The CRISPR/Cas Machinery Evolution and Gene Flow in the Hot Spring Cyanobacterium Thermostichus

Drivers of the speciation in bacteria, including geographical isolation and horizontal gene transfer, are still poorly understood. Here, we characterized a new lineage within an anciently diverged thermophilic cyanobacteria. We sequenced the whole genome of a strain Thermostichus vulcanus isolated from the Rupite spring (Bulgaria), which is closely related to Theromstichus strains JA-2-3Aa and JA-2-3B′a(2-13). We performed phylogenetic inference, horizontal gene transfer estimation, and CRISPR/Cas system characterization. We found that the Rupite strain is a distinct species from strains JA-2-3Aa, and JA-2-3B′a(2-13). Furthermore, the horizontal gene transfer seemed to be more frequent among the geographically distant species than between the two species within the same hot spring. The CRISPR/Cas system had variable complexity among the species of Thermostichus in terms of both the number of spacers and genes. The Rupite strain had the highest, and JA-2-3Aa the lowest number of spacers among the analyzed strains, and the CRISPR spacers were only rarely shared among the strains. We conclude that the CRISPR/Cas system size varied among the lineages as well as the gene flow

The CRISPR/Cas Machinery Evolution and Gene Flow in the Hot Spring Cyanobacterium <i>Thermostichus</i>

Drivers of the speciation in bacteria, including geographical isolation and horizontal gene transfer, are still poorly understood. Here, we characterized a new lineage within an anciently diverged thermophilic cyanobacteria. We sequenced the whole genome of a strain Thermostichus vulcanus isolated from the Rupite spring (Bulgaria), which is closely related to Theromstichus strains JA-2-3Aa and JA-2-3B′a(2-13). We performed phylogenetic inference, horizontal gene transfer estimation, and CRISPR/Cas system characterization. We found that the Rupite strain is a distinct species from strains JA-2-3Aa, and JA-2-3B′a(2-13). Furthermore, the horizontal gene transfer seemed to be more frequent among the geographically distant species than between the two species within the same hot spring. The CRISPR/Cas system had variable complexity among the species of Thermostichus in terms of both the number of spacers and genes. The Rupite strain had the highest, and JA-2-3Aa the lowest number of spacers among the analyzed strains, and the CRISPR spacers were only rarely shared among the strains. We conclude that the CRISPR/Cas system size varied among the lineages as well as the gene flow

Intact cell MALDI-TOF mass spectrometric analysis of Chroococcidiopsis cyanobacteria for classification purposes and identification of possible marker proteins.

Cyanobacteria represent a bacterial phyllum characteristic by the ability to photosynthesize. They are potentially applicable for the production of useful compounds but may also cause poisoning or at least health problems as they can produce cyanotoxins. The introduction of a fast methodology is important not only for fundamental taxonomic purposes, but also for reliable identifications in biological studies. In this work, we have used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of intact cells to study Chroococcidiopsis strains. A library of the obtained reference mass spectra containing characteristic peptide/protein profiles was examined by software tools to characterize similarities and differences applicable for diagnostics and taxonomy. Both a similarity tree and heat map constructed from the mass spectrometric data proved consistent with 16S rRNA sequencing results. We show as novelty that a binary matrix combining ferulic and sinapinic acids performs well in acquiring reproducible mass spectra of cyanobacteria. Using the matrix solvent, a protein extraction from cells was done. After polyacrylamide gel electrophoresis, the separated protein fractions were in-gel digested and the resulting peptides analyzed by liquid chromatography coupled with tandem mass spectrometry. For the first time, photosystem protein components, phycobilisome proteins, electron transport proteins, nitrogen-metabolism and nucleic acids binding-proteins, cytochromes plus other enzymes and various uncharacterized proteins could be assigned to characteristic peaks in the mass spectrometric profiles and some of them suggested as markers in addition to 30S and 50S ribosomal proteins known from previous studies employing intact cell mass spectrometry of microorganisms

Difference without distinction? Gaps in cyanobacterial systematics; when more is just too much

Cyanobacteria are amongst the most abundant, ubiquitous, ecologically and evolutionarily significant microbes on Earth. Unique among the Bacteria in their capacity to be identified using morphology, understanding the evolutionary relationships and describing the diversity of this lineage is both important and challenging. The advent of modern sequencing technology has proven a boon to those studying cyanobacterial systematics as it has provided copious amounts of sequence data (mainly of the 16S rRNA gene sequence). However, this influx of data has also led to taxonomic confusion and recognition of polyphyly in many genera. Thus, the purpose of this paper is to describe this apparent paradox of increasing data yet poor phylogenetic resolutions by employing the Poisson Tree Process (PTP) algorithm and to propose some ameliorative efforts

<i>Elainella</i> gen. nov.: a new tropical cyanobacterium characterized using a complex genomic approach

<p>Cyanobacteria represent an ancient, monophyletic lineage of bacteria with the ability to undertake oxygenic photosynthesis. Although they possess a relatively high degree of morphological variability compared with other prokaryotes and there is a wealth of molecular data, there are still significant gaps in our knowledge of cyanobacterial diversity, especially in tropical areas. Here, we present a novel, filamentous, tropical cyanobacterium, which could be classified as <i>Pseudophormidium</i> based on morphological criteria. A total evidence investigation employing ecological, morphological and genomic data, indicated that our strains form a new and ancient evolutionary lineage among cyanobacteria unrelated to <i>Pseudophormidium</i>. Based on this polyphasic assessment, our strains represent a novel, monospecific genus: <i>Elainella</i>. This new genus represents an example of phenotypic convergence, which seems to be a prevalent macroevolutionary pattern in cyanobacteria, a likely cause of the frequently cited polyphyly within a majority of genera.</p