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Sterols are essential components of the yeast membrane and their synthesis requires oxygen. Yet, Saccharomyces cerevisiae has developed the ability to take up sterols from the medium under anaerobiosis. Here we investigated sterol uptake efficiency and the expression of genes related to sterol import in Saccharomyces and non-Saccharomyces wine yeast species fermenting under anaerobic conditions. The sterol uptake efficiency of 39 strains was evaluated by flow cytometry (with 25-NBD Cholesterol, a fluorescent cholesterol probe introduced in the medium) and we found an important discrepancy between Saccharomyces and non- Saccharomyces wine yeast species that we correlated to a lower final cell population and a lower fermentation rate. A high uptake of sterol was observed in the various Saccharomyces strains. Spot tests performed on 13 of these strains confirmed the differences between Saccharomyces and non-Saccharomyces strains, suggesting that the presence of the sterol uptake transporters AUS1 and PDR11 could cause these discrepancies. Indeed, we could not find any homologue to these genes in the genome of Hanseniaspora uvarum , H. guillermondii, Lachancea thermotolerans, Torulaspora delbreueckii, Metschnikowia pulcherrima , or Starmarella bacillaris species. The specialization of sterol import function for post genome-duplication species may have favored growth under anaerobiosis. [ABSTRACT FROM AUTHOR]

The yeast Lachancea thermotolerans is of significant biotechnological interest, and selected strains of this species have become commonly used starter cultures in wine fermentation. However, the impact of this species on wine is frequently limited by the rapid dominance of Saccharomyces cerevisiae strains which are better adapted to wine alcoholic fermentation conditions. Previous studies have shown that the major limiting factor for L. thermotolerans competitive performance in the wine ecosystem is oxygen availability, and not ethanol levels as had been previously suggested. Here we investigated the transcriptional response of L. thermotolerans to anaerobiosis in wine fermentation conditions. The data show that L. thermotolerans broadly redirects gene expression towards genes involved in central carbon metabolism, lipid metabolism, remodeling of the cell wall as well as autophagy. Furthermore, the induction of genes that are likely involved in the generation of lactate indicates a redirection of metabolic flux towards this metabolite. The data provide the first insight into the oxygen-dependent response of L. thermotolerans and suggest potential genetic targets to improve lactate production and/or anaerobic fermentation performance of this yeast. [ABSTRACT FROM AUTHOR]

Next generation biofuels including longer-chain alcohols such as butanol are attractive as renewable, high-energy fuels. A barrier to microbial production of butanols is the increased toxicity compared to ethanol; however, the cellular targets and microbial defense mechanisms remain poorly understood, especially under anaerobic conditions used frequently in industry. Here we took a comparative approach to understand the response of Saccharomyces cerevisiae to 1-butanol, isobutanol, or ethanol, across three genetic backgrounds of varying tolerance in aerobic and anaerobic conditions. We find that strains have different growth properties and alcohol tolerances with and without oxygen availability, as well as unique and common responses to each of the three alcohols. Our results provide evidence for strain-byalcohol- by-oxygen interactions that moderate how cells respond to alcohol stress. [ABSTRACT FROM AUTHOR]

Running events range from 60-m sprints to ultra-marathons covering 100 miles or more, which presents an interesting diversity in terms of the parameters for successful performance. Here, we review the physiological and biomechanical variations underlying elite human running performance in sprint to ultramarathon distances. Maximal running speeds observed in sprint disciplines are achieved by high vertical ground reaction forces applied over short contact times. To create this high force output, sprint events rely heavily on anaerobic metabolism, as well as a high number and large cross-sectional area of type II fibers in the leg muscles. Middle distance running performance is characterized by intermediates of biomechanical and physiological parameters, with the possibility of unique combinations of each leading to high-level performance. The relatively fast velocities in mid-distance events require a high mechanical power output, though ground reaction forces are less than in sprinting. Elite mid-distance runners exhibit local muscle adaptations that, along with a large anaerobic capacity, provide the ability to generate a high power output. Aerobic capacity starts to become an important aspect of performance in middle distance events, especially as distance increases. In distance running events, V̇O2 is an important determinant of performance, but is relatively homogeneous in elite runners. V̇O2 and velocity at lactate threshold have been shown to be superior predictors of elite distance running performance. Ultramarathons are relatively new running events, as such, less is known about physiological and biomechanical parameters that underlie ultra-marathon performance. However, it is clear that performance in these events is related to aerobic capacity, fuel utilization, and fatigue resistance. [ABSTRACT FROM AUTHOR]

Cultivation in a bioreactor of immobilized deep-sea hydrothermal microbial community was tested in order to assess the stability and reactivity of this new system. A community composed of eight hydrothermal strains was entrapped in a polymer matrix that was used to inoculate a continuous culture in a gas-lift bioreactor. The continuous culture was performed for 41 days at successively 60°C, 55°C, 60°C, 85°C and 60°C, at pH 6.5, in anaerobic condition and constant dilution rate. Oxic stress and pH variations were tested at the beginning of the incubation. Despite these detrimental conditions, three strains including two strict anaerobes were maintained in the bioreactor. High cell concentrations (3 × 108 cells mL-1) and high ATP contents were measured in both liquid fractions and beads. Cloning-sequencing and qPCR revealed that Bacillus sp. dominated at the early stage, and was later replaced by Thermotoga maritima and Thermococcus sp. Acetate, formate and propionate concentrations varied simultaneously in the liquid fractions. These results demonstrate that these immobilized cells were reactive to culture conditions. They were protected inside the beads during the stress period and released in the liquid fraction when conditions were more favorable. This confirms the advantage of immobilization that highlights the resilience capacity of certain hydrothermal microorganisms after a stress period. [ABSTRACT FROM AUTHOR]

Aims The aims of this study were (i) to develop a protocol for the entrapment of anaerobic (hyper)thermophilic marine micro-organisms; (ii) to test the use of the chosen polymers in a range of physical and chemical conditions and (iii) to validate the method with batch cultures. Methods and Results The best conditions for immobilization were obtained at 80°C with gellan and xanthan gums. After 5-week incubation, beads showed a good resistance to all tested conditions except those simultaneously including high temperature (100°C), low NaCl (<0∙5 mol l−1) and extreme pH (4/8). To confirm the method efficiency, batch cultures with immobilized Thermosipho sp. strain AT1272 and Thermococcus kodakarensis strain KOD1 showed an absence of detrimental effect on cell viability and a good growth within and outside the beads. Conclusion This suggests that entrapment in a gellan-xanthan matrix could be employed for the culture of anaerobic (hyper)thermophilic marine micro-organisms. Significance and Impact of the Study (Hyper)thermophilic marine micro-organisms possess a high biotechnological potential. Generally microbial cells are grown as free-cell cultures. The use of immobilized cells may offer several advantages such as protection against phage attack, high cell biomass and better production rate of desired metabolites. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis has become a major health hazard. It is necessary to study its metabolism and hopefully uncover therapeutic targets. Cultivating S. epidermidis at increasing oxygen concentration [O2] enhanced growth, while inhibiting biofilm formation. Respiratory oxidoreductases were differentially expressed, probably to prevent reactive oxygen species formation. Under aerobiosis, S. epidermidis expressed high oxidoreductase activities, including glycerol-3-phosphate dehydrogenase, pyruvate dehydrogenase, ethanol dehydrogenase and succinate dehydrogenase, as well as cytochromes bo and aa3; while little tendency to form biofilms was observed. Under microaerobiosis, pyruvate dehydrogenase and ethanol dehydrogenase decreased while glycerol-3-phosphate dehydrogenase and succinate dehydrogenase nearly disappeared; cytochrome bo was present; anaerobic nitrate reductase activity was observed; biofilm formation increased slightly. Under anaerobiosis, biofilms grew; low ethanol dehydrogenase, pyruvate dehydrogenase and cytochrome bo were still present; nitrate dehydrogenase was the main terminal electron acceptor. KCN inhibited the aerobic respiratory chain and increased biofilm formation. In contrast, methylamine inhibited both nitrate reductase and biofilm formation. The correlation between the expression and/or activity or redox enzymes and biofilm-formation activities suggests that these are possible therapeutic targets to erradicate S. epidermidis. [ABSTRACT FROM AUTHOR]

Spontaneous rifampicin-resistant (RFMR) mutants were isolated from Bacillus subtilis 168 cultivated in the presence or absence of oxygen. By DNA sequencing, the mutations were located within Cluster I of the rpoB gene encoding the β subunit of RNA polymerase. The spectrum of RFMR rpoB mutations isolated from B. subtilis cells grown anaerobically differed from aerobically grown cells, not only with respect to the location of mutations within Cluster I but also in the class of mutation observed (transition versus transversion). In the absence of RFM, RFMR mutants exhibited poorer growth under anaerobic conditions than did the wild-type strain, indicating their lower fitness in the absence of antibiotic selection. [ABSTRACT FROM AUTHOR]

Aims The aim of the study is to gain further insight on the respiratory behaviour of Lactobacillus plantarum and its consequences on stress tolerance. Methods and Results We investigated the effect of temperature and respiration on the growth and stress (heat, oxidative, freezing, freeze-drying) response of Lact. plantarum C17 during batch cultivations. Temperature as well as respiration clearly affected the physiological state of cells, and generally, cultures grown under respiratory conditions exhibited improved tolerance of some stresses (heat, oxidative, freezing) compared to those obtained in anaerobiosis. Our results revealed that the activities in cell-free extracts of the main enzymes related to aerobic metabolism, POX (pyruvate oxidase) and NPR ( NADH peroxidase), were significantly affected by temperature. POX was completely inhibited at 37°C, while the activity of NPR slightly increased at 25°C, indicating that in Lact. plantarum, the temperature of growth may be involved in the activation and modulation of aerobic/respiratory metabolism. Conclusions We confirmed that respiration confers robustness to Lact. plantarum cells, allowing a greater stress tolerance and advantages in the production of starter and probiotic cultures. Significance and Impact of the Study This is the first study on respiratory metabolism on a strain other than the model strains WCFS1; novel information on the role of temperature in the modulation of aerobic/respiratory metabolism in Lact. plantarum is presented. [ABSTRACT FROM AUTHOR]

The article focuses on a study that was conducted to examine the impact of Anaerobiosis on strain-dependent phenotypic variations in Pseudomonas aeruginosa bacterium. It mentions the procedure where phenotypes were compared between Pseudomonas aeruginosa and clinical isolates under anaerobic conditions. It reveals the result showing that aeruginosa is strain dependent.

In diatoms, metabolic activity during long dark periods leads to a chlororespiratory electron flow, which is accompanied by the build-up of a proton gradient strong enough to activate the diadinoxanthin (Ddx) de-epoxidation reaction of the Ddx cycle. In the present study, the impact of chlororespiration on non-photochemical quenching (NPQ) of chlorophyll fluorescence and the regulation of the Ddx cycle in the diatom Thalassiosira pseudonana was investigated by manipulation of the redox state of the photosynthetic electron transport chain during darkness. The response of a transfer of T. pseudonana cells from growth light conditions to 60 min darkness was found to depend on oxygen: in its presence there was no significant reduction of the PQ pool and no de-epoxidation of Ddx to diatoxanthin (Dtx). Under anaerobic conditions a high reduction state of the electron transport chain and a slow but steady de-epoxidation of Ddx was observed, which resulted in a significant accumulation of Dtx after 60 min of anaerobiosis. Unexpectedly, this high concentration of Dtx did not induce a correspondingly high NPQ as it would have been observed with Dtx formed under high light conditions. However, the sensitivity of NPQ to Dtx in cells kept under dark anaerobic conditions increased during reoxygenation and far-red (FR) light illumination. The results are discussed with respect to the activation of the de-epoxidation reaction and the formation of NPQ and their dependence on the extent of the proton gradient across the thylakoid membrane. [ABSTRACT FROM PUBLISHER]

Despite the scientific and applied interest in the anaerobic metabolism of Saccharomyces cerevisiae, not all genes whose transcription is upregulated under anaerobic conditions have yet been linked to known transcription factors. Experiments with a reporter construct in which the promoter of the anaerobically upregulated TIR1 gene was fused to lacZ revealed a loss of anaerobic upregulation in an snf7Δ mutant. Anaerobic upregulation was restored by expression of a truncated allele of RIM101 that encodes for a constitutively active Rim101p. Analysis of lacZ expression in several deletion mutants confirmed that the effect of Snf7p on anaerobic upregulation of TIR1 involved Rim101p. Further studies with deletion mutants in NRG1, NRG2 and SMP1, which were previously shown to be regulated by Rim101p, could not totally elucidate the TIR1 regulation, suggesting the involvement of a more complex regulation network. However, the aerobic repression mechanism of TIR1 involved the general repressor Ssn6p–Tup1p. Transcriptome analysis in anaerobic chemostat cultures revealed that 26 additional genes exhibited an Snf7p/Rim101p-dependent anaerobic upregulation, among which, besides TIR1, are four other anaerobic genes SML1, MUC1, AAC3 and YBR300C. These results provide new evidence on the implication of the Rim101p cascade in the transcriptional regulation of anaerobic metabolism in S. cerevisiae. [ABSTRACT FROM AUTHOR]

Analysis reveals that there is limited overlap in the sets of transcripts that show significant changes in abundance during anaerobiosis in different plant species. This may be due to the fact that a combination of primary effects, changes due to the presence or absence of oxygen, and secondary effects, responses to primary changes or tissue and developmental responses, are measured together and not differentiated from each other. In order to dissect out these responses, the effect of the presence or absence of oxygen was investigated using three different experimental designs using rice (Oryza sativa) as a model system. A total of 110 metabolites and 9,596 transcripts were found to change significantly in response to oxygen availability in at least one experiment. However, only one-quarter of these showed complementary responses to oxygen in all three experiments, allowing the core response to oxygen availability to be defined. A total of 10 metabolites and 1,136 genes could be defined as aerobic responders (up-regulated in the presence of oxygen and down-regulated in its absence), and 13 metabolites and 730 genes could be defined as anaerobic responders (up-regulated in the absence of oxygen and down-regulated in its presence). Defining core sets of transcripts that were sensitive to oxygen provided insights into alterations in metabolism, specifically carbohydrate and lipid metabolism and the putative regulatory mechanisms that allow rice to grow under anaerobic conditions. Transcript abundance of a specific set of transcription factors was sensitive to oxygen availability during all of the different experiments conducted, putatively identifying primary regulators of gene expression under anaerobic conditions. Combined with the possibility of selective transcript degradation, these transcriptional processes are involved in the core response of rice to anaerobiosis. [ABSTRACT FROM AUTHOR]

Background and Aims: The lack of knowledge about key traits in field environments is a major constraint to germplasm improvement and crop management because waterlogging-prone environments are highly diverse and complex, and the mechanisms of tolerance to waterlogging include a large range of traits. A model is proposed that waterlogging tolerance is a product of tolerance to anaerobiosis and high microelement concentrations. This is further evaluated with the aim of prioritizing traits required for waterlogging tolerance of wheat in the field. [ABSTRACT FROM PUBLISHER]

A successful pathogen manipulates its host for its own benefit. After ingestion, on reaching the intestine Salmonella encounters the resident tissue macrophages. Rather than being destroyed by these professional phagocytes after internalization, Salmonella survives intracellularly. Invasive Salmonella has been reported to induce apoptosis of macrophages as a part of its infection process, which may allow it to avoid detection by the innate immune system. However, the induction of apoptosis under different host environments, including the anaerobic stress encountered by the pathogen in the gut, remains to be examined. The present study is aimed at investigating the apoptotic potential of S. enterica serovar Typhi ( S. typhi) grown under anaerobic conditions simulating the in vivo situation encountered by the pathogen. Apoptotic cell death was determined by assessment of nucleosomal DNA and flow cytometric analysis. Evaluation of the data revealed that anaerobically grown S. typhi could induce apoptosis in significantly more number of macrophages compared to the bacterial cells grown under aerobic conditions. A significantly enhanced generation of reactive nitrogen intermediates and caspase-3 activity during macrophage apoptosis induced by anaerobic S. typhi correlated with the increased generation of tumour necrosis factor-α, interleukin (IL)-1α and IL-6. The results indicate that reactive nitrogen intermediates and monokines induce caspase-3 mediated apoptosis of macrophages by S. typhi under anaerobic conditions. These findings may be relevant for clearer understanding of the Salmonella–macrophage interactions and may be of clinical importance in the development of preventive intervention against the infection. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa causes chronic infections in the lungs of cystic fibrosis (CF) individuals and remains the leading cause of morbidity and mortality associated with the disease. Biofilm growth and phenotypic diversification are factors thought to contribute to this organism's persistence. Most studies have focused on laboratory isolates such as strain PAO1, and there are relatively few reports characterizing the properties of CF strains, especially under decreased oxygen conditions such as occur in the CF lung. This study compared the phenotypic and functional properties of P. aeruginosa from chronically infected CF adults with those of strain PAO1 and other clinical non-CF isolates under aerobic and anaerobic culture conditions. The CF isolates overall displayed a reduced ability to form biofilms in standard in vitro short-term models. They also grew more slowly in culture, and exhibited decreased adherence to glass and decreased motilities (swimming, swarming and twitching). All of these characteristics were markedly accentuated by anaerobic growth conditions. Moreover, the CF strain phenotypes were not readily reversed by culture manipulations designed to encourage planktonic growth. The CF strains were thus inherently different from strain PAO1 and most of the other non-CF clinical P. aeruginosa isolates tested. In vitro models used to research CF isolate biofilm growth need to take the above properties of these strains into account. [ABSTRACT FROM AUTHOR]

The KlAAC gene, encoding the ADP/ATP carrier, has been assumed to be a single gene in Kluyveromyces lactis, an aerobic, petite-negative yeast species. The Klaac null mutation, which causes a respiratory-deficient phenotype, was fully complemented by AAC2, the Saccharomyces cerevisiae major gene for the ADP/ATP carrier and also by AAC1, a gene that is poorly expressed in S. cerevisiae. In this study, we demonstrate that the Klaac null mutation is partially complemented by the ScAAC3 gene, encoding the hypoxic ADP/ATP carrier isoform, whose expression in S. cerevisiae is prevented by oxygen. Once introduced into K. lactis, the AAC3 gene was expressed both under aerobic and under partial anaerobic conditions but did not support the growth of K. lactis under strict anaerobic conditions. [ABSTRACT FROM AUTHOR]

Polymorphisms in rpoS are common in Escherichia coli. rpoS status influences a trade-off between nutrition and stress resistance and hence fitness across different environments. To analyze the selective pressures acting on rpoS, measurement of glucose transport rates in rpoS+ and rpoS bacteria was used to estimate the role of Fnc, the fitness gain due to improved nutrient uptake, in the emergence of rpoS mutations in nutrient-limited chemostat cultures. Chemostats with set atmospheres, temperatures, pH's, antibiotics, and levels of osmotic stress were followed. Fnc was reduced under anaerobiosis, high osmolarity, and with chloramphenicol, consistent with a reduced rate of rpoS enrichment in these conditions. Fnc remained high, however, with alkaline pH and low temperature but rpoS sweeps were diminished. Under these conditions, Fsp, the fitness reduction due to lowered stress protection, became significant. We also estimated whether the fitness need for the gene was related to its regulation. No consistent pattern emerged between the level of RpoS and the loss of rpoS function in particular environments. This dissection allows an unprecedented view of the genotype-by-environment interactions controlling a mutational sweep and shows that both Fnc and Fsp are influenced by individual stresses and that additional factors contribute to selection pressure in some environments. [ABSTRACT FROM AUTHOR]

Expression of the anaerobically inducible focA-pfl operon in Escherichia coli was activated nearly sevenfold relative to wild-type under aerobic growth conditions by increasing the dosage of the fnr gene on a pBR322-based plasmid (pCH21). No effect on anaerobic expression levels was observed, suggesting that operon expression under these conditions is maximal. Examination of the complex transcript pattern of the focA-pfl operon confirmed that in strains bearing pCH21 all transcripts, with the exception of the promoter 7 transcript, were up-regulated aerobically. Western analysis of strains bearing pCH21 revealed that the fumarate nitrate regulator (FNR) level was increased approximately ninefold relative to the level in strains bearing a single copy of the fnr gene aerobically, but was only overproduced threefold anaerobically. Analysis of an fnr-lacZ fusion indicated that fnr expression was more strongly negatively autoregulated in anaerobic cells compared with aerobic cells when pCH21 was present. Taken together, these findings suggest that high-level overproduction of FNR is prevented anaerobically by active FNR repressing expression of the fnr gene. Furthermore, transcription from promoter 7 of the focA-pfl operon, which depends on both ArcA-P and FNR, cannot be activated aerobically by overproduction of FNR alone, while promoter 6, which is less dependent on ArcA-P, can be activated under these conditions. [ABSTRACT FROM AUTHOR]

Aims: To establish the influence of oxygen on Escherichia coil before, during and after exposure to UVA or simulated sunlight. Methods and Results: Bacterial suspensions were exposed either to UVA or simulated sunlight. Conventional aerobic plate counts of illuminated cell suspensions were consistently lower than those obtained under conditions where reactive oxygen species (ROS) were neutralized, either (i) by the addition of the peroxide scavenger sodium pyruvate (0·05% w/v) to the medium with subsequent incubation in an anaerobic jar or (ii) by culturing on a prereduced medium within an anaerobic cabinet, indicating that a substantial proportion of such cells are sublethally injured. While the presence of oxygen during the growth period resulted in a greater resistance of aerobically grown cells to simulated sunlight compared with their anaerobic counterparts, the extent of inactivation during illumination was directly related to the dissolved oxygen content of the water. Conclusions: The results show that, at each stage, oxygen has a marked influence on the observed colony count. Significance and Impact of the Study: Overall, the results indicate that future studies of bacteria exposed to UVA or sunlight should consider the effects of oxygen at every stage in the procedure, and especially during enumeration, where the inhibitory effects of ROS must be neutralized in order to obtain a valid count. An investigation of the effects of ROS neutralization on the counts of faecal bacteria under field conditions in natural waters is now required to establish the significance of these finding to solar water treatment. [ABSTRACT FROM AUTHOR]

The abundance of 4Fe–4S motifs of the form CX2CX2CX3C was analyzed in the open reading frames (ORFs) of 120 prokaryotic genomes. The abundance of ORFs containing the CX2CX2CX3C motif or isORFs correlated (r=0.82) with methanogenesis (p=0.0001), archaea (p=0.0173), anaerobiosis (p<0.0001) and genome size (p<0.0001). Optimal growth temperature (hyperthermophily) did not correlate with the number of isORFs (p=0.6283). Large numbers of CX2CX2CX3C motifs may be associated with unique physiologies: methanogenic archaea contained the greatest number of CX2CX2CX3C motifs found among the prokaryotic groups; however, only about 15% of the motifs were in genes directly involved in methanogenesis. Large numbers of CX2CX2CX3C motifs may also be associated with generalists such as Desulfitobacterium hafniense, which is an anaerobic bacterium containing multiple reductases. [Copyright &y& Elsevier]

Although grape berries have been classified as non-climacteric fruits, ongoing studies on grape ethylene signalling challenge the role of ethylene in their ripening. One of the significant molecular changes in berries is the up-regulation of ADH (alcohol dehydrogenase, EC 1.1.1.1) enzyme activity at the inception of fruit ripening and of VvADH2 transcript levels. This paper shows that the ethylene signal transduction pathway could be involved in the control of VvADH2 expression in grapevine berries and in cell suspensions. The induction of VvADH2 transcription, either in berries at the inception of ripening or in cell suspensions, was found to be partly inhibited by 1-methylcyclopropene (1-MCP), an inhibitor of ethylene receptors. Treatment of cell suspensions with 2-chloroethylphosphonic acid (2-CEPA), an ethylene-releasing compound, also resulted in a significant increase in ADH activity and VvADH2 transcription under anaerobiosis, showing that concomitant ethylene and anaerobic treatments in cell suspensions could result in changes in VvADH2 expression. All these results associated with the presence in the VvADH2 promoter of regulatory elements for ethylene and anaerobic response, suggest that the ethylene transduction pathway and anaerobic stress could be, in part, involved in the regulation of VvADH2 expression in ripening berries and cell suspensions. These data open new aspects of the expression control of a ripening-related gene in a non-climacteric fruit. [ABSTRACT FROM PUBLISHER]

DNA insertional transformants of Chlamydomonas reinhardtii were screened chemochromically for attenuated H2 production. One mutant, displaying low H2 gas photoproduction, has a nonfunctional copy of a gene that shows high homology to the family of isoamylase genes found in several photosynthetic organisms. DNA gel blotting and gene complementation were used to link this isoamylase gene to previously characterized nontagged sta7 mutants. This mutant is therefore denoted sta7-10. In C. reinhardtii, the STA7 isoamylase gene is important for the accumulation of crystalline starch, and the sta7-10 mutant reported here contains <3% of the glucose found in insoluble starch when compared with wild-type control cells. Hydrogen photoproduction rates, induced after several hours of dark, anaerobic treatment, are attenuated in sta7 mutants. RNA gel blot analysis indicates that the mRNA transcripts for both the HydA1 and HydA2 [Fe]-hydrogenase genes are expressed in the sta7-10 mutant at greater than wild-type levels 0.5 h after anaerobic induction. However, after 1.5 h, transcript levels of both HydA1 and HydA2 begin to decline rapidly and reach nearly undetectable levels after 7 h. In wild-type cells, the hydrogenase transcripts accumulate more slowly, reach a plateau after 4 h of anaerobic treatment, and maintain the same level of expression for >7 h under anaerobic incubation. Complementation of mutant cells with genomic DNA corresponding to the STA7 gene restores both the starch accumulation and H2 production phenotypes. The results indicate that STA7 and starch metabolism play an important role in C. reinhardtii H2 photoproduction. Moreover, the results indicate that mere anaerobiosis is not sufficient to maintain hydrogenase gene expression without the underlying physiology, an important aspect of which is starch metabolism. [ABSTRACT FROM AUTHOR]

The ability to evolve hydrogen using methyl viologen as an electron donor was assayed in the nitrogen-fixing actinomycetes Frankia sp. R43 and Frankia sp. KB5. To further examine the nature of hydrogen-evolving enzymes that may be present in these organisms immunological studies were performed. Under anaerobic conditions (both nitrogen-limiting and nitrogen-containing) Frankia sp. R43 but not Frankia sp. KB5 evolved hydrogen,which was not linked to NAD-reducing activity. Immunological analysis of total protein from Frankia sp. R43 and Frankia sp. KB5 using an antiserum raised against Ralstonia eutropha HoxF, recognized an antigen in Frankia sp. R43 but not in Frankia sp. KB5. Immunogold labeling using antibodies raised against the R. eutropha HoxH recognized sites in both hyphae and vesicles of Frankia sp. R43, but not in Frankia sp. KB5. Based on these physiological and immunological findings, we conclude that Frankia sp. R43 has a hydrogen-evolving hydrogenase. [Copyright &y& Elsevier]

Stem segments from terminal leaders of Douglas‐fir, Pseudotsuga menziesii (Mirb.) Franco, seedlings were sampled in mid‐December when cambial cells were dormant. The residual, debudded leaders were resampled again in early May when the cambium was metabolically active. May stems had higher constitutive ethanol concentrations than December stems. This was not the result of cambial hypoxia generated by rapid spring respiration rates, because when aerobic respiration was stimulated by incubating the stems in air at 30 °C ethanol production was induced in December, but not in May. Rapid respiration rates at 30 °C may have depleted O2 supplies and induced ethanol production in December stems because dormant, thick‐walled cambial cells may be less permeable to CO2 and O2, compared with metabolically active, thin‐walled cambial cells in May. December stem segments incubated in a N2 atmosphere at 30 °C synthesized 1.8 times more ethanol than segments from May, most likely because spring growth had reduced the soluble sugars available for fermentation. CO2 efflux from May stems (after 5.5 h of incubation at 30 °C) was equal to December stems per unit volume, but greater than December stems per unit surface area. N2‐induced ethanol concentrations were positively related with CO2 efflux per unit volume, indicating that rapidly respiring leaders can maintain rapid fermentation rates, provided soluble sugars are readily available. N2‐induced ethanol and CO2 efflux per unit volume declined with increasing leader diameter in both seasons, whereas there were no relationships between CO2 efflux per unit surface area and diameter. Cambium physiology and phenology influence the induction of fermentation and concentrations of ethanol produced in terminal leaders of Douglas‐fir, and probably other conifers as well. This needs to be considered when comparing fermentation among species, or comparing individuals from different seasons, or disparate ages within a species. [ABSTRACT FROM PUBLISHER]

Abstract Aims: Yeast strains, used in the brewing industry, experience distinctive physiological conditions. During a brewing fermentation, yeast are exposed to anaerobic conditions, high pressure, high specific gravity and low temperatures. The purpose of this study was to examine the global gene expression profile of yeast subjected to brewing stress. Methods and Results: We have carried out a microarray analysis of a typical brewer's yeast during the course of an 8-day fermentation in 15°P wort. We used the probes derived from Saccharomyces cerevisiae genomic DNA on the chip and RNA isolated from three stages of brewing. This analysis shows a high level of expression of genes involved in fatty acid and ergosterol biosynthesis early in fermentation. Furthermore, genes involved in respiration and mitochondrial protein synthesis also show higher levels of expression. Conclusions: Surprisingly, we observed a complete repression of many stress response genes and genes involved in protein synthesis throughout the 8-day period compared with that at the start of fermentation. Significance and Impact of the Study: This microarray data set provides an analysis of gene expression under brewing fermentation conditions. The data provide an insight into the various metabolic processes altered or activated by brewing conditions of growth. This study leads to future experiments whereby selective alterations in brewing conditions could be introduced to take advantage of the changing transcript profile to improve the quality of the brew. [ABSTRACT FROM AUTHOR]

Aims: In order to clarify the involvement of an energy-yielding system in the antibacterial action of surfactants, the effects of carbon source and anaerobiosis during the growth period on the surfactant sensitivity of Escherichia coli cells were investigated. Methods and Results: Cetyltrimethylammonium bromide (CTAB) and N -dodecyl-N ,N -dimethylglycine, at relatively low concentrations, caused a delay in growth of E. coli cells. Cells grown in M9 medium supplemented with glycerol, succinate or acetate as a carbon source were more sensitive to surfactants and had a higher respiratory activity than those grown with glucose. Cultivation under anaerobiosis made cells resistant to CTAB. Conclusions: Bacterial sensitivity to surfactants was affected by carbon source and anaerobiosis. Significance and Impact of the Study: The results obtained should be helpful in determining suitable conditions of treatment in the practical use of surfactants for bacterial decontamination. [ABSTRACT FROM AUTHOR]

This study explored the geochemistry and microbial diversity of a Superfund site containing trichloroethene (TCE) and an unusual co-pollutant, tetrakis(2-ethylbutoxy)silane. Geochemical analysis of contaminated groundwater indicated subsurface anaerobiosis, reductive dechlorination of TCE to predominantly cis-1,2-dichloroethene, and (transient) accumulation of 2-ethylbutanol and 2-ethylbutyrate as a result of tetrakis(2-ethylbutoxy)silane breakdown. Comparative analysis of 106 16S rDNA and 61 16S–23S rDNA intergenic spacer region sequences – obtained from pristine and contaminated groundwater via DNA extraction, PCR amplification, cloning and sequencing – revealed that the contaminated groundwater featured (i) a distinct microbial community, (ii) reduced species diversity, (iii) various anaerobes, and (iv) bacteria closely related to the TCE-dechlorinating, dichloroethene-accumulating genus Dehalobacter, whereas (v) the TCE-dechlorinating, ethene-producing species Dehalococcoides ethenogenes was not detectable. Thus, geochemical and molecular biological results were in excellent agreement in this first ecological field study linking in situ reductive dechlorination of TCE to metabolism of tetraalkoxysilanes. [Copyright &y& Elsevier]

Several kinds of signal may be generated when roots are exposed to an environmental stress. Some, but not all, are conveyed to shoots in the transpiration stream. Principles are summarized that may help establish experimentally the presence or intensity of root signals transported by transpiration. In many dryland species, flooding of the soil induces developmental responses in the shoot such as epinastic leaf curvature, stomatal closure and slowing of leaf expansion. These reactions compensate for diminished input of resources from the roots. They lend themselves to the study of root-to-shoot signalling by commencing after a time lag of only a few hours, by persisting for several days and by being highly reproducible. Evidence implicating chemical and hydraulic signals in promoting stomatal closure and epinastic curvature in flooded plants is reviewed. Further progress will depend upon examining a wider range of putative signals, accounting for any interactions between them and improving methods for the evaluation of signal durability in transit, and effectiveness at target sites. [ABSTRACT FROM AUTHOR]

Sterol esterification in Saccharomyces cerevisiae is catalyzed by two acyl-CoA:sterol acyltransferases encoded by the genes ARE1 and ARE2. Using double mutants in the HEM1 gene and individual ARE genes we demonstrated that the relative contribution of these two enzymes to sterol esterification was dependent on cellular heme status. Observed changes in sterol esterification could be explained by a different effect of heme on the transcription of both genes: while the ARE1 transcript level was elevated in heme-deficient and anaerobic cells, the ARE2 gene transcript was more abundant in aerobic cells competent for heme synthesis. Our results indicate that transcriptional regulation of ARE genes by heme and specific substrate preferences of Are1p and Are2p may be involved in the adaptation of yeast sterol metabolism to hypoxia. [Copyright &y& Elsevier]

Shoot elongation of arrowhead (Sagittaria pygmaea Miq.) tubers was stimulated in anaerobic conditions. The anaerobic elongation was attributed to stimulation of cell elongation in the middle of the shoots. The anaerobic elongation of the shoots was severely inhibited by ethylene glycol bis(β-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). The EGTA inhibition was completely nullified by exogenous CaCl2, which acts as an enhancer of anaerobic elongation. Moreover, calcium channel blockers, varapamil, diltiazem and LaCl3, inhibited the anaerobic elongation enhanced by CaCl2. These results showed that calcium plays an important role in stimulating the elongation in anaerobic conditions. Incorporation of 45Ca into the shoot tissues was measured to determine the involvement of calcium uptake in anaerobic elongation. Incorporation of 45Ca into the cell sap, which was collected from frozen and thawed shoots after thorough washing with LaCl3, was significantly stimulated in anaerobic conditions. Verapamil and diltiazem prevented the stimulation of 45Ca incorporation in anaerobic conditions. These results suggest that calcium uptake from the medium serves to enhance shoot elongation of arrowhead tubers under anaerobic conditions. [ABSTRACT FROM AUTHOR]

Nine out of ten industrially important strains of Saccharomyces cerevisiae did not grow in minimal media under anaerobic conditions even when ergosterol and unsaturated fatty acids were provided. Anaerobiosis was maintained either by flushing the culture flasks with prepurified nitrogen or by incubating the flasks in an anaerobic chamber. Traces of oxygen present in 'prepurified nitrogen gas' were sufficient to initiate yeast growth and on removal of the oxygen by catalytic means the yeasts failed to grow. The yeast grew very well anaerobically if the medium was supplemented with a mixture of amino acids or with a mixture of purines and pyrimidines. The growth initiated by including a mixture of amino acids was further enhanced when the medium was supplemented with ergosterol and an unsaturated fatty acid. Since no oxygen requirement for the synthesis of amino acids or purines and pyrimidines has been demonstrated, growth promotion by these compounds under anaerobic conditions is most likely not by eliminating the need for oxygen for their synthesis. We suggest that the amino acids and the nucleic acid bases yielded, through some hitherto unknown reactions, small amounts of a molecular or usable form of oxygen which allowed key reactions essential for 'anaerobic' growth to proceed. [ABSTRACT FROM AUTHOR]

N-acetylchitooIigosaccharides (fragments of chitin) elicit defense responses, including phytoalexin production, in suspension-cultured rice cells. They induced rapid and transient membrane depolarization accompanied by a transient increase in net CP−-efflux. The membrane depolarization was not affected by anaerobiosis or azide, suggesting that the major part of the depolarization was mediated by ion channels, not by energy-dependent ion pumps. Depolarization was partly inhibited in the presence of Ca2+- or Cl−-channel blockers and highly inhibited by depletion of Ca2+ in the extracellular medium. A calcium ionophore, A23187, caused a transient depolarization but not an increase in Cl− efflux, while it did not inhibit the elic-itor-induced transient depolarization and Cl− efflux. These suggest that the influx of Ca2+ from the extracellular space to the cytoplasm is necessary as an initial trigger but not sufficient for membrane depolarization, Cl− efflux, and the following signaling processes. [ABSTRACT FROM AUTHOR]

The tolerance of germinating rice seedlings to anaerobiosis cannot be fully accounted for by ethanolic fermentation alone. Nitrate metabolism (nitrate reduction to NH4+ plus its subsequent assimilation) may provide an additional sink mechanism for excess protons and NADH produced during anaerobiosis. To follow the fate of nitrate, 15N-labelled nitrate and ammonium incorporation in aerobic and anaerobic rice coleoptiles was examined using 15N-edited 1H NMR and gas chromatography-mass spectrometry methods. After 22 h of treatments, protein-free Ala, Glu, Gln, and γ-aminobutyrate were the main 15N-labelled products for both nitrate and ammonium-treated anaerobic rice coleoptiles, with Gln, Glu, and Ala being the most enriched. The total amount of 15N label incorporation into Ala and GAB increased significantly in response to anaerobiosis. The 15N-labelling pattern of Glu and Gln suggests that the GS/GOGAT system was primarily involved in ammonium assimilation whereas Glu dehydrogenase may play a role in nitrate assimilation. 15N incorporation into protein-dried amino acids was also significant and was more substantial in anaerobic than in aerobic rice coleoptiles, which indicate that protein biosynthesis remained active in anaerobic rice coleoptiles. Thus, anaerobic assimilation of inorganic N into amino acids, particularly Ala and Glu/GAB, may serve to supplement ethanolic fermentation in sustaining glycolysis and energy production in rice coleoptiles. [ABSTRACT FROM PUBLISHER]

We sought to investigate how far the growth of Saccharomyces cerevisiae under full anaerobiosis is dependent on the widely used anaerobic growth factors (AGF) ergosterol and oleic acid. A continuous cultivation setup was employed and, even forcing ultrapure N2 gas through an O2 trap upstream of the bioreactor, neither cells from S. cerevisiae CEN.PK113–7D (a lab strain) nor from PE-2 (an industrial strain) washed out after an aerobic-to-anaerobic switch in the absence of AGF. S. cerevisiae PE-2 seemed to cope better than the laboratory strain with this extremely low O2 availability, since it presented higher biomass yield, lower specific rates of glucose consumption and CO2 formation, and higher survival at low pH. Lipid (fatty acid and sterol) composition dramatically altered when cells were grown anaerobically without AGF: saturated fatty acid, squalene and lanosterol contents increased, when compared to either cells grown aerobically or anaerobically with AGF. We concluded that these lipid alterations negatively affect cell viability during exposure to low pH or high ethanol titers. [ABSTRACT FROM AUTHOR]

The imposition of anaerobic conditions resulted in a rapid (2 min) and transient increase in cGMP content in root and coleoptile of rice seedlings. The level of cAMP was strongly decreased by anoxia. These data are discussed in relation to the anaerobic response. [ABSTRACT FROM AUTHOR]

Presents an abstract of the study `Sulfide Induced Anaerobiosis in Urechis caupo,' by V.G. Encomio and A.J. Arp submitted for presentation during the 1996 Society for Integrative and Comparative Biology Annual Meeting.

Presents an abstract of the study `Facultative Anaerobiosis in Larvae of the Tiger Beetle, Cicindela togata,' by W.W. Hoback, David W. et al submitted for presentation during the 1996 Society for Integrative and Comparative Biology Annual Meeting.