Structure-based method for analyzing protein–protein interfaces

Hydrogen bond, hydrophobic and vdW interactions are the three major non-covalent interactions at protein–protein interfaces. We have developed a method that uses only these properties to describe interactions between proteins, which can qualitatively estimate the individual contribution of each interfacial residue to the binding and gives the results in a graphic display way. This method has been applied to analyze alanine mutation data at protein–protein interfaces. A dataset containing 13 protein–protein complexes with 250 alanine mutations of interfacial residues has been tested. For the 75 hot-spot residues (G1.5 kcal mol^-1), 66 can be predicted correctly with a success rate of 88%. In order to test the tolerance of this method to conformational changes upon binding, we utilize a set of 26 complexes with one or both of their components available in the unbound form. The difference of key residues exported by the program is 11% between the results using complexed proteins and those from unbound ones. As this method gives the characteristics of the binding partner for a particular protein, in-depth studies on protein–protein recognition can be carried out. Furthermore, this method can be used to compare the difference between protein–protein interactions and look for correlated mutation. Figure Key interaction grids at the interface between barnase and barstar. Key interaction grid for barnase and barstar are presented in one figure according to their coordinates. In order to distinguish the two proteins, different icons were assigned. Crosses represent key grids for barstar and dots represent key grids for barnase. The four residues in ball and stick are Asp40 in barstar and Arg83, Arg87, His102 in barnase.     

Characterization of eubacterial and archaeal community diversity in the pit mud of Chinese Luzhou-flavor liquor by nested PCR–DGGE

The aim of this study was to investigate and compare the microbial community structures of eubacteria and archaea in the pit mud of Chinese Luzhou-flavor liquor from the wall (C_w) and bottom (C_b) of cellar through nested PCR–denaturing gradient gel electrophoresis (DGGE). The Shannon–Wiener index (H) calculated from the DGGE profiles showed that the community diversities of eubacteria and archaea in samples from C_b were almost higher than that from C_w. In addition, cluster analysis of the DGGE profiles revealed that some differences were found in the microbial community structure in samples from different locations. The closely relative microorganisms of all eubacterial 16S rRNA gene sequences fell into four phyla (Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria), including 12 genera and 2 uncultured eubacteria. Moreover, 37.1 % eubacteria were affiliated with Clostridium. Particularly, genus Acinetobacter was absent in all samples from C_b but present in all samples from C_w. The closely relative microorganisms of all archaeal 16S rRNA gene sequences fell into four genera, which included Methanobrevibacter, Methanoculleus, Methanobacterium and Methanosaeta, while the dominant archaea in samples from C_w and C_b were similar. Results presented in this study provide further understanding of the spatial differences in microbial community structure in the pit mud, and is of great importance for the production and quality improvement of Luzhou-flavor liquor.     

A method for measuring depth-integrated community metabolism in experimental planktonic–benthic ecosystems

We developed incubation chambers of clear acrylic extending the full depth of the water column to measure in situ depth-integrated community metabolism in experimental planktonic–benthic ecosystems. A magnetic stirring apparatus at mid-depth in each chamber prevented stratification and maintained a vertical turnover time that roughly matched turnover in the experimental ecosystems. We used the chambers in light–dark experiments to partition the depth-integrated photosynthesis and respiration occurring in the experimental ecosystems among water column, wall periphyton, and benthic communities. Community metabolism measured with the chambers was positively related to the chlorophyll-a concentrations of the water column and wall periphyton communities. We also found close agreement between the summation of partitioned community metabolism measured in the chambers and an independent measure of total ecosystem metabolism in the experimental ecosystems. Incubations in chambers and in light–dark bottles revealed similar patterns of water column and wall periphyton community metabolism, but magnitudes differed. In addition to application in experimental ecosystems, this incubation chamber technique may provide an efficient and realistic measure of water column and benthic metabolism and nutrient uptake in lakes and estuaries.     

Changes in the bacterial community in the fermentation process of kôso,a Japanese sugar-vegetable fermented beverage

Kôso is a Japanese fermented beverage made with over 20 kinds of vegetables, mushrooms, and sugars. The changes in the bacterial population of kôso during fermentation at 25 °C over a period of 10 days were studied using 454 pyrosequencing of the 16S rRNA gene. The analysis detected 224 operational taxonomic units (OTUs) clustered from 8 DNA samples collected on days 0, 3, 7, and 10 from two fermentation batches. Proteobacteria were the dominant phylum in the starting community, but were replaced by Firmicutes within three days. Seventy-eight genera were identified from the 224 OTUs, in which Bifidobacterium, Leuconostoc, Lactococcus, and Lactobacillus dominated, accounting for over 96% of the total bacterial population after three days’ fermentation. UniFrac–Principal Coordinate Analysis of longitudinal fermented samples revealed dramatic changes in the bacterial community in kôso, resulting in significantly low diversity at the end of fermentation as compared with the complex starting community.     

Inputs of seabird guano alter microbial growth,community composition and the phytoplankton–bacterial interactions in a coastal system

Seabird guano enters coastal waters providing bioavailable substrates for microbial plankton, but their role in marine ecosystem functioning remains poorly understood. Two concentrations of the water soluble fraction (WSF) of gull guano were added to different natural microbial communities collected in surface waters from the Ría de Vigo (NW Spain) in spring, summer, and winter. Samples were incubated with or without antibiotics (to block bacterial activity) to test whether gull guano stimulated phytoplankton and bacterial growth, caused changes in taxonomic composition, and altered phytoplankton–bacteria interactions. Alteromonadales, Sphingobacteriales, Verrucomicrobia and diatoms were generally stimulated by guano. Chlorophyll a (Chl a) concentration and bacterial abundance significantly increased after additions independently of the initial ambient nutrient concentrations. Our study demonstrates, for the first time, that the addition of guano altered the phytoplankton–bacteria interaction index from neutral (i.e. phytoplankton growth was not affected by bacterial activity) to positive (i.e. phytoplankton growth was stimulated by bacterial activity) in the low-nutrient environment occurring in spring. In contrast, when environmental nutrient concentrations were high, the interaction index changed from positive to neutral after guano additions, suggesting the presence of some secondary metabolite in the guano that is needed for phytoplankton growth, which would otherwise be supplied by bacteria.     

Participation of algal–bacterial community in the formation of modern stromatolites in Cock Soda Lake,Altai Region

Modern dolomite stromatolites are found in Cock Soda Lake (Kulunda Steppe) at a salinity of 100–200 g/L and pH of 10. The mineralogical analysis has revealed the presence in the stromatolites of Ca?Mg-carbonates of various compositions. The organisms–edificators of the phototrophic community developing in the lake are determined. They are identified as a part of the mineralized biota (cyanobacteria, bacteria, and eukaryotic alga Ctenocladus circinnatus). Morphological and ultrastructural features of exopolysaccharides secreted by cyanobacteria and bacteria dominant in the phototrophic community are characterized. It is shown that polysaccharides secreted primarily by cyanobacteria have the utmost importance for the formation of stromatolites in Petukhovskoe Soda Lake.     

Analysis of bacterial communities on aging flue-cured tobacco leaves by 16S rDNA PCR–DGGE technology

Many microorganisms, growing on aging flue-cured tobacco leaves, play a part in its fermentation process. These microflora were identified and described by culture-dependent methods earlier. In this study we report the identity of the microflora growing on the tobacco leaf surface by employing culture-independent methods. We have amplified microbial 16S rDNA sequences directly from the leaf surface and used denaturing gradient gel electrophoresis (DGGE) to identify bacterial community on the tobacco leaves. Our culture-independent methods for the study of microbial community on tobacco leaves showed that microbial community structures on leaves of variety Zhongyan 100, NC89 and Zhongyan 101 were similar between 0 and 6 months aging, and between 9 and 12 months aging, while the similarity is low between 0 and 6, and between 9 and 12 months aging, respectively. There were certain similarities of bacterial communities (similarity up to 63%) among the three tobacco varieties for 0 to 6 months aging. Five dominant 16S rDNA DGGE bands A, B, C, D and E were isolated, cloned, and sequenced. They were most similar to two cultured microbial species Bacteriovorax sp. EPC3, Bacillus megaterium, and three uncultured microbial species, respectively.     

A preliminary fish assemblage index for a transitional river–reservoir system in southeastern Brazil

Large river–reservoir systems are some of the most difficult aquatic ecosystems to assess because: (1) they typically lack minimally disturbed reference sites; (2) the reservoirs are not natural systems to begin with; and (3) reservoirs with high exchange rates are transitional systems between rivers and lakes. These features are further complicated in Brazil where fish species taxonomy is incomplete (let alone fully described ecologically), where waters naturally have high organic and thermal loadings, and where dams and reservoirs provide most of the nation's electricity and water supplies. As a first step towards generating a biological tool for assessing the effects of reservoirs on rivers, we developed a preliminary River–Reservoir Fish Assemblage Index (RRFAI) in a transitional river–reservoir system in southeastern Brazil. To do so, we gill-netted fish monthly between October 2006 and September 2007 (excluding May and July 2007) immediately upriver of the reservoir, in the upper reservoir, in the lower reservoir, and immediately downriver of the reservoir. In developing our RRFAI we sought fish assemblage metrics to represent ecological characteristics including richness, habitat, trophic, tolerance, and resilience guilds. Despite clear differences in fish assemblage composition between river and reservoir sites, we found 9 metrics common to both systems that were nonredundant and had low sampling variability (number of native species, number of characiform species, number of siluriform species, % omnivorous individuals, % invertivorous individuals, % non-native carnivorous individuals, % intolerant individuals, % tolerant individuals, number of tolerant species). Fish assemblage condition was significantly and consistently lower in the lower reservoir. There was no significant difference between the dry and wet season in RRFAI scores, suggesting that a single season sample should usually suffice. Further research is needed along distinct disturbance gradients in multiple river–reservoir systems in Brazil to confirm the sensitivity of our preliminary RRFAI for assessing the physical and chemical habitat disturbances common to such systems.     

TRUNCATULIX – a data warehouse for the legume community

Background  

Databases for either sequence, annotation, or microarray experiments data are extremely beneficial to the research community, as they centrally gather information from experiments performed by different scientists. However, data from different sources develop their full capacities only when combined. The idea of a data warehouse directly adresses this problem and solves it by integrating all required data into one single database – hence there are already many data warehouses available to genetics. For the model legume Medicago truncatula, there is currently no such single data warehouse that integrates all freely available gene sequences, the corresponding gene expression data, and annotation information. Thus, we created the data warehouse TRUNCATULIX, an integrative database of Medicago truncatula sequence and expression data.     

Beyond the continuum: a multi-dimensional phase space for neutral–niche community assembly

Neutral and niche processes are generally considered to interact in natural communities along a continuum, exhibiting community patterns bounded by pure neutral and pure niche processes. The continuum concept uses niche separation, an attribute of the community, to test the hypothesis that communities are bounded by pure niche or pure neutral conditions. It does not accommodate interactions via feedback between processes and the environment. By contrast, we introduce the Community Assembly Phase Space (CAPS), a multi-dimensional space that uses community processes (such as dispersal and niche selection) to define the limiting neutral and niche conditions and to test the continuum hypothesis. We compare the outputs of modelled communities in a heterogeneous landscape, assembled by pure neutral, pure niche and composite processes. Differences in patterns under different combinations of processes in CAPS reveal hidden complexity in neutral–niche community dynamics. The neutral–niche continuum only holds for strong dispersal limitation and niche separation. For weaker dispersal limitation and niche separation, neutral and niche processes amplify each other via feedback with the environment. This generates patterns that lie well beyond those predicted by a continuum. Inferences drawn from patterns about community assembly processes can therefore be misguided when based on the continuum perspective. CAPS also demonstrates the complementary information value of different patterns for inferring community processes and captures the complexity of community assembly. It provides a general tool for studying the processes structuring communities and can be applied to address a range of questions in community and metacommunity ecology.     

Plant–pollinator interactions in a Mexican Acacia community

Competition for pollination is thought to be an important factor structuring flowering in many plant communities, particularly among plant taxa with morphologically similar and easily accessible flowers. We examined the potential for heterospecific pollen transfer (HPT) in a community of four Acacia species in a highly seasonal tropical habitat in Mexico. Partitioning of pollen flow among sympatric species appears to be achieved, in part, through segregation of flowering in seasonal time, and interspecific differences in pollinator guilds. However, two coflowering species (Acacia macracantha and Acacia angustissima) shared multiple flower visitors, raising the possibility of HPT. Each of these coflowering species showed high intraspecific daily synchrony in pollen release, but dehisce at different times of day. Pollinators rapidly harvested available pollen from one species before abandoning it to visit the flowers of the second later in the day. The activity of shared pollinators, predominantly bees, is thus structured throughout the day, and potential for HPT reduced. Suggestive evidence in favour of a resource partitioning explanation for this pattern is provided by the fact that A. macracantha showed significantly greater intraspecific synchrony when coflowering with a potential competitor (A. angustissima) than when flowering alone. We discuss our results in light of previous work on coflowering acacia assemblages in Tanzania and Australia. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.

Differentiated free-living and sediment-attached bacterial community structure inside and outside denitrification hotspots in the river–groundwater interface

This study assessed the functional significance of attached and free-living bacterial communities involved in the process of denitrification in a shallow aquifer of a riparian zone (Garonne River, SW France). Denitrification enzyme activity (DEA), bacterial density (BD) and bacterial community composition (BCC) were measured in two aquifer compartments: the groundwater and the sandy fraction of the sediment deposit. Samples were collected in wells located inside (IHD) and outside (OHD) identified hotspots of denitrification. Despite high BD values (up to 1.14 × 10¹² cells m⁻³), DEA was not detected in the water compartment (< 0.32 mg N–N₂O m⁻³ d⁻¹). The sandy fraction showed detectable DEA (up to 1,389 mg N–N₂O m⁻³ d⁻¹) and, consistent with BD pattern, higher DEA values were measured in IHD zones than in OHD zones. The BCC assessed by 16S rDNA polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) partly supported this result: attached and free-living communities were significantly different (< 30% similarity) but patterns of BCC did not cluster according to IHD and OHD zones. Targeting the denitrifying communities by means of a culture enrichment step prior to 16S rDNA PCR-DGGE showed that the free-living and sediment attached communities differed. Most sequences obtained from DGGE profiles of denitrifying communities were affiliated to Proteobacteria and showed low genetic distance with taxa that have already been detected in aquifers (e.g., Azoarcus sp., Acidovorax sp. and Pseudomonas spp.). This study confirms that in the aquifer the sediment-attached fraction exhibits different functions (DEA) from free-living communities and suggests that this functional difference is related to the communities’ structure.     

Development of LNA oligonucleotide–PCR clamping technique in investigating the community structures of plant-associated bacteria

Simultaneous extraction of plant organelle (mitochondria and plastid) genes during the DNA extraction step is major limitation in investigating the community structures of plant-associated bacteria. Although locked nucleic acid (LNA) oligonucleotides was designed to selectively amplify the bacterial small subunit rRNA genes by applying the PCR clamping technique, those for plastids were applicable only for particular plants, while those for mitochondria were available throughout most plants. To widen the applicable range, new LNA oligonucleotides specific for plastids were designed, and the efficacy was investigated. PCR without LNA oligonucleotides predominantly amplified the organelle genes, while bacterial genes were predominantly observed in having applied the LNA oligonucleotides. Denaturing gradient gel electrophoresis (DGGE) analysis displayed additional bacterial DGGE bands, the amplicons of which were prepared using the LNA oligonucleotides. Thus, new designed LNA oligonucleotides specific for plastids were effective and have widened the scope in investigating the community structures of plant-associated bacteria.     

Is there a role for replication fork asymmetry in the distribution of genes in bacterial genomes?

Replication generates bacterial chromosomes with strands that differ in the number of genes and base composition. It has been suggested that in bacteria such as Bacillus subtilis, PolC is responsible for the synthesis of the leading strand and DnaE for the lagging strand, whereas in many other bacteria DnaE is responsible for the synthesis of both strands. Here, I show that the possession of PolC correlates with leading strands that contain an average of 78% of genes compared with 58% for genomes that do not contain PolC. This suggests that asymmetrical replication forks could have a major role in defining and constraining the structure of the bacterial chromosome. The presence of PolC is not correlated with compositional strand bias, suggesting that the two biases result from different types of structural asymmetry.     

Phosphate-limited ocean regions select for bacterial populations enriched in the carbon–phosphorus lyase pathway for phosphonate degradation

In tropical and subtropical oceanic surface waters phosphate scarcity can limit microbial productivity. However, these environments also have bioavailable forms of phosphorus incorporated into dissolved organic matter (DOM) that microbes with the necessary transport and hydrolysis metabolic pathways can access to supplement their phosphorus requirements. In this study we evaluated how the environment shapes the abundance and taxonomic distribution of the bacterial carbon–phosphorus (C–P) lyase pathway, an enzyme complex evolved to extract phosphate from phosphonates. Phosphonates are organophosphorus compounds characterized by a highly stable C–P bond and are enriched in marine DOM. Similar to other known bacterial adaptions to low phosphate environments, C–P lyase was found to become more prevalent as phosphate concentrations decreased. C–P lyase was particularly enriched in the Mediterranean Sea and North Atlantic Ocean, two regions that feature sustained periods of phosphate depletion. In these regions, C–P lyase was prevalent in several lineages of Alphaproteobacteria (Pelagibacter, SAR116, Roseobacter and Rhodospirillales), Gammaproteobacteria, and Actinobacteria. The global scope of this analysis supports previous studies that infer phosphonate catabolism via C–P lyase is an important adaptive strategy implemented by bacteria to alleviate phosphate limitation and expands the known geographic extent and taxonomic affiliation of this metabolic pathway in the ocean.     

Characteristics of the mean radiant temperature in high latitude cities—implications for sensitive climate planning applications

Knowledge of how the mean radiant temperature (T _mrt ) is affected by factors such as location, climate and urban setting contributes to the practice of climate sensitive planning. This paper examines how T _mrt varies within an urban setting and how it is influenced by cloudiness. In addition, variations of T _mrt in three high latitude cities are investigated in order to analyse the impact of geographical context and climate conditions. Results showed large spatial variations between sunlit and shaded areas during clear weather conditions, with the highest values of T _mrt close to sunlit walls and the lowest values in the areas shaded by buildings and vegetation. As cloudiness increases, the spatial pattern is altered and the differences are reduced. The highest T _mrt under cloudy conditions is instead found in open areas where the proportion of shortwave diffuse radiation from the sky vault is high. A regional comparison between three Swedish coastal cities showed that T _mrt during summer is similar regardless of latitudinal location. On the other hand, large differences in T _mrt during winter were found. Shadows, both from buildings and vegetation are the most effective measure to reduce extreme values of T _mrt . However, extensive areas of shadow are usually not desired within outdoor urban environments at high latitude cities. One solution is to create diverse outdoor urban spaces in terms of shadow and also ventilation. This would provide individuals with access to a choice of thermal environments which they can use to assist their thermal regulation, based on personal needs and desires.     

Trichoderma affects the physiochemical characteristics and bacterial community composition of saline–alkaline maize rhizosphere soils in the cold-region of Heilongjiang Province

Plant and Soil - Heilongjiang Province in China experiences extremely cold weather, and its soil is saline–alkaline. Salinity and alkalinity severely restrain the growth of maize. Although...