Stable isotope fractionation of strontium in coccolithophore calcite: Influence of temperature and carbonate chemistry

Marine calcifying eukaryotic phytoplankton (coccolithophores) is a major contributor to the pelagic production of CaCO₃ and plays an important role in the biogeochemical cycles of C, Ca and other divalent cations present in the crystal structure of calcite. The geochemical signature of coccolithophore calcite is used as palaeoproxy to reconstruct past environmental conditions and to understand the underlying physiological mechanisms (vital effects) and precipitation kinetics. Here, we present the stable Sr isotope fractionation between seawater and calcite (Δ^88/86Sr) of laboratory cultured coccolithophores in individual dependence of temperature and seawater carbonate chemistry. Coccolithophores were cultured within a temperature and a pCO₂ range from 10 to 25°C and from 175 to 1,240 μatm, respectively. Both environmental drivers induced a significant linear increase in coccolith stable Sr isotope fractionation. The temperature correlation at constant pCO₂ for Emiliania huxleyi and Coccolithus braarudii is expressed as Δ^88/86Sr = ?7.611 × 10^?3 T + 0.0061. The relation of Δ^88/86Sr to pCO₂ was tested in Emiliania huxleyi at 10 and 20°C and resulted in Δ^88/86Sr = ?5.394 × 10^?5 pCO₂ – 0.0920 and Δ^88/86Sr = ?5.742 × 10^?5 pCO₂ – 0.1351, respectively. No consistent relationship was found between coccolith Δ^88/86Sr and cellular physiology impeding a direct application of fossil coccolith Δ^88/86Sr as coccolithophore productivity proxy. An overall significant correlation was detected between the elemental distribution coefficient (D_Sr) and Δ^88/86Sr similar to inorganic calcite with a physiologically induced offset. Our observations indicate (i) that temperature and pCO₂ induce specific effects on coccolith Δ^88/86Sr values and (ii) that strontium elemental ratios and stable isotope fractionation are mainly controlled by precipitation kinetics when embedded into the crystal lattice and subject to vital effects during the transmembrane transport from seawater to the site of calcification. These results provide an important step to develop a coccolith Δ^88/86Sr palaeoproxy complementing the existing toolbox of palaeoceanography.     

The effects of different light–dark cycles on the metabolism of the diazotrophic,unicellular cyanobacteria Cyanothece sp. ATCC 51142, and Cyanothecesp. PCC 7822

The diazotrophic unicellular cyanobacterium Cyanothece sp. ATCC 51142 demonstrates circadian patterns in nitrogenase activity, H₂ production and glycogen storage when grown under nitrogen‐fixing, 12:12 light:dark (L:D) conditions. In this study, we grew Cyanothece sp. ATCC 51142, and another strain in this genus, Cyanothece sp. PCC 7822, under long‐day (16:8 L:D) and short‐day (8:16 L:D) nitrogen‐fixing conditions to determine if they continued to display circadian rhythms. Both strains demonstrated similar circadian patterns for all three metabolic parameters when grown under long‐day conditions. However, the strains responded differently to short‐day growth conditions. Cyanothece sp. ATCC 51142 retained reasonable circadian patterns under 8:16 L:D conditions, whereas Cyanothece sp. PCC 7822 had quite damped patterns without a clear circadian pattern. In particular, glycogen storage changed very little throughout the day and we ascribe this to the difference in the type of glycogen granules in Cyanothece sp. PCC 7822 which has small β‐granules, compared to the large, starch‐like granules in Cyanothece sp. ATCC 51142. The results suggested that both mechanistic and regulatory processes play a role in establishing the basis for these metabolic oscillations.     

Asteriscus v. lapillus: comparing the chemistry of two otolith types and their ability to delineate riverine populations of common carp Cyprinus carpio

The chemical composition of common carp Cyprinus carpio asteriscus (vaterite) and lapillus (aragonite) otoliths from the same individual and reflecting the same growth period was measured to (1) determine whether there are differences in the uptake of trace metals (Mg:Ca, Mn:Ca, Sr:Ca and Ba:Ca ) and Sr isotope ratios (⁸⁷Sr:⁸⁶Sr) in co‐precipitating lapilli and asterisci and (2) compare the ability of multi‐element and isotopic signatures from lapilli, asterisci and both otolith types combined to discriminate C. carpio populations over a large spatial scale within a river basin. Depth profile analyses at the otolith edge using laser‐ablation inductively coupled plasma mass spectrometry showed that asterisci were enriched in Mg and Mn and depleted in Sr and Ba relative to lapilli, whilst ⁸⁷Sr:⁸⁶Sr values were nearly identical in both otolith types. Significant spatial differences among capture locations were found when all trace element and Sr isotope ratio data were aggregated into a multi‐element and isotopic signature, regardless of which otolith type was used or if they were used in combination. Discriminatory power was enhanced, however, when data for both otolith types were combined, suggesting that analysis of multiple otolith types may be useful for studies attempting to delineate C. carpio populations at finer spatial or temporal scales.     

Coupling of Cellular Processes and Their Coordinated Oscillations under Continuous Light in Cyanothece sp. ATCC 51142, a Diazotrophic Unicellular Cyanobacterium

Unicellular diazotrophic cyanobacteria such as Cyanothece sp. ATCC 51142 (henceforth Cyanothece), temporally separate the oxygen sensitive nitrogen fixation from oxygen evolving photosynthesis not only under diurnal cycles (LD) but also in continuous light (LL). However, recent reports demonstrate that the oscillations in LL occur with a shorter cycle time of ~11 h. We find that indeed, majority of the genes oscillate in LL with this cycle time. Genes that are upregulated at a particular time of day under diurnal cycle also get upregulated at an equivalent metabolic phase under LL suggesting tight coupling of various cellular events with each other and with the cell’s metabolic status. A number of metabolic processes get upregulated in a coordinated fashion during the respiratory phase under LL including glycogen degradation, glycolysis, oxidative pentose phosphate pathway, and tricarboxylic acid cycle. These precede nitrogen fixation apparently to ensure sufficient energy and anoxic environment needed for the nitrogenase enzyme. Photosynthetic phase sees upregulation of photosystem II, carbonate transport, carbon concentrating mechanism, RuBisCO, glycogen synthesis and light harvesting antenna pigment biosynthesis. In Synechococcus elongates PCC 7942, a non-nitrogen fixing cyanobacteria, expression of a relatively smaller fraction of genes oscillates under LL condition with the major periodicity being 24 h. In contrast, the entire cellular machinery of Cyanothece orchestrates coordinated oscillation in anticipation of the ensuing metabolic phase in both LD and LL. These results may have important implications in understanding the timing of various cellular events and in engineering cyanobacteria for biofuel production.     

Evaluation of four fresh-water unicellular cyanobacteria as potential hosts for mosquitocidal toxins

Summary For biocontrol of mosquitoes, mosquitocidal toxin genes from Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus have been cloned into a number of cyanobacteria. However, little is known about the persistence of such recombinant cyanobacteria in mosquito larval habitats. Four fresh water unicellular cyanobacteria, Synechococcus PCC6301, PCC7425, PCC7942 and Synechocystis PCC 6803, were evaluated under laboratory conditions related to mosquito breeding environments. Results indicated that Synechococcus PCC6301 was potentially the most suitable organism for use in the natural mosquito habitat as it could tolerate a wide range of temperatures, salinities, and biological and chemical insecticides. Moreover, strain PCC6301 could be ingested and digested by Culex quinquefasciatus larvae and could support the development of larvae to full insect maturity.     

Allometric constraints on Sr/Ca and Ba/Ca partitioning in terrestrial mammalian trophic chains

In biological systems, strontium (Sr) and barium (Ba) are two non-essential elements, in comparison to calcium (Ca) which is essential. The Sr/Ca and Ba/Ca ratios tend to decrease in biochemical pathways which include Ca as an essential element, and these processes are termed biopurification of Ca. The quantitative pathway of the biopurification of Ca in relation to Sr and Ba between two biological reservoirs (R _n and R_{n -1}) is measured with an observed ratio (OR) expressed by the (Sr/Ca) _Rn /(Sr/Ca) _Rn-1 and (Ba/Ca) _Rn /(Ba/Ca) _Rn-1 ratios. For a mammalian organism, during the whole biopurification of Ca starting with the diet to the ultimate reservoir of Ca which is the bone, the mean values for OR_Sr and OR_Ba are 0.25 and 0.2, respectively. In this study, published Sr/Ca and Ba/Ca ratios are used for three sets of soils, plants, and bones of herbivorous and carnivorous mammals, each comprising a trophic chain, to illustrate the biopurification of Ca at the level of trophic chains. Calculated OR_Sr and OR_Ba of herbivore bones in relation to plants and of bones of carnivores in relation to bones of herbivores give OR_Sr=0.30±0.08 and OR_Ba=0.16±0.08, thus suggesting that trophic chains reflect the Sr/Ca and Ba/Ca fluxes that are prevalent at the level of a mammalian organism. The slopes of the three regression equations of log(Sr/Ca) vs. log(Ba/Ca) are similar, indicating that the process of biopurification of Ca with respect to Sr and Ba is due to biological processes and is independent of the geological settings. Modifications of the logarithmic expression of the Sr/Ca and Ba/Ca relationship allow a new formula of the biopurification process to be deduced, leading to the general equation OR_Ba=OR_Sr^1.79±0.33,where the allometric coefficient is the mean of the slopes of the three regression equations. Some recent examples are used to illustrate this new analysis of predator-prey relations between mammals. This opens up new possibilities for the utilization of Ba/Ca and Sr/Ca in addition to stable isotope ratios (¹³C and ¹⁵N) for the determination of the relative contribution of different food sources to an animals diet.     

An isotopic and geochemical study of carbonate-clay mineralization in basaltic caves: abiotic versus microbial processes

Stable isotope and geochemical data are used here to differentiate between contemporaneous abiotic and microbial processes leading to formation of modern carbonate‐ (calcite, aragonite and magnesite) and silicate‐rich (kerolite) mineralization in basaltic sea caves on the island of Kauai, Hawaii. Strontium isotope and Ca/Sr ratios in meteoric water and cave carbonates suggest that the majority of Sr and Ca are derived from rock–water interaction within the host basalts situated above the caves. Oxygen and hydrogen isotope ratios and chemical compositions of cave and surface waters indicate that evaporation does not control cave‐water composition. However, evaporation of drops and thin films of water in microenvironments can lead to precipitation of some phases. This behaviour is suggested by the covariance in δ¹⁸O and δ¹³C values of some carbonates, especially magnesite, which is considered to be a late‐stage evaporative precipitate. Modelling of water evolution suggests that evaporation can be a cause of supersaturation for magnesite, kerolite and some Ca carbonates. However, the highly elevated δ¹³C values (up to +8.2) of some Ca carbonates, compared to average dissolved inorganic carbon δ¹³C values (~?12), are best explained as the product of microbial photosynthesis, in particular by cyanobacteria, present in the upper layers of active microbial mats on cave surfaces. The preferential uptake of ¹²C by cyanobacteria is recorded in the low δ¹³C values (?29.1 to ?22.6) of organic matter in mats and mineralized microbialites. The resulting ¹³C‐enrichment of dissolved inorganic carbon is recorded in the elevated δ¹³C values of these Ca carbonates. A positive correlation exists between the δ¹³C values of the carbonates and coexisting organic matter. The large enrichment in ¹³C of carbonate minerals, relative to dissolved inorganic carbon, and its covariance with the δ¹³C values of coexisting organic matter are useful for identification of carbonate‐rich mineralization resulting from autotrophic microbial activity.     

FraC/FraD-dependent intercellular molecular exchange in the filaments of a heterocyst-forming cyanobacterium, Anabaena sp

The filamentous, heterocyst‐forming cyanobacteria are multicellular organisms in which two different cell types, the CO₂‐fixing vegetative cells and the N₂‐fixing heterocysts, exchange nutrients and regulators. In Anabaena sp. strain PCC 7120, inactivation of sepJ or genes in the fraC operon (fraC, fraD and fraE) produce filament fragmentation. SepJ, FraC and FraD are cytoplasmic membrane proteins located in the filament's intercellular septa that are needed for intercellular exchange of the fluorescent tracer calcein (622 Da). Transmission electron microscopy showed an alteration in the heterocyst cytoplasmic membrane at the vegetative cell‐heterocyst septa in ΔfraC and ΔfraD mutants. Immunogold labelling of FraD confirmed its localization in the intercellular septa and clearly showed the presence of part of the protein between the cytoplasmic membranes of the adjacent cells. This localization seemed to be affected in the ΔfraC mutant but was not impaired in a ΔsepJ mutant. Intercellular transfer of a smaller fluorescent tracer, 5‐carboxyfluorescein (374 Da), was largely impaired in ΔfraC, ΔfraD and double ΔfraC‐ΔfraD mutants, but much less in the ΔsepJ mutant. These results show the existence in the Anabaena filaments of a FraC/FraD‐dependent intercellular molecular exchange that does not require SepJ.     

Changes in Sr/Ca, Ba/Ca and 87Sr/86Sr ratios between trophic levels in two forest ecosystems in the northeastern U.S.A.

The variability and biologicalfractionation of Sr/Ca, Ba/Ca and ⁸⁷Sr/⁸⁶Srratios were studied in a soil–plant–invertebrate–bird food chain in two forested ecosystems withcontrasting calcium availability in the northeasternU.S.A. Chemical measurements were made of the soilexchange pool, leaves, caterpillars, snails, and boththe femurs and eggshells of breeding insectivorousmigratory songbirds. ⁸⁷Sr/⁸⁶Sr values weretransferred up the food chain from the soil exchangepool to leaves, caterpillars, snails and eggshellswithout modification. Adult birds were the oneexception; their ⁸⁷Sr/⁸⁶Sr values generallyreflected those of lower trophic levels at each site,but were lower and more variable, probably becausetheir strontium was derived in part from foods intropical winter habitats where lower⁸⁷Sr/⁸⁶Sr ratios are likely to predominate. Sr/Ca and Ba/Ca ratios decreased at each successive trophiclevel, supporting previous suggestions that Sr/Ca andBa/Ca ratios can be used to identify the trophic levelat which an organism is primarily feeding. The changesin Sr/Ca and Ba/Ca ratios we measured for vegetationand insects were comparable to similar measurementsmade previously (but based on single samples of eachorganism) in an alpine ecosystem. Changes in Sr/Ca andBa/Ca ratios between birds and their food have notpreviously been measured, but the values we obtainedwere similar to those for herbivorous and carnivorousmammals at similar trophic levels. Our results provideevidence that supports the use of Sr/Ca ratios in thedetermination of human paleodiets and suggests thatSr/Ca ratios may also provide a useful tool in studiesof modern food webs. Furthermore, our findings suggestthat ⁹⁰Sr from nuclear fallout will notbioaccumulate in forests and that changes in Sr/Caratios between trophic levels will need to beconsidered in some cases when using⁸⁷Sr/⁸⁶Sr as a tracer of calciumbiogeochemistry.     

Profiles of trace elements and stable isotopes derived from giant long-lived Tridacna gigas bivalves: Potential applications in paleoclimate studies

This study investigates the environmental and biological controls on trace element partitioning and stable isotope composition of modern giant long-lived bivalves (Tridacna gigas) with the aim to use these archives for paleoclimatic reconstructions. Firstly, the intra-shell variability is studied by measuring time equivalent profiles in the different shell layers characterised by different growth rates. Secondly, the inter-site variability is studied by comparing profiles derived from three modern specimens collected in sites across the Indo-Pacific region characterised by different ranges of temperature and productivity.These results show that δ¹⁸O profiles are highly reproducible across the shell regardless of significantly different growth rates. Shell δ¹⁸O is primarily controlled by water δ¹⁸O and temperature. Comparison of intra shell Mg/Ca profiles shows a clear and systematic partitioning where inner layer Mg/Ca values are a least 2–3 times higher than outer layer and hinge areas. Inner layer Mg/Ca shows seasonal oscillations but superimposed on an ontogenetic trend with increasing values and increasing amplitude Mg/Ca oscillations with age. The Sr/Ca profiles do not show clear reproducible seasonal trends in the different shell zones. It is concluded that Mg/Ca and Sr/Ca profiles appear to reflect a combination of biological and environmental controls that will need to be disentangled before using these proxies in paleoclimatic studies.Finally, intra shell Ba/Ca profiles are reproducible in great detail for all modern specimens studied. Inter-site comparison shows that the amplitude and the timing of the Ba/Ca peaks appear to reflect the timing and the amplitude of the chlorophyll peaks associated with phytoplankton blooms at each locality making this tracer a potential paleoproductivity indicator.     

High myristic acid content in the cyanobacterium Cyanothece sp. PCC 8801 results from substrate specificity of lysophosphatidic acid acyltransferase

Analysis of fatty acids from the cyanobacterium Cyanothece sp. PCC 8801 revealed that this species contained high levels of myristic acid (14:0) and linoleic acid in its glycerolipids, with minor contributions from palmitic acid (16:0), stearic acid, and oleic acid. The level of 14:0 relative to total fatty acids reached nearly 50%. This 14:0 fatty acid was esterified primarily to the sn-2 position of the glycerol moiety of glycerolipids. This characteristic is unique because, in most of the cyanobacterial strains, the sn-2 position is esterified exclusively with C16 fatty acids, generally 16:0. Transformation of Synechocystis sp. PCC 6803 with the PCC8801_1274 gene for lysophosphatidic acid acyltransferase (1-acyl-sn-glycerol-3-phosphate acyltransferase) from Cyanothece sp. PCC 8801 increased the level of 14:0 from 2% to 17% in total lipids and the increase in the 14:0 content was observed in all lipid classes. These findings suggest that the high content of 14:0 in Cyanothece sp. PCC 8801 might be a result of the high specificity of this acyltransferase toward the 14:0-acyl-carrier protein.     

Analysis of carbohydrate storage granules in the diazotrophic cyanobacterium Cyanothece sp. PCC 7822

The unicellular diazotrophic cyanobacteria of the genus Cyanothece demonstrate oscillations in nitrogenase activity and H₂ production when grown under 12 h light–12 h dark cycles. We established that Cyanothece sp. PCC 7822 allows for the construction of knock-out mutants and our objective was to improve the growth characteristics of this strain and to identify the nature of the intracellular storage granules. We report the physiological and morphological effects of reduction in nitrate and phosphate concentrations in BG-11 media on this strain. We developed a series of BG-11-derived growth media and monitored batch culture growth, nitrogenase activity and nitrogenase-mediated hydrogen production, culture synchronicity, and intracellular storage content. Reduction in NaNO₃ and K₂HPO₄ concentrations from 17.6 and 0.23 to 4.41 and 0.06 mM, respectively, improved growth characteristics such as cell size and uniformity, and enhanced the rate of cell division. Cells grown in this low NP BG-11 were less complex, a parameter that related to the composition of the intracellular storage granules. Cells grown in low NP BG-11 had less polyphosphate, fewer polyhydroxybutyrate granules and many smaller granules became evident. Biochemical analysis and transmission electron microscopy using the histocytochemical PATO technique demonstrated that these small granules contained glycogen. The glycogen levels and the number of granules per cell correlated nicely with a 2.3 to 3.3-fold change from the minimum at L0 to the maximum at D0. The differences in granule morphology and enzymes between Cyanothece ATCC 51142 and Cyanothece PCC 7822 provide insights into the formation of large starch-like granules in some cyanobacteria.     

Identifying calcium sources at an acid deposition-impacted spruce forest: a strontium isotope,alkaline earth element multi-tracer approach

Depletion of calcium from forest soils has important implications for forest productivity and health. Ca is available to fine feeder roots from a number of soil organic and mineral sources, but identifying the primary source or changes of sources in response to environmental change is problematic. We used strontium isotope and alkaline earth element concentration ratios of trees and soils to discern the record of Ca sources for red spruce at a base-poor, acid deposition-impacted watershed. We measured ⁸⁷Sr/⁸⁶Sr and chemical compositions of cross-sectional stemwood cores of red spruce, other spruce tissues and sequential extracts of co-located soil samples. ⁸⁷Sr/⁸⁶Sr and Sr/Ba ratios together provide a tracer of alkaline earth element sources that distinguishes the plant-available fraction of the shallow organic soils from those of deeper organic and mineral soils. Ca/Sr ratios proved less diagnostic, due to within-tree processes that fractionate these elements from each other. Over the growth period from 1870 to 1960, ⁸⁷Sr/⁸⁶Sr and Sr/Ba ratios of stemwood samples became progressively more variable and on average trended toward values that considered together are characteristic of the uppermost forest floor. In detail the stemwood chemistry revealed an episode of simultaneous enhanced uptake of all alkaline earth elements during the growth period from 1930 to 1960, coincident with reported local and regional increases in atmospheric inputs of inorganic acidity. We attribute the temporal trends in stemwood chemistry to progressive shallowing of the effective depth of alkaline earth element uptake by fine roots over this growth period, due to preferential concentration of fine roots in the upper forest floor coupled with reduced nutrient uptake by roots in the lower organic and upper mineral soils in response to acid-induced aluminum toxicity. Although both increased atmospheric deposition and selective weathering of Ca-rich minerals such as apatite provide possible alternative explanations of aspects of the observed trends, the chemical buffering capacity of the forest floor-biomass pool limits their effectiveness as causal mechanisms.     

Strontium stratigraphy of the upper Miocene Lithothamnion Limestone in the Majella Mountain,central Italy,and its palaeoenvironmental implications

The ⁸⁷Sr/⁸⁶Sr isotope ratio has been widely used as a physical tool to date and correlate carbonate successions due to the long Sr residence time in comparison with the ocean mixing time. If this method works on oceanic successions, marginal basins may show different Sr isotope records in comparison with the coeval ocean one due to sea‐level variations, continental run‐off and restricted water exchanges. In this work, we present the ⁸⁷Sr/⁸⁶Sr isotope record of the upper Miocene carbonate ramp of the Lithothamnion Limestone (Majella Mountain, central Apennines), as an example of the onset of restricted water exchanges between a marginal basin and the ocean water masses. The overall latemost Tortonian–early Messinian Sr isotope record of the Lithothamnion Limestone fits below the global reference line. This deviation has been interpreted as due to the strong control that freshwater input and enhanced continental run‐off, linked to the migration of the Apennine accretionary wedge and foredeep system, have had on the central Adriatic water chemistry. These results imply that an accurate oceanographic and geodynamic framework along with diagenetic overprint investigation has to be taken into consideration prior to apply SIS on carbonate successions on marginal basins, even when facies analyses indicate fully marine conditions. This seems to be the case for the upper Miocene Central Mediterranean carbonate successions, but may have more general validity and be extended to other recent or past marginal basins.     

Spatial and temporal variations in otolith chemistry and relationships with water chemistry: a useful tool to distinguish Atlantic salmon Salmo salar parr from different natal streams

Otolith elemental (Sr:Ca, Ba:Ca, Mn:Ca, Mg:Ca and Rb:Ca) and isotopic (⁸⁷Sr:⁸⁶Sr) profiles from several annual cohorts of juvenile Atlantic salmon Salmo salar were related to the physico‐chemical characteristics (chemical signatures, flow rate, temperature and conductivity) of their natal rivers over an annual hydrological cycle. Only Sr:Ca, Ba:Ca and ⁸⁷Sr:⁸⁶Sr in otoliths were determined by their respective ratios in the ambient water. Sr:Ca ratios in stream waters fluctuated strongly on a seasonal basis, but these fluctuations, mainly driven by water flow regimes, were not recorded in the otoliths. Otolith Sr:Ca ratios remained constant during freshwater residency at a given site and were exclusively related to water Sr:Ca ratios during low flow periods. While interannual differences in otolith elemental composition among rivers were observed, this variability was minor compared to geographic variability and did not limit classification of juveniles to their natal stream. Success in discriminating fish from different sites was greatest using Sr isotopes as it remained relatively constant across years at a given location.     

Simultaneous increases in the levels of compatible solutes by cost-effective cultivation of Synechocystis sp. PCC 6803

Synechocystis sp. PCC 6803, a cyanobacterium widely used for basic research, is often cultivated in a synthetic medium, BG-11, in the presence of 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES) or 2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid buffer. Owing to the high cost of HEPES buffer (96.9% of the total cost of BG-11 medium), the biotechnological application of BG-11 is limited. In this study, we cultured Synechocystis sp. PCC 6803 cells in BG-11 medium without HEPES buffer and examined the effects on the primary metabolism. Synechocystis sp. PCC 6803 cells could grow in BG-11 medium without HEPES buffer after adjusting for nitrogen sources and light intensity; the production rate reached 0.54 g cell dry weight·L⁻¹·day⁻¹, exceeding that of commercial cyanobacteria and Synechocystis sp. PCC 6803 cells cultivated under other conditions. The exclusion of HEPES buffer markedly altered the metabolites in the central carbon metabolism; particularly, the levels of compatible solutes, such as sucrose, glucosylglycerol, and glutamate were increased. Although the accumulation of sucrose and glucosylglycerol under high salt conditions is antagonistic to each other, these metabolites accumulated simultaneously in cells grown in the cost-effective medium. Because these metabolites are used in industrial feedstocks, our results reveal the importance of medium composition for the production of metabolites using cyanobacteria.     

Respiration accumulates Calvin cycle intermediates for the rapid start of photosynthesis in Synechocystis sp. PCC 6803

We tested the hypothesis that inducing photosynthesis in cyanobacteria requires respiration. A mutant deficient in glycogen phosphorylase (?GlgP) was prepared in Synechocystis sp. PCC 6803 to suppress respiration. The accumulated glycogen in ΔGlgP was 250–450% of that accumulated in wild type (WT). The rate of dark respiration in ΔGlgP was 25% of that in WT. In the dark, P700⁺ reduction was suppressed in ΔGlgP, and the rate corresponded to that in (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone)-treated WT, supporting a lower respiration rate in ?GlgP. Photosynthetic O₂-evolution rate reached a steady-state value much slower in ?GlgP than in WT. This retardation was solved by addition of d-glucose. Furthermore, we found that the contents of Calvin cycle intermediates in ?GlgP were lower than those in WT under dark conditions. These observations indicated that respiration provided the carbon source for regeneration of ribulose 1,5-bisphosphate in order to drive the rapid start of photosynthesis.     

DEWFALL AS A WATER SOURCE FREQUENTLY ACTIVATES THE ENDOLITHIC CYANOBACTERIAL COMMUNITIES IN THE GRANITES OF TAYLOR VALLEY,ANTARCTICA1

Endolithic photosynthetic microorganisms like cyanobacteria and algae are well known from savannas and deserts of the world, the high Arctic, and also Antarctic habitats like the Dry Valleys in the Ross Dependency. These endolithic microbial communities are thought to be at the limits of life with reported ages in the order of thousands of years. Here we report on an extensive chasmoendolithic cyanobacterial community inside granite rocks of Mt. Falconer in the lower Taylor Valley, Dry Valleys. On average, the cyanobacterial community was 4.49 ± 0.95 mm below the rock surface, where it formed a blue‐green layer. The community was composed mainly of the cyanobacterium Chroococcidiopsis sp., with occasional Cyanothece cf. aeruginosa (Nägeli) Komárek and Nostoc sp. Mean biomass was 168 ± 44 g carbon · m^?2, and the mean chl a content was 24.3 ± 34.2 mg · m^?2. In situ chl fluorescence measurements—a relative measure of photosynthetic activity—showed that they were active over long periods each day and also showed activity the next day in the absence of any moisture. Radiocarbon dating gave a relatively young age (175–280 years) for the community. Calculations from microclimate data demonstrated that formation of dew or rime was possible and could frequently activate the cyanobacteria and may explain the younger age of microbial communities at Mt. Falconer compared to older and less active endolithic microorganisms reported earlier from Linnaeus Terrace, a higher altitude region that experiences colder, drier conditions.     

Otolith chemistry of prey fish consumed by a fish predator: does digestion hinder Russian doll techniques?

The effect of digestion by a predatory fish (largemouth bass Micropterus salmoides) on stable isotopic (δ¹³C and δ¹⁸O) and trace elemental (Sr:Ca and Ba:Ca) compositions of prey fish (bluegill Lepomis macrochirus) otoliths was investigated in a laboratory experiment. Trace element and stable‐isotopic signatures of L. macrochirus otoliths were not significantly altered for up to 16 h after L. macrochirus were consumed by M. salmoides. Prey fish otoliths recovered from predator digesta can retain environmental stable isotopic and trace elemental signatures, suggesting that determination of environmental history for prey fishes by stable‐isotope and trace‐element analysis of otoliths recovered from stomachs of piscivorous fishes will be feasible.     

The γ‐aminobutyric acid shunt contributes to closing the tricarboxylic acid cycle in Synechocystis sp. PCC 6803

A traditional 2‐oxoglutarate dehydrogenase complex is missing in the cyanobacterial tricarboxylic acid cycle. To determine pathways that convert 2‐oxoglutarate into succinate in the cyanobacterium Synechocystis sp. PCC 6803, a series of mutant strains, Δsll1981, Δslr0370, Δslr1022 and combinations thereof, deficient in 2‐oxoglutarate decarboxylase (Sll1981), succinate semialdehyde dehydrogenase (Slr0370), and/or in γ‐aminobutyrate metabolism (Slr1022) were constructed. Like in Pseudomonas aeruginosa, N‐acetylornithine aminotransferase, encoded by slr1022, was shown to also function as γ‐aminobutyrate aminotransferase, catalysing γ‐aminobutyrate conversion to succinic semialdehyde. As succinic semialdehyde dehydrogenase converts succinic semialdehyde to succinate, an intact γ‐aminobutyrate shunt is present in Synechocystis. The Δsll1981 strain, lacking 2‐oxoglutarate decarboxylase, exhibited a succinate level that was 60% of that in wild type. However, the succinate level in the Δslr1022 and Δslr0370 strains and the Δsll1981/Δslr1022 and Δsll1981/Δslr0370 double mutants was reduced to 20–40% of that in wild type, suggesting that the γ‐aminobutyrate shunt has a larger impact on metabolite flux to succinate than the pathway via 2‐oxoglutarate decarboxylase. ¹³C‐stable isotope analysis indicated that the γ‐aminobutyrate shunt catalysed conversion of glutamate to succinate. Independent of the 2‐oxoglutarate decarboxylase bypass, the γ‐aminobutyrate shunt is a major contributor to flux from 2‐oxoglutarate and glutamate to succinate in Synechocystis sp. PCC 6803.