Osmoregulation and cell composition in salt-adaptation of Nostoc muscorum

Adaptation to salt in the cyanobacterium Nostocmuscorum, is composed of a few mechanisms which together lead to the generation of a salt-tolerant cell. The initial mechanism combines a stimulation of photosynthetic activity with the accumulation of sucrose as an osmoregulator. The secondary mechanism involves the adaptation of N₂ fixation activity and protein biosynthesis. The adaptation is most efficient in response to NaCl-induced stress and functions only partially under stress induced by either KCl or a nonionic osmoticum such as mannitol.     

Structural aspects of the adaptation of Nostoc muscorum to salt

The physiological and biochemical changes during the adaptation of Nostoc muscorum to salt are accompanied by specific structural changes. Cells of Nostoc muscorum exposed to saline medium vary in size and envelope organization. There are also drastic changes in the intracellular organization of the thylakoidal assembly. The heterocysts exhibit a preferential tolerance to NaCl rather than mannitol. These findings suggest that Nostoc muscorum is equipped with a specific physiological capacity for NaCl tolerance.     

Biotransformation of hydrocortisone by a natural isolate of Nostoc muscorum

Hydrocortisone was converted in the culture of an isolated strain of the cyanobacterium Nostoc muscorum PTCC 1636 into some androstane and pregnane derivatives. The microorganism was, isolated during a screening program from soil samples collected from paddy fields of north of Iran. The bioproducts obtained were purified using chromatographic methods and identified as 11beta-hydroxytestosterone, 11beta-hydroxyandrost-4-en-3,17-dione and 11beta,17alpha,20beta,21-tetrahydroxypregn-4-en-3-one on the basis of their spectroscopic features.     

Metabolic changes associated with cyanophage N-1 infection of the cyanobacterium Nostoc muscorum

The development of cyanophage N-1 in the N₂-fixing cyanobacterium Nostoc muscorum is dependent on light. The redox state of thioredoxin m was altered in phage infected cells, with the proportion of reduced thioredoxin increasing during the eclipse period. In one step growth experiments, the specific activity of glucose-6-phosphate dehydrogenase increased transiently during the eclipse period, whereas that of glutamine synthetase increased towards the end of the eclipse period (2–4h after infection) then remained high until the end of the latent period (about 7 h after infection). The rate of respiratory O₂ uptake was maintained until the end of the latent period. In contrast, the specific activity of phosphoribulokinase and the rate of photosynthetic O₂ evolution began to decrease towards the end of the eclipse period and later than the level of extractable protein began to decrease. Nitrogenase activity remained high throughout the eclipse period then decreased rapidly after 5 h. The level of glutamine synthetase protein decreased in parallel with the decrease in total extractable protein, whereas the level of thioredoxin m protein decreased more slowly.     

Effect of cyanophage N-1 development on nitrogen metabolism of cyanobacterium Nostoc muscorum

Abstract The impact of cyanophage N-1 development on nitrogenase, glutamine synthetase (GS) and aminotransferases activities in the diazotrophic cyanobacterium Nostoc muscorum was investigated during its latent period. The nitrogenase activity was inhibited after 2 h of infection, suggesting that phage development does not require the product of nitrogenase activity. GS activity was not inhibited until 4 h of infection; however, a decline in activity was subsequently observed. Glutamate oxaloacetate transaminase was inhibited after 1 h of infection and no activity was detectable during the entire latent period. In contrast, glutamate pyruvate transaminase activity increased 2-fold by 4 h of infection and remained higher than the background level until the end of the latent period. The results suggested that under nitrogen fixing conditions, N-1 multiplication proceeds in the absence of nitrogen fixation and that the metabolism of amino acids is altered in favour of phage multiplication.     

Antibiotic production by the cyanobacteriumNostoc muscorum

A broad spectrum antimicrobial antibiotic is produced byNostoc muscorum (Lancashire Polytechnic Culture Collection 23) during the post-exponential phase of growth. The antibiotic inhibits the growth of bacteria, notably multiple-resistantStaphylococcus aureus, and a biocide resistantPseudomonas aeruginosa: fungi such as the biodeteriogens,Cladosporium herbarum andHormoconis resinae and yeasts such asCandida albicans andC. pseudotropicalis. The antibiotic has an apparent molecular weight of 2000–3000 Daltons. Production appears to be dependent upon the limitation of one or more nutrients in the medium. author for correspondence     

Effect of four heavy metals on the biology of Nostoc muscorum

Summary This study presents the effects of Cr, Pb, Ni and Ag on growth, pigments, protein, DNA, RNA, heterocyst frequency, uptake of NH₄ ⁺ and N0₃ ^–, loss of electrolytes (Na⁺ and K⁺), nitrate reductase and glutamine synthetase activities ofNostoc muscorum. The statistical tests revealed a direct positive correlation between the metal concentration and inhibition of different processes. Ni was found to be more toxic against growth, pigments and heterocyst differentiation compared to the other metals. Inhibition of pigment showed the following trend: chlorophyll > phycocyanin > carotenoid. No generalized trend for inhibition of macromolecules was observed. The loss of K⁺ and Na⁺ as affected by Cr, Ni and Pb was similar but more pronounced for K⁺ than Na⁺. The inhibition of physiological variables depicted the following trend: Na⁺ loss > K⁺ loss > glutamine synthetase > NH₄ uptake > growth > N0₃ ^– uptake > nitrate reductase > heterocyst frequency. This study therefore suggests that loss of electrolytes can be used as a first signal of metal toxicity in cyanobacteria. However, further study is needed to confirm whether the abnormality induced by nickel (branch formation) is a physiological or genetic phenomenon.     

Effects of cyclic nucleotides on morphogenesis inNostoc muscorum

The filamentous cyanophyteNostoc muscorum A grew aseriately in light in a mineral salts (sugar-free) culture medium supplemented with adenosine 3:5-cyclic-monophosphate or N⁶, O²-dibutyryl adenosine 3:5-cyclic-monophosphate (1 mM). The aseriate morphology thus formed in the light on the 10th day following inoculation was similar to that formed in the dark after 20–30 days growth in cAMP-free medium containing glucose or sucrose. Inoculum previously grown in sucrose- or glucose-containing medium displayed aseriate morphology with lesser proliferation of coccoid cells as compared to inoculum grown in the absence of glucose or sucrose. cGMP, ADP, AMP and inhibitors of phosphodiesterase (theophylline and caffeine) did not have any effect on the persistence of aseriate morphology. However they stimulated cell division at the aseriate stage and delayed the release of hormogonia.Abbreviations cAMP adenosine 3:5-cyclic-monophosphate - db cAMP N⁶, O²-dibutyryl adenosine 3:5-cyclic-monophosphate - cGMP guanosine 3:5-cyclic-monophosphate - ATP adenosine 5-triphosphate - ADP adenosine5-diphosphate - AMP adenosine 5-monophosphate     

Reconstitution of electron transport by cytochrome c-553 in a cell-free system of Nostoc muscorum

Treatment of spheroplasts of Nostoc museorum with hypotonic buffer results in membranes depleted of cytochrome c-553, but still active in photosynthetic and respiratory electron transport. These membranes retain full photosystem II activity (H₂ODAD_ox). Complete linear electron transport (H₂ONADP⁺), however, is decreased as compared with untreated spheroplasts. Addition of basic Nostoc cytochrome c-553 to depleted membranes reconstitutes NADP⁺ reduction and redox reactions of the photosystem I region as well.Using NADPH as electron donor, respiration of depleted membranes is also stimulated by adding cytochrome c-553, indicative of its function in respiratory electron transport.Cytochrome c-553 from Bumilleriopsis filiformis, Spirulina platensis (acidic types), Phormidium foveolarum (basic type), and mitochondrial horse-heart cytochrome c-550 are not effective in reconstituting both photosynthetic and respiratory electron transport, which points to a specific role of Nostoc cytochrome c-553.Abbreviations BSA bovine serum albumin - DAD 3,6-diaminodurene - DAD_ox 3,6-diaminodurene oxidized by potassium ferricyanide - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DCIP 2,6-dichlorophenolindophenol - DPC 1,5-diphenylcarbazide - Fd ferredoxin - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - MES 2(-N-morpholino)-ethanesulfonic acid - MV methylviologen (1,1-dimethyl-4,4-bipyridylium dichloride) - PS I photosystem I - PS II photosystem II - Tris tris-(hydroxymethyl)-aminomethane     

Nitrogenase activity in the non-heterocystous cyanobacterium Oscillatoria sp. grown under alternating light-dark cycles

The non-heterocystous cyanobacterium Oscillatoria sp. strain 23 fixes nitrogen under aerobic conditions. If nitrate-grown cultures were transferred to a medium free of combined nitrogen, nitrogenase was induced within about 1 day. The acetylene reduction showed a diurnal variation under conditions of continuous light. Maximum rates of acetylene reduction steadily increased during 8 successive days. When grown under alternating light-dark cycles, Oscillatoria sp. fixes nitrogen preferably in the dark period. For dark periods longer than 8 h, nitrogenase activity is only present during the dark period. For dark periods of 8 h and less, however, nitrogenase activity appears before the beginning of the dark period. This is most pronounced in cultures grown in a 20 h light – 4 h dark cycle. In that case, nitrogenase activity appears 3–4 h before the beginning of the dark period. According to the light-dark regime applied, nitrogenase activity was observed during 8–11 h. Oscillatoria sp. grown under 16 h light and 8 h dark cycle, also induced nitrogenase at the usual point of time, when suddenly transferred to conditions of continuous light. The activity appeared exactly at the point of time where the dark period used to begin. No nitrogenase activity was observed when chloramphenicol was added to the cultures 3 h before the onset of the dark period. This observation indicated that for each cycle, de novo nitrogenase synthesis is necessary.     

Caesium uptake and toxicity in the cyanobacterium <Emphasis Type="Italic">Nostoc muscorum</Emphasis>

A NH₄⁺ transport-defective mutant and a K⁺ transport-defective mutant of the cyanobacterium Nostoc muscorum were analysed with regard to percentage survival as a function of CsCl toxicity and Cs⁺ uptake activity. Neither survival nor Cs⁺ uptake was affected in either of the two mutants when compared with the wild type. The results indicate that the toxicity of Cs⁺ is determined at more than one cellular site in this organism.     

Glyoxylate decreases the oxygen sensitivity of nitrogenase activity and photosynthesis in the cyanobacterium Anabaena cylindrica

Raising the pO₂ reduced nitrogenase activity (C₂H₂ reduction) of Anabaena cylindrica for both glyoxylate-treated (5 mM) and untreated cells. The stimulation caused by glyoxylate, however, increased with increases of pO₂ from 2 to 99 kPa. As the pO₂ increased the net CO₂ fixation was lowered (Warburg effect) while the CO₂ compensation point increased. Glyoxylate partly relieved this sensitivity of net photosynthesis to oxygen and reduced the compensation point considerably. The cells used were preincubated in the dark to exhaust photosynthetic pools. A more pronounced reduction in sensitivity of nitrogenase to oxygen for glyoxylate-treated cells was evident when a preincubation in air with reduced pCO₂ (13 l l^-1) was used. This was, however, not evident until after a 10-h incubation in air. Before this point 2 kPa O₂ sustained the highest nitrogenase activity. Addition of 0.5 and 5 mM of HCO ₃ ^- to Anabaena cultures preincubated at low CO₂ levels (29 l l^-1) abolished the stimulatory effect of glyoxylate on the nitrogenase. Thus, the results sustain the suggestion that glyoxylate may act as an inhibitor of photorespiratory activities in cyanobacteria and can be used as a means of increasing their nitrogen and CO₂ fixation capacities.Abbreviation RuBP ribulose 1,5-bisphosphate     

Calcium requirement in nitrogen fixation in the cyanobacterium Synechococcus RF-1

Under diurnal 16/8-h light-dark cycles, ethyleneglycol-bis-(-aminoethyl ether)-N,N,N,N-tetraacetic acid (EGTA) at 1 mM completely blocked the appearance of rhythmic N₂-fixing activity in Synechococcus RF-1. Ca²⁺ at 2 mM, when supplied either together with or several hours after the EGTA application, restored the nitrogenase activity, whereas, when Ca²⁺ was supplied several hours later, the peak of nitrogenase activity was shifted from the dark to the light period in which the activity is normally suppressed. Sr²⁺ also reversed the inhibition by EGTA, but only partially. When O₂ in the gas phase above the culture was below 1%, the inhibition of nitrogenase activity by EGTA was reduced to less than 20% of the control value without EGTA. Thus Ca²⁺ appears to be required by the cell to protect its nitrogenase from inactivation by O₂. In media without EGTA, a close correlation between nitrogenase activity and concentrations of Ca²⁺ was also observed.Abbreviation EGTA ethyleneglycol-bis-(-aminoethyl ether)-N,N,N,N-tetraacetic acid     

Pathways of hydrogen uptake in the cyanobacterium Nostoc muscorum

Two pathways of hydrogen uptake in Nostoc muscorum are apparent using either oxygen or nitrogen as electron acceptor. Hydrogen uptake (under argon with some oxygen as electron acceptor assayed in the dark; oxyhydrogen reaction) is found to be more active in dense, light-limited cultures than in thin cultures when light is not limiting. Addition of bicarbonate inhibits this hydrogen uptake, because photosynthesis is stimulated. In a cell-free hydrogenase assay, a 10-fold increase of the activity can be measured, after the cells having been kept under lightlimiting conditions. After incubation under light-saturating conditions, no hydrogen uptake is found, when filaments are assayed under argon plus some oxygen. Assaying these cells under a nitrogen atmosphere, a strong hydrogen uptake occurs. The corresponding cell-free hydrogenase assay exhibits low hydrogenase activity. Furthermore, the hydrogen uptake by intact filaments under nitrogen in the light apparently is correlated with nitrogenase activity. These studies give evidence that, under certain physiological conditions, hydrogen uptake of heterocysts proceeds directly via nitrogenase, with no hydrogenase involved.Abbreviations Chl chlorophyll - DCMU (diuron) 3-3,4-dichlorophenyl)-1,1-dimethylurea - pev packed cell volume     

Fructose utilization by the cyanobacterium Anabaena variabilis studied using whole filaments and isolated heterocysts

We have investigated the utilization of [¹⁴C]-fructose by whole filaments and isolated heterocysts of Anabaena variabilis ATCC 29413, a strain which is capable of fructose-dependent heterotrophic growth. The experimental conditions were chosen such that both transport and subsequent metabolism were studied. The apparent K_m for fructose was 60 mM, close to the results of previous studies. Rates of fructose utilization were the same in light and darkness. When photosynthetic CO₂ fixation was possible, almost all the label appeared as cell-carbon. In darkness or in the presence of DCMU appreciable amounts of label were released as CO₂. Isolated heterocysts with high rates of endogenous metabolism were not capable of utilizing added fructose at significant rates. The effects of oxygen concentration on the metabolism of added fructose in darkness showed that uptake was saturated at low pO₂ values. Increasing the pO₂ values lead to an increase in the ratio between the lable released as CO₂ and that recovred as cell-carbon. These results suggest that fructose is taken up only by the vegetative cells but carbon derived from added fructose can be released as CO₂ as a result of respiration in the heterocysts. Fructose utilization was inhibited by uncouplers. The greatest inhibition was found when both (delta) (psi) and (delta) pH were abolished. High concentrations of erythrose inhibited fructose utilization. None of the other potential analogs tested had any effect.     

Inorganic nitrogen regulation of glutamate uptake in the cyanobacterium Nostoc muscorum

In Nostoc muscorum (Anabaena ATCC 27893) glutamate was not metabolised as a fixed nitrogen source, rather it functioned as an inhibitor of growth. The latter effect was nitrogen source specific and occurred in N₂-fixing cultures but not in cultures assimilating nitrate or ammonium. NO₃^--grown cultures lacked heterocysts and nitrogenase activity and showed a nearly 50% reduction in glutamate uptake rates, as well as in the final extent of glutamate taken up, compared to N₂-fixing or nitrogen-limited control cultures. NH₄⁺-grown cultures showed a similar response, except that the reduction in glutamate uptake rates and the final exten of glutamate taken up was over 80%. The present results suggest a relation between nitrate/ammounium nitrogen-dependent inhibition of glutamate uptake, probably via repression of the glutamate transport system, and glutamate toxicity.     

Lysogeny in the blue-green algaNostoc muscorum

Lysogens were isolated after confluent lysis of the blue-green algaNostoc muscorum by N-1 virus and characterised. The spontaneous liberation of active virus particles in lysogens was not lost after treating them with viricidal concentration of EDTA and virus titre obtained in their cultures was >5×10³ PFU/ml. The virus adsorbed on the lysogens with a slower rate than on parent alga, although it followed the pattern of a first order reaction. The heat treatment (45°C for 14h) of the lysogens failed to induce lysis, whereas mitomycin C (1–2 g/ml) was effective in inducing lysis.List of Abbreviations EDTA ethylene diamine tetra-acetic acid - IM input multiplicity - k adsorption rate constant - k _v specific constant for virus production - MC mitomycin C - PFU plaqueforming units     

Ethylenediamine uptake and metabolism in the cyanobacterium Anabaena variabilis

The cyanobacterium Anabaena variabilis showed a pH dependent uptake of ethylenediamine. No uptake of ethylenediamine was detected at pH 7.0. At higher pH values (e.g. pH 8.0 and pH 9.0) accumulation did occur and was attributed to diffusion of uncharged ethylenediamine in response to a pH gradient. A biphasic pattern of uptake was observed at these higher pH values. Treatment with l-methionine-d,l-sulphoximine (MSX) to inactivate glutamine synthetase (GS) inhibited the second slower phase of uptake without any significant alteration of the initial uptake. Therefore for sustained uptake, metabolism of ethylenediamine via GS was required. NH ₄ ⁺ did not alter the uptake of ethylenediamine. Ethylenediamine was converted in the second phase of uptake to an analogue of glutamine which could not be detected in uptake experiments at pH 7.0 or in uptake experiments at pH 9.0 following pretreatment of cells with MSX. Ethylenediamine treatment inhibited nitrogenase activity and this inhibition was greatest at high pH values.Abbreviations EDA 1,2-diaminoethane (ethylenediamine) - GS glutamine synthetase - HEPES 4-(2-hydroxyethyl)-1 piperazine ethanesulphonic acid - MSX l-methionine-dl-sulphoximine - membrane potential - Tricine N-tris(hydroxymethyl) methylglycine     

Photoinhibition and its wavelength dependence in the cyanobacterium Anabaena variabilis

In the cyanobacterium Anabaena variabilis the dependence of photoinhibition on fluence rate, duration and wavelength of irradiation were studied by measurements of oxygen production and fluorescence emission spectra. The analysis of the photosynthetic activity revealed that photoinhibition affects exclusively photosystem II (PS II), whereas photosystem I (PS I) remained largely unimpaired. Furthermore, PS II fluorescence emission decreased much faster in bleached than in unbleached controls.Studying the wavelength dependence of photoinhibition it was found that only radiation between 520 and 680 nm causes photoinhibition. This is about the same range of wavelengths which causes photobleaching. Fluorescence emission spectra of samples exposed to high fluence rates of 582 and 662 nm, respectively, essentially agree with those samples exposed to high fluence rates of white light, whereas the fluorescence emission spectra of samples exposed to blue light resemble those exposed to dim white light.NaN₃, a substance which prevents photobleaching, inhibits the photosynthetic O₂ production of Anabaena and, hence, enhances the photoinhibitory effect.     

Distribution and ecology of the edible cyanobacterium Ge-Xian-Mi (Nostoc) in rice fields of Hefeng County in China

Ge-Xian-Mi (an edible species of Nostoc) grows insome mountain paddy fields in China during winter and forms macroscopicallyvisible subspherical colonies. The geology and climate at one of its locations,Hefeng County, were investigated, and the present-day situation of Ge-Xian-Miwas assessed in order to raise awareness that it may be endangered. There wereformerly 796 ha of rice fields suited to its growth in HefengCounty and the maximum annual yield ever reached was 25 t. Theannual mean temperature is about 12.2 °C, and the annualrainfall is 1934 mm with mean relative humidity of 78–87%.The distribution of Ge-Xian-Mi was found to be associated with the source ofwater and the pH values of water suited to its growth were 6.2–6.3. Soilsin its habitats were enriched with phosphorus and contained more microbes thanthose without its distribution. With changing agricultural techniques most ofthe habitats are endangered or already extinct. The widespread use ofherbicides, pesticides and fertilizers containing chlorine had been suggestedtobe an important factor limiting its distribution. The taxonomic identity ofGe-Xian-Mi is discussed.