Genetic diversity of Rhodopirellula strains.

Rhodopirellula baltica SH1(T) is a marine planctomycete with 7,325 genes in its genome. Ten strains of the genus Rhodopirellula were studied in whole genome microarray experiments to assess the extent of their genetic relatedness to R. baltica SH1(T). DNA of strains which were previously affiliated with the species R. baltica (OTU A) hybridized with 3,645-5,728 genes of the type strain on the microarray. Strains SH398 and 6C (OTU B), representing a closely related species with an average nucleotide identity of 88 %, showed less hybridization signals: 1,816 and 3,302 genes gave a hybridization signal, respectively. Comparative genomics of eight permanent draft genomes revealed the presence of over 4,000 proteins common in R. baltica SH1(T) and strains of OTU A or B. The genus Rhodopirellula is characterized by large genomes, with over 7,000 genes per genome and a core genome of around 3000 genes. Individual Rhodopirellula strains have a large portion of strain-specific genes.

Expression of sulfatases in Rhodopirellula baltica and the diversity of sulfatases in the genus Rhodopirellula.

The whole genome sequence of Rhodopirellula baltica SH1(T), published nearly 10years ago, already revealed a high amount of sulfatase genes. So far, little is known about the diversity and potential functions mediated by sulfatases in Planctomycetes. We combined in vivo and in silico techniques to gain insights into the ecophysiology of planktomycetal sulfatases. Comparative genomics of nine recently sequenced Rhodopirellula strains detected 1120 open reading frames annotated as sulfatases (Enzyme Commission number (EC) 3.1.6.*). These were clustered into 173 groups of orthologous and paralogous genes. To analyze the functional aspects, 708 sulfatase protein sequences from these strains were aligned with 67 sulfatase reference sequences of reviewed functionality. Our analysis yielded 22 major similarity clusters, but only five of these clusters contained Rhodopirellula sequences homologous to reference sequences, indicating a surprisingly high diversity. Exemplarily, R. baltica SH1(T) was grown on different sulfated polysaccharides, chondroitin sulfate, λ-carrageenan and fucoidan. Subsequent gene expression analyses using whole genome microarrays revealed distinct sulfatase expression profiles based on substrates tested. This might be indicative for a high structural diversity of sulfated polysaccharides as potential substrates. The pattern of sulfatases in individual planctomycete species may reflect ecological niche adaptation.

Substrate-controlled succession of marine bacterioplankton populations induced by a phytoplankton bloom.

Phytoplankton blooms characterize temperate ocean margin zones in spring. We investigated the bacterioplankton response to a diatom bloom in the North Sea and observed a dynamic succession of populations at genus-level resolution. Taxonomically distinct expressions of carbohydrate-active enzymes (transporters; in particular, TonB-dependent transporters) and phosphate acquisition strategies were found, indicating that distinct populations of Bacteroidetes, Gammaproteobacteria, and Alphaproteobacteria are specialized for successive decomposition of algal-derived organic matter. Our results suggest that algal substrate availability provided a series of ecological niches in which specialized populations could bloom. This reveals how planktonic species, despite their seemingly homogeneous habitat, can evade extinction by direct competition.

Practical application of self-organizing maps to interrelate biodiversity and functional data in NGS-based metagenomics.

Next-generation sequencing (NGS) technologies have enabled the application of broad-scale sequencing in microbial biodiversity and metagenome studies. Biodiversity is usually targeted by classifying 16S ribosomal RNA genes, while metagenomic approaches target metabolic genes. However, both approaches remain isolated, as long as the taxonomic and functional information cannot be interrelated. Techniques like self-organizing maps (SOMs) have been applied to cluster metagenomes into taxon-specific bins in order to link biodiversity with functions, but have not been applied to broad-scale NGS-based metagenomics yet. Here, we provide a novel implementation, demonstrate its potential and practicability, and provide a web-based service for public usage. Evaluation with published data sets mimicking varyingly complex habitats resulted into classification specificities and sensitivities of close to 100% to above 90% from phylum to genus level for assemblies exceeding 8 kb for low and medium complexity data. When applied to five real-world metagenomes of medium complexity from direct pyrosequencing of marine subsurface waters, classifications of assemblies above 2.5 kb were in good agreement with fluorescence in situ hybridizations, indicating that biodiversity was mostly retained within the metagenomes, and confirming high classification specificities. This was validated by two protein-based classifications (PBCs) methods. SOMs were able to retrieve the relevant taxa down to the genus level, while surpassing PBCs in resolution. In order to make the approach accessible to a broad audience, we implemented a feature-rich web-based SOM application named TaxSOM, which is freely available at http://www.megx.net/toolbox/taxsom. TaxSOM can classify reads or assemblies exceeding 2.5 kb with high accuracy and thus assists in linking biodiversity and functions in metagenome studies, which is a precondition to study microbial ecology in a holistic fashion.

Life cycle analysis of the model organism Rhodopirellula baltica SH 1(T) by transcriptome studies.

The marine organism Rhodopirellula baltica is a representative of the globally distributed phylum Planctomycetes whose members exhibit an intriguing lifestyle and cell morphology. The analysis of R. baltica's genome has revealed many biotechnologically promising features including a set of unique sulfatases and C1-metabolism genes. Salt resistance and the potential for adhesion in the adult phase of the cell cycle were observed during cultivation. To promote the understanding of this model organism and to specify the functions of potentially useful genes, gene expression throughout a growth curve was monitored using a whole genome microarray approach. Transcriptional profiling suggests that a large number of hypothetical proteins are active within the cell cycle and in the formation of the different cell morphologies. Numerous genes with potential biotechnological applications were found to be differentially regulated, revealing further characteristics of their functions and regulation mechanisms. More specifically, the experiments shed light on the expression patterns of genes belonging to the organism's general stress response, those involved in the reorganization of its genome and those effecting morphological changes. These transcriptomic results contribute to a better understanding of thus far unknown molecular elements of cell biology. Further, they pave the way for the biotechnological exploitation of R. baltica's distinctive metabolic features as a step towards sourcing the phylum Planctomycetes at large.

Transcriptional response of the model planctomycete Rhodopirellula baltica SH1(T) to changing environmental conditions.

BACKGROUND: The marine model organism Rhodopirellula baltica SH1(T) was the first Planctomycete to have its genome completely sequenced. The genome analysis predicted a complex lifestyle and a variety of genetic opportunities to adapt to the marine environment. Its adaptation to environmental stressors was studied by transcriptional profiling using a whole genome microarray. RESULTS: Stress responses to salinity and temperature shifts were monitored in time series experiments. Chemostat cultures grown in mineral medium at 28 degrees C were compared to cultures that were shifted to either elevated (37 degrees C) or reduced (6 degrees C) temperatures as well as high salinity (59.5 per thousand) and observed over 300 min. Heat shock showed the induction of several known chaperone genes. Cold shock altered the expression of genes in lipid metabolism and stress proteins. High salinity resulted in the modulation of genes coding for compatible solutes, ion transporters and morphology. In summary, over 3000 of the 7325 genes were affected by temperature and/or salinity changes. CONCLUSION: Transcriptional profiling confirmed that R. baltica is highly responsive to its environment. The distinct responses identified here have provided new insights into the complex adaptation machinery of this environmentally relevant marine bacterium. Our transcriptome study and previous proteome data suggest a set of genes of unknown functions that are most probably involved in the global stress response. This work lays the foundation for further bioinformatic and genetic studies which will lead to a comprehensive understanding of the biology of a marine Planctomycete.

Model based substrate set point control of yeast cultivation processes based on FIA measurements.

In this contribution a model based substrate control system for Saccharomyces cerevisiae fed batch cultivations is presented. The intention is to keep the concentration of the substrate glucose at a fixed selected set point during the process run. Set points of 0.07 g L(-1) and 0.5 g L(-1) are chosen, as the cells change their metabolism from pure oxidative to oxidative-reductive depending on the glucose concentration. The precise control of glucose concentration during cultivations still poses a challenge as the analysis with available on-line measurement systems still has the problem of noise and a time delay of at least 6 min. To compensate these effects a control system based on an ordinary FIA system for glucose measurements complemented by an extended Kalman filter is employed. The Kalman filter could handle the dynamics of the process accurately. Based on the glucose measurement every 3 min it estimated the biomass and glucose concentration as well as the growth rate factor and the volume of the culture broth. Utilising the estimated values of the process variables a feed forward controller was complemented by a PI controller to adjust the glucose concentration at the desired set points. During the control phase the standard deviation of the measurements are 0.002 g L(-1) and 0.022 g L(-1) for the set points of 0.07 g L(-1) and 0.5 g L(-1), respectively.

In vivo regulation of glucose transporter genes at glucose concentrations between 0 and 500 mg/L in a wild type of Saccharomyces cerevisiae.

When the yeast Saccharomyces cerevisiae consumes glucose, the expression of the genes for the glucose transport is controlled via signal transduction pathways and sensor molecules. Most publications describe the behavior of deletion strains while little is published about the in vivo regulation of glucose transporters in a wild type of S. cerevisiae. Here a global gene expression analysis via microarray experiments from cultivations with glucose concentrations of 50, 70, 100 and 500 mg/L is presented. This permits the observation of the fine-tuning of gene expression in dependency on the glucose concentration. We detected indications that the transport system for high glucose concentrations is activated at glucose concentrations between 50 and 100 mg/L. The regulation of genes coding enzymes for the signal pathways and of those encoding the transporters themselves supports this assumption. The expression of sensor-, signal- and transporter genes will be discussed in detail. In addition, new information about the behavior of the so far little described carriers HXT8, HXT12, HXT13, HXT17 and GAL2 will be given. According to our findings, HXT13 is active during starvation. HXT12, HXT17 and GAL2 are used at low glucose concentrations. The carrier HXT8 supports the glucose transport both during starvation and at low glucose concentrations.