquatic food web, few data are available addressing the element stoichiometry of flagellate consumers. Ochromonas danica, a mixotrophic bacterivore, was used as a model organism to study the relationships among temperature, growth rate and element stoichiometry. Ochromonas danica was grown in chemostats at dilution rates ranging between 0.03 and 0.10 h(-1) and temperatures ranging between 15 and 28 °C. Cells accumulated elements as interactive functions of temperature and growth rate, with the highest element concentrations corresponding to cells grown at a low temperature and high growth rates. The highest concentrations of elements were associated with small cells. Temperature and growth rate affected the element stoichiometry (as C:N, C:P and N:P) of O. danica in a complex manner, but the growth rate had a greater effect on ratios than did temperature.     

Effect of Wetting Agents on Permeability of Ochromonas danica

SYNOPSIS. The effect of 10 detergents on the inhibitory activity of neomycin was tested against Ochromonas danica. All 4 anionic detergents used were synergistic with neomycin. Evidence is presented that the synergisms were due to the effect of the detergents on the cell membrane. In the presence of Tergitol 7, O. danica is more susceptible to changes in osmotic pressure by increasing concentrations of KCl.     

Sterols of Soil Amoebas and Ochromonas danica: Phylogenetic Approach

SYNOPSIS. Sterols of Ochromonas danica, Mayorella palestinensis, Acanthamoeba sp. and Hartmannella rhysodes were examined by thin-layer and gas-liquid chromatography. The sterol patterns of O. danica and the amoebas were strikingly similar. Two of the sterols present in both groups, were identified as ergosterol and stigmasterol. This similarity is construed as evidence that "soil" amoebas evolved from phytoflagellates having sterol patterns similar to that of O. danica. An explanation is suggested for the role of ergosterol in phytoflagellates, as originally an ultra-violet-protective agent, then a membrane component.     

Cytoskeleton organization during the cell cycle in two stramenopile microalgae, Ochromonas danica (Chrysophyceae) and Heterosigma akashiwo (Raphidophyceae), with special reference to F-actin organization and its role in cytokinesis

F-actin organization during the cell cycle was investigated in two stramenopile microalgae, Ochromonas danica (Chrysophyceae; UTEX LB1298) and Heterosigma akashiwo (Raphidophyceae; NIES-6) using FITC-phalloidin. In the interphase cell of O. danica, F-actin bundles were localized forming a network structure in the cortical region, which converged from the anterior region to the posterior, whereas in the interphase cell of H. akashiwo, F-actin bundles were observed forming a network structure in the cortical region without any polarity. In both O. danica and H. akashiwo, at the initial stage of mitosis the cortical F-actin disappeared, and then during cytokinesis assembly of an actin-based ring-like structure occurred in the cell cortex in the plane of cytokinesis. The ring-like structure initiated from aster-like structures was composed of F-actin in both O. danica and H. akashiwo. Different from animal cells, later stages of cytokinesis of O. danica seemed to be promoted by microtubules, although the early stages of cytokinesis progressed with a constriction of the ring-like structure, whereas cytokinesis of H. akashiwo was apparently completed by constriction of the cell mediated by the F-actin ring, as in animal cells.     

39 Phylogeny of aulacoseira (bacillariophyta)

Ochromonas sensu lato is the largest genus in the Chrysophyceae, containing over 100 names. Ochromonas species are biflagellate, naked, plastid-bearing single cells, distinguished from loricate, scaled, colonial and colorless genera. Most, if not all, species of Ochromonas are mixotrophic, i.e., they photosynthesize but they also engulf bacteria and other small prey. Preliminary evidence from SSU rRNA sequences show that Ochromonas is a polyphyletic genus. Ochromonas tuberculata is the most distinct from all other Ochromonas species. The other Ochromonas species (examined thus far) are scattered in three clades. For example, O. danica and O. sphaerocystis are sister to Poterioochromonas stipitata and P. malhamensis . Four additional species (identified by light microscopy as O. elegans Doflein, O. globosa Skuja, O. ovalis Dolfein, O. sociabilis Pringheim ) have SSU rRNA sequences identical to P. malhamensis . Of these, only O. sociabilis has been transferred to Poterioochromonas . Thus, at least some species may be synonymous with others. Two clades of marine species are also known, one containing coastal species and the other containing open ocean species. A number of genera (some also polyphyletic) are interspersed amongst the Ochromonas species (e.g., Chrysolepidomonas, Chrysonephele, Chrysoxys, Cyclonexis, Dinobryon, Epipyxis, Uroglena, Uroglenopsis ). The goal of this research (just beginning) is to establish a monophyletic Ochromonas , probably by assigning some species to other genera (existing or new). One major problem is that the type species, O. triangulata Vysotskii , hasn't been observed in over 100 years, and it is unclear which of several clades of Ochromonas contains the type. Results will be discussed.     

Sulfolipid in Ochromonas danica

SYNOPSIS. Cells of Ochromonas danica were grown under photoautotrophic as well as heterotrophic conditions in the presence of ³⁵SO₄=, and the content of sulfolipid was studied using the technics of paper chromatography and paper electrophoresis. Photoauto-trophic cells of O. danica contained 5 to 6 times as much sulfolipid, sulfoquinovosyldiglyceride, as did cells grown under heterotrophic conditions.     

Properties of an Ethionine-Resistant (ER) Mutant of Ochromonas danica

SYNOPSIS. On prolonged incubation of ethionine-sensitive (ES) cells of Ochromonas danica in L-ethionine-containing media, growth was resumed by an ethionine-resistant (ER) mutant. Such mutants arise at random and are selected by the ethionine-containing medium. Ethionine resistance is not lost on repeated transfers thru ethionine-less media. ES cells incubated with ethionine form a large posterior vacuole before they disintegrate. Inhibition of reserve substance utilization is suggested to underlie growth inhibition of O. danica by ethionine. In ES cells incubated with ethionine, ¹⁴C uptake from labeled methionine, ethionine or serine is reduced by 65%. In ER cells the decrease in ¹⁴C uptake is 90%. This decrease in uptake of ethionine seems to be how ER O. danica evades growth inhibition by ethionine.     

Metabolism of phenols by Ochromonas danica

Abstract This study investigated the catabolic potential of a eukaryotic alga to degrade one of the most common organic pollutants, phenol. The alga, Ochromonas danica (993/28), was selected for study after screening for its heterotrophic capabilities. The catabolic versatility of the alga was elucidated by incubating with a variety of phenolic compounds. The alga removed phenol, all the cresol isomers and 3,4-xylenol from its incubation media, with phenol being removed more rapidly than any of its methylated homologues. Consequently, the alga was found to have a greater specificity for phenol than for o - or p -cresols. This study shows that O. danica could catabolize phenol and its methylated homologues.     

ENDOCYTOSIS OF MICROCYSTIS AERUGINOSA BY OCHROMONAS DANICA1

Ochromonas danica Prings., a chrysomonad alga which demonstrates a high degree of nutritional versatility, is capable of feeding on the toxic blue-green alga Microcystis aeruginosa Kuetz. In this paper light microscopic, electron microscopic, and cytochemical examinations of endocytosis in O. danica are reported, with particular emphasis on the vicissitudes of endocytic and lysosomal activities during intra-cellular digestion. An interpretation of the function of organelles associated with endocytosis is presented.     

Cytochemical Changes in Aging Ochromonas; Evidence for an Alkaline Phosphatase

SYNOPSIS. Changes accompanying aging in light-grown stationary cultures of Ochromonas danica were examined cytochemically. Succinate dehydrogenase activity increased during the log phase and decreased steadily during stationary and later phases. Acid phosphatase, alkaline phosphatase and lipase activities increased during the several phases of growth, as did accumulation of lipid. These results imply loss of mitochondrial activity and a gain in lysosomal activity with aging of the cell population. Alkaline phosphatase, widely distributed in animals and believed absent from most photosynthetic organisms and bacteria, is here reported in the photosynthetic genus Ochromonas.     

Heavy water (D2O)-induced shape changes, movements and F-actin redistribution in human neutrophil granulocytes

Heavy water (D2O) induces characteristic shape changes and a distinct type of movement in human neutrophil granulocytes. In contrast to front-tail polarity as evoked by chemotactic peptides and microtubule-disassembling agents, D2O-based media produce non-polar neutrophils with many small or long surface projections. This phenotype is similar to that elicited by both phorbol myristate acetate and diacylglycerols, but the surface projections are smaller and more densely placed and are often associated with a single large projection. D2O-induced non-polar cells with surface projections perform continuous shape changes without front-tail polarity and without the unidirectional movement and cytoplasmic streaming seen in cells with front-tail polarity. Some of the cells show circus movements of a large projection indicating circular polarity. In neutrophils suspended in D2O, F-actin is shifted to the cell periphery, mainly into the surface projections of activated cells. The D2O-induced effects are reversed in H2O-based medium. D2O is dominant over the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), colchicine and taxol in that the combined action of D2O with any of these agents results in the D2O-induced phenotype. In contrast, cytochalasin B alone and in combination with fMLP induces a considerable decrease of non-polar cells and an increase of spherical cells similar to non-stimulated cells in H2O-based medium. Earlier studies indicated that D2O acts on microtubules. Our results suggest that D2O may act on the microfilament system. Neutrophils suspended in D2O-based medium may represent a useful model to study the relationship between shapes, movements, and particular functions of these cells.     

Biodegradation of phenols by the alga Ochromonas danica.

The eukaryotic alga Ochromonas danica, a nutritionally versatile, mixotrophic chrysophyte, grew on phenol as the sole carbon source in axenic culture and removed the phenol carbon from the growth medium. Respirometric studies confirmed that the enzymes involved in phenol catabolism were inducible and that the alga oxidized phenol; the amount of oxygen consumed per mole of oxidized substrate was approximately 65% of the theoretical value. [U-14C]phenol was completely mineralized, with 65% of the 14C label appearing as 14CO2, approximately 15% remaining in the aqueous medium, and the rest accounted for in the biomass. Analysis of the biomass showed that 14C label had been incorporated into the protein, nucleic acid, and lipid fractions; phenol carbon is thus unequivocally assimilated by the alga. Phenol-grown cultures of O. danica converted phenols to the corresponding catechols, which were further metabolized by the meta-cleavage pathway. This surprising result was rigorously confirmed by taking the working stock culture through a variety of procedures to check that it was axenic and repeating the experiments with algal extracts. This is, as far as is known, the first definitive identification of the meta-cleavage pathway for aromatic ring degradation in a eukaryotic alga, though its incidence in other eukaryotes has been (infrequently) suggested.     

Identification of the Mitochondrial Genome in the Chrysophyte Alga Ochromonas danica

ABSTRACT. Analysis of total DNA isolated from the Chrysophyte alga Ochromonas danica revealed, in addition to nuclear DNA, two genomes present as numerous copies per cell. The larger genome (?120 kilobase pairs or kbp) is the plastid DNA, which is identified by its hybridization to plasmids containing sequences for the photosynthesis genes rbcL, psbA, and psbC. The smaller genome (40 kbp) is the mitochondrial genome as identified by its hybridization with plasmids containing gene sequences of plant cytochrome oxidase subunits I and II. Both the 120- and 40-kbp genomes contain genes for the small and large subunits of rDNA. The mitochondrial genome is linear with terminal inverted repeats of about 1.6 kbp. Two other morphologically similar species were examined, Ochromonas minuta and Poteriochromonas malhamensis. All three species have linear mitochondrial DNA of 40 kbp. Comparisons of endonuclease restriction-fragment patterns of the mitochondrial and chloroplast DNAs as well as those of their nuclear rDNA repeats failed to reveal any fragment shared by any two of the species. Likewise, no common fragment size was detected by hybridization with plasmids containing heterologous DNA or with total mitochondrial DNA of O. danica; these observations support the taxonomic assignment of these three organisms to different species. The Ochromonas mitochondrial genomes are the first identified in the chlorophyll a/c group of algae. Combining these results with electron microscopic observations of putative mitochondrial genomes reported for other chromophytes and published molecular studies of other algal groups suggests that all classes of eukaryote algae may have mitochondrial genomes < 100 kbp in size, more like other protistans than land plants.     

Ethionine and Methionine Metabolism by the Chrysomonad Flagellate Ochromonas danica

SYNOPSIS. Growth of Ochromonas danica is competitively inhibited by ethionine. Inhibition can be reversed by methionine. Inhibition indexes of the effect of ethionine on growth and methionine incorporation into proteins are 1 and 4, respectively. Inside the cell, methionine is partially de-methylated and metabolized to form cysteine. Ethionine is partially de-ethylated, and the homocysteine moiety is either re-methylated to form methionine or further metabolized to form cysteine. Ethionine is also incorporated into proteins of O. danica. The kind of metabolic interference, expressed by inhibition of growth, and correlated with incorporation of ethionine, is yet unknown.     

Comparative ultrastructural study of four reticuloendothelias viruses.

The morphology and development of four members of the reticuloendotheliosis virus group were studied by transmission electron microscopy. Virions of duck spleen necrosis virus, duck infectious anemia virus, chicken syncytial virus, and reticuloendotheliosis virus strain T are sperical with a diameter of approximately 110 nm. They are covered with surface projections about 6 nm long and 10 nm in diameter. The center-to-center distance of surface projections is about 14 nm. The budding virions contain crescent-shaped electron-dense cores 73 nm in diameter with electron-lucent centers. After release of the virions the cores apparently become condensed to 67 nm in diameter. Virions were found budding at the plasma membrane and into smooth-walled, intracytoplasmic vesicles of productively infected cells. The distribution of budding reticuloendotheliosis viruses on cells appeared random over the cell surface, and occasionally aberrant multiple forms of budding virions were observed. The virions appear to resemble mammalian leukemia and sarcoma viruses more closely than avian leukosis-sarcoma viruses.     

金黄滴虫细胞同步分裂的诱导

无菌培养的金黄滴虫在25℃及连续通气的条件下,经12小时光照(2500Lux)和12小时黑暗的相间处理,可连续出现三次同步分裂,同步分裂率最高可达92％,即细胞数在一小时内可陡增92％。 细胞核动态的观察表明,在黑暗处理期中细胞被阻止于细胞分裂的前期。当黑暗处理11小时后,前期细胞突破阻拦,实现了同步分裂。     

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. Received: 22 November 1996 / Accepted: 14 February 1997