The growth stimulating effect of 5-bromouracil and uracil on chlorococcal algae

The growth stimulating effect of 5-bromouracil and uracil on two strains of chlorococcal algae has been found in cell colonies grown from single cells, which were inoculated onto an agar medium. Analyses of the effect recorded in the cell cycles after treatment have revealed that the growth stimulating effect required four, or more, cell cycles to become evident. This has been proved by the number of autospores released from the treated cells and by the length of the lag phase after inoculation. The differences between the control and the treated population in some experiments with 5-bromouracil inChlorella and with uracil inScenedesmus obliquus have been visible by the unaided eye, whereas in some other experiments, they have been proved statistically. Growth stimulation has not been found only in a small number of experiments. The inhibition of growth induced by 5-bromouracil has been recorded in one experiment withScenedesmus quadricauda.     

Influence of pyrocatechol on characteristics of photosynthesis in an algae culture

Reduced intensity of delayed fluorescence of chlorophyll and suppressed growth recorded in an experimental population of Scenedesmus quadricauda under the influence of pyrocatechol demonstrate reduction in efficiency of light reactions of photosynthesis, which, in turn, determines deterioration of algae productivity.     

Comparison of N-ethyl-N-nitrosourea Effects in three strains of chlorococcal algae

Three strains of three different species of chlorococcal algae(Chlorella kesslerii Scenedesmus quadricauda andScenedesmus obliquus) were compared in their physiological and genetical responses to N-ethyl-N-nitrosourea (NEU). The mutagen was applied in the basic doses corresponding to their specific tolerance to it. Exponentially-graded doses derived from the basic ones acted for the same period of time and single basic doses acted at linearly-increased periods of time in parallel experiments. The influence of the mutagen on the cell cycle just in progress was characterized by the increase in the frequency of cell divisions which yielded a lower number of autospores. The conditions of the cells during the first three days after treatment were determined according to the average increase of their lag phase. The relative toxicity of the mutagen was expressed as the survival frequencies. The mutation effects were evaluated according to the frequency of permanent changes observed in the growing cell colonies : lethal, morphological and pigmentation. NEU was found to be a relatively suitable mutagen for the chlorococcal algae studied.     

Variability in characteristics of production and macromolecule content within microalgae cells during growth period

Variability in primary production and destruction during growth of an experimental population of Scenedesmus quadricauda has been determined. Accumulation of proteins, total lipids, and carbohydrates in algae cells was depending on phases of an algae population lifecycle.     

Sorption of some ions by algae related to their trophic conditions

The unicellular algaeScenedesmus obliquus (125),Chlorella pyrenoidosa (82) andCoccomyxa solorinae saccatae (111) were studied with respect to the form of uptake of potassium, phosphate, calcium and zinc ions and to the energy sources involved: light under autotrophic conditions, glucose under mixotrophic or heterotrophic conditions (in light and in darkness or together with yeast extract as an auxotrophic substrate). We respected the trophic conditions of algae when preparing the experimental material (precultivation). The following conditions were reached:

1. (1)
   The three algae grow faster in a glucose medium under mixotrophic conditions and are capable of growing on it also heterotrophically:Ch. pyrenoidosa andSc. obliquus grow substantially better thanC. solorinae saccatae. The first two algae grow more intensively in a glucose medium containing yeast extract whileCoccomyxa does not. After cultivation under mixotrophic conditions the first two diminish endogenous respiration, the third raises it. Glucose stimulates respiration in the first two when grown autotrophically, while after mixotrophic cultivation the effect of glucose is suppressed inCh. pyrenoidosa and in the other two only after growth on glucose with yeast extract.     
   
   

The Gas Exchange of Hydrogen-adapted Algae as Followed by Mass Spectrometry

A mass spectrometer inlet and an oxygen electrode in the same vessel allowed the continuous recording of the gases exchanged (H₂, CO₂, O₂) by hydrogenase-containing anaerobically adapted Scenedesmus obliquus strain D₃ (Gaffron) and Chlorella fusca Shihira et Krauss (= pyrenoidosa) 211-15. A light intensity which produces more photosynthetic oxygen than the cells can re-reduce to water leads to de-adaptation and the substitution of normal photosynthesis for photoreduction. The sequence of these metabolic events was recorded in a matter of a few minutes. Upon exposure of these adapted algae to light, an evolution of hydrogen lasting up to 60 seconds preceded any other light-dependent gas exchange. In the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea, this initial hydrogen production was inhibited approximately 50%, pointing to a contribution of electrons by photosystem II. At very low hydrogen tensions (0.1 microliter per milliliter), a balance between light-induced production and absorption of hydrogen was observed in normal, unpoisoned algae. Addition of either glucose or inhibitors of phosphorylation increased the release of hydrogen in the light very considerably. When the light was turned off the algae consumed the remaining amount of hydrogen, only to release it again upon illumination. This reversible hydrogen exchange persisted even when any concomitant carbon dioxide exchange had been abolished.     

The Combined Effect of Shungite and Heavy Metals on the Growth of Microalgae Роpulation

The combined effect of 3 mg/L potassium dichromate, 1.5 mg/L cadmium sulfate, and 100 g/L shungite on the growth of chlorococcales green microalgae culture Scenedesmus quadricauda (Turp.) Bréb. is studied. The toxic effect of potassium dichromate and cadmium sulfate on S. quadricauda is estimated by calculating the share of living and dead cells and physiological parameters. The toxic effect of heavy metals does not manifest itself under the combined action of potassium dichromate or cadmium sulfate and shungite on S. quadricauda. The best growth of the algae culture occurred only when only shungite was added to the culture medium. Shungite can be used to neutralize the toxic effect of heavy metals.     

6种微藻对氯霉素和硫酸新霉素敏感性研究

目的:探讨3种真眼点藻(点状魏氏藻(Visderia punctata)、波氏真眼点藻(Eustigmatos polyphem)、魏氏真眼点藻(Eustigmatos vischeri))和3种绿藻(栅藻(Scnedesmus sp.)、斜生栅藻(Scenedesmus obliqulis)、爪哇栅藻(Scenedesmus jaoaensis))对2种抗生素的敏感性.方法:采用藻液细胞计数法和藻细胞固体平板培养法研究了氯霉素和硫酸新霉素对6种微藻生长的影响.结果:液体培养,3种绿藻对氯霉素敏感性均高于硫酸新霉素,10μg·mL-1氯霉素即可明显抑制3种绿藻的生长(P＜0.05),而硫酸新霉素在浓度为200 μg·mL-1时才显示出明显抑制作用:3种真眼点藻对2种抗生素都不敏感.固体培养,除波氏真眼点藻外,其它5种微藻对氯霉素的致死浓度均为50μg·mL-1;波氏真眼点藻、栅藻、斜生栅藻和爪哇栅藻对硫酸新霉素的致死浓度分别为100 μg·mL-1、200μg· mL-1、50μg·mL-1和50μg·mL-1.结论:氯霉素可作为选育6种微藻抗性突变株的筛选剂.     

Synchronization of the cultures ofScenedesmus quadricauda by optimalizing the length of the light period

Experiments revealed that the synchronized cultures ofScenedesmus quadricauda, strainGreifswald/15, must be kept under continuous illumination throughout the whole growth phase (from the release of daughter coenobia up to the moment when all cells are capable of further reproduction). The length of the light period is among others the function of the suspension density. With regard to these facts a method of synchronization was worked out on the basis of a rhythmical repetition of the calculated light regime, corresponding to the physiological parameters of the synchronized strain under given cultivation conditions. This method enables to maintain synchronized cultures in adequate cycles for any required period, even in sufficiently dense, linearly growing suspensions. It may be applied to any chlorococcal algae; however, if coenobial types are studied, it is necessary to consider special peculiarities, which result from the regular arrangement of the cells in the coenobium.     

光、温限制后铜绿微囊藻和斜生栅藻的超补偿生长与竞争效应

研究了铜绿微囊藻(Microcystis aeruginosa)和斜生栅藻(Scenedesmus obliquus)低温和低光照限制后的超补偿效应,以及共培养条件下的竞争效应。结果表明,低温和低光照均显著抑制微藻的生长发育,但低温对铜绿微囊藻的抑制效应更强,而斜生栅藻则对低光胁迫更敏感。经过低光和低温培养后,铜绿微囊藻和斜生栅藻在恢复正常培养时藻细胞密度短期内都表现出超补偿增长效应,但不同藻类超补偿模式不同,斜生栅藻补偿生长时间不超过1周,而铜绿微囊藻的补偿效应可以持续10天;此外,统计结果表明铜绿微囊藻细胞密度对低温限制解除表现出更显著的补偿生长,斜生栅藻则在低光解除后表现出更强的超补偿效应。微藻叶绿素a指标在光恢复条件下都表现出显著的补偿效应,但温度恢复过程中叶绿素a含量与藻密度增长不同步,低温胁迫对恢复正常培养后微藻叶绿素a的形成产生了一定的负效应;铜绿微囊藻产毒株(912)在两种恢复模式下脱氢酶活性显著高于对照,产毒株(912)脱氢酶活性的补偿响应明显高于其它两种材料。共培养实验结果表明斜生栅藻同铜绿微囊藻产毒株(912)相比处于竞争劣势,而在同无毒株(469)的共培实验中,尽管连续正常培养情况下两者竞争能力差异不显著,但在恢复培养条件下斜生栅藻竞争能力显著高于后者。因此产毒型铜绿微囊藻低温和低光后的补偿生长效应以及对斜生栅藻的竞争优势可能是蓝藻爆发的内源性机制之一。     

Biomass production,total protein,chlorophylls, lipids and fatty acids of freshwater green and blue-green algae under different nitrogen regimes

Two green algae (Chlorella vulgaris and Scenedesmus obliquus) and four blue-green algae (Anacystis nidulans, Microcystis aeruginosa, Oscillatoria rubescens and Spirulina platensis) were grown in 81 batch cultures at different nitrogen levels. In all the algae increasing N levels led to an increase in the biomass (from 8 to 450 mg/l), in protein content (from 8 to 54 %) and in chlorophyll. At low N levels, the green algae contained a high percentage of total lipids (45 % of the biomass). More than 70 % of these were neutral lipids such as triacylglycerols (containing mainly 16:0 and 18:1 fatty acids) and trace amounts of hydrocarbons. At high N levels, the percentage of total lipids dropped to about 20 % of the dry weight. In the latter case the predominant lipids were polar lipids containing polyunsaturated C₁₆ and C₁₈ fatty acids. The blue-green algae, however, did not show any significant changes in their fatty acid and lipid compositions, when the nitrogen concentrations in the nutrient medium were varied. Thus the green but not the blue-green algae can be manipulated in mass cultures to yield a biomass with desired fatty acid and lipid compositions. The data may indicate a hitherto unrecognized distinction between prokaryotic and eukaryotic organisms.     

On the mass culture of microalgae: Areal density as an important factor for achieving maximal productivity

Scenedesmus obliquus was grown in a bioreactor under constant light and at a temperature of 24°C. From measurements of biomass production, it was possible to determine the influence of areal density on productivity in open as well as closed cultures. Maximal productivity occurred at an areal density of 40 to 45 g(dw) M⁻², which was in good agreement with predicted values from a model. The optimal areal density for maximal productivity was influenced by factors such as culture depth, algal species, turbulence, available light, etc., and hence the establishment and maintenance of an optimal areal density is seen as one of the most important operational procedures for the mass culture of algae. Supersaturated concentrations of oxygen resulted in lower productivities due to photorespiration and/or oxidation.     

Influence of barley straw (Hordeum vulgare L.) extract on phytoplankton dominated by Scenedesmus species in laboratory conditions: the importance of the extraction duration

The response of a natural phytoplankton assemblage dominated by algae of the genus Scenedesmus to the addition of barley straw extract was studied in a laboratory experiment. The aim of the study was to compare the inhibiting effect of water extracts obtained by soaking the straw for 1, 2 and 3 months. We analysed the response of four species, Scenedesmus subspicatus, Scenedesmus ecornis, Scenedesmus quadricauda and Scenedesmus acuminatus, during 14 days of their exposure to different types of barley straw extract. S. subspicatus and S. ecornis responded with decreasing numbers only to the addition of the 3-month solution (ANOVA; F?=?290.1, p <0.001; and F?=?11.8, p <0.01, respectively); the two other species were inhibited by all types of extracts. The results indicate the need for more research on the importance of extraction duration to the inhibitory abilities of barley straw which can be applied in the management of water quality in water bodies.     

Influence of endophytic strains of the bacterium Bacillus subtilis on cell number in monocultures of green algae

Effect of several endophytic bacterial strains of Bacillus subtilis on growth of Scenedesmus quadricauda and Chlorella vulgaris algae was investigated. Introduction of bacillus strains into the algal cultures diminished the cell number of microalgae. The cells of C. vulgaris exhibited equal sensitivity to all the endophyte strains examined. The number of S. quadricaudas cells reduced to larger extent by the 7th day of algal culturing in the presence of B. subtilis 11V or 11VM strains than in the presence of B. subtilis 26D strain, the latter strain being the main component of the commercial biofungicide Fitosporin-M. Washed bacterial cells inactivated by autoclaving did not reduce the number of C. vulgaris cells. The bacterial cells that were autoclaved without separation from the culture liquid, as well as the autoclaved supernatant of cell suspension exerted the same inhibitory effect as living bacterial cells. The decrease in the algal cell number under the action of endophytic bacillus strains is presumably due to thermostable biologically active substances synthesized and secreted by the bacillus strains.     

Effect of pH on Inorganic Carbon Uptake in Algal Cultures

Biomass production by the green algae Scenedesmus obliquus and Chlorella vulgaris in intensive laboratory continuous cultures was considerably affected by the pH at which the cultures were maintained. Carbon photoassimilation experiments revealed that pH values in the range of 8 to 9 were important for determining the free CO₂ concentrations in the medium. With higher pH values, additional pH effects were observed involving a decrease in the relative high affinity of low CO₂-adapted algae to free CO₂. The carbon uptake rate by high CO₂-adapted algae after transfer to low free CO₂ medium was characterized by a lag period of about 30 min, after which the affinity of the algae to CO₂ increased considerably. Both continuous growth and carbon uptake experiments indicated that artificially maintained high free CO₂ concentrations are recommended for maximal production in intensive outdoor algal cultures.     

Light Structures Phototroph,Bacterial and Fungal Communities at the Soil Surface

The upper few millimeters of soil harbour photosynthetic microbial communities that are structurally distinct from those of underlying bulk soil due to the presence of light. Previous studies in arid zones have demonstrated functional importance of these communities in reducing soil erosion, and enhancing carbon and nitrogen fixation. Despite being widely distributed, comparative understanding of the biodiversity of the soil surface and underlying soil is lacking, particularly in temperate zones. We investigated the establishment of soil surface communities on pasture soil in microcosms exposed to light or dark conditions, focusing on changes in phototroph, bacterial and fungal communities at the soil surface (0–3 mm) and bulk soil (3–12 mm) using ribosomal marker gene analyses. Microbial community structure changed with time and structurally similar phototrophic communities were found at the soil surface and in bulk soil in the light exposed microcosms suggesting that light can influence phototroph community structure even in the underlying bulk soil. 454 pyrosequencing showed a significant selection for diazotrophic cyanobacteria such as Nostoc punctiforme and Anabaena spp., in addition to the green alga Scenedesmus obliquus. The soil surface also harboured distinct heterotrophic bacterial and fungal communities in the presence of light, in particular, the selection for the phylum Firmicutes. However, these light driven changes in bacterial community structure did not extend to the underlying soil suggesting a discrete zone of influence, analogous to the rhizosphere.     

Utilization of Organic Nitrogen Sources by Two Phytoplankton Species and a Bacterial Isolate in Pure and Mixed Cultures

Algal production of dissolved organic carbon and the regeneration of nutrients from dissolved organic carbon by bacteria are important aspects of nutrient cycling in the sea, especially when inorganic nitrogen is limiting. Dissolved free amino acids are a major carbon source for bacteria and can be used by phytoplankton as a nitrogen source. We examined the interactions between the phytoplankton species Emiliania huxleyi and Thalassiosira pseudonana and a bacterial isolate from the North Sea. The organisms were cultured with eight different amino acids and a protein as the only nitrogen sources, in pure and mixed cultures. Of the two algae, only E. huxleyi was able to grow on amino acids. The bacterium MD1 used all substrates supplied, except serine. During growth of MD1 in pure culture, ammonium accumulated in the medium. Contrary to the expectation, the percentage of ammonium regenerated from the amino acids taken up showed no correlation with the substrate C/N ratio. In mixed culture, the algae grew well in those cultures in which the bacteria grew well. The bacterial yields (cell number) were also higher in mixed culture than in pure culture. In the cultures of MD1 and T. pseudonana, the increase in bacterial yield (number of cells) over that of the pure culture was comparable to the bacterial yield in mixed culture on a mineral medium. This result suggests that T. pseudonana excreted a more-or-less-constant amount of carbon. The bacterial yields in mixed cultures with E. huxleyi showed a smaller and less consistent difference than those of the pure cultures of MD1. It is possible that the ability of E. huxleyi to use amino acids influenced the bacterial yield. The results suggest that interactions between algae and bacteria influence the regeneration of nitrogen from organic carbon and that this influence differs from one species to another.     

The effect of synchronizing dark period on populations ofScenedesmus quadricauda

Populations ofScenedesmus quadricauda grown in a continuous chemostatic regime and synchronized by light and dark intervals were exposed to continuous illumination. The effect of light on synthetic and reproduction processes during the time of the omitted dark period and in the subsequent cell cycle was studied. In general, the sequence of cellular processes and their mutual coupling remain the same as in the darkened population. Synthetic processes and photosynthetic activity are depressed during the period of protoplast fission also in the light. The synchronizing effect of the dark period in chlorococcal algae consists in reducing the developmental variability in the population. The developmental state of daughter cells at the end of the dark interval varies in the span of two (as a rule) or four (at maximum) genomic cycles, while at the end of the light period this variability comprises up to eight genomic states. The more advanced autospores start the next cycle with a greater lag.     

Effect of culture medium on hydrogen production by sulfur-deprived marine green algae Platymonas subcordiformis

To study the effect of culture medium on hydrogen production by the marine green algae, Platymonas subcordiformis under sulfur deprivation, cell growth, hydrogen production, and starch and protein catabolism was investigated in the work. Algae cells cultured only in optimized medium required 6～8 days to reach the late logarithmic at the approximate density of (2.00 ± 0.18) × 10⁶ cells/mL, which in traditional medium needed 18～22 days to reach (1.85 ± 0.20) × 10⁶ cells/mL. Increased levels of Chlorophyll (10.74 ± 0.20 μg/mL), starch (149.50 ± 6.15 μg/mL), and protein (213.00 ± 7.36 μg/mL) were accumulated in optimized medium, which were 1.06, 1.47, and 1.87-fold of the algae cells cultured in traditional medium, respectively. The sealed culture of algae cells in sulfur-deprived optimized medium shifted to anaerobic conditions after 96 h of light illumination and produced 0.45 ± 0.12 mL H₂, but in traditional medium maintained aerobic condition and no hydrogen was produced. In addition, changes in starch and protein content during continuous light illumination indicated that more endogenous substrate was consumed in the sulfur-deprived optimized medium than that in the sulfur-deprived traditional medium.     

重金属铅与两种淡水藻的相互作用

为了研究重金属铅与淡水藻类之间的相互作用,采用不同Pb2+浓度处理铜绿微囊藻(Microcysis aeruginosa Kutz.)和斜生栅藻[Scenedesmus obliquus(Turp.)Kutz.],分别对两种藻的生物量、藻液电导率、O-·2含量、过氧化物酶(POD)、过氧化氢酶(CAT)活性以及藻对Pb2+的吸收作用等进行了测定,并通过扫描电镜观察了不同Pb2+浓度处理下两种藻细胞的表面结构。结果显示:(1)Pb2+浓度低于3 mg/L促进铜绿微囊藻生长,高于9 mg/L抑制其生长;但在3—12 mg/L范围内,Pb2+均明显抑制了斜生栅藻的生长,说明斜生栅藻对Pb2+毒性的敏感程度要高于铜绿微囊藻。(2)受到铅离子的胁迫,两种藻细胞膜通透性均有一定改变,扫描电子显微镜的照片观察,两种藻细胞表面的絮状物随着Pb2+的升高而增多,尤其是斜生栅藻细胞结构改变明显,多数细胞变形破裂;同时,O-·2含量升高,POD、CAT活性早期均可随Pb2+的增加而上升,表明氧自由基的产生增多以及由其引起的细胞生理生化改变可能是铅离子作用于藻细胞的主要机制。(3)两种淡水藻对Pb2+均有吸收作用,单位量藻细胞内,斜生栅藻对Pb2+的吸收能力好于铜绿微囊藻。所有结果提示:斜生栅藻不仅可以作为对重金属敏感的指示生物来监测水体Pb2+污染程度;同时由于斜生栅藻比铜绿微囊藻具有更好的Pb2+吸收能力,因此还可以利用斜生栅藻作为处理水体Pb2+的生物材料。