The Effect of Deleterious Concentrations of Mercury on the Photosynthesis and Growth of Chlorella pyrenoidosa

The effect of mercury ions on the photosynthesis and growth of Chlorella pyrenoidosa in a balanced medium has been studied and compared with the effect of copper. In many ways Hg treated algae behave like algae treated with Cu in concentrations of the same molarity, but important deviations occur. Hg acts at a lower and in a more narrow range of concentrations than does Cu due to a more specific binding. The depressing effect of Hg is not counteracted by other cations such as potassium and sodium, and iron has only a slight effect. Cell division is stopped after Hg addition and there is no accumulation of assimilation products. On the contrary the cells become pale yellow. Preliminary studies indicate a light-independent leakage in the cytoplasmatic membrane leading to an outflow of potassium ions. This is the primary action of both Hg and Cu poisoning, but the leakage does not seem to be correlated with the decrease of photosynthesis. After a lapse of time — dependent on the mercury/cell concentration ratio — a detoxication takes place probably due to the binding of Hg to “insensitive sites” in the cells. Probable mechanisms for the action of Hg on photosynthesis and growth are discussed.     

The Effect of Deleterious Concentrations of Copper on the Photosynthesis of Chlorella pyrenoidosa

If a single salt solution of CuSO₄ is used, Cu penetrates immediately iuto the plasma of Chlorella cells, reducing the rates of photosynthesis at both high and low illumination. If CuSO₄ is added to ordinary Österlind culture-medium (pH 8) it takes some hours before any influence of deleterious concentrations of Cu is observed and initially only at light saturation. The algae must have been illuminated during the whole period. Maximum influence of CuSO₄ is found duriug the first 24 hours of treatment. A significant deleterious influence of Cu concentrations as low as about 1 μg/l is found. The influence of Cu increases with decreasing concentrations of the alga. If a culture medium at pH 5 is used instead of the ordinary one at pH 8, copper concentrations ahout 10 times as high must he used be order to obtain the same deleterious effect. An increase of the cotncentration of K reduces the influence of Cu to some extent. These facts show that the effect of deleterious concentrations of Cu in halanced solutions is not due to a marked penelration of this ion into the plasma but to a binding to the cytoplasmic membrane whereby the celts i.a. become more or less unable to divide. The cells become saturated with assimilation products which have a depressant effect on the rate of photosynthesis. Other cations compete with Cu for the “active sites” on the membranes.     

自养和兼养条件下蛋白核小球藻昼夜节律的响应

【目的】研究自养和兼养两种培养方式对蛋白核小球藻(Chlorella pyrenoidosa)生长、细胞分裂和生化组分积累的影响,探讨人工培养蛋白核小球藻的昼夜节律响应机制和优化技术。【方法】小球藻自养培养采用BG11培养基,兼养培养基在BG11培养基中添加4种不同浓度(1、5、10、20 g/L)的葡萄糖,培养周期为10 d。血球板计数法测定藻细胞浓度,干重法测定藻细胞生物量。显微观察藻细胞大小和分裂情况。脂染色法测定小球藻总脂的含量,藻细胞的叶绿素、蛋白和淀粉分别采用甲醇、氢氧化钠、硝酸钙浸提后通过紫外分光光度法定量测定。【结果】葡萄糖兼养培养对蛋白核小球藻具有显著的促生长效应,最适浓度为10 g/L。10 d收获时,兼养组(10 g/L葡萄糖)藻细胞浓度和干重分别是自养组的2.57倍和6.73倍。分析一昼夜中的藻细胞增殖规律可知,第2天和第5天时自养组中增殖的新生子细胞约有76.00%在黑暗期分裂产生,而兼养组中第2天和第5天光照期的新细胞增殖量占比分别达到40.90%和67.50%。一昼夜内藻细胞大小的迁移动态监测表明,第2天自养组藻细胞的体积变化静息期为8 h,兼养组只有4 h;第5天两组藻细胞大小迁移动态的昼夜节律明显,但兼养组黑暗结束后较大细胞(D6μm)占比显著高于自养组。第8天时,兼养组藻细胞已处于稳定期,总脂和蛋白含量均显著高于自养组,藻细胞总脂和色素含量在一昼夜中相对稳定,但蛋白和淀粉含量分别在光照8 h和12 h左右达到峰值。从第2天开始,对兼养组细胞每天进行2 h光延长,收获时藻细胞浓度和干重分别比对照组提高13%和11%。【结论】葡萄糖兼养培养能大幅提高蛋白核小球藻的生物量。蛋白核小球藻生长增殖与生化组分积累均受昼夜节律调控,自养条件下藻细胞以光照期生长黑暗期增殖为主。兼养培养提高藻细胞生物量的机制在于缩短藻细胞生长静息期,在昼夜节律中加速藻细胞生长并显著提高通过细胞周期检查点的细胞比例,光照期效应尤其明显。藻细胞蛋白和淀粉含量昼夜节律明显,最佳收获时间分别在光照8 h和12 h后。     

The Effect of Chromium on the Photosynthesis and Growth of Diatoms and Green Algae

The effect of hexavalent chromium ions on the photosynthesis and growth of the diatom Nitzschia palea and the green alga Chlorella pyrenoidosa has been studied and compared with the effect of Cu and Hg. The effect of chromium is very similar to that of Cu and Hg, but it is more than one hundred times less toxic for the algae than copper. The toxicity of chromium is not counteracted by adding Fe to the culture solution. The problems regarding cleaning glass-ware in dichromate-sulphuric acid are discussed and it is strongly recommended to use other cleaning agents.     

EFFECTS OF HYDROXAMATE SIDEROPHORES (STRONG Fe (III) CHELATORS) ON THE GROWTH OF ALGAE1

Varying concentrations of Fe were tested with three hydroxamate siderophores to demonstrate the interactions affecting growth of Chlamydomonas reinhardtii and Chlorella vulgaris. Schizokinen was purified from the excretions of the blue-green alga Anabaena sp. grown in low-Fe medium. Chlamydomonas reinhardtii was inhibited by schizokinen when in molar excess of the Fe concentration; the inhibition was overcome with excess Fe. The growth of C. vulgaris was not affected by this chelator. Results with desferrioxamine were similar. A weaker chelator, rhodorulic acid, did not inhibit the growth of either alga. Low concentrations of the chelators may stimulate algal growth when Fe precipitates are hydrolyzed. Since different algae respond differently to the presence of the chelators, the observed interactions could be important in determining competitive relationships when Fe is limiting. If an alga can excrete a strong chelating agent, as does Anabaena, algae lacking the ability to compete with the chelator may not grow.     

Observations on tolerance to heavy metals of four green algae in relation to pH

Abstract

The inhibition of growth by different concentration of eight heavy metals: Cd, Cr, Co, Cu, Hg, Ni, Se and Tl, in inorganic medium at pH 3 and 6.5, was studied in four green algae: Chlorella protothecoides Krüger, Chlorella saccharophila (Krüger) Migula, Coenochloris sp. and Stichococcus bacillaris Naegeli.

The results suggest that pH has an important effect on heavy metal toxicity in algae although it is difficult to establish a relationship between pH and heavy metals.     

Responses of Two Coastal Algae (Skeletonema costatum and Chlorella vulgaris) to Changes in Light and Iron Levels1

Iron (Fe) is essential for phytoplankton growth and photosynthesis, and is proposed to be an important factor regulating algal blooms under replete major nutrients in coastal environments. Here, Skeletonema costatum, a typical red-tide diatom species, and Chlorella vulgaris, a widely distributed Chlorella, were chosen to examine carbon fixation and Fe uptake by coastal algae under dark and light conditions with different Fe levels. The cellular carbon fixation and intracellular Fe uptake were measured via ¹⁴C and ⁵⁵Fe tracer assay, respectively. Cell growth, cell size, and chlorophyll-α concentration were measured to investigate the algal physiological variation in different treatments. Our results showed that cellular Fe uptake proceeds under dark and the uptake rates were comparable to or even higher than those in the light for both algal species. Fe requirements per unit carbon fixation were also higher in the dark resulting in higher Fe: C ratios. During the experimental period, high Fe addition significantly enhanced cellular carbon fixation and Fe uptake. Compared to C. vulgaris, S. costatum was the common dominant bloom species because of its lower Fe demand but higher Fe uptake rate. This study provides some of the first measurements of Fe quotas in coastal phytoplankton cells, and implies that light and Fe concentrations may influence the phytoplankton community succession when blooms occur in coastal ecosystems.     

The relationship of protein synthesis to cell division and oral development in synchronized Tetrahymena pyriformis GL-C: An analysis employing cycloheximide

The effects of cycloheximide on synchronized Tetrahymena pyriformis strain GL-C were investigated at concentrations ranging from 0.01 to 10 μg/ml. The initial inhibition of protein synthesis was nearly total (>85%) at 1 μg/ml and above, partial (50–80%) at 0.2 to 0.05 μg/ml, and slight (<30%) at 0.02 μg/ml. Eventual recovery of protein synthesis to a rate approaching that of the controls took place at concentrations of 1 μg/ml and less. When the drug was added before a “transition point” at 55 minutes after the end of the synchronizing treatment (EST), cell division was blocked by 10 μg/ml, and delayed at concentrations of 1 μg/ml or less. The duration of delay was related to the degree of initial inhibition, and to the time required for recovery of protein synthesis; it also depended on the time after EST at which the drug was added. At a given concentration, maximum division delay was observed just prior to the “transition point;” this maximum delay was correlated with resorption of differentiating oral primordia, followed by the appearance of new primordia. The lesser delays observed at earlier times were correlated with temporary blockage of development of primordia in the “stomatogenic field” stage. Resumption of oral primordium development was, in both cases, temporally correlated with a substantial recovery of protein synthesis. After the “transition point,” cell division, and completion of oral development, was delayed slightly at the lower concentrations, and more substantially at 1 and 10 μg/ml, with some division-arrest at the latter concentration. Except for the recovery phenomenon, the developmental responses elicited by cycloheximide were similar to those observed earlier with puromycin. The bearing of these findings on the mechanism of synchronization in Tetrahymena is considered in the Discussion.     

Physiologische Untersuchungen an einer ungewöhnlich säureresistenten Chlorella

Summary The influence of the hydrogen-ion concentration on the growth and metabolism of a highly acid-resistant green alga, Chlorella ellipsoidea (strain Marburg St), was studied. Chlorella pyrenoidosa (Emerson strain) served as a normal control organism. Growth of Chlorella ellipsoidea occurs in the entire range from Ph 2.0 to Ph 10, whereas for Chlorella pyrenoidosa the limits were found to be Ph 3.5 and Ph 10. Respiration is much less sensitive to hydrogen-ion concentration in the acid-resistant as compared to the normal strain. Thus an increase in acidity from Ph 4.0 to Ph 2.0 increases the respiratory oxygen uptake by 120% in Chlorella pyrenoidosa and by 25% in Chlorella ellipsoidea. In addition, only the less resistant Chlorella pyrenoidosa shows an accumulation of nitrite in the dark in acid culture media, indicating a disturbance of the normal course of nitrate reduction under these conditions. On the other hand, the rate of photosynthesis of both organisms was found to be almost independent of acidity between Ph 4.0 and Ph 2.0. At the acid and alkaline limits of growth in both algae, an inhibition of cell division leads to an increase of cell size and dry weight per cell, frequently connected with the occurrence of bizarre giant cells. — In addition, adaptation phenomena were found to play a role in determining the acid limit of growth. Cells of Chlorella ellipsoidea, after inoculation from normal medium (Ph about 6) into a solution of Ph 2.0, begin growth at a high rate only after a lag of about two weeks. Cells grown previously in an acid medium, however, immediately resume growth upon inoculation into a medium of Ph 2.0. This adaptation involves a considerable reduction of cell size.     

The effect of indomethacin on the growth and metabolism of green alga <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> Beijerinck

The aim of the present work was to study the effect of indomethacin (IM), a pleiotropic therapeutic substance commonly used in animal systems, at concentration range of 10⁻⁸ to 10⁻³ M on the growth and metabolism of single-celled Chlorella vulgaris (Chlorophyceae). Because of the presence of the indole ring in its molecule, IM is characterized by structural similarity with natural auxins, e.g. IAA. It was found that IM influenced algal growth, macromolecular synthesis and metabolism in dose-dependent manner. IM had the highest stimulating effect on algae at 10⁻⁷ M on the 5th day of culture resulting in the increase in cell number and dry mass, DNA, RNA, proteins, phosphates, monosaccharides, photosynthetic pigments and glycolic acid content as well as protein extracellular secretion to the environment. Specific proteins from the region 20–139 kDa appeared during 10⁻⁷ M IM treatment on the 5th day of cultivation as analysed by SDS-PAGE. IM-induced photosynthetic oxygen exchange in green alga was also noted. In contrast, the treatment with IM at the highest concentration of 10⁻³ M suppressed cell division, dry mass production and decreased the level of the analysed parameters during the whole 7-day period of cultivation. Therefore, it could be speculated that IM functioned as a plant growth regulator affecting cell division and metabolism of green alga C. vulgaris.     

Fatty acid production by heterotrophic Chlorella saccharophila

The fatty acid composition of the alga Chlorella saccharophila was investigated under different growth conditions. Using glucose as the sole carbon source, heterotrophically-grown Chlorella saccharophila produced a greater proportion of the polyunsaturated fatty acids (C18: 2 and C18: 3) than photosynthetic cultures, with linoleic acid (C18: 2) predominating. An unexpected discovery was the observation that at the lowest glucose concentration (2.5 gl^–1) the lipid content of the algae increased to between 36–47% of the cell weight, depending on the temperature. At glucose concentrations of 5 g l^–1 or more, the lipid content fell to 10–12% of the cell, although total fatty acid yield was higher due to higher biomass concentrations. Aeration of heterotrophic cultures promoted the production of unsaturated fatty acids compared to non-aerated cultures.     

Metal Tolerance and Accumulation Ability of the Ni Hyperaccumulator Streptanthus polygaloides Gray (Brassicaceae)

High concentrations of metals occur in some plant species (termed hyperaccumulators), such as the Ni hyperaccumulator Streptanthus polygaloides. We determined the tolerance of S. polygaloides to, and its accumulation abilities for, six metals (Ni, Zn, Cu, Co, Mn, and Pb). Potting mix concentrations used for all metals ranged from 0 to 1200 μg/g dry weight. For Ni, a treatment of 1600 μg/g was included. For Mn, treatments of 1600, 2000, and 2500 μg/g also were used, and for Pb these concentrations plus 3500 μg/g were included. Germination, plant number per pot, and size at days 30 and 39, number of plants at the end of the experiment (day 49), flower production, and metal concentration in the aboveground biomass were documented. Lead and Ni showed no consistent effects on plant performance, but yielded increased tissue metal concentrations. Streptanthus polygaloides was more sensitive to Co, Cu, and Zn, as ≥ 400 mg/g significantly suppressed plant growth, survival, and flower production. Tissue metal concentrations also were increased to maxima of 1500 μg Co/g, 120 μg Cu/g, and 6000 μg Zn/g. Manganese affected S. polygaloides less markedly, as ≥ 800 mg/kg decreased growth, survival, and flower production. Maximum tissue Mn concentration was 2900 μg/g. We concluded that S. polygaloides would be an appropriate phytoextractor for soils contaminated with Ni or low levels of Co but would not be useful for Cu, Zn, Mn, and Pb.     

The Influence of Cu on Photosynthesis and Growth in Diatoms

Cu in ionic form in a balanced medium affects the rate of photosynthesis and growth in the diatom Nitzschia palea in about the same way as in the green alga Chlorella pyrenoidosa. Remarkable differences are found, however. Small concentrations of Cu influence the rate of photosynthesis considerably more in the diatom than in the green alga whereas the opposite is the case concerning growth. The latter is most likely a consequence of excretion of organic matter by the diatom in presence of Cu whereas this does not take place in Chlorella. Some of the excreted organic matter may bind Cu and thus make the medium suitable for growth. Although pre-treatment with Cu in the dark has no influence on the subsequent rate of photosynthesis in the light, organic matter is excreted in the dark immediately after the addition of Cu. The influence of Cu on the rate of photosynthesis varies by a factor of about 30 according to the stage of growth in a synchronized culture.     

Effects of iron,copper, zinc,calcium, and magnesium on human lymphocytes in culture

The effects of simultaneous changes of calcium, magnesium, iron, copper, and zinc concentrations were evaluated in normal human T and B lymphocytes, cultured in cation-depleted media. Optimal concentrations for thymidine incorporation (TI) in both cell populations were Fe and Zn 15 μM and Cu 5 μM; for t cells Ca 2 mM and Mg 4 mM; for B cells Ca 4 mM and Mg 6 mM. TI decreased with increasing molarity of cations and the decrease was particularly apparent with Cu. Minimal amounts of Ca and Mg (0.5 mM) were necessary for growth, even in presence of optimal concentrations of Fe, Cu, and Zn. Fe and Cu showed synergistic stimulatory effects at low concentrations and synergistic inhibitory effects at high concentrations. Antagonism between Fe and Zn, Cu and Zn, and Ca and Zn was also demonstrated. CD4/CD8 increased with PHA stimulation in presence of Zn, and decreased with ConA stimulation in presence of Zn or Fe. The results demonstrate: (1) the relationship and interdependence of Fe, Cu, and Zn concentrations in modulating the growth of normal lymphocytes; (2) the stimulatory effects of Fe on B cells and Zn on CD8 positive cells; (3) the inhibitory effect of Cu at concentrations lower than those of Fe and Zn; (4) the requirement of Ca and Mg in certain concentration and ratio for the action of the other cations; and (5) the Ca and Mg requirement for the growth of B cells higher than T cells.     

The Demand for Iodine and Bromine of three Marine Brown Algae grown in Bacteria-free Cultures

Three marine brown algae have been cultivated with different additions of iodine and bromine in bacteria-free cultures. Ectocarpus jasciculatus appeared to have an absolute demand for iodine and was inhibited by a concentration of 64 μmol of KJ per 1. Lithosiphon pusillus had the best growth in the highest concentration tested (64 μmol/1) but there was always some growth in the series without iodine. Additions could be made either as inorganic iodine or as organically bound iodine. Additions of KJ to a culture medium consisting of vitamin-free Asp 6 F with B₁₂ (1 μg/1) and kinetin (20 μmol/1) remarkably increased the growth of the zoospores of Pylaiella litoralis. Lithosiphon pusilius proved to be indifferent to bromide additions in media containing KJ. In media lacking KJ addition of 1 μmol of KBr per 1 is stimulating but higher concentrations of KBr are inhibiting. The inhibiting effect is overcome by iodide addition.     

Effects of cytochalasin B on cell division and vacuole formation in Tetrahymena pyriformis GL

Cytochalasin-B (Cyt-B) was tested for its effect on cell division and vacuole formation in Tetrahymena. Little effect was found on cell division in the synchronous cell system in concentrations up to 37 μg/ml; however, slight delay was caused by 71 μg/ml. As measured by particle uptake, much lower concentrations, 7–8 μg/ml, caused significant inhibition of vacuole formation in exponentially multiplying and in starved cells, 16.6 or 37 μg/ml caused strong inhibition. This effect was immediate and completely reversible. The presence of Cyt-B caused starvation of Tetrahymena. The essential absence of inhibition of cell division by Cyt-B may reflect that the drug can enter the cell only by way of vacuoles.     

Influence of the external redox state on heavy metal adsorption by whole cells and isolated cell walls of Chlorella fusca

Summary A reduction of the external redox potential from 343 mV (atmospheric O₂ saturation) to 143 mV had only a slight effect on heavy metal uptake by whole cells of Chlorella fusca but caused an increased copper adsorption and a threefold increase of lead accumulation by isolated cell walls.     

Cr3+和Cd2+对普通小球藻生长及抗氧化酶活性的影响

[目的] 为探究重金属对淡水绿藻生长的影响。[方法] 选取对水质检测具有明显指示作用的普通小球藻（Chlorella vulgaris）为实验材料,CdCl₂·2H₂O和CrCl₃·7H₂O提供重金属离子,探究不同浓度Cr³⁺和Cd²⁺在单一和复合胁迫下对藻细胞浓度、叶绿素a及相关抗氧化酶活性的影响。[结果] 随着Cr³⁺和Cd²⁺浓度不断增加,藻细胞浓度呈先增长后下降趋势;叶绿素a含量呈现先下降后升高再下降的现象,浓度为1 mg/L的单一和复合胁迫下有最大值,且毒性作用表现为Cr³⁺ < Cd²⁺ < Cr³⁺+Cd²⁺;与藻细胞膜相关的丙二醛（MDA）和过氧化氢（H₂O₂）含量随着重金属离子浓度的增大而增长;重金属离子浓度低于10 mg/L时对藻细胞内抗氧化酶系统中的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和过氧化物酶（POD）表现为促进作用,而大于10 mg/L时具有抑制作用。[结论] 结果表明在单一或复合重金属胁迫下,普通小球藻会充分调动与抗逆性相关的酶来维持自身的正常生长。     

Integration of algae cultivation as biodiesel production feedstock with municipal wastewater treatment: strains screening and significance evaluation of environmental factors

The objectives of this study are to find the robust strains for the centrate cultivation system and to evaluate the effect of environmental factors including light intensity, light–dark cycle, and exogenous CO₂ concentration on biomass accumulation, wastewater nutrient removal and biodiesel production. The results showed that all 14 algae strains from the genus of Chlorella, Haematococcus, Scenedesmus, Chlamydomonas, and Chloroccum were able to grow on centrate. The highest net biomass accumulation (2.01 g/L) was observed with Chlorella kessleri followed by Chlorella protothecoides (1.31 g/L), and both of them were proved to be capable of mixotrophic growth when cultivated on centrate. Environmental factors had significant effect on algal biomass accumulation, wastewater nutrients removal and biodiesel production. Higher light intensity and exogenous CO₂ concentration with longer lighting period promote biomass accumulation, biodiesel production, as well as the removal of chemical oxygen demand and nitrogen, while, lower exogenous CO₂ concentration promotes phosphorus removal.     

Cell wall and cell growth characteristics of giant‐celled algae

Because of their large sizes and simple shapes, giant‐celled algae have been used to study how the structural and mechanical properties of cell walls influence cell growth. Here we review known relationships between cell wall and cell growth properties that are characteristic of three representative taxa of giant‐celled algae, namely, Valonia ventricosa, internodal cells of characean algae, and Vaucheria frigida. Tip‐growing cells of the genus Vaucheria differ from cells undergoing diffuse growth in V. ventricosa and characean algae in terms of their basic architectures (non‐lamellate vs. multilamellate) and their dependence upon pH and Ca²⁺ for cell wall extensibility. To further understand the mechanisms controlling cell growth by cell walls, comparative analyses of cell wall structures and/or associated growth modes will be useful. The giant‐celled algae potentially serve as good models for such investigations because of their wide variety of developmental processes and cell shapes exhibited.