Effects of Metabolic Inhibitors on the Calcification of a Freshwater Green Alga Gloeotaenium loitlesbergarianum Hansgirg. 2. Effects of Some Inhibitors of Protein Synthesis

The effects of five inhibitors of protein synthesis, viz. streptomycin,aurin tricarboxylic acid, tetracycline, chloramphenicol, andcycloheximide, on the calcification of Gloeotaenium loitlesbergarianumHansgirg, a freshwater green alga were studied. Streptomycinhad no effect while aurin tricarboxylic acid at 50 µgml^–1 and tetracyline, chloramphenicol and cycloheximideat 20 µg ml^–1 completely inhibited calcificationin the alga. High concentrations of chloramphenicol and cycloheximidewere not completely inhibitory when added 26 h and 32 h respectivelyafter the material was incubated in the induction medium. Itis concluded that the effects by these substrates are the resultsof inhibition of protein synthesis, which is directly or indirectlylinked to calcification. calcification, Gloeotaenium loitlesbergarianum Hansgirg, green alga, chlorophyceae, protein synthesis inhibitors     

Effects of Some Metabolic Inhibitors on Heterocyst Formation in the Blue-green Alga, Anabaena doliolum

3, p-chlorophenyl urea, dinitrophenol, iodoacetic acid, andsodium azide inhibit heterocyst formation, whereas maleic hydrazidedoes not. Glucose promotes the frequency of heterocysts andis very effective in counteracting the inhibition by CMU andazide. The DNP inhibition is not reversed by glucose but ATPcauses a partial reversal. The iodoacetic acid inhibition isreversed partially by pyruvic acid; ATP and glucose are ineffectivein this case. Red light increases heterocyst frequency; greenand blue lights are inhibitory. It is concluded in the light of the mechanisms of action ofthe inhibitors that heterocyst formation depends on the availabilityof carbon intermediates and ATP. The former is supplied by photosynthesisand the latter most probably by oxidative metabolism.     

Effect of Metabolic Inhibitors on the Formation of Cell Walls in a Green Alga, Micrasterias crux-melitensis

The effects of cytochalasin B, N-ethylmaleimide (NEM), colchicine,vinblastine and cycloheximide on the formation of birefringentcell wall layers were studied. Birefringent layers accumulatedoutside the plasma membrane of daughter semicells when cellswere cultured in a 0.16 M mannitol solution without any inhibitors.In cells treated with 2 x 10^–5 M cytochalasin B, 3 x 10^–5M NEM, 10^–4 M vinblastine or 10^–5 M cycloheximidefor 24 hr, birefringent layers were not observed outside theplasma membrane, but were present in cells treated with 10^–2M colchicine. The possibility is discussed that substances necessaryfor wall synthesis could be transported from the cytoplasm tothe outside of the plasma membrane by a system associated withmicrofilaments, microtubules and myosin-like structures. (Received June 26, 1981; Accepted September 24, 1981)     

Effects of Phosphorus Limitation on Respiratory Metabolism in the Green Alga Selenastrum minutum

The effects of phosphorus nutrition on several physiological and biochemical parameters of the green alga, Selenastrum minutum, have been examined. Algal cells were cultured in chemostats under conditions of either Pi limitation or nutrient sufficiency. Pi limitation resulted in: (a) a 5-fold lower rate of respiration, (b) a 3-fold decline in rates of photosynthetic carbon dioxide fixation and oxygen evolution, (c) a 3-fold higher rate of dark carbon dioxide fixation, (d) significant increases in activities of phosphoenolpyruvate (PEP) carboxylase and PEP phosphatase (128% and 158% of nutrient sufficient activities, respectively), (e) significant reductions in activities of nonphosphorylating NADP-glyceraldehyde-3-phosphate dehydrogenase and NAD malic enzyme, and (f) no change in levels of ATP:fructose-6-phosphate 1-phosphotransferase, phosphorylating NAD-glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, and pyruvate kinase. The intracellular concentrations of Pi, ATP, AMP, soluble protein, and chlorophyll were also significantly reduced in response to Pi limitation. As well, the level of ADP was about 11-fold lower in the Pi-limited cells as compared to the nutrient sufficient controls. It was predicted that because of this low level of ADP, pyruvate kinase catalyzed conversion of PEP to pyruvate may be restricted in Pi-limited cells. During Pi limitation, PEP carboxylase and PEP phosphatase may function to “bypass” the ADP dependent pyruvate kinase, as well as to recycle Pi for its reassimilation into cellular metabolism.     

Rapid Biotransformation of Arsenate into Oxo-Arsenosugars by a Freshwater Unicellular Green Alga,Chlamydomonas reinhardtii

We examined the short-term metabolic processes of arsenate for 24 h in a freshwater unicellular green alga, Chlamydomonas reinhardtii wild-type strain CC-125. The arsenic species in the algal extracts were identified by high-performance liquid chromatography/inductively coupled plasma mass spectrometry after water extraction using a sonicator. Speciation analyses of arsenic showed that the levels of arsenite, arsenate, and methylarsonic acid in the cells rapidly increased for 30 min to 1 h, and those of dimethylarsinic acid and oxo-arsenosugar-glycerol also tended to increase continuously for 24 h, while that of oxo-arsenosugar-phosphate was quite low and fluctuated throughout the experiment. These results indicate that this alga can rapidly biotransform arsenate into oxo-arsenosugar-glycerol for at least 10 min and then oxo-arsenosugar-phosphate through both reduction of incorporated arsenate to arsenite and methylation of arsenite and/or arsenate retained in the cells to dimethylarsinic acid via methylarsonic acid as an possible intermediate.     

Photosynthetic Eukaryotes of Freshwater Wetland Biofilms: Adaptations and Structural Characteristics of the Extracellular Matrix in the Green Alga, Cosmarium reniforme (Zygnematophyceae, Streptophyta)

ABSTRACT. Cosmarium reniforme (Zygnematophyceae, Streptophyta) is a green alga that is commonly found in biofilms of wetlands of the Adirondack region, NY (USA). Two distinctive characteristics that are critical to this alga's survival in a benthic biofilm are its elaborate cell morphology and extracellular matrix (ECM). In this study, ultrastructural, immunocytochemical, and experimental methodologies were employed in order to elucidate the cellular characteristics that are critical for survival in a biofilm. The ECM consists of a thick, outwardly lobed cell wall (CW), which contains a patterned network of structurally complex pores. Each pore consists of a narrow channel, terminating internally at a bulb that invaginates localized regions of the plasma membrane. The outer region of the pore contains arabinogalactan protein-like and extensin epitopes that are likely involved in adhesion mechanisms of the cell. External to the CW is the extracellular polymeric substance that is employed in ensheathment of the cell to the substrate and in gliding motility. The architectural design/biochemical make-up of the CW and a secretory system that encompasses the coordinated activities of the endomembrane and cytomotile/cytoskeletal systems provide the organism with effective mechanisms to support life within the biofilm complex.     

Chlorophyll and Primary Productivity of Some Green,Freshwater Sponges

Relationships between ash-free dry weight, viable chlorophyll a and pheopigments a were examined by linear regression analysis for non-gemmulating and gemmulated Spongilla lacustris and for an unidentified, non-gemmulating species. The primary productivity of S. lacustris was determined by both oxygen and ¹⁴C techniques and expressed on a biomass, chlorophyll a, and area basis, and the photosynthetic efficiency was calculated. The proportion of algal photosynthate excreted by the sponge was investigated, and the transfer of algal photosynthate to the sponge was examined by autoradiography.     

The Role of a Green Alga in the Precipitation of Calcite and the Coprecipitation of Phosphate in Freshwater

The precipitation of calcite from a calcium bicarbonate solution, similar in ionic strength to natural hardwaters, was observed in a series of experiments utilizing an automated culture apparatus. Seeded growth experiments, using calcite seed crystals, were performed at a range of phosphate concentrations to observe inhibitory effects. These experiments demonstrated a linear relationship of increasing inhibition with increasing initial phosphate concentration. A further series of experiments was performed in which an actively photosynthesizing culture of a unicellular green alga (Chlorococcum sp.) was added to the culture vessel in order to initiate precipitation. Experiments to observe spontaneous precipitation, occurring in the absence of both seed and alga additions, were carried out to compare with precipitation rates in the algal experiments. A control experiment was also performed to investigate whether precipitation occurrred in algal cultures maintained in darkness. The carbonate site mechanistic model, developed for calcite precipitation in abiotic conditions, was used to analyse the results of the algal experiments and found to be applicable.     

Phosphorus-Limited Growth of a Green Alga and a Blue-Green Alga

The phosphorus-limited growth kinetics of the chlorophyte Scenedesmus quadricauda and the cyanophyte Synechococcus Nägeli were studied by using batch and continuous culturing techniques. The steady-state phosphate transport capability and the phosphorus storage capacity is higher in S. Nägeli than in S. quadricauda. Synechococcus Nägeli can also deplete phosphate to much lower levels than can S. quadricauda. These results, along with their morphological characteristics, were used to construct partial physiological profiles for each organism. The profiles indicate that this unicellular cyanophyte (cyanobacterium) is better suited for growth in phosphorus-limited oligotrophic niches than is this chlorophyte (green alga).     

Effect of pH on the Activity of Some Respiratory Inhibitors

Inhibition of respiration of cultured cells of Petunia hybrida by NaF, NaN₃, malonic acid, and salicylhydroxamic acid increased at low pH. This increase could be partially reversed by raising the pH of the medium. Uptake experiments show that the greater inhibition of malonic acid at low pH was not the result of greater uptake. The results suggest that the increase in inhibition at low pH could not be attributed to greater cell penetration.     

Selenium Accumulation in Unicellular Green Alga Chlorella vulgaris and Its Effects on Antioxidant Enzymes and Content of Photosynthetic Pigments

The aim of the present study was to investigate selenite effects in the unicellular green algae Chlorella vulgaris as a primary producer and the relationship with intracellular bioaccumulation. The effects of selenite were evaluated by measuring the effect of different selenite concentrations on algal growth during a 144 h exposure period. It was found that lower Se concentrations (≤75 mg L⁻¹) positively promoted C. vulgaris growth and acted as antioxidant by inhibiting lipid peroxidation (LPO) and intracellular reactive oxygen species (ROS). The antioxidative effect was associated with an increase in guaiacol peroxidase (GPX), catalase (CAT), superoxide dismutase (SOD) and photosynthetic pigments. Meanwhile, significant increase in the cell growth rate and organic Se content was also detected in the algae. In contrast, these changes were opposite in C. vulgaris exposed to Se higher than 100 mg L⁻¹. The antioxidation and toxicity appeared to be correlated to Se bioaccumulation, which suggests the appropriate concentration of Se in the media accumulation of C. vulgaris should be 75 mg L⁻¹. Taken together, C. vulgaris possesses tolerance to Se, and Se-Chlorella could be developed as antioxidative food for aquaculture and human health.     

179Conjugation Process of a Freshwater Green Alga, Spirogyra Varians (Zygnematales, Charophyceae Examined by Fitc-Lectins)

The conjugation processes of filamentous freshwater green alga Spirogyra varians were examined by the use of FITC-lectins. Conjugation comprised of five steps: 1) array with adjacent filaments, 2) formation of conjugation protrusion (papilla), 3) fusion of the protrusions, 4) formation of conjugation tube, and 5) formation of zygotes. Three lectins, ConA, RCA and UEA, showed considerable labeling during the progression of conjugation. FITC-ConA labeled the surfaces of filaments throughout the whole conjugation processes. No labeling of FITC-RCA was detected on the surface of vegetative filaments. FITC-RCA labeling was observed at the conjugation protrusions only after the papilla formation. Strong labeling continued until formation of zygotes even in hollow area between the conjugation tube. The labeling decreased gradually over time and disappeared when zygotes were formed. FITC-UEA showed similar labeling pattern with FITC-RCA except that the labeling did not disappear even after zygote formation. Inhibition experiments using D-galactose, L-fucose and D-mannose, which are complementary carbohydrates for the above lectins, showed considerable decrease of conjugation (<50% vs. 83% in control). Hamagglutination experiment using crude extract of Spyrogyra varians revealed existence of lectins specific for the above carbohydrates. These results suggested that the lectin-carbohydrate recognition system might be involved in the conjugation of Spirogyra varians .     

Effects of Some Metabolic Inhibitors on the Pigments of Euglena gracilis in an Acidic Medium

SYNOPSIS. Darkness and O₂-lack promote formation of a pheophytin-like pigment from chlorophyll in Euglena gracilis in an acidic medium. Dinitrophenol (DNP) produces similar but more drastic pigment alterations in aerated and illuminated cells. The extent of pigment decay was dependent on DNP concentration and external pH. Since volume expansion and inhibition of the contractile vacuole were also noted, it is suggested that pheophytin formation is secondary to hydrogen ion influx from the external medium. Major structural alterations of the chloroplast accompanied the pigment changes.
Pentachlorophenol, iodoacetamide, Na fluoride, and Na azide produced similar pigment changes. Malonic acid and fluoroacetate were ineffective under the conditions described. However, in the dark, cells treated with fluoroacetate formed pheophytin rapidly. These effects are discussed in relation to the maintenance of intracellular [H⁺].     

RNAi Knock-Down of LHCBM1, 2 and 3 Increases Photosynthetic H2 Production Efficiency of the Green Alga Chlamydomonas reinhardtii

Single cell green algae (microalgae) are rapidly emerging as a platform for the production of sustainable fuels. Solar-driven H₂ production from H₂O theoretically provides the highest-efficiency route to fuel production in microalgae. This is because the H₂-producing hydrogenase (HYDA) is directly coupled to the photosynthetic electron transport chain, thereby eliminating downstream energetic losses associated with the synthesis of carbohydrate and oils (feedstocks for methane, ethanol and oil-based fuels). Here we report the simultaneous knock-down of three light-harvesting complex proteins (LHCMB1, 2 and 3) in the high H₂-producing Chlamydomonas reinhardtii mutant Stm6Glc4 using an RNAi triple knock-down strategy. The resultant Stm6Glc4L01 mutant exhibited a light green phenotype, reduced expression of LHCBM1 (20.6% ±0.27%), LHCBM2 (81.2% ±0.037%) and LHCBM3 (41.4% ±0.05%) compared to 100% control levels, and improved light to H₂ (180%) and biomass (165%) conversion efficiencies. The improved H₂ production efficiency was achieved at increased solar flux densities (450 instead of ∼100 µE m⁻² s⁻¹) and high cell densities which are best suited for microalgae production as light is ideally the limiting factor. Our data suggests that the overall improved photon-to-H₂ conversion efficiency is due to: 1) reduced loss of absorbed energy by non-photochemical quenching (fluorescence and heat losses) near the photobioreactor surface; 2) improved light distribution in the reactor; 3) reduced photoinhibition; 4) early onset of HYDA expression and 5) reduction of O₂-induced inhibition of HYDA. The Stm6Glc4L01 phenotype therefore provides important insights for the development of high-efficiency photobiological H₂ production systems.     

UV-Induced Effects on Growth,Photosynthetic Performance and Sunscreen Contents in Different Populations of the Green Alga Klebsormidium fluitans (Streptophyta) from Alpine Soil Crusts

Members of the green algal genus Klebsormidium (Klebsormidiales, Streptophyta) are typical components of biological soil crust communities worldwide, which exert important ecological functions. Klebsormidium fluitans (F. Gay) Lokhorst was isolated from an aeroterrestrial biofilm as well as from four different biological soil crusts along an elevational gradient between 600 and 2350 m in the Tyrolean and South Tyrolean Alps (Austria, Italy), which are characterised by seasonally high solar radiation. Since the UV tolerance of Klebsormidium has not been studied in detail, an ecophysiological and biochemical study was applied. The effects of controlled artificial ultraviolet radiation (UVR; <9 W m^–2 UV-A, <0.5 W m^–2 UV-B) on growth, photosynthetic performance and the capability to synthesise mycosporine-like amino acids (MAAs) as potential sunscreen compounds were comparatively investigated to evaluate physiological plasticity and possible ecotypic differentiation within this Klebsormidium species. Already under control conditions, the isolates showed significantly different growth rates ranging from 0.42 to 0.74 μm day^?1. The UVR effects on growth were isolate specific, with only two strains affected by the UV treatments. Although all photosynthetic and respiratory data indicated strain-specific differences under control conditions, UV-A and UV-B treatment led only to rather minor effects. All physiological results clearly point to a high UV tolerance in the K. fluitans strains studied, which can be explained by their biochemical capability to synthesize and accumulate a putative MAA after exposure to UV-A and UV-B. Using HPLC, a UV-absorbing compound with an absorption maximum at 324 nm could be identified in all strains. The steady-state concentrations of this Klebsormidium MAA under control conditions ranged from 0.09 to 0.93 mg g^?1 dry weight (DW). While UV-A led to a slight stimulation of MAA accumulation, exposure to UV-B was accompanied by a strong but strain-specific increase of this compound (5.34–12.02 mg^?1 DW), thus supporting its function as UV sunscreen. Although ecotypic differences in the UVR response patterns of the five K. fluitans strains occurred, this did not correlate with the altitude of the respective sampling location. All data indicate a generally high UV tolerance which surely contributes to the aeroterrestrial lifestyle of K. fluitans in soil crusts of the alpine regions of the European Alps.