Limiting Carbon Source Research Articles

Evolution of an Escherichia coli PTS- strain: a study of reproducibility and dynamics of an adaptive evolutive process.

Adaptive laboratory evolution (ALE) has been used to study and solve pressing questions about evolution, especially for the study of the development of mutations that confer increased fitness during evolutionary processes. In this contribution, we investigated how the evolutionary process conducted with the PTS- mutant of Escherichia coli PB11 in three parallel batch cultures allowed the restoration of rapid growth with glucose as the carbon source. The significant findings showed that genomic sequence analysis of a set of newly evolved mutants isolated from ALE experiments 2-3 developed some essential mutations, which efficiently improved the fast-growing phenotypes throughout different fitness landscapes. Regulator galR was the target of several mutations such as SNPs, partial and total deletions, and insertion of an IS1 element and thus indicated the relevance of a null mutation of this gene in the adaptation of the evolving population of PB11 during the parallel ALE experiments. These mutations resulted in the selection of MglB and GalP as the primary glucose transporters by the evolving population, but further selection of at least a second adaptive mutation was also necessary. We found that mutations in the yfeO, rppH, and rng genes improved the fitness advantage of evolving PTS- mutants and resulted in amplification of leaky activity in Glk for glucose phosphorylation and upregulation of glycolytic and other growth-related genes. Notably, we determined that these mutations appeared and were fixed in the evolving populations between 48 and 72 h of cultivation, which resulted in the selection of fast-growing mutants during one ALE experiments in batch cultures of 80 h duration.Key points• ALE experiments selected evolved mutants through different fitness landscapes in which galR was the target of different mutations: SNPs, deletions, and insertion of IS.• Key mutations in evolving mutants appeared and fixed at 48-72 h of cultivation.• ALE experiments led to increased understanding of the genetics of cellular adaptation to carbon source limitation.

Mechanism of nutrient removal enhancement in low carbon/nitrogen wastewater by a novel high-frequency micro-aeration/anoxic (HMOA) mode

In this study, a novel high-frequency micro-aeration/anoxic (HMOA) mode with a high aeration frequency (15 times/h) and short aeration duration (Taer = 1 h/cycle) was proposed. Compared with continuous aeration modes, the highest nitrogen and phosphorus removal efficiencies were achieved in the sequencing batch reactor (SBR) under HMOA mode when treating wastewater with carbon/nitrogen (C/N) ratios of 4.5 (85% and 97%, respectively) and 3 (77% and 75%, respectively). Metagenomic analysis was utilized to analyse the microbial metabolic mechanism under the HMOA mode. The results showed that under the HMOA mode, the enhanced transduction and metabolism pathways of nitrate, nitrite, oxygen, phosphorus and acetate provided favourable nutritional conditions for the proliferation of denitrifiers and phosphorus accumulating organisms (PAOs), and simultaneously strengthened the survival capacity of nitrifiers under low dissolved oxygen (DO) conditions. In addition, genes involved in carbon metabolism were upregulated by the HMOA mode, which further increased the utility of carbon sources for denitrifier and PAO metabolism. Consequently, the limited carbon source could be fully utilized in nitrogen and phosphorus removal, which improved the efficiency of treating low C/N wastewater. This study proposed a potential aeration mode for microbial metabolism regulation to enhance nutrient removal in biological wastewater treatment processes.

Physiological Capabilities of Cryoconite Hole Microorganisms.

Cryoconite holes are miniature freshwater aquatic ecosystems that harbor a relatively diverse microbial community. This microbial community can withstand the extreme conditions of the supraglacial environment, including fluctuating temperatures, extreme and varying geochemical conditions and limited nutrients. We analyzed the physiological capabilities of microbial isolates from cryoconite holes from Antarctica, Greenland, and Svalbard in selected environmental conditions: extreme pH, salinity, freeze-thaw and limited carbon sources, to identify their physiological limits. The results suggest that heterotrophic microorganisms in cryoconite holes are well adapted to fast-changing environmental conditions, by surviving multiple freeze-thaw cycles, a wide range of salinity and pH conditions and scavenging a variety of organic substrates. Under oxic and anoxic conditions, the communities grew well in temperatures up to 30°C, although in anoxic conditions the community was more successful at colder temperatures (0.2°C). The most abundant cultivable microorganisms were facultative anaerobic bacteria and yeasts. They grew in salinities up to 10% and in pH ranging from 4 to 10.5 (Antarctica), 2.5 to 10 (Svalbard), and 3 to 10 (Greenland). Their growth was sustained on at least 58 single carbon sources and there was no decrease in viability for some isolates after up to 100 consecutive freeze-thaw cycles. The elevated viability of the anaerobic community in the lowest temperatures indicates they might be key players in winter conditions or in early melt seasons, when the oxygen is potentially depleted due to limited flow of meltwater. Consequently, facultative anaerobic heterotrophs are likely important players in the reactivation of the community after the polar night. This detailed physiological investigation shows that despite inhabiting a freshwater environment, cryoconite microorganisms are able to withstand conditions not typically encountered in freshwater environments (namely high salinities or extreme pH), making them physiologically more similar to arid soil communities. The results also point to a possible resilience of the most abundant microorganisms of cryoconite holes in the face of rapid change regardless of the location.

The Distribution Patterns of Timberline and Its Response to Climate Change in the Himalayas

Himalayan region represents the highest and most diverse treeline over the world. As one of the most conspicuous boundaries between montane forests and alpine vegetation, the alpine timberline attracted the interest of researchers for many decades. However, timberline in the Himalayas is understudied compared with European counterparts due to remoteness. Here we review the distribution pattern of timberline and its climatic condition, the carbon and nutrient supply mechanism for treeline formation, and treeline shift and treeline tree recruitment under climate change scenarios. Growth limitation, rather than carbon source limitation is the physiological cause of timberline under the low temperature condition. Nutrient limitation and water stress are not the direct cause of timberline formation. However, more clear local limitation factors are need to integrate in order to enable us to predict the potential impacts and changes caused by human activity and related global change in this sensitive region.

Long-Term Impacts of Defoliator Outbreaks on Larch Xylem Structure and Tree-Ring Biomass.

Defoliator insects are a major disturbance agent in many forests worldwide. During outbreaks, they can strongly reduce photosynthetic carbon uptake and impact tree growth. In the Alps, larch budmoth (Zeiraphera diniana) outbreaks affect European larch (Larix decidua) radial growth over several years. However, immediate and legacy effects on xylem formation, structure, and functionality are still largely unknown. In this study, we aimed at assessing the impact of budmoth defoliations on larch xylem anatomical features and tree-ring structure. Analyses were performed in the Lötschental (Swiss Alps) within (1,900 m a.s.l.) and above (2,200 m a.s.l.) the optimum elevational range of larch budmoth. We investigated variability of xylem anatomical traits along century-long tree-ring series of larch (host) and Norway spruce (non-host) trees. We identified eight outbreaks affecting larch xylem anatomy during the 20th century, particularly at 1,900 m a.s.l. Tracheid number always showed a higher percent reduction than properties of individual cells. Cell lumen size was slightly reduced in the first 2–3 years of outbreaks, especially in the early part of the ring. The more carbon-demanding cell wall was thinned along the entire ring, but more evidently in the last part. Theoretical tree-ring hydraulic conductivity was reduced for several years (up to 6), mostly due to cell number decrease. Reduced cell wall area and cell number resulted in a strong reduction of the tree-ring biomass, especially in the first year of outbreak. Our study shows that, under carbon source limitations caused by natural defoliation, cell division is more impacted than wall thickening and cell enlargement (the least affected process). Consequences on both xylem hydraulic properties and tree-ring biomass should be considered when assessing long-term defoliator effects on xylem functioning, forest dynamics, and terrestrial carbon cycle.

Nutrient factor-dependent performance of bacterial quorum sensing system during population evolution.

Bacterial quorum sensing (QS) system regulates the production of most costly but sharable extracellular products (public goods) in a growth-phase-dependent manner, and the development of this energy-intensive process is susceptible to environmental changes. However, the role of nutrient factors in dominating the QS-mediated cooperative interaction and intracellular metabolism still remains less understood. Here we studied the performance of QS system by growing Pseudomonas aeruginosa under different nutrient and culture conditions. The results of comparative-transcriptomic analyses revealed that carbon source-limitation was the main factor suppressing the activation of QS system, and a substantial number of public-good-encoding genes were induced when phosphorus is limiting in short-term culture. By contrast, although the QS regulation of P. aeruginosa in all the cultures was generally decreased along with the enrichment of QS-deficient individuals during evolution, limitation of different nutrient factors had discrepant effects in directing the formation of population structure by coordinating the production of public goods and primary metabolism, especially the starch and sucrose metabolism. These findings demonstrate the pleiotropy of QS regulation in balancing the development of cooperative behavior and metabolism, and provide a reference for further understanding the role of QS system in causing persistent infections.

Evaluation of Product Distribution in Chemostat and Batch Fermentation in Lactic Acid-Producing Komagataella phaffii Strains Utilizing Glycerol as Substrate.

Lactic acid is the monomeric unit of polylactide (PLA), a bioplastic widely used in the packaging, automotive, food, and pharmaceutical industries. Previously, the yeast Komagataella phaffii was genetically modified for the production of lactate from glycerol. For this, the bovine L-lactate dehydrogenase- (LDH)-encoding gene was inserted and the gene encoding the pyruvate decarboxylase (PDC) was disrupted, resulting in the GLp strain. This showed a yield of 67% L-lactic acid and 20% arabitol as a by-product in batches with oxygen limitation. Following up on these results, the present work endeavored to perform a detailed study of the metabolism of this yeast, as well as perturbing arabitol synthesis in an attempt to increase lactic acid titers. The GLp strain was cultivated in a glycerol-limited chemostat at different dilution rates, confirming that the production of both lactic acid and arabitol is dependent on the specific growth rate (and consequently on the concentration of the limiting carbon source) as well as on the oxygen level. Moreover, disruption of the gene encoding arabitol dehydrogenase (ArDH) was carried out, resulting in an increase of 20% in lactic acid and a 50% reduction in arabitol. This study clarifies the underlying metabolic reasons for arabitol formation in K. phaffii and points to ways for improving production of lactic acid using K. phaffii as a biocatalyst.

Absence of the highly expressed small carbohydrate-binding protein Cgt improves the acarbose formation in Actinoplanes sp. SE50/110

Actinoplanes sp. SE50/110 (ATCC 31044) is the wild type of industrial producer strains of acarbose. Acarbose has been used since the early 1990s as an inhibitor of intestinal human α-glucosidases in the medical treatment of type II diabetes mellitus. The small secreted protein Cgt, which consists of a single carbohydrate-binding module (CBM) 20-domain, was found to be highly expressed in Actinoplanes sp. SE50/110 in previous studies, but neither its function nor a possible role in the acarbose formation was explored, yet. Here, we demonstrated the starch-binding function of the Cgt protein in a binding assay. Transcription analysis showed that the cgt gene was strongly repressed in the presence of glucose or lactose. Due to this and its high abundance in the extracellular proteome of Actinoplanes, a functional role within the sugar metabolism or in the environmental stress protection was assumed. However, the gene deletion mutant ∆cgt, constructed by CRISPR/Cas9 technology, displayed no apparent phenotype in screening experiments testing for pH and osmolality stress, limited carbon source starch, and the excess of seven different sugars in liquid culture and further 97 carbon sources in the Omnilog Phenotypic Microarray System of Biolog. Therefore, a protective function as a surface protein or a function within the retainment and the utilization of carbon sources could not be experimentally validated. Remarkably, enhanced production of acarbose was determined yielding into 8–16% higher product titers when grown in maltose-containing medium.

Activation of RelA by pppGpp as the basis for its differential toxicity over ppGpp in Escherichia coli

The nucleotide derivatives (p)ppGpp, comprising ppGpp and pppGpp, are important signalling molecules that control various facets of gene regulation and protein synthesis in Escherichia coli. Their synthesis is catalysed by RelA (in response to amino acid limitation) and by SpoT (in response to the limitation of carbon source or fatty acids). SpoT is also a hydrolase for degradation of both ppGpp and pppGpp, while GppA catalyses the conversion of pppGpp to ppGpp. Here we provide evidence to show that pppGpp exerts heightened toxicity compared to that by ppGpp. Thus, gppA spoT double mutants exhibited lethality under conditions in which the single mutants were viable. The extent of RelA-catalysed (p)ppGpp accumulation in the gppA spoT strain was substantially greater than that in its isogenic gppA+ derivative. The data is interpreted in terms of a model in which toxicity of pppGpp in the gppA spoT mutants is mediated by its activation of RelA so as to result in a vicious cycle of (p)ppGpp synthesis.

Metabolomic insights into polyhydroxyalkanoates production by halophilic bacteria with acetic acid as carbon source.

A metabolomics method was established to analyze changes of intracellular metabolites and study the mechanism for enhancing polyhydroxyalkanoates production by halotolerant bacteria, Bacillus cereus strain HY-3, using acetic acid as carbon source. Maximum poly(3-hydroxybutyrate) (PHB) contents for the medium with 0.5 g/L and 5.0 g/L of acetic acid were 41.0 ± 0.415% and 49.2 ± 1.21%. Principal components analysis revealed clear metabolic differences in different growth stages and different concentrations of carbon source. According to statistical analysis, 3-hydroxybutyrate (3-HB), serine, threonine, malate, and pyruvate were determined as potential biomarkers for PHB production. Moreover, metabolic pathways analysis indicated that high level of 3-HB in death phase was due to the limitation of carbon source. Metabolism of glycine, serine, and threonine was influential pathway for PHB production among amino acid metabolisms. High levels of organic acids from the TCA cycle could stimulate the carbon source flux into PHB biosynthetic pathway.

Accelerated corrosion of pipeline steel in the presence of Desulfovibrio desulfuricans biofilm due to carbon source deprivation in CO2 saturated medium.

The chemistry of bacterial biofilms as well as the nutritional composition of culture environments may differ at any time within a growth process, especially for SRB consortia within oil wells with limited carbon sources. In the oilfield, the presence of SRB biofilms on surfaces of steel substrates leads to microbiologically influenced corrosion and compromised material integrity. In this work, the survival of SRB cells and their impact on the pipeline steel corrosion within simulated CO2-saturated oilfield-produced water with different concentrations of organic carbon source have been investigated. Cell counts reduced with the level of carbon source reduction (CSR) after incubation but more sessile cells survived at 80% CSR (moderate carbon starvation) compared to 100% CSR (extreme carbon starvation). The energy needed for cellular survival as well as biological support toward MIC could have been harnessed by a combination of extracellular Feo oxidation and intracellular sulfate reduction even after carbon source starvation. Severe anodic steel dissolution was observed at the end of the culture period within the simulated CO2-saturated oilfield-produced water, and this is attributed to SRB-led MIC and CO2 corrosion. Pipeline steel corroded more when cultured within 80% CSR compared to the medium with both lactate and citrate. Steel substrate corroded less with 100% CSR due to severely weakened SRB biofilms from nutrient deprivation.

Enhanced Natamycin production by Streptomyces natalensis in shake-flasks and stirred tank bioreactor under batch and fed-batch conditions

BackgroundNatamycin is an antifungal polyene macrolide antibiotic with wide applications in health and food industries. Currently, it is the only antifungal food additive with the GRAS status (Generally Regarded as Safe).ResultsNatamycin production was investigated under the effect of different initial glucose concentrations. Maximal antibiotic production (1.58 ± 0.032 g/L) was achieved at 20 g/L glucose. Under glucose limitation, natamycin production was retarded and the produced antibiotic was degraded. Higher glucose concentrations resulted in carbon catabolite repression. Secondly, intermittent feeding of glucose improved natamycin production due to overcoming glucose catabolite regulation, and moreover it was superior to glucose-beef mixture feeding, which overcomes catabolite regulation, but increased cell growth on the expense of natamycin production. Finally, the process was optimized in 7.5 L stirred tank bioreactor under batch and fed-batch conditions. Continuous glucose feeding for 30 h increased volumetric natamycin production by about 1.6- and 1.72-folds in than the batch cultivation in bioreactor and shake-flasks, respectively.ConclusionsGlucose is a crucial substrate that significantly affects the production of natamycin, and its slow feeding is recommended to alleviate the effects of carbon catabolite regulation as well as to prevent product degradation under carbon source limitation. Cultivation in bioreactor under glucose feeding increased maximal volumetric enzyme production by about 72% from the initial starting conditions.

Inhibiting Biocorrosion of Pipeline Steel in the Presence of Zinc Molybdate Nanocrystals within a CO2 Saturated Desulfovibrio Desulfuricans bacterial Culture Under Carbon Source Deprivation

Even with limited organic carbon sources in oilfields, the presence of sulphate reducing bacteria (SRB) intensifies microbiologically influenced corrosion (MIC), consequently leading to compromised material integrity. On pipeline steel substrates with active biofilms, Feo is subsequently utilized as an alternative donor by SRBs during energy metabolism. In this work, the survival of Desulfovibrio desulfuricans (ATCC 27774) SRB cells and their impact on the pipeline steel (API 5L X70) corrosion within simulated CO2-saturated oilfield-produced water with different concentrations of organic carbon source have been investigated. Cell counts reduced with the level of carbon source reduction after incubation, but more cells survived at moderate carbon starvation compared to extreme conditions. To combat MIC, we have introduced different concentrations of zinc molybdate (ZnM) crystals against bacterial growth, synthesized from a new chemical route. The rate of MIC reduces more than 80 times in the presence of ZnM crystals under anaerobic conditions, and this has been attributed to the cellular growth inhibitive effect of ZnM. The extent of SRB colonization of steel was monitored within duration of study using confocal laser scanning microscopy after staining with appropriate Live/Dead dyes. Steel biocorrosion mechanism is elucidated by means of electrochemical impedance spectroscopy, potentiodynamic polarization and surface analyses. Adhering corrosion products on steel in the presence of D. desulfuricans biofilm are predominantly FeO films and FeS/MnS aggregates; this has been confirmed by X-ray photoelectron spectroscopy. Severe anodic steel dissolution is observed on steel within the test culture without ZnM, and this is attributed to SRB-led MIC and CO2 corrosion. Here, the energy needed for cellular survival was harnessed by a combination of Feo oxidation and sulfate reduction even after carbon source starvation. The degree of biocorrosion inhibition increases with ZnM crystal concentration and as more bacterial cells die off within the culture media.

Nutrient sink limitation constrains growth in two barley species with contrasting growth strategies.

Mineral nutrients exert important limitations on plant growth. Growth is limited by the nutrient source when it is constrained by nutrient availability and uptake, which may simultaneously limit investment in photosynthetic proteins, leading to carbon source limitation. However, growth may also be limited by nutrient utilization in sink tissue. The relative importance of these processes is contested, with crop and vegetation models typically assuming source limitations of carbon and mineral nutrients (especially nitrogen). This study compared the importance of source and sink limitation on growth in a slower‐growing wild perennial barley (Hordeum bulbosum) and a faster‐growing domesticated annual barley (Hordeum vulgare), by applying a mineral nutrient treatment and measuring nitrogen uptake, growth, allocation, and carbon partitioning. We found that nitrogen uptake, growth, tillering, shoot allocation, and nitrogen storage were restricted by low nutrient treatments. Multiple lines of evidence suggest that low nutrient levels do not limit growth via carbon acquisition: (a) Carbohydrate storage does not increase at high nutrient levels. (b) Ratio of free amino acids to sucrose increases at high nutrient levels. (c) Shoot allocation increases at high nutrient levels. These data indicate that barley productivity is limited by the capacity for nutrient use in growth. Models must explicitly account for sink processes in order to properly simulate this mineral nutrient limitation of growth.

Pattern evolution characterizes the mechanism and efficiency of CVD graphene growth

Growing large-area, high-quality monolayers is the holy grail of graphene synthesis. In this work, the efficiency of graphene growth and the quality of their continuous films are explored through the time evolution of individual domains and their surface coverage on the substrate. Our phase-field modeling results and experimental characterization clearly demonstrate the critical roles of the deposition flux, edge-reaction kinetics and the surface diffusion of active carbon sources in modulating the pattern evolution and rate of growth. We find that the contrast between the edge-kinetics-limited and surface-diffusion-limited regimes is remarkable, which can be characterized by the evolution of domain patterns and considered as an indicator of the growth regime. However, common features exist in these two regimes, showing that the growth rate scales with time as t2 in the early stage of growth and is regime-independent, which is explained by the coarsen profiles of carbon concentration for both the compact and dendritic domains. The rate decays rapidly in the final stage of growth due to the competition between neighboring domains on the limited carbon sources diffusing on the substrate, which is highly regime-sensitive and extremely low in the surface-diffusion-limited regime with narrow gaps between the domains to be filled. Based on these findings, synthesis strategies to improve the growth efficiency and film quality are discussed.

Limited carbon source retards inorganic arsenic release during roxarsone degradation in Shewanella oneidensis microbial fuel cells

Directly relevant to the toxicity, mobility, and fate of arsenic, the biotransformation of inorganic and organic arsenicals has been extensively concerned, including roxarsone, a widely applied organoarsenical feed additive in poultry industry. Yet, little is known about the transformation details of roxarsone in microbial fuel cells (MFC). In this study, a two-chambered Shewanella oneidensis MR-1 microbial fuel cell was employed to investigate the transformation processes of roxarsone at various carbon source levels. Results show that limited carbon source remarkably inhibited inorganic arsenic release along roxarsone transformation, whereas numerous arsenical species were detected to be released into systems with sufficient carbon source supply, including trivalent and pentavalent inorganic arsenics, monomethylarsonous acid (MMA), and 4-hydroxy-3-aminobenzene arsonic acid (HAPA). Shewanella oneidensis MR-1 was able to cleave the C-As bond of trivalent HAPA yielding inorganic arsenics and MMA, even in the absence of the arsI gene encoding ArsI C-As lyase. We proposed a two-step nitro- and pentavalent-arsenate group reduction pathway for the roxarsone bioelectrochemical transformation. In addition, results indicated that the attached cells onto the electrode surface played a key function in the two-step reduction of roxarsone to trivalent HAPA, whereas planktonic cells were most likely responsible for the C-As bond breakage and the following dearylation. With these qualitative and quantitative estimations, it provides new insights into the mechanistic understanding of the roxarsone biotransformation process in microbial fuel cells, which is important for the biogeochemical cycling of arsenic.

Vitis vinifera berry metabolic composition during maturation: Implications of defoliation.

Leaves are an important contributor toward berry sugar and nitrogen (N) accumulation, and leaf area, therefore, affects fruit composition during grapevine (Vitis vinifera) berry ripening. The aim of this study was to investigate the impact of leaf presence on key berry quality attributes in conjunction with the accumulation of primary berry metabolites. Shortly after the start of véraison (berry ripening), potted grapevines were defoliated (total defoliation and 25% of the control), and the accumulation of berry soluble solids, N and anthocyanins were compared to that of a full leaf area control. An untargeted approach was undertaken to measure the content in primary metabolites by gas chromatography/mass spectrometry. Partial and full defoliation resulted in reduced berry sugar and anthocyanin accumulation, while total berry N content was unaffected. The juice yeast assimilable N (YAN), however, increased upon partial and full defoliation. Remobilized carbohydrate reserves allowed accumulation of the major berry sugars during the absence of leaf photoassimilation. Berry anthocyanin biosynthesis was strongly inhibited by defoliation, which could relate to the carbon (C) source limitation and/or increased bunch exposure. Arginine accumulation, likely resulting from reserve translocation, contributed to increased YAN upon defoliation. Furthermore, assessing the implications on various products of the shikimate pathway suggests the C flux through this pathway to be largely affected by leaf source limitation during fruit maturation. This study provides a novel investigation of impacts of leaf C and N source presence during berry maturation, on the development of key berry quality parameters as underlined by alterations in primary metabolism.

Some Mechanism Seaweeds Employ to Cope with Salinity Stress in the Harsh Euhaline Oceanic Environment

In order to prevent salt damage because seaweed enzymes can only operate under hypohaline conditions (salinity ≈ 6‰ - 12‰) but also obtain for photosynthesis an in the aquatic environment—due to a 10,000 fold strongly limited carbon source—seaweeds developed several mechanisms to meet these vital demands for survival in the harsh euhaline oceanic environment (salinity range: 32‰ - 35‰), we tested this range of adaptation mechanisms in the euhaline oceanic collected water in combination with the seaweed moisture. We obtained under laboratory conditions at 10 bar mechanical pressure for four seaweed species: Ulva lactuca, Caulerpa sertularioides, Caulerpa cf. brachypus (all three green) and Undaria pinnatifidia (brown). Oceanic water and seaweed moisture were measured for salinity, pH and by Inductively Coupled Plasma Spectroscopy (ICP)-techniques concentrations for macro-elements: (Ca, Fe, K, Mg, Mn, Na, P, & S), micro-elements ≈ [HM]: (Al, As, Cd, Co, Cr, Cu, Mo, Ni, Pb & Zn) and nutrients (N-total & P-total). The [seawater compound X]/[oceanic compound X] ration is a reflection of an inward (uptake) or excretion mechanism over the seaweed cellular membrane which is operative. Our observations gave a clear dispersion to salinity stress with on one hand the green seaweed U. lactuca and on the other the brown seaweed U. pinnatifidia. Both Caulerpa spp. took in an intermediate position. Observed in compensatory responses to salinity stress was ranging Ulva sp. both Caulerpa spp.-Undaria sp.: 1) amount pressed seaweed moisture: [ml/g Fresh Weight]; 2) salinity: (in ‰); 3) Na+ storage vacuole volume; 4) Na+:K+ ratio (reflection of K+ as osmolyticum); 5) ∑[HM] (as osmolyticum); 6) pH (seaweed moisture); 7) Nutrients (N & P); 8) availability of essential metal elements for plants (Cu, Fe, Zn, Mn, Mo, Ni); 9) transport direction of micro- and macro-elements. Finally, the role of brown vs. green seaweeds in the evolutionary eukaryotic tree of life in relation to the ability of the brown seaweeds to produce their own osmolyticum will be discussed.

Spatial heterogeneity of glycogen and its metabolizing enzymes in Aspergillus nidulans hyphal tip cells

Glycogen is a homopolymer of glucose and a ubiquitous cellular-storage carbon. This study investigated which Aspergillus nidulans genes are involved in glycogen metabolism. Gene disruptants of predicted glycogen synthase (gsyA) and glycogenin (glgA) genes accumulated less cellular glycogen than the wild type strain, indicating that GsyA and GlgA synthesize glycogen similarly to other eukaryotes. Meanwhile, gene disruption of gphA encoding glycogen phosphorylase increased the amount of glycogen to a higher degree than wild type during the stationary phase that accompanies carbon-source limitation. GFP-tagged GsyA and GphA were distributed in the cytosol and formed punctate and filamentous structures, respectively. Carbon starvation resulted in elongated GphA-GFP filaments and increased numbers of filaments. These structures were more frequently located in the basal regions of tip cells and adjacent cells than in the apical regions of tip cells. Cellular glycogen visualized by incorporation of a fluorescent glucose analog accumulated in cytoplasmic puncta that were more prevalent in the basal regions, a pattern similar to that seen for GsyA. The colocalization of glycogen and GsyA at punctate structures in tip and sub-apical cells likely represents the cellular machinery for synthesizing glycogen. More frequent colocalization in the basal, rather than tip cell apical regions indicated that tip cells have differentiated subcellular regions for glycogen synthesis. Our findings regarding glycogen, GsyA and GphA distribution evoke the spatial heterogeneity of glycogen metabolism in fungal hyphae.

The Establishment and Characteristics of Dominant Syntrophic Propionate Oxidation Bacteria and Sulfate-Reducing Bacteria in a Mixed Culture

The anaerobic bio-treatment of lower COD/SO42- ratio wastewater constitutes a bottle-aqneck due to the limited carbon source. In this type of environment, interactions between acidogenic fermentation bacteria and methane-producing bacteria (MPB) are aided by syntrophic fatty acid oxidation bacteria present in the functional niche; MPB and sulfate-reducing bacteria (SRB) compete with each other due to the availability of sulfate. Therefore, introduction of syntrophic fatty acid oxidation bacteria into an SRB dominated culture can achieve maximum utilization of the substrate, weakening the competition of MPB. In this study, the mixed culture of dominant syntrophic propionate oxidation bacteria (SPOB) and SRB was established and characterized at 35°C. The dominant SPOB and SRB were enriched in a continuous flow stirred-tank reactor and in batch experiments, respectively. Subsequently, an equal proportion of the enriched sludge was mixed and incubated in conical flasks with propionate as the sole substrate. Six parallel experiments (SP1–SP6) were conducted and two of them (SP2 and SP4) were chosen. The results revealed that the sulfate removal efficiencies of SP2 and SP4 were 91.83% and 93.03%, respectively. Furthermore, the outcome of the microbial analysis showed that the dominant microbial communities included Uncultured Smithella spp., Uncultured Desulfobacteraceae bacterium and sequences related to members of the family Desulfovibrio. Excess propionate was mainly syntrophically decomposed via sulfate reduction. Findings from this study could provide valuable information for the establishment of new processes and the regulation of parameters in the treatment of lower COD/SO42- ratio wastewater.