Effects of calcium ion on the colony formation,growth, and photosynthesis in the edible cyanobacterium Nostoc sphaeroides

Journal of Applied Phycology - Calcium ions are crucial for the growth and development of photosynthetic organisms, whereas its eco-physiological significance on the edible cyanobacterium Nostoc...     

Acclimation to low ultraviolet-B radiation increases photosystem I abundance and cyclic electron transfer with enhanced photosynthesis and growth in the cyanobacterium Nostoc sphaeroides

Ultraviolet-B radiation is known to harm most photosynthetic organisms with the exception of several studies of photosynthetic eukaryotes in which UV-B showed positive effects. In this study, we investigated the effect of acclimation to low UV-B radiation on growth and photosynthesis of the cyanobacterium Nostoc sphaeroides. Exposure to 0.08 W m⁻² UV-B plus low visible light for 14 d significantly increased the growth rate and biomass production by 16% and 30%, respectively, compared with those under visible light alone. The UV-B acclimated cells showed an approximately 50% increase in photosynthetic efficiency (α) and photosynthetic capacity (P_max), a higher PSI/PSII fluorescence ratio, an increase in PSI content and consequently enhanced cyclic electron flow, relative to those of non-acclimated cells. Both the primary quinone-type acceptor and plastoquinone pool re-oxidation were up-regulated in the UV-B acclimated cells. In parallel, the UV-B acclimated colonies maintained a higher rate of D1 protein synthesis following exposure to elevated intensity of UV-B or visible light, thus functionally mitigating photoinhibition. The present data provide novel insight into photosynthetic acclimation to low UV-B radiation and suggest that UV-B may act as a positive ecological factor for the productivity of some photosynthetic prokaryotes, especially during twilight periods or in shaded environments.     

Cytochemical changes in the developmental process of Nostoc sphaeroides (cyanobacterium)

There are several apparent developmental stages in the life cycle of Nostoc sphaeroides Kützing, an edible cyanobacterium found mainly in paddy fields in central China. The cytochemical changes in developmental stages such as hormogonia, aseriate stage, filamentous stage and colony in N. sphaeroides were examined using fluorescent staining and colorimetric methods. The staining of acidic and sulfated polysaccharides increased with development when hormogonia were used as the starting point. Acidic polysaccharides (AP) were most abundant at the aseriate stage and then decreased, while the sulfated polysaccharides (SP) were highest at the colony stage. Quantitatively, along the developmental process from hormogonia to colony, total carbohydrates first increased, then became stable, and then reached their highest level at the colony stage, while reducing sugars were highest at the hormogonia stage and then decreased sharply once development began. SP were not detectable in the hot water soluble polysaccharides (HWSP), and hormogonia had the lowest content of AP, while old colonies had the highest. The AP content of the aseriate stage, filamentous stage and young colony stage were very similar. The evolutionary relationships reflected in the developmental stages of N. sphaeroides are discussed.     

UV-B radiation and phosphorus limitation interact to affect the growth,pigment content,and photosynthesis of the toxic cyanobacterium Microcystis aeruginosa

In this study, Microcystis aeruginosa was cultivated in a P-limited and P-replete culture medium and exposed to artificial UV-B radiation to investigate the interactive effect of UV-B exposure and phosphorus limitation on this harmful alga. After 15 days, both UV-B exposure and phosphorus limitation led to a significant decline in pigment content (phycocyanin and carotene) and photosynthetic activity (F _v/F _m and ETR_max), and the impact was most pronounced when the two conditions were combined. Due to the interactive effect, P-limited M. aeruginosa under UV-B exposure exhibited the lowest cell density compared to the other treatments. These results suggest that phosphorus limitation increases the stress of UV-B radiation in Microcystis. In other words, high-level UV-B radiation has higher growth inhibitory on Microcystis in P-limited lakes than in P-replete lakes.     

Na+ requirement for growth, photosynthesis, and pH regulation in the alkalotolerant cyanobacterium Synechococcus leopoliensis

We have found that Na+ is required for the alkalotolerance of the cyanobacterium Synechococcus leopoliensis. Cell division did not occur at any pH in the absence of Na+, but cells inoculated into Na+-free growth medium at pH 6.8 did continue metabolic activity, and over a period of 48 h, the cells became twice their normal size. Many of these cells remained viable for at least 59 h and formed colonies on Na+ -containing medium. Cells grown in the presence of Na+ and inoculated into Na+ -free growth medium at pH 9.6 rapidly lost viability. An Na+ concentration of ca. 0.5 milliequivalents X liter-1 was required for sustained growth above pH 9.0. The Na+ requirement could be only partially met by Li+ and not at all by K+ or Rb+. Cells incubated in darkness in growth medium at pH 6.8 had an intracellular pH near neutrality in the presence or absence of Na+. When the external pH was shifted to 9.6, only cells in the presence of Na+ were able to maintain an intracellular pH near 7.0. The membrane potential, however, remained high (-120 mV) in the absence or presence of Na+ unless collapsed by the addition of gramicidin. Thus, the inability to maintain a neutral intracellular pH at pH 9.6 in the absence of Na+ was not due to a generalized disruption of membrane integrity.Even cells containing Na+ still required added Na+ to restore photosynthetic rates to normal after the cells had been washed in Na+ -free buffer at pH 9.6. This requirement was only partially met by Li+ and was not met at all by K+, Rb+, Cs+ Mg2+, or Ca2+. The restoration of photosynthesis by added Na+ occurred within 30 s and suggests a role for extracellular Na+. Part of our results can be explained in terms of the operation of an Na+/H+ antiporter activity in the plasma membrane, but some results would seem to require other mechanisms for Na+ action.     

Utilization of inorganic carbon in the edible cyanobacterium Ge-Xian-Mi (Nostoc) and its role in alleviating photo-inhibition

The present work investigated the inorganic carbon (C_i) uptake, fluorescence quenching and photo‐inhibition of the edible cyanobacterium Ge‐Xian‐Mi (Nostoc) to obtain an insight into the role of CO₂ concentrating mechanism (CCM) operation in alleviating photo‐inhibition. Ge‐Xian‐Mi used HCO₃^– in addition to CO₂ for its photosynthesis and oxygen evolution was greater than the theoretical rates of CO₂ production derived from uncatalysed dehydration of HCO₃^–. Multiple transporters for CO₂ and HCO₃^– operated in air‐grown Ge‐Xian‐Mi. Na⁺‐dependent HCO₃^– transport was the primary mode of active C_i uptake and contributed 53–62% of net photosynthetic activity at 250 µmol L^?1 KHCO₃ and pH 8.0. However, the CO₂‐uptake systems and Na⁺‐independent HCO₃^– transport played minor roles in Ge‐Xian‐Mi and supported, respectively, 39 and 8% of net photosynthetic activity. The steady‐state fluorescence decreased and the photochemical quenching increased in response to the transport‐mediated accumulation of intracellular C_i. Inorganic carbon transport was a major factor in facilitating quenching during the initial stage and the initial rate of fluorescence quenching in the presence of iodoacetamide, an inhibitor of CO₂ fixation, was 88% of control. Both the initial rate and extent of fluorescence quenching increased with increasing external dissolved inorganic carbon (DIC) and saturated at higher than 200 µmol L^?1 HCO₃^–. The operation of the CCM in Ge‐Xian‐Mi served as a means of diminishing photodynamic damage by dissipating excess light energy and higher external DIC in the range of 100–10000 µmol L^?1 KHCO₃ was associated with more severe photo‐inhibition under strong irradiance.     

The role of Na2S in anoxygenic photosynthesis and H2 production in the cyanobacterium Nostoc muscorum

Na2S is known to support anoxygenic photosynthesis in some strains of cyanobacteria and to stimulate H2 production in N2 fixing filaments of Nostoc muscorum. We have shown electron transfer between Na2S and Photosystem I to be dependent on cytochrome b559 which was detected only in vegetative cells. An electron mediator was required to support Na2S driven nitrogenase activity in isolated heterocysts. Na2S was also found to deplete the ATP pool, probably by inhibiting electron transfer from Photosystem I.     

Purification and properties of an enzyme capable of degrading the polysaccharide of the cyanobacterium,Nostoc commune

A novel Nostoc commune-polysaccharide (NPS)-degrading enzyme with a molecular mass of 128.5 kDa was purified from Paenibacillus glycanilyticus DS-1. The optimum pH and temperature of the enzyme activity were 5.5 and 35 degrees C, respectively. The enzyme completely degraded NPS to oligosaccharides, ranging from tetra to hexasaccharides and could degrade the xylan weakly whereas xanthan, gellan, cellulose, curdlan and p-nitrophenyl-beta-D-xylopyranoside were not degraded. Homology analysis of the N-terminal amino acid sequence of the NPS-degrading enzyme against the PIR and SWISS-PROT databases indicated that the sequence was not homologous to any other polysaccharide-degrading enzyme.     

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.     

Iron limitation in the marine cyanobacterium Trichodesmium reveals new insights into regulation of photosynthesis and nitrogen fixation

* As iron (Fe) deficiency is a main limiting factor of ocean productivity, its effects were investigated on interactions between photosynthesis and nitrogen fixation in the marine nonheterocystous diazotrophic cyanobacterium Trichodesmium IMS101. * Biophysical methods such as fluorescence kinetic microscopy, fast repetition rate (FRR) fluorimetry, and in vivo and in vitro spectroscopy of pigment composition were used, and nitrogenase activity and the abundance of key proteins were measured. * Fe limitation caused a fast down-regulation of nitrogenase activity and protein levels. By contrast, the abundance of Fe-requiring photosystem I (PSI) components remained constant. Total levels of phycobiliproteins remained unchanged according to single-cell in vivo spectra. However, the regular 16-kDa phycoerythrin band decreased and finally disappeared 16-20 d after initiation of Fe limitation, concomitant with the accumulation of a 20-kDa protein cross-reacting with the phycoerythrin antibody. Concurrently, nitrogenase expression and activity increased. Fe limitation dampened the daily cycle of photosystem II (PSII) activity characteristic of diazotrophic Trichodesmium cells. Further, it increased the number and prolonged the time period of occurrence of cells with elevated basic fluorescence (F(0)). Additionally, it increased the effective cross-section of PSII, probably as a result of enhanced coupling of phycobilisomes to PSII, and led to up-regulation of the Fe stress protein IsiA. * Trichodesmium survives short-term Fe limitation by selectively down-regulating nitrogen fixation while maintaining but re-arranging the photosynthetic apparatus.     

Cytotoxic and non-cytotoxic exometabolites of the cyanobacterium Nostoc insulare

The isolation, identification and quantification of exometabolites from culture media of the cyanobacterium Nostoc insulare are described. Besides the known exometabolite 4,4′-dihydroxybiphenyl (I), two more compounds, the β-carboline 9H-pyrido(3,4-b)indole (norharmane, II) and N,N′-(4,5-dimethyl-1,2-phenylene)bis-acetamide (III), were discovered. Concentrations of all three compounds in media and biomass of five 250 L cultures of N. insulare were determined. Culture medium values for I ranged between 200 and 1,250 μg L⁻¹ (1.1–6.7 μmol L⁻¹), for III between 115 and 390 μg L⁻¹ (0.5–1.8 μmol L⁻¹), whereas concentrations of II were conspicuously lower (2–16 μg L⁻¹ or 0.014–0.094 μmol L⁻¹). Amounts of III per volume of culture medium were about tenfold higher than in the biomass contained in an equal culture volume. This difference is an indication for an active excretion of III. Amounts of I and II in biomass and media were of no significant difference. In the neutral red uptake assay, I and II were found to be toxic against eukaryotic cells as follows: I was of considerable cytotoxicity in concentrations from 1,000 to 10 mg L⁻¹ and of lower cytotoxicity (causing a 27 % decrease of cell viability) in a concentration of 1,000 μg L⁻¹, whereas II was merely of considerable cytotoxicity in concentrations from 1,000 to 100 mg L⁻¹. Because of the cytotoxicity of I and because of the many known pharmacological effects of II there is a possibility that a certain amount of risk to humans and livestock comes from cultures or even from biomass- free culture media of N. insulare. The natural function of the examined exometabolites is discussed.     

Biofilm growth and near-infrared radiation-driven photosynthesis of the chlorophyll d-containing cyanobacterium Acaryochloris marina

The cyanobacterium Acaryochloris marina is the only known phototroph harboring chlorophyll (Chl) d. It is easy to cultivate it in a planktonic growth mode, and A. marina cultures have been subject to detailed biochemical and biophysical characterization. In natural situations, A. marina is mainly found associated with surfaces, but this growth mode has not been studied yet. Here, we show that the A. marina type strain MBIC11017 inoculated into alginate beads forms dense biofilm-like cell clusters, as in natural A. marina biofilms, characterized by strong O(2) concentration gradients that change with irradiance. Biofilm growth under both visible radiation (VIS, 400 to 700 nm) and near-infrared radiation (NIR, ～700 to 730 nm) yielded maximal cell-specific growth rates of 0.38 per day and 0.64 per day, respectively. The population doubling times were 1.09 and 1.82 days for NIR and visible light, respectively. The photosynthesis versus irradiance curves showed saturation at a photon irradiance of E(k) (saturating irradiance) >250 μmol photons m(-2) s(-1) for blue light but no clear saturation at 365 μmol photons m(-2) s(-1) for NIR. The maximal gross photosynthesis rates in the aggregates were ～1,272 μmol O(2) mg Chl d(-1) h(-1) (NIR) and ～1,128 μmol O(2) mg Chl d(-1) h(-1) (VIS). The photosynthetic efficiency (α) values were higher in NIR-irradiated cells [(268 ± 0.29) × 10(-6) m(2) mg Chl d(-1) (mean ± standard deviation)] than under blue light [(231 ± 0.22) × 10(-6) m(2) mg Chl d(-1)]. A. marina is well adapted to a biofilm growth mode under both visible and NIR irradiance and under O(2) conditions ranging from anoxia to hyperoxia, explaining its presence in natural niches with similar environmental conditions.     

Phosphorus and nitrogen limitation of phytoplankton growth in eutrophic Lake Inba, Japan

Lake Inba is one of the most eutrophic lakes in Japan. In this study, field sampling and nutrient enrichment bioassays were conducted to determine the seasonal patterns of nutrient limitation for phytoplankton growth in this lake. Phytoplankton biomass increased significantly with the additions of phosphorus (P) on almost all sampling dates, indicating P limitation of phytoplankton growth from spring to autumn. However, nitrogen (N) limitation was also observed during summer (i.e., 19 August). On 10 August, a typhoon struck Lake Inba. After this event, dissolved inorganic nitrogen (DIN) and phosphorus concentrations increased, probably because of increased river discharge. At the same time, phytoplankton growth in the control treatment became relatively high, with the addition of neither P nor N stimulating the growth. However, 10 days after the typhoon, the phytoplankton growth rate in the control treatment decreased, with only the addition of N having a significant positive effect on phytoplankton growth. N limitation during summer is caused by the low concentrations of DIN, as well as changes in the N:P ratio due to allochthonous nutrient loads. These results indicate that a reduction of both P and N input is necessary to control phytoplankton blooms in Lake Inba.     

pH regulation of growth, photosynthesis, glutamine synthetase activity and micronutrient transport in the cyanobacterium Hapalosiphon welwitschii

Abstract Optima for growth, oxygenic photosynthesis and glutamine synthetase activity occurred at pH 10, thus suggesting that the cyanobacterium Hapalosiphon welwitschii is an alkalophile. It produced a Cu-Zn efflux system at pH 9 or 10, but not at pH 7 or 8, to relieve photosynthesis from Cu or Zn inhibition. This finding has a bearing on the ecophysiological competence of the cyanobacterium under natural conditions.     

2 site-specific endonucleases of the cyanobacterium Nostoc linckia]

Two restrictases Nli387/7 I and Nli387/7 II have been isolated from cyanobacterium Nostoc linckia using chromatography on phosphocellulose, "Mono Q" column, and heparin sepharose 4B. The preparations are described by the method of electrophoresis in polyacrylamide gel under denaturing conditions. Catalytic properties of restrictases are determined: optimal pH of the action--9.0--9.5, optimal concentration of Na+--5 mM, that of Mg2+--6 mM, optimal temperature--37 degrees C. The isolated enzymes are isoschizomers of restrictases avaI and AvaII. The point of cutting is determined for enzyme Nli387/7 I. It is shown that restrictase Nli387/7 I is a false isoschizomer Ava I.     

Isolation of a symbiotic cyanobacterium,Nostoc cycadae,and its nitrogen metabolism

Nostoc cycadae isolated from the host Cycas revoluta grew well in medium devoid of combined nitrogen but maximum growth was in medium containing nitrate (4.1 g chlorophyll a ml^-1). Aerated coralloid roots in the dark produced more NH₃ when treated with l-methionine-dl-sulphoximine (MSO), an inhibitor of glutamine synthetase. With cultured N. cycadae and freshly isolated N. cycadae, NH₃ production was enhanced by adding a host-tissue extract in the light or in the dark, whereas it was decreased by adding MSO. Nitrogenase activity was four times higher in coralloid root than in the cultured endophyte N. cycadae. The host-tissue extract may inhibit NH₃ assimilatory pathways, thus inducing production of NH₃ that can be utilized by the host itself.V. Singh, M.R. Goyle and E.R.S. Talpasayi are with the Laboratory of Algal Physiology and Biochemistry and A.K. Srivastava is with the Ecology Research Laboratory, both of the Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi-221005, India.     

Evidence for the nitrate assimilation-dependent nitrite excretion in cyanobacterium Nostoc MAC

Nitrate uptake and nitrite efflux patterns in Nostoc MAC showed a rapid phase followed by their saturation. Nitrite efflux was maximum in nitrate medium whereas the cells incubated in N₂ and NH ₄ ⁺ media exhibited a decreased nitrite efflux activity. The simultaneous presence of NH ₄ ⁺ and nitrate significantly decreased nitrite efflux. L-Methionine-Dl-sulphoximine (MSX) prevented inhibition of nitrite efflux by NH ₄ ⁺ . In the dark there was negligible nitrite efflux, whereas illumination increased the rate of nitrite efflux significantly. The nitrite efflux system was maximally operative at pH 8.0, 30°C and a photon fluence rate of 50 mol m^-2. s^-1. These results confirm that (i) the nitrite efflux system in Nostoc MAC is dependent upon nitrate uptake and assimilation and is repressible by NH ₄ ⁺ ; (ii) NH ₄ ⁺ itself is not the actual repressor of nitrite efflux; a product of NH ₄ ⁺ assimilation via glutamine synthetase (GS) is required for repression to occur; (iii) the catalytic function of GS does not appear to be involved in nitrate assimilation-dependent nitrite efflux, and (iv) the optimum pH, temperature and illumination for maximum nitrite efflux were found to be 8.0, 30°C and 50mol m^-2. s respectively.B.B. Singh, P.K. Pandey and P.S. Bisen are with the Department of Microbiology, Barkatullah University. Bhopal 462026, India. S.Singh is with the Department of Microbiology, School of Life Sciences, North Maharashtra University, Jalgaon, India     

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     

Phosphorus limitation on photosynthesis of two dominant understory species in a lowland tropical forest

Aims Elevated nitrogen (N) deposition in tropical regions may accelerate ecosystem phosphorus (P) limitation. However, it is not explicitly addressed that how changes in soil N and P availability affect foliar nutrients and photosynthesis of plants in tropical forests. In this study, we examined the effects of N and P additions on foliar nutrients and net photosynthesis of two dominant understory species, Randia canthioides (R. canthioides) and Cryptocarya concinna (C. concinna) in an N-saturated old-growth tropical forest (>400-year-old) in southern China.Methods A full factorial NP addition experiment (2×2) was established in 2007 and continued through August 2010. Four treatments, including control, N addition (150kg N ha^-1 year^-1), P addition (150kg P ha^-1 year^-1) and NP addition (150kg N ha^-1 year^-1 plus 150kg P ha^-1 year^-1) were set up in this experiment. Photosynthetic traits (maximum photosynthetic CO₂ assimilation (A max), stomatal conductance (g s), leaf transpiration (E), light saturating point, concentrations of chlorophyll a/b and foliar nutrients (N and P) of the two species were measured with standard methods.Important findings Three years of N addition had no significant effects on any measured photosynthetic parameter of either species. However, N addition significantly elevated foliar N and P concentrations of one species (R. canthioides), resulting in lower photosynthetic nitrogen use efficiency (PNUE). N treatments decreased foliar P concentration of the other (C. concinna), resulting in increased photosynthetic phosphorus use efficiency, which was potentially related to N-induced P shortage. In contrast, positive effects of P treatments on g s of R. canthioides, A max and chlorophyll a+b of C. concinna were observed. P treatments also elevated foliar P and PNUE of both species, implying P induced more efficient use of N. Our results suggested a more important role of P than N on influencing photosynthetic traits of these two understory species. Alleviation of P shortage through P addition may enhance photosynthetic performances of some understory species in N-rich tropical forests.