Effects of iron-, manganese-, or magnesium-deficiency on the growth and morphology of Euglena gracilis

Iron-, manganese-, or magnesium-deficiency has been induced in Euglena gracilis. Each arrests cell proliferation, decreases the intracellular content of the deficient metal, and increases that of several other metals. Light and electron microscopy of stationary phase cells reveal that Fe-deficient (-Fe) cells are similar in size and shape to control organisms. Magnesium-deficient (-Mg) cells, however, are larger, and approximately 14% are multilobed, containing 2 to 12 lobes of equal size emanating from a central region. Individual (-Mg) cells and each lobe of multilobed cells contain a single nucleus. Manganese-deficient (-Mn) organisms are morphologically more heterogeneous than (-Fe) or (-Mg) cells. Most are spherical and larger than controls. Approximately 15% are multilobed but, unlike (-Mg) cells, contain lobes of unequal size with either zero, one, or several nuclei present in each. Nuclei of (-Mn) cells differ in size and shape from those of control, (-Fe), or (-Mg) cells. All three deficient cell types accumulate large quantities of paramylon. Other cytoplasmic structures, however, appear normal. Addition of Fe, Mn, or Mg to the respective deficient stationary phase cultures reverses growth arrest and restores normal morphology. The results suggest that Fe-, Mn-, and Mg-deficiencies affect different stages of the E. gracilis cell cycle.     

Effects of nalidixic acid, chloramphenicol, cycloheximide, and anisomycin on structure and development of plastids and mitochondria in greening Euglena gracilis

The substructure of plastids and mitochondria and the alterations caused by the addition of antibiotics were investigated during light-induced proplastid-to-chloroplast transformation in Euglena gracilis. The organisms were grown in presence of the inhibitors up to 3 days (5 generations). Both 40 μg/ml nalidixic acid and 1–1.5 mg/ml chloramphenicol prevent the formation of chloroplasts of normal size and structure by blocking development during early stages. Under our conditions 2 to 5 straight thylakoids are formed beside 1 to 2 girdle-like thylakoids. The former rarely fuse into bands. Non-crystalline prolamellar bodies of considerable size are formed at the distended ends of the plastids in the presence of both drugs. Chloramphenicol also influences mitochondrial size, shape and internal structure. Giant mitochondria can be observed. Nalidixic acid does not change the size and shape of mitochondria, but the matrix frequently appears highly osmiophilic. Cycloheximide in sublethal doses (2–5 μg/ml) or 50 μg/ml anisomycin inhibits plastid development only in the early period after addition. In later culture periods chloroplasts are found enlarged in size with an increased number of thylakoids and bands per organelle. Insertions of new bands are noted at the inner membrane of the chloroplast envelope. The electronmicroscopic observations agree with the results of chloroplast-specific biochemical activities such as light-induced increase in chlorophyll synthesis and of two chloroplast-bound enzyme activities. The results are discussed with respect to metabolic and biogenetic correlations between the two types of organelles in E. gracilis cells.     

Dr Han L. Golterman,a Limnologist: a tribute on the occasion of his 65th birthday and retirement

The green algae D. tertiolecta, the flagellate I. galbana and the diatom C. gracilis were grown in batch cultures. The organisms were analysed for lipid class composition at the logarithmic and stationary growth phases using the Chromarod-Iatroscan thin layer chromatography with flame ionization detection (TLC-FID) system.There were major differences in lipid class production among the organisms investigated, but few differences in lipid class distribution between log phase and stationary phase cultures of D. tertiolecta and I. galbana. C. gracilis displayed the general trend exhibited in diatom metabolism, which can be characterized by an increase in triacylglycerol synthesis in situations of stress.     

THE ULTRASTRUCTURE OF GLENODINIOPSIS STEINII (DINOPHYCEAE)

The freshwater dinoflagellate Glenodiniopsis steinii Wolsoszyńska was examined using computer-assisted three-dimensional reconstruction of serially sectioned cells observed with the transmission electron microscope and images from the scanning electron microscope. Vegetative cells contain ultrastructure typical of freshwater dinoflagellates including trichocysts, mitochondria, Golgi bodies, starch grains, and lipid bodies. The chloroplast is a single, multilobed structure, not multiple discoid chloroplasts as previously described. The “C” shape of the nucleus is apparently due in part to the size and location of the pusule.     

Isolation and Identification of a New Analogue of Aspergillic Acid Derived from Valine and Isoleucine

The biological activities of zinc and cadmium on Euglena gracilis grown in zinc deficient and sufficient media were examined- Cadmium was neither involved in the normal cell metabolism of E. gracilis under zinc deficient conditions, nor did not replace zinc, which is essential for the normal growth. More cadmium was incorporated into cells grown in zinc deficient media than in zinc sufficient ones, resulting in more toxic effects in zinc deficient media than in zinc sufficient ones. Cadmium effectively provoked abnormal cells under zinc deficient conditions, suggesting that the normal process of cell division was interrupted by cadmium.     

ULTRASTRUCTURE OF THE CARPOSPOROPHYTE AND CARPOSPOROGENESIS IN THE PARASITIC RED ALGA PLOCAMIOCOLAX PULVINATA SETCH, (GIGARTINALES,PLOCAMIACEAE) 1

The ultrastructure of the carposporophyte and carposporogenesis is described for the parasitic red alga Plocamiocolax pulvinata Setch. After presumed fertilization the zygote nucleus is apparently transferred to the auxiliary cell which initiates gonimoblast cell production. These gonimoblast cells differentiate into storage or generative cells. Storage gonimoblast cells (SGC) are large and multinucleate, contain large quantities of starch and are located nearest the auxiliary cell, when compared to the smaller uninucleate, devoid of starch, generative gonimoblast cells (GGC) that form terminal lobes of carpospores. In addition, compressed membrane bodies and annulate lamellae are common in these cells. During carposporophyte maturation the amount of starch in the SGC's decreases and eventually the auxiliary cell, as well as SGC's, degenerate. Generative gonimoblast cells (GGC's) cleave repeatedly to form carpospores which are interconnected by small pit connections. Stage one-carpospores are recognized by their elongated shape, the formation of small     

Glumatic-Oxaloacetic Transaminase in Leaves of Phosphorus-Deficient Citrus Plants

Phosphorus deficiency in citrus leaves resulted in reduced glutamic-oxaloacetic transaminase (GOT) activity and low pyridoxal-phosphate (PLP) content. GOT activity was estimated in crude enzyme extracts by spectrophotometry PLP content was detected colorimetrically in water-alcoholic extracts. K and Cu deficiencies increased; -N, -S and -Zn decreased and -Mg, -Fe and -Mn did not affect GOT activity in citrus leaves. Experiments were conducted to restore enzyme activity either by direct addition of PLP to the reaction mixture or by infiltration of PLP or KH₂PO₄ to detached, intact leaves. The infiltration was carried out in vacuo and the leaves were incubated on wet paper for 26 h, after which enzyme activity was estimated. Transamination activity of -P leaves more restored by PLP than by KH₂PO₄ treatments. In zinc-deficient leaves the enzyme activity was not restored by infiltration of KH₂PO₄.     

Changes of the plastid system of Euglena gracilis induced with streptomycin and dihydrostreptomycin

Summary Streptomycin-like antibiotics cause hereditary and irreversible aplastidity of Euglena gracilis by inhibiting the replication of plastids, while normal cell division is maintained.Therefore, a gradual dilution of plastids takes place in a multiplying culture. Streptomycin was found to be more effective as bleaching agent than dihydrostreptomycin. The cells of Euglena gracilis are totally deprived of plastids by streptomycin treatment after 4.5 cell divisions, while 9 cell divisions are required with dihydrostreptomycin. In addition to the inhibition of plastid replication both antibiotics bring about formation of pathological plastids, both in growing and in stationary cultures. In this latter case pathological plastids are released from cells only after further cell division has taken place.     

LE PLASTIDOME CHEZ EUGLENA GRACILIS Z1

Chloroplast morphology changes cyclically during each cell generation of synchronous photoheterotrophic cultures of Euglena gracilis Z; these cycles persist until the transition to stationary phase. Structural modifications, development of pyrenoids, and increase in chloroplast volume accompany the cessation of cell division. Chloroplasts of stationary phase heterotrophic cells resemble those of exponential phase autotrophic cells, suggesting a relationship between chloroplast morphology and available carbon.     

THE ULTRASTRUCTURE OF GALLS ON THE RED ALGA GRACILARIA EPIHIPPISORA1

A strain of Gracilaria epihippisora Hoyle produces gall-like cell proliferations in culture. These growths can be excised and grown separately, where they retain an undifferentiated morphology and reach 5mm in diameter. The gall tissue consists of a single morphological cell type without any differentiation between surface and internal cells as is characteristic of normal thallus tissue. Gall cells are typically 20–40 μm in diameter and contain the usual complement of organelles and a prominent vacuole, although there are several distinct features. The large multilobed plastids have an extensive proliferation of thylakoid membranes, which form an arrangement of loops and spirals. The thallus outer cell wall layer is highly reduced. The gall growths contain intracellular virus-like particles (ca. 80 nm in diameter) that occur in discrete groups.     

Regulation of catalase synthesis in Saccharomyces cerevisiae by carbon catabolite repression.

Summary A number of strains of Saccharomyces cerevisiae, wild type or respiratory deficient, were grown on glucose, galactose or raffinose. Specific activities of catalase T were about tenfold higher in late stationary wild type cells grown on glucose than in wild type cells harvested when glucose had just disappeared completely from the medium, or in respiratory deficient strains (rho^–, mit^–, pet) grown to stationary phase.Catalase A activity is completely absent in wild type cells grown to zero percent glucose or in respiratory deficient cells grown on glucose to stationary phase. High catalase A activity was detected in derepressed wild type cells and in a strain carrying the op 1 (pet 9) mutation, although this strain is unable to grow on nonfermentable carbon sources. All respiratory deficient strains tested have low, but significant catalase A activities after growth on galactose or raffinose.Wild type cells harvested during growth on glucose and rho^–-cells grown on low glucose to stationary phase contain enzymatically inactive catalase A protein. The apoprotein of the enzyme is apparently accumulated in rho^–-cells whereas glucose-repressed wild type cells seem to contain a mixture of apoprotein and heme-containing catalase A monomer.These results show that a source of chemical energy, probably ATP, is required for derepression of yeast catalase from catabolite repression. At least in the case of catalase A, energy produced by respiration is necessary if catabolite repression is caused by glucose. If less repressing sugars are utilized, ATP derived from fermentation appears sufficient for partial derepression. Formation of the active enzyme can apparently be influenced by carbon catabolite repression at different points: (1) at the level of protein synthesis, (2) at the stage of heme incorporation, (3) at the level of formation of the enzymatically active tetramer.     

Dose-dependent disruptive effects of melatonin,light and temperature on regeneration in the planarian Phagocata gracilis

Many critical cellular processes, including cell proliferation and tissue repair, exhibit melatonin sensitivity. We examined the effects of exogenous melatonin, photoperiod and temperature on regeneration in Phagocata gracilis, a common North American turbellarian. Planarian regeneration involves the proliferation and migration of pluripotent stem cells capable of replacing missing body components. A characteristic mass of cells (blastema) forms during planarian regeneration, and its formation, growth and differentiation provide reliable endpoints for the study of regenerative processes. We tested the effects of exogenous melatonin at two concentrations and three photoperiods on both head and tail regenerates under seasonal diurnal temperature regimes. High temperatures and melatonin treatment increased mortality in P. gracilis, and surviving melatonin-treated subjects exhibited delayed regeneration. Asymmetrical and deficient regeneration was more common in melatonin-treated, high temperature and dark treated subjects. Additionally, high temperature and melatonin induced a novel locomotor dysfunction.     

Hemmung der lichtabhängigen Chloroplastenentwicklung etiolierterEuglena gracilis durch Glucose

Summary In order to examine the inhibitory effect of heterotrophic nutrition on the regreening of etiolatedEuglena gracilis, strain Z, the organisms were cultivated in the light in the presence of glucose or carbon dioxide as carbon source. After about 120 hours of illumination the chlorophyll and carbohydrate contents per cell of the photoheterotrophically cultivatedEuglena differs significantly from that of autotrophically grown cells. Mainly in so far as the addition of glucose diminishes the number and size of the chloroplasts per cell and the amount of the chlorophyll-protein-complex CP II in the thylakoids, whereas the amount of the chlorophyll-protein-complex CP I is not influenced.

     

Water-soluble vitamins in cells and spent culture supernatants of Poteriochromonas stipitata,Euglena gracilis,and Tetrahymena thermophila

Vitamins B₆ and B₁₂, biotin, folates, riboflavin, nicotinate, pantothenate, biopterin, and vitamin C (l-ascorbate) were assayed in Poteriochromonas stipitata, Euglena gracilis, and Tetrahymena thermophila cells grown in defined media and in spent culture supernatants. P. stipitata and E. gracilis synthesized, stored and excreted folates (mainly as 5-methyltetrahydrofolate), B₆, riboflavin, pantothenate, nicotinate, biopterin, and ascorbate. E. gracilis synthesized and stored biotin. T. thermophila did not synthesize the above vitamins except for B₁₂, biopterin, and ascorbate; it excreted biopterin and stored B₁₂ and ascorbate. Thiamin was left of consideration because all 3 organisms are thiamin auxotrophs. Possible ecological implications of these findings are considered.     

Auxospore formation inLicmophora (Bacillariophyta)

InLicmophora gracilis var.anglica two auxospores are produced per pair of mother-cells, through the allogamic fusion of migratory and stationary gametes. Both active gametes are produced from the same mother-cell and hence both zygotes are formed in the other mother-cell. Pairing can occur between two stalked cells, or between a stalked cell and a detached cell; in the latter case the migratory gametes derive from the detached cell. The auxospores expand parallel to one another and to the apical axis of the donor mother-cell. Behavioural anisogamy of this kind, which may be termed thecis-type, seems to be characteristic of most araphid pennates and contrasts with thetrans-type exhibited byCymbella, Gomphonema and some other raphid taxa, where each mother-cell produces one migratory and one stationary gamete.     

Giant mitochondria in chloroplast-deprivedEuglena gracilis late after N-succinimidylofloxacin treatment

Euglena gracilis cells were treated with N-succinimidylofloxacin. After two years of cultivation of chloroplast-free mutants a number of anomalous mitochondria have been observed. Aberrant mitochondria about 5–8 μm long and wide with unusual shape, disfigured mitochondria with enormous size, giant swollen oval-shaped megamitochondria, narrow and very long (50–60 μm) mitochondria localized at the cell perimeter were found in addition to normal mitochondria. The volume percentage of aberrant mitochondria in 58% of cells reached 65%. Swollen mitochondria probably do not fulfil the mitochondrial function and may cause cell death with subsequent elimination of injured mitochondria.     

The pH-related morphological variations of two acidophilic species of Desmidiales (Viridiplantae) isolated from a lowland peat bog,Czech Republic

Morphological variation related to pH was investigated in two acidophilic desmid species (Euastrum binale var. gutwinskii and Staurastrum hirsutum) utilizing geometric morphometric methods. Clones isolated from acidic habitats were cultured using a range of pH values from 3.5 to 6.5. The plasticity of ensuing populations was quantified and illustrated by the general Procrustes superimposition of landmarks placed along the outline of cells and subsequent statistical analyses of shape data. In both species, there was a significant effect of pH on the morphology of cells. In Staurastrum hirsutum, the pH-related morphological change was accompanied by a decrease in the size of cells cultured at a higher pH. However, in Euastrum binale, cell size did not differ in relation to pH, but cell shape was characterized by a deepening of the incisions between cell lobes at higher pH. In both species, cell complexity based on surface-to-volume ratio was positively correlated with increasing pH. We conclude that by manipulating their surface-to-volume ratios, these desmid species can respond to pH variations in their environment.     

The structure of the gas bladder of the spotted gar,Lepisosteus oculatus

We report here on the macroscopic, light microscopic, and electron microscopic structure of the gas bladder (GB) of the spotted gar, Lepisosteus oculatus. The GB opens into the pharynx, dorsal to the opening of the oesophagus, through a longitudinal slit bordered by two glottal ridges. Caudal to the ridges, the GB is an elongated sac divided into a central duct and right and left lobes. The lobes are formed by a cranio‐caudal sequence of large air spaces that open into the central duct. The structure of the GB is that of a membranous sac supported by a system of septa arising from the walls of a central duct. The septa contain variable amounts of striated and smooth muscle might function to maintain the bladder shape and in providing contractile capabilities. The presence of muscle cells, nerves, and neuroepithelial cells in the wall of the GB strongly suggests that GB function is tightly regulated. The central duct and the apical surface of the thickest septa are covered by mucociliated epithelium. Most of the rest of the inner bladder surface is covered by a respiratory epithelium which contains goblet cells and a single type of pneumocyte. These two cell types produce surfactant. The respiratory barrier contains thick areas with fibrillar material and cell prolongations, and thin areas that only contain basement membrane material between the capillary wall and the respiratory epithelium. Lungs and GBs share many anatomical and histological features. There appears to be no clear criterion for structural distinction between these two types of respiratory organs. J. Morphol. 276:90–101, 2015. © 2014 Wiley Periodicals, Inc.     

Cadmium cytotoxicity and variation in nuclear content of DNA in Euglena gracilis

Cultures of Euglena gracilis (strain Z from French CNRS collection) can be made cadmium resistant if grown in a medium with 5x10^-4M cadmium chloride. This resistance is reflected by the appearance of a second exponential growth phase. The development of this resistance was studied at the cellular level by determining the relative content of DNA at different stages of the cell cycle in an asynchronously grown culture. The culture was followed until the second, cadmium resistant, growth phase had reached its stationary state. During the first exponential growth phase, cells were mostly in the late period of DNA synthesis (stage S of the cell cycle), or in the gap preceding mitosis (stage G₂ of the cell cycle). In addition, some cells contained high multiples of the normal amount of DNA. In the beginning of the second exponential growth phase, a few cells were again in G₁ (the post mitotic stage of the cell cycle preceding DNA synthesis). These G₁ cells were predominant at the end of the second growth period. During the second stationary phase the DNA content of the cadmium treated cells was similar to the stationary phase of the control culture. Cells had stopped growing in G₁ with an unreplicated genome. The implications of these data are discussed.