Nitrate Reductase and Chlorate Toxicity in Chlorella vulgaris Beijerinck

A study of the growth-inhibiting effect of chlorate on the Berlin strain of Chlorella vulgaris Beijerinck provided complete confirmation of the theory of chlorate toxicity first proposed by Åberg in 1947. Chlorate was toxic to the cells growing on nitrate, and relatively nontoxic to the cells growing on ammonium. The latter cells contained only 0.01 as much NADH-nitrate reductase as the nitrate-grown cells. Chlorate could substitute for nitrate as a substrate of the purified nitrate reductase with Km = 1.2 mm, and V_max = 0.9V_max for nitrate. Bromate, and to a much smaller extent, iodate, also served as alternate substrates. Nitrate is a reversible competitive inhibitor of chlorate reduction, which accounts for the partial reversal, by high nitrate concentrations, of the observed inhibition of cell growth by chlorate. During the reduction of chlorate by NADH in the presence of purified nitrate reductase, there was a progressive, irreversible inhibition of the enzyme activity, presumably brought about by the reduction product, chlorite. Both the NADH-nitrate reductase activity and the associated NADH-cytochrome c reductase activity were inactivated to the same extent by added chlorite. The spectral properties of the cytochrome b₅₅₇ associated with the purified enzyme were not affected by chlorite. The inactivation of the nitrate reductase by chlorite could account for the toxicity of chlorate to cells grown on nitrate, though the destruction of other cell components by chlorite or its decomposition products cannot be excluded.     

The Effect of Toxic Doses of Copper upon Respiration, Photosynthesis and Growth of Chlorella vulgaris

When cells of Chlorella vulgaris absorb copper under anaerobic conditions, subsequent respiration, photosynthesis and growth of the cells are all severely inhibited. This does not occur when the metal is absorbed under aerobic conditions. When, after aerobic absorption of copper, the cells are exposed to a period of anaerobiosis, respiratory inhibition is as profound as when the uptake is anaerobic. In this case, however, respiration must eventually recover, for growth is not affected so severely as it is when copper is taken up under anaerobic conditions. It is concluded that the extra copper absorbed under anaerobic conditions is directly or indirectly responsible for the greatly increased toxicity to growth, and that this copper is bound to sites not normally available under aerobic conditions. Some aspects of the apparently unique toxic effect of copper suggest that these extra sites are sulphydryl groups.     

Copper tolerance in the green alga, Chlorella vulgaris

Abstract. The effect of sub-lethal concentrations of copper upon tolerant and non-tolerant strains of Chlorella vulgaris was investigated. Copper concentrations of 0.2 and 0.4 mg dm⁻³ increased the lag phase of both strains, the effect being greater in the non-tolerant strain. No difference was observed in the toxicity of copper to the photosynthetic rates of the isolated chloroplasts of either strain. However, significant differences were shown at the whole cell level.
Lower copper uptake was shown by the tolerant cells. In both strains initial uptake of copper was followed by a phase of desorption before cell division occurred. In cultures of both strains the concentration of ionic copper was decreased by complexation with extracellular organic material. Over a 14 day growth period more organic material was produced by the tolerant cells. The organic material produced by the tolerant cell formed organo-copper complexes which had a higher conditional stability constant.
It is proposed that the cell wall acts as a barrier to copper in the tolerant cells and prevents copper from affecting cell metabolism. Organo-copper complexation occurs at this barrier and this complex is then released into the medium.     

Growth of Chlorella vulgaris and associated bacteria in photobioreactors

The aim of this study was to test three flat plate photobioreactor configurations for growth of Chlorella vulgaris under non‐axenic conditions and to characterize and quantify associated bacterial communities. The photobioreactor cultivations were conducted using tap water‐based media to introduce background bacterial population. Growth of algae was monitored over time with three independent methods. Additionally, the quantity and quality of eukaryotes and bacteria were analysed using culture‐independent molecular tools based on denaturing gradient gel electrophoresis (PCR‐DGGE) and quantitative polymerase chain reaction (QPCR). Static mixers used in the flat plate photobioreactors did not generally enhance the growth at the low light intensities used. The maximum biomass concentration and maximum specific growth rate were 1.0 g l^?1 and 2.0 day^?1 respectively. Bacterial growth as determined by QPCR was associated with the growth of C. vulgaris. Based on PCR‐DGGE, bacteria in the cultures mainly originated from the tap water. Bacterial community profiles were diverse but reproducible in all flat plate cultures. Most prominent bacteria in the C. vulgaris cultures belonged to the class Alphaproteobacteria and especially to the genus Sphingomonas. Analysis of the diversity of non‐photosynthetic microorganisms in algal mass cultures can provide useful information on the public health aspects and unravel community interactions.     

Toxicity of bisphenol A and its structural congeners to microalgae Chlorella vulgaris and Desmodesmus armatus

Journal of Applied Phycology - Bisphenol A and its structural congeners are increasingly recognized as emerging contaminants with toxic and estrogenic potential that have been widely used in many...     

Toxicity of carbofuran to soil isolates of Chlorella vulgaris,Nostoc linckia and N. muscorum

A microalga, Chlorella vulgaris, and two diazotrophic cyanobacteria, Nostoc linckia and N. muscorum, all isolated from a rice soil, were compared for their response in terms of growth and metabolic activities to the application of carbofuran. The toxicity criteria included cell constituents (chlorophyll a, total protein, carbohydrate), ¹⁴CO₂ uptake and nitrate reductase, besides nitrogenase activity (acetylene reduction) in the cyanobacteria. C. vulgaris and N. muscorum were more sensitive to carbofuran than was N. linckia. The significant toxicity of the insecticide, observed with higher concentrations of 20 and 50 g ml^–1, to nitrogenase activity in N. linckia was reversed by the addition of ATP at 10 M. Transmission electron microscopy of the cultures, exposed to 50 g carbofuran ml^–1 showed certain cellular abnormalities, indicating interference of the insecticide with membrane properties. Correspondence to: K. Venkateswarlu     

Growth at Low Temperature Mimics High-Light Acclimation in Chlorella vulgaris

Structural and functional alterations to the photosynthetic apparatus after growth at low temperature (5[deg]C) were investigated in the green alga Chlorella vulgaris Beijer. Cells grown at 5[deg]C had a 2-fold higher ratio of chlorophyll a/b, 5-fold lower chlorophyll content, and an increased xanthophyll content compared to cells grown at 27[deg]C even though growth irradiance was kept constant at 150 [mu]mol m-2 s-1. Concomitant with the increase in the chlorophyll a/b ratio was a lower abundance of light-harvesting polypeptides in 5[deg]C-grown cells as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and confirmed by western blotting.The differences in pigment composition were found to be alleviated within 12 h of transferring 5[deg]C-grown cells to 27[deg]C. Furthermore, exposure of 5[deg]C-grown cells to a 30-fold lower growth irradiance (5 [mu]mol m-2 s-1) resulted in pigment content and composition similar to that in cells grown at 27[deg]C and 150 [mu]mol m-2 s-1. Although both cell types exhibited similar measuring-temperature effects on CO2-saturated O2 evolution, 5[deg]C-grown cells exhibited light-saturated rates of O2 evolution that were 2.8-and 3.9-fold higher than 27[deg]C-grown cells measured at 27[deg]C and 5[deg]C, respectively. Steady-state chlorophyll a fluorescence indicated that the yield of photosystem II electron transport of 5[deg]C-grown cells was less temperature sensitive than that of 27[deg]C-grown cells. This appears to be due to an increased capacity to keep the primary, stable quinone electron acceptor of photosystem II (QA) oxidized at low temperature in 5[deg]C- compared with 27[deg]C-grown cells regardless of irradiance. We conclude that Chlorella acclimated to low temperature adjusts its photosynthetic apparatus in response to the excitation pressure on photosystem II and not to the absolute external irradiance. We suggest that the redox state of QA may act as a signal for this photosynthetic acclimation to low temperature in Chlorella.     

Responses of Heterotrophic Cultures of Chlorella vulgaris Beyerinck to Darkness and Light. II. Action Spectrum for and Mechanism of the Light Requirement for Heterotrophic Growth

Chlorella vulgaris Beyerinck (Emerson's strain), fails to grow in the dark even when sugars are provided. This phenomenon was clearly demonstrated in the alga, C. vulgaris, for which the growth rate in darkness on a glucose medium remained constant for 2 days and then declined to approach zero. Pigment concentrations also declined in darkness. Changes in flow rate of 1% CO₂-in-air from zero to 7 ml per minute caused a progressive increase in the dark growth rate over a 5-day period, but did not maintain growth in the dark. Rates above 7 ml per minute produced no changes in growth rates.

White light intensities below the compensation point of the alga maintained heterotrophic growth. The saturation value for this response was 0.8 μw/cm². White light also initiated growth in nongrowing cultures transferred from darkness to light.

The action spectrum for heterotrophic growth indicated a porphyrin as the active pigment. Light in the 425 mμ region was 4 times as effective as white light in stimulating heterotrophic growth. A secondary peak of growth stimulation occurred in the 575 mμ region.

The respiration of glucose by the alga was stimulated by low intensities of white light. This response was not immediate, but was clearly present after the third day of incubation.

Malonate and cyanide were inhibitory to growth of C. vulgaris on inorganic medium or glucose medium under 300 ft-c of white light. These data suggested that succinic dehydrogenase and cytochrome oxidase systems were present.

Substances inhibitory to growth were excreted into the medium under dark-growth conditions, and 2 of these substances were indentified as formic and acetic acids.

The evidence suggested that respiration of glucose cannot proceed for an extended period of time in darkness. The reason for this is postulated to be the lack of a cytochrome or a cytochrome precursor.

     

植物激素在小球藻异养培养中的作用

本文研究了几种植物激素对小球藻异养培养的影响。结果表明,IAA、IBA及6-BA三种植物激素均不同程度地促进了小球藻的异养生长,培养≤36h时,IAA或IBA以20mg／L的促进小球藻异养生长的效应最大,100mg／L IAA或IBA则抑制了藻的生长;>36h时,100mg／L IAA或IBA表现出促进小球藻生长的效应,并最终获最大净A540增长量;6-BA以0.1 mg／L的促进作用最大。IBA与6-BA组合同样表现出促进小球藻异养生长的效应,但并非IBA和6-BA简单的加合效应,5 mg／LIBA与6-BA组合的效应维持6-BA单因子的作用趋势,20mg／LIBA与lmg/L6-BA组合的效应大于与0.1mg／L6-BA组合的,100mg／LIBA与0.1mg／L6-BA组合的效应在≤36h时大于与1mg／L6-BA组合的,>36h时则相反。另外,高浓度IBA(≥20mg／L)与6-BA组合抑制了前中期异养藻对葡萄糖的吸收,但加速了中后期葡萄的吸收。再者,IBA与6-BA组合加速了异养小球藻对No-3的吸收。     

Morphometric and Stereologic Analysis of Chlorella vulgaris Under Heterotrophic Growth Conditions

A computer-supported determination of stereological parameterswas used to study the possible ultrastructural changes of Chlorellavulgaris UAM 101 under photolithotrophic, mixotrophic and photoheterotrophicconditions of growth. Data recording was carried out througha semi-automatic digitizing image analysis system instead ofthe current method of superimposition of an array of short lines. Glucose promoted drastic physiological changes [Martínezand Orús, 1991. Plant Physiology (in press)], which stronglyaffected the size of the cells and volume densities of storagematerials. However, volumetric ratios of the mitochondrion orchloroplast active fraction were not affected by the presenceof glucose, probably indicating that these ratios are characteristicof each species. Cell wall ultrastructure was also analysed and the presenceof sporopollenin demonstrated, in contrast to the thin and sporopollenin-lackingcell wall generally described for Chlorella vulgaris species. Chlorella vulgaris, photoautotrophic, mixotrophic, photoheterotrophic, image analysis, stereology