Characterization of a small endogenous plasmid from the CyanobacteriumPlectonema boryanum

The nonheterocystous filamentous CyanobacteriumPlectonema boryanum strain UTEX 594 contains at least two plasmids. A small 145 kb plasmid was cloned in pBR322. It has no homology with the bigger resident plasmid or with chromosomal DNA. A small fraction of the plasmid is present in the form of multimers or concatemers. Copy number and hybridization patterns of the plasmid were similar under dinitrogen-fixing and non-fixing conditions. Restriction site mapping of the plasmid was done to enable its use in the development of cyanobacterial cloning vectors. It is among the smallest natural plasmids reported from bacteria.     

The occurrence of the hydrogenase in some blue-green algae

Several blue-green algae were surveyed for the occurrence of the hydrogenase which was assayed by the oxyhydrogen or Knallgas reaction in the intact organisms. In aerobically grown cultures, the reaction was detectable in Anabaena cylindrica, Nostoc muscorum and in two Anabaena variabilis species, whereas virtually no activity was observed in Anacystis nidulans and Cyanophora paradoxa. In these latter two algae, the reaction was, however, found after growth under molecular hydrogen for several days, which drastically increased the activity levels with all the algae tested. In the nitrogen fixing species, the activity of the Knallgas reaction was enhanced when all combined nitrogen was omitted from the media. H₂ and hydrogenase could not significantly support the CO₂-fixation in photoreduction experiments with all blue-green algae investigated here. Hydrogenase was assayed by the dithionite and methyl viologen dependent evolution of hydrogen and was found to be present with essentially the same specific activity levels in preparations of both heterocysts and vegetative cells from Anabaena cylindrica. Na₂S₂O₄ as well as H₂ supported the C₂H₂-reduction of the isolated heterocysts. The H₂-dependent C₂H₂-reduction did not require the presence of oxygen but was strictly light-dependent where H₂ served as an electron donor to photosystem I of these cells. It is concluded that hydrogen can be utilized by two different pathways in blue-green algae.Abbreviations Chl chlrophyll - CP creatine phosphate - CP kinase creatine phosphokinase - DCMU N-(3,4-dichlorophenyl)N,N-dimethylurea     

Immunogold localization of ribulose-1,5-bisphosphate carboxylase/oxygenase in the nitrogen-fixing cyanobacterium,Plectonema boryanum

Summary Antiserum against the Calvin cycle enzyme, ribulose-1,5-bisphosphate carobxylase/oxygenase (RuBisCO), was used in conjunction with colloidal gold to localize RuBisCO in nitrogen-fixing (fix+) and nonfixing (fix–)Plectonema boryanum cells. RuBisCO antiserum consistently labeled the cytoplasm and polyhedral bodies (carboxysomes) in both fix+ and fix– cells. Through morphometry, it was determined that significantly less gold label (indicative of RuBisCO) was present in fix+ cells. This decreased RuBisCO content correlated with a decrease in net photosynthetic oxygen evolution also observed in fix+P. boryanum.Abbreviations RuBisCO Ribulose-1,5-bisphosphate carboxylase/oxygenase - fix+ nitrogen-fixing - fix– nonfixing     

Toxic cyanobacteria (blue-green algae) in Finnish fresh and coastal waters

A survey of the occurrence of toxic blooms of cyanobacteria in Finnish fresh and coastal waters was made during 1985 and 1986. Toxicity of the freeze-dried water bloom samples was tested by mouse-bioassay (i.p.). Forty-four per cent (83/188) of the bloom samples were found to be lethally toxic. Hepatotoxic blooms (54) were almost twice as common as neurotoxic ones (29). Anabaena was the most frequently found genus in toxic and non-toxic blooms and it was present in all neurotoxic samples. Statistical associations were found between hepatotoxicity and incidence of Microcystis aeruginosa, M. viridis, M. wesenbergii, Anabaena flos-aquae and Anabaena spiroides. Neurotoxicity was statistically associated with Anabaena lemmermannii, Anabaena flos-aquae and Gomphosphaeria naegeliana. Isolation of strains of cyanobacteria confirmed the occurrence of hepatotoxic and neurotoxic strains of Anabaena, as well as hepatotoxic strains of Microcystis and Oscillatoria species.Toxic blooms caused cattle poisonings at three different lakes during the study period. Toxic blooms also occurred in drinking water sources. Our study shows that toxic cyanobacteria are more common in Finnish lakes than would be expected on the basis of animal poisonings. The results of this study show the existence of toxic cyanobacteria in Finnish water supplies and the need for their continued study as agents of water based disease.     

Combination and positional distribution of fatty acids in lipids from blue-green algae

The main glycerolipids (monogalactosyl-, digalactosyl-, sulphoquinovosyl diacylglycerol, phosphatidylglycerol) from five blue-green algae (Microcystis, Anabaena, Nostoc, Oscillatoria, Tolypothrix) were analyzed for fatty acid composition, occurrence of diglyceride species and positional distribution of fatty acids between thesn-1- andsn-2-position of glycerol. In contrast to eucaryotic plants biosynthetically closely related lipids (monogalactosyl-, digalactosyl-, trigalactosyl diacylglycerol) show nearly identical diglyceride moieties, whereas sulphoquinovosyl diacylglycerol and phosphatidylglycerol are separated from galactolipids by composition as well as occurrence of fatty acids. On the other hand the positional distribution of fatty acids in all lipids is controlled exclusively by chain length and not by degree of unsaturation with C₁₈-fatty acids at thesn-1- and C₁₆-fatty acids at thesn-2-position. These results show that in procaryotic organisms the diversity in diglyceride portions of lipids is reduced as compared to eucaryotic organisms, but nevertheless does exist.Abbreviations MGD, DGD, TGD, SQD monogalactosyl-, digalactosyl-, trigalactosyl-, sulphoquinovosyl diacylglycerol - PG phosphatidyl glycerol     

Functional 70S hybrid ribosomes from blue-green algae and bacteria

Hybrid 70S ribosomes were produced by combining Anacystis nidulans and Escherichia coli 30S and 50S subunits. Both the A. nidulans 30S-E. coli 50S and E. coli 30S- A. nidulans 50S hybrids were functional in synthesizing protein when tested in a standard in vitro amino acid incorporating system. Both 70S hybrids were inhibited by streptomycin but the degree of inhibition was dependent upon the source of the 30S subunit. The ability to form functional 70S ribosomes from subunits of blue-green algae and bacteria is further evidence of the procaryotic nature of blue-greens and of the functional homology of the two protein synthesizing systems.     

The relationship between heat-stress and photobleaching in green and blue-green algae

Two characteristic temperatures were identified from measurements of the temperature dependence of O₂ evolution by Chlorella vulgaris and Anacystis nidulans: T₁, the threshold temperature for inhibition of O₂ evolution under saturating light conditions, and T₂, the upper temperature limit for O₂ evolution. Measurement of delayed light emission from photosystem II (PSII) showed that it passed through a maximum at T₁ and was virtually eliminated on heating the samples to T₂. Related changes were observed in low-temperature (77K) fluoresence emission spectra. Heat-stress had little effect on the absorption properties of the cells at temperatures below T₁ but incubation at higher temperatures, particularly under high-light conditions, resulted in extensive absorption losses. An analysis of these measurements suggests that this increased susceptibility to photobleaching is triggered by an inhibition of the flow of reducing equivalents from PSII that normally serves to protect the light-harvesting apparatus of the cells from photo-oxidation. Adaptation to higher growth temperatures resulted in increases in the values of T₁ and T₂ for Anacystis nidulans but not for Chlorella vulgaris.Abbreviations PSI photosystem I - PSII photosystem II - Chl a chlorophyll a - Chl b chlorophyll b - DCMU 3-(3 4 dichlorophenyl)-11-dimethylurea - PC plastocyanin - APC allophycocyanin CIW-DPB Publication No. 887.     

Heterotrophic capacities of Plectonema boryanum

Acquisition of the dark heterotrophic growth capacity on glucose in Plectonema boryanum involves both adaptation and enrichment of a fast-growing genotype. The adaptation includes induction of functions involved in glucose incorporation and increase in glucose-6-phosphate dehydrogenase activity. Photosynthetic products are implicated in the control of both systems. Efficient energy conversion in the dark, as measured by cyanophage multiplication, correlates in time with the increase in potential for glucose incorporation while heterotrophic growth capacity correlates with the increase in glucose-6-phosphate dehydrogenase activity. The lower efficiency of heterotrophic growth compared to photoautotrophic growth is discussed in light of the conservation of the photosynthetic potency in the heterotrophic cells.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DTT dithiothreitol - G6P glucose-6-phosphate - NADP nicotinamide adenine dinucleotide phosphate - NTG N-methyl-N-nitro-N-nitrosoguanidine - RUDP ribulose-1,5-diphosphate - TCA trichloroacetic acid Dedicated to Prof. R. Y. Stanier on the occasion of his 60th birthday     

A study of methods for in situ X-ray energy dispersive analysis of polyphosphate bodies in Plectonema boryanum

A variety of preparative methods for in situ X-ray energy dispersive analysis were tested to determine their effects on the elemental composition of polyphosphate bodies in P. boryanum. The bodies were found to contain large amounts of P and K and small amounts of Ca and Mg. Air drying, freeze-drying and freeze-drying from a liquid nitrogen slush all gave similar results. Fixation of the cells in glutaraldehyde and/or OsO₄ resulted in loss of the K and enhancement of the Ca peak. Magnesium was lost during embedding in epoxy.     

Studies on 9-amino-6-chloro-2-methoxy acridine fluorescence quenching and enhancement in relation to energy transduction in spheroplasts and intact cells of the cyanobacterium Plectonema boryanum

The fluorescent probe 9-amino-6-chloro-2-methoxy acridine was used to study the energy transduction in the thylakoid and cell membranes of the cyanobacterium Plectonema boryanum. Apart from light-driven electron transfer, the dark endogenous respiration also leads to energization resulting in an ACMA fluorescence response, that is sensitive to the electron flow inhibitor 2, 5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, to the energy transfer inhibitors dicyclohexylcarbodiimide and venturicidine and to the uncoupler 5-chloro-3-t-butyl-2-chloro-4-nitrosalicylanilide.In spheroplasts, in which the cell membranes have lost their capacity to maintain a proton gradient, the respiration-and light-induced ACMA fluorescence changes (quenching) are similar to those in chloroplasts. In intact cells a combination of reversible quenching and enhancement of ACMA fluorescence was found. This dualistic behaviour is supposedly caused by an opposite orientation of the thylakoid and cell membranes. ACMA quenching at the level of the thylakoids was obtained either by respiratory or photosynthetic electron transfer and gave similar responses to those obtained in the spheroplasts. The slower ACMA fluorescence enhancement, only observed in cells with intact cell membranes, also evoked by both respiration and light-induced energization is sensitive to the compounds mentioned above and in addition to KCN.Our results support the view [8] that dark oxidation of substrates by O₂ proceeds via the thylakoid membrane and terminates at a CN^- sensitive oxidase located in the cell membrane which requires the involvement of a mobile cytoplasmic redox mediator.Abbreviations ACMA 9-amino-6-chloro-2-methoxy acridine - chl a chlorophyll a - DBMIB 2, 5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCCD dicyclohexylcarbodiimide - DNP dinitrophenol - DNP-INT dinitrophenyl ether of 2-iodo-4-nitrothymol - FCCP carbonylcyanide-p-trifluoro-methoxy phenylhydrazone - S-13 5-chloro-3-t-butyl-2-chloro-4-nitrosalicylanilide - tricine N-2 (2-Hydroxy-1, 1-bis (hydroxymethyl) ethyl)-glycine - Tris Tris (hydroxymethyl) amino methane     

The effect of Daphnia on filament length of blue-green algae

Water from a hypertrophic lake rich in filamentous blue-green algae was passed through a continuous-flow system of aquaria containing Daphnia magna, and a control system without Daphnia. Daphnia caused a significant decrease in the blue-green algal density, and a two-fold reduction in filament length. It is suggested that feeding activity of Daphnia may result in an increase in the availability of blue-green filaments to filter-feeding cladocerans.     

Photoreactivation of the UV light effects on growth of Scots pine (Pinus sylvestris L.) seedlings

Summary Ultraviolet-B light (UV-B) and ultraviolet-A light (UV-A) at higher doses exert a strong inhibitory (toxic) effect on axis growth in Scots pine (Pinus sylvestris L.) seedlings. This effect is unrelated to control of growth rate by phytochrome. Rather, after a toxic UV dose growth of the pine seedling no longer responded to phytochrome. Both, the effect of UV-B as well as the inhibiting effect of UV-A could be photoreactivated by blue light (B). The action of UV-A was 2 fold: (i) it exerted a toxic effect which could be photoreactivated by B, and (ii) applied after UV-B it photoreactivated to some extent the toxic UV-B effect. Obviously, the UV-A range causes a toxic effect, and at the same time is capable of photoreactivating the toxic UV effect. At higher doses the toxic effect prevails.     

Light-induced proton translocation through thylakoid and cytoplasmic membranes of Plectonema boryanum

Light-induced fluorescence changes of 9-aminoacridine, an indicator of proton gradient in energy-transducing membranes, were studied in Plectonema boryanum and other cyanobacteria. The fluorescence changes observed in cell suspensions resulted from a superposition of fluorescence quenching and enhancement as the analysis of the kinetic data shows. Both components of the fluorescence changes are abolished by 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU) and m-chlorocarbonylcyanide phenylhydrazone. The inhibitory effect of DCMU is removed by 2,3,5,6- or N,N,N′,N′-tetramethyl-p-phenylenediamine. The fluorescence quenching sensitive to substrates and uncouplers of the photophosphorylation is only observed in membrane vesicles obtained by osmotic shock of P. boryanum spheroplasts. Presumably, light-induced quenching of the dye fluorescence in the cells of cyanobacteria is due to the proton transport from the cytoplasm in the inner space of thylakoids while fluorescence enhancement is due to the proton efflux from the cytoplasm into the incubation medium.     

感染丝状蓝藻的噬藻体的裂解周期和释放量的测定

近年来 ,随着浮游病毒的认识的深入 ,人们认识到浮游病毒对水体中初级生产力的影响是巨大的[1] ,其主要证据就是发现噬藻体在海洋蓝藻的种群控制上发挥着重要作用[2 ] 。噬藻体的释放量和裂解周期是衡量噬藻体感染力的重要指标 ,很多重要的生态指标如病毒在生态系统中对宿主的致死率、病毒种群得以维持的阈浓度等都需要使用病毒的释放量和裂解周期来加以推算[3,4 ] ,因此准确地测定这两个基本参数是十分重要的。在自然界 ,很多丝状蓝藻 ,如颤藻、鱼腥藻、螺旋藻、席藻等是能够形成水华的 ,其中有些还具有产毒的功能[5] 。丝状蓝藻的形态特征…     

Ultrastructure of the nitrogen-fixing,filamentous, nonheterocystous cyanobacterium,Plectonema boryanum

Summary The ultrastructure of fructose-supplemented and unsupplemented nitrogen-fixing (fix +) and nonfixing (fix –)Plectonema boryanum UTEX 581 cells was examined by transmission electron microscopy. The most prominent structural differences included the arrangement and morphology of the thylakoids and alterations in the appearance of the interthylakoidal spaces. These ultrastructural differences, together with other observations such as glycogen content and presence of nitrogenase (using acetylene reduction assay and immunogold localization), readily distinguished nonfixingP. boryanum from nitrogen-fixing cells.     

Hydrogen metabolism in the facultative anoxygenic cyanobacteria (blue-green algae) Oscillatoria limnetica and Aphanothece halophytica

Two facultative anoxygenic photoautotrophic cyanobacteria, Oscillatoria limnetica and Aphanothece halophytica were found capable of CO₂ photoassimilation using molecular hydrogen as electron donor in a photosystem I driven reaction. A. halophytica was also capable of evolving hydrogen from Na-dithionite reduced methylviologen in a light independent reaction.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DSPD Disallcylidenepropanediamine - FCCP Carbonylcyanide p-trifluoromethoxyphenyl hydrazone - Tricine N-tris(hydroxy methyl)-methylglycine     

Isolation and characterization of rapidly-growing,marine, nitrogen-fixing strains of blue-green algae

Five strains of heterocystous blue-green algae capable of high rates of growth and nitrogenase activity were isolated from shallow coastal environments. Growth of the organisms was characterized with respect to temperature, NaCl concentration in the medium, and nitrogen source. The temperature optima ranged from 35–42°C, and all but one of the strains displayed a requirement for added NaCl. The generation times under N₂-fixing conditions were 5.1–5.9 h, and were as low as 3.4 h for growth on NH₄Cl. Nitrogenase activity (C₂H₂ reduction) was high throughout the logarithmic growth phase of each strain. The maximum value observed for one strain was 65.5 nmoles C₂H₄ produced/mg protein x min, and the average values for the five strains ranged from 24.5–46.7 nmoles C₂H₄/mg protein x min. The organisms all belong to the genusAnabaena. The growth and nitrogenase activity of these strains are much higher than those of the heterocystous blue-green algae commonly used for investigation of nitrogen metabolism, and they thus should prove to be useful physiological tools. Their prevalence, as judged by the ease of their enrichment and isolation, in bay and estuarine environments suggests that they are important contributors of combined nitrogen.     

Consequences of iron deficiency on photosynthetic and respiratory electron transport in blue-green algae

Growth of Aphanocapsa in low iron media resulted in a decrease of the endogenous iron pool. Below a critical concentration photosynthetic electron transfer was specifically depressed. This was caused by a strong inhibition of the synthesis of cytochromes b-559 of PSII, cytochromes b-563, f-557, and the Rieske Fe-S center of the cytochrome complex and especially the Fe-S centers of PSI. The influence of iron limitation on respiration and chlorphyll formation was negligible.Paper presented at the FESPP meeting (Strasbourg, 1984)     

Bioactive compound production by thermophilic and thermotolerant cyanobacteria (blue-green algae)

A thermotolerant species of Phormidium produced extracellular anti-microbial material during batch culture. Although this material was inactive when screened against a number of other cyanobacteria, it inhibited the growth of a wide range of Gram-positive and Gram-negative heterotrophic bacteria, Candida albicans and Cladosporium resinae.The authors are with the Department of Biological Sciences, University of Dundee, Dundee DD1 4HN, UK     

Growth interactions among blue-green (Anabaena Oscillarioides,Microcystis aeruginosa) and green (Chlorella sp.) algae

The growth interactions amongst the blue-green algal species Anabaena oscillarioides, Microcystis aeruginosa and the green alga, Chlorella sp. were studied both in mixed cultures and in filter cultures separated by a membrane filter in the two arms of an interaction U-tube. The role of nutrients especially phosphate upon the interaction has also been studied. Anabaena and Microcystis both inhibited the growth of Chlorella while Microcystis also inhibited the growth of Anabaena. The inhibitory effect of Microcystis was found to be dependent on high concentrations of the initial algal inocula and independent of the initial concentration of nutrients such as inorganic phosphate, indicating that the nature of the inhibition is probably due to the production of inhibitory extracellular products by Microcystis. On the other hand, the inhibitory effect of Anabaena on Chlorella is the consequence of nutrient competition with Anabaena competing more effectively for the available phosphate.