Carbon Substrate Utilization Research Articles

Regulation of Carbon Substrate Utilization by Cardiac Mitochondria

Mitochondria play key roles in central metabolism, not only by synthesizing ATP via oxidative phosphorylation, but also by synthesizing key intermediates that serve as anaplerotic entry points in numerous pathways. Because reactions producing and/or consuming several of the intermediates in the tricarboxylic acid cycle that are transported across the mitochondrial inner membrane (pyruvate, α-ketoglutarate, citrate) are regulated in part by calcium concentration, we hypothesize the calcium is a key regulator of entry and exit of substrates into and out of the TCA cycle. We set out to first refine this hypothesis through following time courses of consumption of various carbohydrate based substrates (including complex I substrates such as pyruvate and citrate, and the complex II substrate succinate) and matching data on rates of oxidation and NADH production to simulations of mitochondrial metabolic kinetics. Specifically, model simulations were fit to data on total NAD(P)H and oxygen consumption flux at different respiratory states and with varying inorganic phosphate concentrations. The resulting identified model is used to predict how calcium regulates carbon substrate utilization by mitochondria. These hypotheses are tested using in vitro experiments with purified mitochondria. By comparing the experimental data with citrate alone to combination of pyruvate, citrate and malate as the substrates, we conclude that transport of citrate into the matrix is limited in the absence of malate. The rate of reduction of NAD is slower with pyruvate alone as substrate compared to when malate is present. The role of glutamate dehydrogenase in the context of cardiac energetics is investigated.

High-Throughput Carbon Substrate Profiling of Mycobacterium ulcerans Suggests Potential Environmental Reservoirs.

BackgroundMycobacterium ulcerans is a close derivative of Mycobacterium marinum and the agent of Buruli ulcer in some tropical countries. Epidemiological and environmental studies pointed towards stagnant water ecosystems as potential sources of M. ulcerans, yet the ultimate reservoirs remain elusive. We hypothesized that carbon substrate determination may help elucidating the spectrum of potential reservoirs.Methodology/Principal findingsIn a first step, high-throughput phenotype microarray Biolog was used to profile carbon substrates in one M. marinum and five M. ulcerans strains. A total of 131/190 (69%) carbon substrates were metabolized by at least one M. ulcerans strain, including 28/190 (15%) carbon substrates metabolized by all five M. ulcerans strains of which 21 substrates were also metabolized by M. marinum. In a second step, 131 carbon substrates were investigated, through a bibliographical search, for their known environmental sources including plants, fruits and vegetables, bacteria, algae, fungi, nematodes, mollusks, mammals, insects and the inanimate environment. This analysis yielded significant association of M. ulcerans with bacteria (p = 0.000), fungi (p = 0.001), algae (p = 0.003) and mollusks (p = 0.007). In a third step, the Medline database was cross-searched for bacteria, fungi, mollusks and algae as potential sources of carbon substrates metabolized by all tested M. ulcerans; it indicated that 57% of M. ulcerans substrates were associated with bacteria, 18% with alga, 11% with mollusks and 7% with fungi.ConclusionsThis first report of high-throughput carbon substrate utilization by M. ulcerans would help designing media to isolate and grow this pathogen. Furthermore, the presented data suggest that potential M. ulcerans environmental reservoirs might be related to micro-habitats where bacteria, fungi, algae and mollusks are abundant. This should be followed by targeted investigations in Buruli ulcer endemic regions.

Removal of pyrene in simulated wetland by joint application of Kyllinga brevifolia Rottb. and immobilized microbes

The objectives of this study were to evaluate the effects of plant and immobilized microbes on pyrene removal and soil microbial functional diversity of co-contaminated soil. The results indicated that the dissipation ratios of pyrene in the soil with plant (P), soil with immobilized microbes (IM) and soil with both of them(PIM) were 51.5 ± 0.69%, 55.2 ± 3.8%, 63.2 ± 1.29% respectively, and were higher than CK (31.2 ± 1.5%) with neither of them. Soil microbial functional diversity was assessed by the community-level physiological profiles (CLPP) using Biolog Eco-plates. The Biolog results revealed that the metabolic intensity in the rhizospheric soil marked P-R and PIM-R was significantly higher than that of non-rhizospheric soil and the CK during the incubation period. A similar variation in the diversity indexes (Shannon, Simpson and McIntosh) was observed. Principal component analysis (PCA) of the distribution of carbon substrate utilization for all treatments suggests that the microbial community catabolic diversity is different. Among the carbon utilization profile, carbohydrates were the main carbon source of the sample soil and IM increased the potential utilization of two substrate groups (phenols and carbohydrates) more strongly than that of the other groups.

Effect of Biofumigants on Soil Microbial Communities and Ecotoxicology of Earthworms (Eisenia andrei)

Biofumigation is considered a good alternative to chemical fumigation because it can control crop pathogens and diseases with lower health and environmental risks than chemical fumigants. Glucosinolates are volatile compounds found in most Brassica species, and when hydrolysed, it forms a range of natural toxins including isothiocyanates that act as biofumigants. However, the effect of glucosinolates and their breakdown products on non-target and beneficial soil organisms is not well documented. Three biofumigants, broccoli, mustard and oilseed radish, were evaluated for their effect on earthworms (Eisenia andrei) and the soil microbial community. Sub-lethal endpoints, including growth and reproductive success of the earthworms, were monitored. Genotoxicity of the biofumigants towards earthworms was evaluated by means of the comet assay. Broccoli reduced earthworm reproduction while mustard induced more DNA strand breaks in earthworm cells compared to the control. Soil microbial community function and structure were evaluated by means of community level physiological profiling and phospholipid fatty acid analyses. The effects exerted by the biofumigants on the microbial community were the most pronounced within the first 14 days after application. Carbon substrate utilisation was most affected by the oilseed radish treatment and microbial community structure by the mustard treatment.

Utilization of carbon substrates by heterotrophic bacteria through vertical sediment profiles in coastal and estuarine seagrass meadows.

Coastal vegetated ecosystems play an important role in carbon cycling and bacterial communities inhabiting coastal sediments are responsible for the remineralization and processing of organic carbon (OC). We collected 1 m-long sediment cores in Posidonia seagrass meadows from coastal and estuarine sites in Australia that differed in their sedimentary organic and inorganic carbon, nitrogen and mud contents. The metabolic diversity of sediment heterotrophic bacterial communities was characterized at different sediment depths, based on the utilization pattern of31 individual carbon substrates using Biolog EcoPlatesTM . High metabolic diversity was recorded at both sites, but the carbon substrate utilization rates and the use of carbohydrates were higher at the coastal site compared to the estuarine site. The heterotrophic bacterial community in the coastal sediment appeared to metabolize a more diverse OC pool compared to the estuarine site, which might partly explain the differences in OC storage among the seagrass habitats studied. The Biolog EcoPlatesTM provided a useful tool for characterising the sediment heterotrophic bacterial communities in the meadows and sediment characteristics and biochemical composition of the organic matter played an important role in shaping heterotrophic bacterial communities and their carbon utilization rates, potentially affecting carbon accumulation and preservation within seagrass sediments.

The effect of biochar loading rates on soil fertility, soil biomass, potential nitrification, and soil community metabolic profiles in three different soils

Biochar is increasingly being used as a soil amendment to both increase soil carbon storage and improve soil chemical and biological properties. To better understand the shorter-term (10 months) impacts of biochar on selected soil parameters and biological process in three different textured soils, a wide range of loading rates was applied. Biochar derived from eucalypt green waste was mixed at 0, 2.5, 5, 10 % (wt/wt) with a reactive black clay loam (BCL), a non-reactive red loam (RL) and a brown sandy loam (BSL) and placed in pots exposed to the natural elements. After 10 months of incubation, analysis was performed to determine the impacts of the biochar rates on the different soil types. Also, microbial biomass was estimated by the total viable counts (TVC) and DNA extraction. Moreover, potential nitrification rate and community metabolic profiles were assayed to evaluate microbial function and biological process in biochar-amended soils. The results showed that biochar additions had a significant impact on NH4 and NO3, total C and N, pH, EC, and soil moisture content in both a soil type and loading-dependent manner. In the heavier and reactive BCL, no significant impact was observed on the available P and K levels, or the total exchangeable base cations (TEB) and CEC. However, in the other lighter soils, biochar addition had a significant effect on the exchangeable Al, Ca, Mg, and Na levels and CEC. There was a relatively limited effect on microbial biomass in amended soils; however, biochar additions and its interactions with different soils reduced the potential nitrification at the higher biochar rate in the two lighter soils. Community metabolic profile results showed that the effect of biochar on carbon substrate utilization was both soil type and loading dependent. The BCL and BSL showed reduced rates of substrate utilization as biochar loading levels increased while the opposite occurred for the RL. This research shows that biochar can improve soil carbon levels and raise pH but varies with soil type. High biochar loading rates may also influence nitrification and the function and activity of microbial community in lighter soils.

Diversity of Ralstonia solanacearum Strains on the Andaman Islands in India.

Fourteen Ralstonia solanacearum strains from solanaceous vegetables on the Andaman Islands, India, were characterized using a polyphasic approach. The strains wilted their respective hosts within 1 to 3 weeks postinoculation. Virulence assays on tomato (Solanum lycopersicum), brinjal (eggplant; S. melongena), and chili pepper (Capsicum annuum) revealed that all strains were infective on all three hosts. However, tomato was more susceptible than eggplant and chili pepper. Strains were identified as R. solanacearum based on carbon substrate utilization profiling with Biolog similarity coefficients >0.82. Species identity was further confirmed by 16S ribosomal RNA and recN gene sequence analysis. Intraspecific identification of strains revealed the presence of race 1 biovar 3 and race 1 biovar 4. Both biovars wilted plants with similar aggressiveness. All strains were identified as phylotype I, and multilocus sequence typing revealed that the strains belong to a small number of clonal complexes that also comprise strains from mainland India, especially West Bengal state and Kerala.

Metabolic effects of azoxystrobin and kresoxim-methyl against Fusarium kyushuense examined using the Biolog FF MicroPlate

Azoxystrobin and kresoxim-methyl are strobilurin fungicides, and are effective in controlling many plant diseases, including Fusarium wilt. The mode of action of this kind of chemical is inhibition of respiration. This research investigated the sensitivities of Fusarium kyushuense to azoxystrobin and kresoxim-methyl, and to the alternative oxidase inhibitor salicylhydroxamic acid (SHAM). The Biolog FF MicroPlate is designed to examine substrate utilization and metabolic profiling of micro-organisms, and was used here to study the activity of azoxystrobin, kresoxim-methyl and SHAM against F. kyushuense. Results presented that azoxystrobin and kresoxim-methyl strongly inhibited conidial germination and mycelial growth of F. kyushuense, with EC50 values of 1.60 and 1.79μgml−1, and 6.25 and 11.43μgml−1, respectively; while not for SHAM. In the absence of fungicide, F. kyushuense was able to metabolize 91.6% of the tested carbon substrates, including 69 effectively and 18 moderately. SHAM did not inhibit carbon substrate utilization. Under the selective pressure of azoxystrobin and kresoxim-methyl during mycelial growth (up to 100μgml−1) and conidial germination (up to 10μgml−1), F. kyushuense was unable to metabolize many substrates in the Biolog FF MicroPlate; while especially for carbon substrates in glycolysis and tricarboxylic acid cycle, with notable exceptions such as β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, α-d-glucose-1-phosphate, d-saccharic acid and succinic acid in the mycelial growth stage, and β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, tween-80, arbutin, dextrin, glycerol and glycogen in the conidial germination stage. This is a new finding for some effect of azoxystrobin and kresoxim-methyl on carbon substrate utilization related to glycolysis and tricarboxylic acid cycle and other carbons, and may lead to future applications of Biolog FF MicroPlate for metabolic effects of other fungicides and other fungi, as well as providing a carbon metabolic fingerprint of F. kyushuense that could be useful for identification.

Average nucleotide identity of genome sequences supports the description of Rhizobium lentis sp. nov., Rhizobium bangladeshense sp. nov. and Rhizobium binae sp. nov. from lentil (Lens culinaris) nodules.

Rhizobial strains isolated from effective root nodules of field-grown lentil (Lens culinaris) from different parts of Bangladesh were previously analysed using sequences of the 16S rRNA gene, three housekeeping genes (recA, atpD and glnII) and three nodulation genes (nodA, nodC and nodD), DNA fingerprinting and phenotypic characterization. Analysis of housekeeping gene sequences and DNA fingerprints indicated that the strains belonged to three novel clades in the genus Rhizobium. In present study, a representative strain from each clade was further characterized by determination of cellular fatty acid compositions, carbon substrate utilization patterns and DNA-DNA hybridization and average nucleotide identity (ANI) analyses from whole-genome sequences. DNA-DNA hybridization showed 50-62% relatedness to their closest relatives (the type strains of Rhizobium etli and Rhizobium phaseoli) and 50-60% relatedness to each other. These results were further supported by ANI values, based on genome sequencing, which were 87-92% with their close relatives and 88-89% with each other. On the basis of these results, three novel species, Rhizobium lentis sp. nov. (type strain BLR27(T) = LMG 28441(T) = DSM 29286(T)), Rhizobium bangladeshense sp. nov. (type strain BLR175(T) = LMG 28442(T) = DSM 29287(T)) and Rhizobium binae sp. nov. (type strain BLR195(T) = LMG 28443(T) = DSM 29288(T)), are proposed. These species share common nodulation genes (nodA, nodC and nodD) that are similar to those of the symbiovar viciae.

Early Changes in Soil Metabolic Diversity and Bacterial Community Structure in Sugarcane under Two Harvest Management Systems

Preharvest burning is widely used in Brazil for sugarcane cropping. However, due to environmental restrictions, harvest without burning is becoming the predominant option. Consequently, changes in the microbial community are expected from crop residue accumulation on the soil surface, as well as alterations in soil metabolic diversity as of the first harvest. Because biological properties respond quickly and can be used to monitor environmental changes, we evaluated soil metabolic diversity and bacterial community structure after the first harvest under sugarcane management without burning compared to management with preharvest burning. Soil samples were collected under three sugarcane varieties (SP813250, SP801842 and RB72454) and two harvest management systems (without and with preharvest burning). Microbial biomass C (MBC), carbon (C) substrate utilization profiles, bacterial community structure (based on profiles of 16S rRNA gene amplicons), and soil chemical properties were determined. MBC was not different among the treatments. C-substrate utilization and metabolic diversity were lower in soil without burning, except for the evenness index of C-substrate utilization. Soil samples under the variety SP801842 showed the greatest changes in substrate utilization and metabolic diversity, but showed no differences in bacterial community structure, regardless of the harvest management system. In conclusion, combined analysis of soil chemical and microbiological data can detect early changes in microbial metabolic capacity and diversity, with lower values in management without burning. However, after the first harvest, there were no changes in the soil bacterial community structure detected by PCR-DGGE under the sugarcane variety SP801842. Therefore, the metabolic profile is a more sensitive indicator of early changes in the soil microbial community caused by the harvest management system.

Rapid quantification of mutant fitness in diverse bacteria by sequencing randomly bar-coded transposons.

ABSTRACTTransposon mutagenesis with next-generation sequencing (TnSeq) is a powerful approach to annotate gene function in bacteria, but existing protocols for TnSeq require laborious preparation of every sample before sequencing. Thus, the existing protocols are not amenable to the throughput necessary to identify phenotypes and functions for the majority of genes in diverse bacteria. Here, we present a method, random bar code transposon-site sequencing (RB-TnSeq), which increases the throughput of mutant fitness profiling by incorporating random DNA bar codes into Tn5 and mariner transposons and by using bar code sequencing (BarSeq) to assay mutant fitness. RB-TnSeq can be used with any transposon, and TnSeq is performed once per organism instead of once per sample. Each BarSeq assay requires only a simple PCR, and 48 to 96 samples can be sequenced on one lane of an Illumina HiSeq system. We demonstrate the reproducibility and biological significance of RB-TnSeq with Escherichia coli, Phaeobacter inhibens, Pseudomonas stutzeri, Shewanella amazonensis, and Shewanella oneidensis. To demonstrate the increased throughput of RB-TnSeq, we performed 387 successful genome-wide mutant fitness assays representing 130 different bacterium-carbon source combinations and identified 5,196 genes with significant phenotypes across the five bacteria. In P. inhibens, we used our mutant fitness data to identify genes important for the utilization of diverse carbon substrates, including a putative d-mannose isomerase that is required for mannitol catabolism. RB-TnSeq will enable the cost-effective functional annotation of diverse bacteria using mutant fitness profiling.

Variable Responses to Carbon Utilization between Planktonic and Biofilm Cells of a Human Carrier Strain of Salmonella enterica Serovar Typhi

Salmonella enterica serovar Typhi (S. Typhi) is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC) biofilm inoculator (96-well peg lid) and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates) and D-threonine (amino acid) were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among different S. Typhi strains has suggested the possible involvement of various metabolic pathways that might be related to the virulence and pathogenesis of this host-restricted human pathogen. The data serve as a caveat for future in-vivo studies to investigate the carbon metabolic activity to the pathogenesis of S. Typhi.

The effects of sediment depth and oxygen concentration on the use of organic matter: An experimental study using an infiltration sediment tank

Water flowing through hyporheic river sediments or artificial recharge facilities promotes the development of microbial communities with sediment depth. We performed an 83-day mesocosm infiltration experiment, to study how microbial functions (e.g., extracellular enzyme activities and carbon substrate utilization) are affected by sediment depth (up to 50cm) and different oxygen concentrations. Results indicated that surface sediment layers were mainly colonized by microorganisms capable of using a wide range of substrates (although they preferred to degrade carbon polymeric compounds, as indicated by the higher β-glucosidase activity). In contrast, at a depth of 50cm, the microbial community became specialized in using fewer carbon substrates, showing decreased functional richness and diversity. At this depth, microorganisms picked nitrogenous compounds, including amino acids and carboxyl acids. After the 83-day experiment, the sediment at the bottom of the tank became anoxic, inhibiting phosphatase activity. Coexistence of aerobic and anaerobic communities, promoted by greater physicochemical heterogeneity, was also observed in deeper sediments. The presence of specific metabolic fingerprints under oxic and anoxic conditions indicated that the microbial community was adapted to use organic matter under different oxygen conditions. Overall the heterogeneity of oxygen concentrations with depth and in time would influence organic matter metabolism in the sediment tank.

Pichia pastoris regulates its gene-specific response to different carbon sources at the transcriptional, rather than the translational, level.

BackgroundThe methylotrophic, Crabtree-negative yeast Pichia pastoris is widely used as a heterologous protein production host. Strong inducible promoters derived from methanol utilization genes or constitutive glycolytic promoters are typically used to drive gene expression. Notably, genes involved in methanol utilization are not only repressed by the presence of glucose, but also by glycerol. This unusual regulatory behavior prompted us to study the regulation of carbon substrate utilization in different bioprocess conditions on a genome wide scale.ResultsWe performed microarray analysis on the total mRNA population as well as mRNA that had been fractionated according to ribosome occupancy. Translationally quiescent mRNAs were defined as being associated with single ribosomes (monosomes) and highly-translated mRNAs with multiple ribosomes (polysomes). We found that despite their lower growth rates, global translation was most active in methanol-grown P. pastoris cells, followed by excess glycerol- or glucose-grown cells. Transcript-specific translational responses were found to be minimal, while extensive transcriptional regulation was observed for cells grown on different carbon sources. Due to their respiratory metabolism, cells grown in excess glucose or glycerol had very similar expression profiles. Genes subject to glucose repression were mainly involved in the metabolism of alternative carbon sources including the control of glycerol uptake and metabolism. Peroxisomal and methanol utilization genes were confirmed to be subject to carbon substrate repression in excess glucose or glycerol, but were found to be strongly de-repressed in limiting glucose-conditions (as are often applied in fed batch cultivations) in addition to induction by methanol.ConclusionsP. pastoris cells grown in excess glycerol or glucose have similar transcript profiles in contrast to S. cerevisiae cells, in which the transcriptional response to these carbon sources is very different. The main response to different growth conditions in P. pastoris is transcriptional; translational regulation was not transcript-specific. The high proportion of mRNAs associated with polysomes in methanol-grown cells is a major finding of this study; it reveals that high productivity during methanol induction is directly linked to the growth condition and not only to promoter strength.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1393-8) contains supplementary material, which is available to authorized users.

Effects of nitrogen addition on soil microbes and their implications for soil C emission in the Gurbantunggut Desert, center of the Eurasian Continent

Nitrogen (N) deposition can influence carbon cycling of terrestrial ecosystems. However, a general recognition of how soil microorganisms respond to increasing N deposition is not yet reached. We explored soil microbial responses to two levels of N addition (2.5 and 5gNm−2yr−1) in interplant soil and beneath shrubs of Haloxylon ammodendron and their consequences to soil respiration in the Gurbantunggut Desert, northwestern China from 2011 to 2013. Microbial biomass and respiration were significantly higher beneath H. ammodendron than in interplant soil. The responses of microbial biomass carbon (MBC) and microbial respiration (MR) showed opposite responses to N addition in interplant and beneath H. ammodendron. N addition slightly increased MBC and MR in interplant soil and decreased them beneath H. ammodendron, with a significant inhibition only in 2012. N addition had no impacts on the total microbial physiological activity, but N addition decreased the labile carbon substrate utilization beneath H. ammodendron when N addition level was high. Phospholipid fatty acid (PLFA) analysis showed that N addition did not alter the soil microbial community structure as evidenced by the similar ratios of fungal to bacterial PLFAs and gram-negative to gram-positive bacterial PLFAs. Microbial biomass and respiration showed close correlations with soil water content and dissolved carbon, and they were independent of soil inorganic nitrogen across three years. Our study suggests that N addition effects on soil microorganisms and carbon emission are dependent on the respiratory substrates and water availability in the desert ecosystem.

Microbial carbon metabolism associated with electrogenic sulphur oxidation in coastal sediments.

Recently, a novel electrogenic type of sulphur oxidation was documented in marine sediments, whereby filamentous cable bacteria (Desulfobulbaceae) are mediating electron transport over cm-scale distances. These cable bacteria are capable of developing an extensive network within days, implying a highly efficient carbon acquisition strategy. Presently, the carbon metabolism of cable bacteria is unknown, and hence we adopted a multidisciplinary approach to study the carbon substrate utilization of both cable bacteria and associated microbial community in sediment incubations. Fluorescence in situ hybridization showed rapid downward growth of cable bacteria, concomitant with high rates of electrogenic sulphur oxidation, as quantified by microelectrode profiling. We studied heterotrophy and autotrophy by following 13C-propionate and -bicarbonate incorporation into bacterial fatty acids. This biomarker analysis showed that propionate uptake was limited to fatty acid signatures typical for the genus Desulfobulbus. The nanoscale secondary ion mass spectrometry analysis confirmed heterotrophic rather than autotrophic growth of cable bacteria. Still, high bicarbonate uptake was observed in concert with the development of cable bacteria. Clone libraries of 16S complementary DNA showed numerous sequences associated to chemoautotrophic sulphur-oxidizing Epsilon- and Gammaproteobacteria, whereas 13C-bicarbonate biomarker labelling suggested that these sulphur-oxidizing bacteria were active far below the oxygen penetration. A targeted manipulation experiment demonstrated that chemoautotrophic carbon fixation was tightly linked to the heterotrophic activity of the cable bacteria down to cm depth. Overall, the results suggest that electrogenic sulphur oxidation is performed by a microbial consortium, consisting of chemoorganotrophic cable bacteria and chemolithoautotrophic Epsilon- and Gammaproteobacteria. The metabolic linkage between these two groups is presently unknown and needs further study.

Cyanobacteria drive community composition and functionality in rock-soil interface communities.

Most ecological research on hypoliths, significant primary producers in hyperarid deserts, has focused on the diversity of individual groups of microbes (i.e. bacteria). However, microbial communities are inherently complex, and the interactions between cyanobacteria, heterotrophic bacteria, protista and metazoa are likely to be very important for ecosystem functioning. Cyanobacterial and heterotrophic bacterial communities were analysed by pyrosequencing, while metazoan and protistan communities were assessed by T-RFLP analysis. Microbial functionality was estimated using carbon substrate utilization. Cyanobacterial community composition was significant in shaping community structure and function in hypoliths. Ecological network analysis showed that most significant co-occurrences were positive, representing potential synergistic interactions. There were several highly interconnected associations (modules), and specific cyanobacteria were important in driving the modular structure of hypolithic networks. Together, our results suggest that hypolithic cyanobacteria have strong effects on higher trophic levels and ecosystem functioning.

Microbial production of lactic acid

Lactic acid is an important commodity chemical having a wide range of applications. Microbial production effectively competes with chemical synthesis methods because biochemical synthesis permits the generation of either one of the two enantiomers with high optical purity at high yield and titer, a result which is particularly beneficial for the production of poly(lactic acid) polymers having specific properties. The commercial viability of microbial lactic acid production relies on utilization of inexpensive carbon substrates derived from agricultural or waste resources. Therefore, optimal lactic acid formation requires an understanding and engineering of both the competing pathways involved in carbohydrate metabolism, as well as pathways leading to potential by-products which both affect product yield. Recent research leverages those biochemical pathways, while researchers also continue to seek strains with improved tolerance and ability to perform under desirable industrial conditions, for example, of pH and temperature.

Metabolic variation in natural populations of wild yeast.

Ecological diversification depends on the extent of genetic variation and on the pattern of covariation with respect to ecological opportunities. We investigated the pattern of utilization of carbon substrates in wild populations of budding yeast Saccharomyces paradoxus. All isolates grew well on a core diet of about 10 substrates, and most were also able to grow on a much larger ancillary diet comprising most of the 190 substrates we tested. There was substantial genetic variation within each population for some substrates. We found geographical variation of substrate use at continental, regional, and local scales. Isolates from Europe and North America could be distinguished on the basis of the pattern of yield across substrates. Two geographical races at the North American sites also differed in the pattern of substrate utilization. Substrate utilization patterns were also geographically correlated at local spatial scales. Pairwise genetic correlations between substrates were predominantly positive, reflecting overall variation in metabolic performance, but there was a consistent negative correlation between categories of substrates in two cases: between the core diet and the ancillary diet, and between pentose and hexose sugars. Such negative correlations in the utilization of substrate from different categories may indicate either intrinsic physiological trade-offs for the uptake and utilization of substrates from different categories, or the accumulation of conditionally neutral mutations. Divergence in substrate use accompanies genetic divergence at all spatial scales in S. paradoxus and may contribute to race formation and speciation.

Effects of cultivation of OsrHSA transgenic rice on functional diversity of microbial communities in the soil rhizosphere

With the widespread cultivation of transgenic crops, there is increasing concern about unintended effects of these crops on soil environmental quality. In this study, we used the Biolog method and ELISA to evaluate the possible effects of OsrHSA transgenic rice on soil microbial utilization of carbon substrates under field conditions. There were no significant differences in average well-color development (AWCD) values, Shannon–Wiener diversity index (H), Simpson dominance indices (D) and Shannon–Wiener evenness indices (E) of microbial communities in rhizosphere soils at eight samplings between OsrHSA transgenic rice and its non-transgenic counterpart. The main carbon sources utilized by soil microbes were carbohydrates, carboxylic acids, amino acids and polymers. The types, capacities and patterns of carbon source utilization by microbial communities in rhizosphere soils were similar throughout the detection period. We detected no OsrHSA protein in the roots of OsrHSA transgenic rice. We concluded that OsrHSA transgenic rice and the rHSA protein it produced did not alter the functional diversity of microbial communities in the rhizosphere.