Cation-exchange capacity of algae and cyanobacteria: A parameter of their metal sorption abilities

Summary The potential metal sorption abilities of algae and cyanobacteria were estimated as cation-exchange capacities, using a potentiometric titration method. Unicellular cyanobacteriaAnacystis nidulans, Synechocystis aquatilis, and the green microalgaStichococcus bacillaris revealed a higher maximal capacity (205–825 eq g^–1 dry wt) than filamentous macroalgaVaucheria sp. (Xanthophyceae, 41 eq g^–1 dry wt). The cation-exchange capacity decreased when external pH decreased. Different ion-exchange properties of cell surfaces of cyanobacteria and algae were observed.     

Observations on naturally and artificially diseased tropical corals: A scanning electron microscope study

Scanning electron microscopic (SEM) observations of naturally and artificially diseased corals reveal that the disease is characterized by a filamentous matrix of cyanobacterial andBeggiatoa filaments. Spiral bacteria are commonly embedded in the matrix. The artificial disease is not manifested as the characteristic black line disease and does not contain filaments of cyanobacteria. This suggests that cyanobacteria are necessary for the black line phenomenon. The colorless, sulfide-oxidizing bacteriumBeggiatoa, however, is always associated with the disease.     

Partial sequence of ribulose-1,5-bisphosphate carboxylase/oxygenase and the phylogeny of Prochloron and Prochlorococcus (Prochlorales)

The prochlorophytes, oxygenic photosynthetic prokaryotes having no phycobiliprotein but possessing chlorophylls a and b, have been proposed to have a common ancestry with green chloroplasts, yet this is still controversal. We report here that partial sequence comparisons of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, including sequence data from two prochlorophytes, Prochlorococcus and Prochloron, indicate that Prochlorococcus is more closely related to a photosynthetic bacterium, Chromatium vinosum (-purple bacteria), than to cyanobacteria, while Prochloron is closely related to the prochlorophyte Prochlorothrix and to cyanobacteria. The molecular phylogenetic tree indicates that a common ancestor of Prochlorococcus and -purple bacteria branched off from the land plant lineage earlier than Prochloron, Prochlorothrix, and cyanobacteria.Correspondence to: A. Shimada     

Preliminary studies of cyanobacteria, picoplankton, and virioplankton in the Salton Sea with special attention to phylogenetic diversity among eight strains of filamentous cyanobacteria

Cyanobacterial diversity in the Salton Sea, a high-salinity, eutrophic lake in Southern California, was investigated using a combination of molecular and morphological approaches. Representatives of a total of 10 described genera (Oscillatoria, Spirulina, Arthrospira, Geitlerinema, Lyngbya, Leptolyngbya, Calothrix, Rivularia, Synechococcus, Synechocystis) were identified in the samples; additionally, the morphology of two cultured strains do not conform to any genus recognized at present by the bacteriological system. Genetic analysis, based on partial 16S rRNA sequences suggested considerable cryptic genetic variability among filamentous strains of similar or identical morphology and showed members of the form-genus Geitlerinema to be distributed among three major phylogenetic clades of cyanobacteria. Cyanobacterial mats, previously described from the Sea were, in fact, composed of both filamentous cyanobacteria and a roughly equivalent biomass of the sulfur-oxidizing bacterium Beggiatoa, indicating their formation in sulfide rich regions of the lake. Flow cytometric analysis of the water samples showed three striking differences between samples from the Salton Sea and representative marine waters: (1) phycoerythrin-containing unicells, while abundant, were much less abundant in the Salton Sea than they were in typical continental shelf waters, (2) Prochlorococcus appears to be completely absent, and (3) small (3–5 m) eukaryotic algae were more abundant in the Salton Sea than in typical neritic waters by one-to-two orders-of-magnitude. Based on flow cytometric analysis, heterotrophic bacteria were more than an order of magnitude more abundant in the Salton Sea than in seawater collected from continental shelf environments. Virus particles were more abundant in the Salton Sea than in typical neritic waters, but did not show increases proportionate with the increase in bacteria, picocyanobacteria, or eukaryotic algae.     

Prochlorococcus marinus nov. gen. nov. sp.: an oxyphototrophic marine prokaryote containing divinyl chlorophyll a and b

Several years ago, prochlorophyte picoplankton were discovered in the N. Atlantic. They have since been found to be abundant within the euphotic zone of the world's tropical and temperate oceans. The cells are extremely small, lack phycobiliproteins, and contain divinyl chlorophyll a and b as their primary photosynthetic pigments. Phylogenies constructed from DNA sequence data indicate that these cells are more closely related to a cluster of marine cyanobacteria than to their prochlorophyte relatives Prochlorothrix and Prochloron. Several strains of this organism have recently been brought into culture, and herewith are given the name Prochlorococcus marinus.     

Circadian oscillators,cell cycles,and singularities: light perturbations of the free-running rhythm of cell division inEuglena

Summary The free-running circadian rhythm of cell division in the algal flagellate,Euglena gracilis (Z) was perturbed by 3-h light signals of varying intensities imposed at different circadian times (CT). Light pulses within the range of 700 to 7,500 lux were found to yield the same strong (Type 0) phase response curve (PRC) comprising both advance and delaye phase shifts as great as 15 h. Dark signals generated a PRC of reduced amplitude with very little, if any, phase advance being observed. Light perturbations of lower intensity, however, elicited quite different responses if applied at a quite specific circadian time: A 40- to 400-lux pulse given at approximately CT 0 (late subjective night) induced total arrhythmicity, and the culture reverted to asynchronous, exponential growth. Different degrees of arryhtmicity were induced by the same low-intensity perturbations (I ^*) given slightly before or after this sensitive phase point (T ^*), but if imposed at other circadian times, they generated normal type 0 phase resetting. The demonstration of the existence of this critical pulse (T ^*,I ^*) provides further evidence that the cell division cycle ofEuglena (and presumably other microorganisms) is regulated by a circadian oscillator and, in particular, by one having limit cycle dynamics.Abbreviations LL continuous illumination - DD continuous darkness - LD light-dark cycle - LD x, y, light-dark cycle comprisingx h of light andy h of dark - t period of a LD cycle - CO circadian oscillator - CR circadian rhythm - period of a freerunning circadian rhythm in constant conditions (taken here to be the time between onsets of cell division in a population of cells - _R phase marker, or phase reference point (here, the onset of the division burst) - phase of the rhythm - change in phase (phase shift) - new phase attained after phase shift - CT circadian time (CT 0 indicates the phase point of a free-running rhythm that has been normalized to 24 h which corresponds to that occurring at the onset of light in aLD:12, 12 reference cycle) - PRC phase response curve (plot of phase shift engendered by a perturbation as a function of the circadian time of its application) - T ^*,I ^*) coordinates of an annihilating (light) stimulus given at a critical circadian time (T ^*, corresponding to the singularity point) and having a critical strength (I ^*) - CDC cell division cycle - average generation (doubling) time of a cell population - average step-size, or factorial increase in cell titer (plateau to plateau) after a phased division burst Dedicated to Prof. Colin S. Pittendrigh on only his 65th birthday     

Processing of antennular input in the brain of the spiny lobster, Panulirus argus

Neurons in the olfactory deutocerebrum of the spiny lobster, Panulirus argus, were recorded intracellularly and filled with biocytin. Recorded neurons arborized in the olfactory lobe (OL), a glomerular neuropil innervated by olfactory and some presumptive mechanosensory antennular afferents. The neurons responded to chemosensory input from the lateral antennular flagellum bearing the olfactory sensilla but not the medial flagellum bearing many non-olfactory chemosensory sensilla. Many neurons received additional mechanosensory input. Thus the OL integrates specifically olfactory with mechanosensory input. OL neurons had multiglomerular arborizations restricted to one or two of the three horizontal layers of the columnar glomeruli. OL local interneurons comprised core neurons with tree-like neurites and terminals in the base of the glomeruli and rim neurons with neurites surrounding the OL and terminals in the cap/subcap. The somata of OL local interneurons lay in the medial soma cluster (100000 somata). OL projection neurons arborized in the base of the glomeruli and ascended via the olfactory glomerular tract to the lateral protocerebrum. A parallel projection pathway is constituted by projection neurons of the accessory lobe, a glomerular neuropil without afferent innervation but intimate links to the OL. The projection neuron somata constituted the lateral soma cluster (200000 somata).Abbreviations AC anterior cluster (cluster 6,7) - AL accessory lobe - aMC anterior subcluster of medial cluster (cluster 9) - A _lNv main antenna I (antennular) nerve - A _lNM antenna I (antennular) motor nerve - A _llNv main antenna II (antennal) nerve - CB central body - CL central layer of accessory lobe - DC deutocerebral commissure - DCN deutocerebral commissure neuropil - dDUMC dorsal subcluster of dorsal unpaired median cluster (cluster 17) - dMC dorsal subcluster of medial cluster (cluster 11) - dVPALC dorsal subcluster of ventral paired anterolateral cluster (cluster 8) G glomerulus - IDUMC lateral subcluster of dorsal unpaired median cluster (cluster 16) - LC lateral cluster (cluster 10) - LF lateral flagellum of antenna I (antennule) - LL lateral layer of accessory lobe - MF medial flagellum of antenna I (antennule) - ML medial layer of accessory lobe - MPN anterior and posterior median protocerebral neuropils - OGT olfactory globular tract - OGTN olfactory globular tract neuropil - OL olfactory lobe - OLALT olfactory lobe-accessory lobe tract - PB protocerebral bridge - pMC posterior subcluster of medial cluster (cluster 9) - PT protocerebral tract - TNv tegumentary nerve - VPMC ventral paired medial cluster (cluster 12) - VUMC ventral unpaired medial cluster (cluster 13) - vVPALC ventral subcluster of ventral paired anterolateral cluster (cluster 8) - ASW artificial sea water - M3 mixture 3 - PRO L-proline - TM TetraMarin extract     

Ingestion of the cyanobacterium Trichodesmium spp. by pelagic harpacticoid copepods Macrosetella,Miracia and Oculosetella

Trichodesmium is a filamentous, colonial nitrogen fixing cyanobacteria, ubiquitous in tropical and subtropical regions of the world's oceans. Trichodesmium fixes atmospheric nitrogen and can comprise a significant fraction of total primary production in oceanic surface waters. Therefore, the consumption and fate of Trichodesmium has important consequences for understanding carbon and nitrogen cycling in the open ocean. The pelagic harpacticoid copepod Macrosetella gracilis uses Trichodesmium not only as a physical substrate for juvenile development, but also as a food source. Several different types of pelagic copepods (including several species of calanoids, harpacticoids and a poecilostomatoid species) were tested for ingestion of Trichodesmium by labelling the cyanobacteria with ¹⁴C. Only the pelagic harpacticoids ingested Trichodesmium. Here we report the first grazing rates based on ¹⁴C-uptake measurements for Macrosetella gracilis (0.173 µg C copepod^–1 h^–1), and the first quantitative measurements of both Miracia efferata (0.402 µg C copepod^–1 h^–1) and Oculosetella gracilis (0.126 µg C copepod^–1 h^–1) ingesting this cyanobacteria. Ingestion rates of M. gracilis and M. efferata on the two different species of Trichodesmium, T. thiebautii and T. erythraeum, as well as the two different colonial morphologies of T. thiebautii, spherical-shaped (puffs) and fusiform (tufts), were also compared. Both Miracia and Macrosetella had higher ingestion rates on the puff colonies than the tuft colonies of T. thiebautii.. Both also had higher ingestion rates of T. erythraeum than T. thiebautii. Trichodesmium thiebautii contains a previously reported neurotoxin which may be an important factor in determining trophodynamic interactions. Our results suggest that pelagic harpacticoid copepods can be quantitatively important in determining the fate of Trichodesmium carbon and nitrogen.     

Cyanobacterial toxicity and migration in a mesotrophic lake in western Washington, USA

In fall 1997, the toxic cyanobacterium Microcystis aeruginosa was documented in Lake Sammamish (western Washington, U.S.A.) for the first time. Cyanobacterial activity and environmental conditions that may promote toxic cyanobacteria were investigated during summer and fall 1999. Development of toxic Microcystis was hypothesized to be due to runoff of nutrients from the watershed (external loading hypothesis) or from vertical migration of dormant cyanobacteria from the nutrient-rich sediments into the water column (cyanobacterial migration hypothesis). Microcystins were detected using an enzyme-linked immunosorbent assay during late August and early September 1999 despite low cyanobacterial abundance. Microcystin concentrations ranged between 0.19–3.8 g l^–1 throughout the lake and at all depths with the exception of the boat launch where concentrations reached 43 g l^–1. Comparison of the conditions associated with the toxic episodes in 1997 and 1999 indicate that Microcystis is associated with a stable water column, increased surface total phosphorus concentrations (> 10 g l^–1), surface temperatures greater than 22°C, high total nitrogen to phosphorus ratios (> 30), and increased water column transparency (up to 5.5 m). Migration of the cyanobacteria, Microcystis and Anabaena, occurred in both the deep and shallow portions of the lake. Microcystis dominated (89–99%) the migrating cyanobacteria with greater migration from the shallow station. External loading of nutrients due to the large rainfall preceding the 1997 toxic episode may have provided the nutrients needed to fuel that bloom. However, toxic Microcystis occurred in 1999 despite the lack of rain and subsequent external runoff. The migration of Microcystis from the nutrient-rich sediments may have been the inoculum for the toxic population detected in 1999.     

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     

A phycourobilin-containing phycoerythrin from the cyanobacterium Oscillatoria sp.

A filamentous cyanobacterium Oscillatoria sp. was isolated from a thermal spring of the Kamchatka peninsula. It contained a phycoerythrin unusual for cyanobacteria in that it had a phycourobilin prosthetic group. The absorption spectrum of the native purified phycoerythrin displayed maxima at 498 and 567 nm. The phycoerythrin comprised - and -subunits of molecular weights 18,700 and 19,800, respectively, in 1:1 stoichiometry. Polyacrylamide gel isoelectric focusing revealed one protein band at pI 4.6. The - and -subunits differed in their chromophore composition and content: -subunit carried two phycoerythrobilins while the -subunit had three phycoerythrobilins and one phycourobilin. The chromophore composition of all known phycoerythrins of cyanobacteria and red algae were compared, and on the basis of this comparative study designations C₁- to C₅-phycoerythrin were proposed for cyanobacterial red pigments.     

Is bigger better? A possibility for adaptation of <Emphasis Type="Italic">Daphnia</Emphasis> to filamentous cyanobacteria in the face of global warming

Body size is a key determinant of fitness in Daphnia. Bigger size means higher feeding efficiency and reproduction. However, filamentous cyanobacteria have a more detrimental effect on larger daphnids. Predicted global warming is expected to reduce size of Daphnia and facilitate frequent occurrence of cyanobacteria. Therefore, we asked two questions: (i) does elevated temperature cause reduced size of daphnids, and (ii) is temperature-dependent size reduction adaptive in an environment dominated by filamentous cyanobacteria? To address these issues, we obtained 8 clones of the Daphnia longispina complex from artificially heated lakes and 8 clones from control lakes, and exposed them to a favorable food Scenedesmus obliquus and a mixture of S. obliquus and the filamentous cyanobacterium Cylindrospermopsis raciborskii in life table experiments at 16, 20, and 24°C. Individuals from heated lakes attained larger body size than those from control lakes. Moreover, exposure to the filamentous cyanobacterium led to reduced fecundity of Daphnia from the non-heated lakes and did not affect reproduction of Daphnia from the heated lakes. We conclude that Daphnia display some evolutionary adaptations to cope with filamentous cyanobacteria, linked to long-term exposition to elevated temperature. Our results violate broadly accepted assumptions that ectotherms are smaller in warmer environments.     

In vivo fluorometric method for early detection of cyanobacterial waterblooms

A specific method was developed for monitoring the concentration of cyanobacteria (blue-green algae) before waterblooms, based on their characteristics ofin vivo fluorescence. The excitation and emission spectra of cyanobacteria are very different from those of eukaryotic algae, due to the importance of phycocyanin, rather than chlorophylla, in determining the fluorescence characteristics. Our results, based on four cyanobacteria:Microcystis aeruginosa, Anabaena cylindrica, Phormidium tenue andSpirulina platensis, indicate that excitation at 620 nm and its emission at 645 nm is a sensitive and specific method for their detection. Furthermore, the addition of 10 M photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) induced only 3% increase in phycocyanin fluorescence, suggesting that this measurement is almost independent of the ongoing rate of photosynthesis.Author for correspondence     

Bacterial zonation,photosynthesis, and spectral light distribution in hot spring microbial mats of Iceland

The zonation and structure of phototrophic microbial mats were studied along two thermal gradients in sulfide-rich hot springs of southwest Iceland. The green, filamentous bacteriumChloroflexus and the unicellular, high-temperature form (HTF) ofMastigocladus formed mats growing up to a temperature limit of 62–66°C. The dominant phototrophs wereChloroflexus sp.,Mastigocladus laminosus, andPhormidium laminosum, respectively, at the three temperature intervals: >60°C, 60°C to 55–50°C, and <55–50°C. AChloroflexus mat growing at 60°C under 60M H₂S was anoxic in the light with the exception of a 0.5 mm thick band of HTFMastigocladus which produced oxygen. The oxygenic photosynthesis of these H₂S-sensitive cyanobacteria was probably dependent on a preceding sulfide depletion by the anoxygenicChloroflexus. Measurements of spectral radiance gradients with a fiberoptic microprobe showed maximum light attenuation by carotenoids and bacteriochlorophyllC. AM. laminosus mat growing at 52°C was oxic throughout and showed maximum light attenuation by carotenoids, chlorophyllA, and phycocyanin, but no detectable phycoerythrocyanin absorption.     

Circadian Input Kinases and Their Homologs in Cyanobacteria: Evolutionary Constraints Versus Architectural Diversification

The circadian input kinase A (cikA) gene encodes a protein relaying environmental signal to the central circadian oscillator in cyanobacteria. The CikA protein has a variable architecture and usually consists of four tandemly arrayed domains: GAF, histidine kinase (HisKA), histidine kinase-like ATPase (HATPase_c), and a pseudo-receiver (REC). Among them, HisKA and HATPase_c are the least polymorphic, and REC is not present in heterocystic filamentous cyanobacteria. CikA contains several conserved motifs that are likely important for circadian function. There are at least three types of circadian systems, each of which possesses a different set of circadian genes. The originally described circadian system (kaiABC system) possesses both cikA and kaiA, while the others lack either only cikA (kaiABC ^Δ) or both (kaiBC). The results we obtained allowed us to approximate the time of the cikA origin to be about 2600–2200 MYA and the time of its loss in the species with the kaiABC ^Δ or kaiBC system between 1100 and 600 MYA. Circadian specialization of CikA, as opposed to its non-circadian homologs, is a result of several factors, including the unique conserved domain architecture and high evolutionary constraints of some domains and regions, which were previously identified as critical for the circadian function of the gene.     

Efferent neurotransmission of circadian rhythms inLimulus lateral eye

Summary We investigated efferent neurotransmission in theLimulus lateral eye by studying the action of pharmacological agents on responses of photoreceptor cells in vitro. We recorded transmembrane potentials from single cells in slices of retina that were excised during the day and maintained for several days in a culture medium. Potentials recorded in the absence of pharmacological agents resemble those recorded from cells in vivo during the day.Octopamine, a putative efferent neurotransmitter, induced changes in photoreceptor potentials that mimicked in part those generated at night by a circadian clock located in the brain. Specifically, octopamine (100 to 500 M) decreased the frequency of occurrence of quantum bumps in the dark and increased the amplitude of photoreceptor responses to intermediate and high light intensities. Similar actions were produced by naphazoline (25 to 100 (M, potent agonist of octopamine), forskolin (8 to 400 M, activator of adenylate cyclase), IBMX (1 mM, inhibitor of phosphodiesterase), and 8-bromo-cAMP (500 M, analogue of cAMP). 8-bromo-cGMP (500 M, analogue of cGMP) decreased the rate of spontaneous quantum bumps only.Our results support the hypothesis that (1) octopamine is an efferent neurotransmitter of circadian rhythms in theLimulus eye and that (2) it activates adenylate cyclase to increase levels of the second messenger, cAMP, in photoreceptor cells. Circadian changes in photoreceptor responses to moderate intensities may be a specific action of cAMP, since cGMP has no effect. Circadian changes in the rate of spontaneous quantum bumps may involve a less specific intermediate, since both cAMP and cGMP reduce bump rate. Characteristics of the retinal slice preparation precluded a detailed study of the effects of pharmacological agents on retinal morphology.Abbreviations cAMP adenosine 35-cyclic monophosphate - IBMX 3-isobutyl-1-methyl xanthine - LPF large potential fluctuation - SPF small potential fluctuation     

Organization of genes required for gellan polysaccharide biosynthesis in<Emphasis Type="Italic"> Sphingomonas elodea</Emphasis> ATCC 31461

Sphingomonas elodea ATCC 31461 produces gellan, a capsular polysaccharide that is useful as a gelling agent for food and microbiological media. Complementation of nonmucoid S. elodea mutants with a gene library resulted in identification of genes essential for gellan biosynthesis. A cluster of 18 genes spanning 21 kb was isolated. These 18 genes are homologous to genes for synthesis of sphingan polysaccharide S-88 from Sphingomonas sp. ATCC 31554, with predicted amino acid identities varying from 61% to 98%. Both polysaccharides have the same tetrasaccharide repeat unit, comprised of [4)--l-rhamnose-(13)--d-glucose-(14)--d-glucuronic acid-(14)--d-glucose-(1]. Polysaccharide S-88, however, has mannose or rhamnose in the fourth position and has a rhamnosyl side chain, while gellan has no sugar side chain but is modified by glyceryl and acetyl substituents. Genes for synthesis of the precursor dTDP-l-rhamnose were highly conserved. The least conserved genes in this cluster encode putative glycosyl transferases III and IV and a gene of unknown function, gelF. Three genes (gelI, gelM, and gelN) affected the amount and rheology of gellan produced. Four additional genes present in the S-88 sphingan biosynthetic gene cluster did not have homologs in the gene cluster for gellan biosynthesis. Three of these gene homologs, gelR, gelS, and gelG, were found in an operon unlinked to the main gellan biosynthetic gene cluster. In a third region, a gene possibly involved in positive regulation of gellan biosynthesis was identified.     

The Cell Wall Teichoic Acids of Streptomycetes from the “Streptomyces cyaneus” Cluster

The cell wall anionic polymers of the 13 species of the Streptomyces cyaneus cluster have a similar structure and contain -glucosylated 1,5-poly(ribitol phosphate) and 1,3-poly(glycerol phosphate). In the degree of glucosylation of the ribitol phosphate units of their teichoic acids, the cluster members can be divided into two groups. The streptomycetes of the first group (S. afghaniensis, S. janthinus, S. purpurascens, S. roseoviolaceus, and S. violatus) are characterized by a very similar structure of their cell walls, the completely glucosylated 1,5-poly(ribitol phosphate) chains, and a high degree of DNA homology (67–88% according to literature data). The cell wall teichoic acids of the second group (S. azureus, S. bellus, S. caelestis, S. coeruleorubidus, S. curacoi, and S. violarus) differ in the degree of -glucosylation of their 1,5-poly(ribitol phosphate) chains and have a lower level of DNA homology (54–76% according to literature data). Two streptomycetes of the cluster (S. cyaneus and S. hawaiiensis) are genetically distant from the other cluster members but have the same composition and structure of the cell wall teichoic acids as the second-group streptomycetes. The data obtained confirm the genetic relatedness of the S. cyaneus cluster members and suggest that the structure of the cell wall teichoic acids may serve as one of the taxonomic criteria of the species-level status of streptomycetes.