Alexander V. Pinevich

Cyanobacteria of the Genus Prochlorothrix

Green cyanobacteria differ from the blue–green cyanobacteria by the possession of a chlorophyll-containing light-harvesting antenna. Three genera of the green cyanobacteria namely Acaryochloris, Prochlorococcus, and Prochloron are unicellular and inhabit marine environments. Prochlorococcus marinus attracts most attention due to its prominent role in marine primary productivity. The fourth genus Prochlorothrix is represented by the filamentous freshwater strains. Unlike the other green cyanobacteria, Prochlorothrix strains are remarkably rare: to date, living isolates have been limited to two European locations. Taking into account fluctuating blooms, morphological resemblance to Planktothrix and Pseudanabaena, and unsuccessful attempts to obtain enrichments of Prochlorothrix, the most successful strategy to search for this cyanobacterium involves PCR with environmental DNA and Prochlorothrix-specific primers. This approach has revealed a broader distribution of Prochlorothrix. Marker genes have been found in at least two additional locations. Despite of the growing evidence for naturally occurring Prochlorothrix, there are only a few cultured strains with one of them (PCC 9006) being claimed to be axenic. In multixenic cultures, Prochlorothrix is accompanied by heterotrophic bacteria indicating a consortium-type association. The genus Prochlorothrix includes two species: P. hollandica and P. scandica based on distinctions in genomic DNA, cell size, temperature optimum, and fatty acid composition of membrane lipids. In this short review the properties of cyanobacteria of the genus Prochlorothrix are described. In addition, the evolutionary scenario for green cyanobacteria is suggested taking into account their possible role in the origin of simple chloroplast.

Diversity at low abundance: The phenomenon of the rare bacterial biosphere

Rare bacterial biosphere (RBB) is a large and probably predominant sector of bacterial diversity, which is specifically represented by small populations. Although some RBB components have been characterized phenotypically (actualistic objects), it has been mainly described as a set of virtual objects, i.e., of the 16S rRNA gene sequences from environmental DNA samples, which are grouped into phylotypes (operational taxonomic units, OTUs). The upper OTU threshold for RBB is presently not standardized. It is usually ~1% of the sum of OTU sequences in the metagenome library, or five sequences per OTU in absolute values. The analyzed RBB objects include (1) virtual and actualistic objects; (2) autochthonous and allochthonous forms; (3) vegetative and differentiated cells; (4) dead bacteria and free DNA; and (5) artifacts and informational gaps. The RBB phenomenon has not been sufficiently explained. According to some concepts, the RBB objects are rare due to restrictive action of unfavorable environmental factors. According to others, they utilize a successful adaptive strategy of low abundance, which facilitates higher genetic diversity, dispersal and colonization of new niches, and microbial conversion of specific substrates. Since RBB was revealed only in the early 2000s and is still poorly studied, its role in organic evolution and its place in the ecosystems should be determined by future research. The information on the RBB composition, distribution, and functions will be important for bacteriology, while some cultured species may be of basic or applied importance.

Proposal to consistently apply the International Code of Nomenclature of Prokaryotes (ICNP) to names of the oxygenic photosynthetic bacteria (cyanobacteria), including those validly published under the International Code of Botanical Nomenclature (ICBN)/International Code of Nomenclature for algae, fungi and plants (ICN), and proposal to change Principle 2 of the ICNP

This taxonomic note was motivated by the recent proposal [Oren & Garrity (2014) Int J Syst Evol Microbiol 64, 309-310] to exclude the oxygenic photosynthetic bacteria (cyanobacteria) from the wording of General Consideration 5 of the International Code of Nomenclature of Prokaryotes (ICNP), which entails unilateral coverage of these prokaryotes by the International Code of Nomenclature for algae, fungi, and plants (ICN; formerly the International Code of Botanical Nomenclature, ICBN). On the basis of key viewpoints, approaches and rules in the systematics, taxonomy and nomenclature of prokaryotes it is reciprocally proposed to apply the ICNP to names of cyanobacteria including those validly published under the ICBN/ICN. For this purpose, a change to Principle 2 of the ICNP is proposed to enable validation of cyanobacterial names published under the ICBN/ICN rules.

Baeocytes in the cyanobacterium Pleurocapsa sp.: Characterization of the differentiated cells produced by multiple fission

Electron microscopy of cyanobacterium Pleurocapsa sp. CALU 1126 revealed that multiple fission proceeds by successive binary fissions. The cultivation conditions were determined when the number of baeocytes (products of multiple fission) was comparable with that of macrocytes (products of binary fission), and cell sorting was achieved for the first time. Juvenile baeocytes were shown to differ from macrocytes in: (1) the absence of sheath; (2) the linear-peripheral configuration of their lamellar system; (3) lower content of phycobiliproteins and higher content of carotenoids; (4) the set of PSII polypeptides. Baeocytes can therefore be considered differentiated cells characterized by the uncoupling between energy and biosynthetic metabolism.

Probing environmental DNA reveals circum-Baltic presence and diversity of chlorophyll a/b-containing filamentous cyanobacteria (genus Prochlorothrix)

Cyanobacteria that possess phycobilisomes, light-harvesting antenna, have been well studied. In contrast, more rare green cyanobacteria (four genera/five species) that instead make use of chlorophyll–protein complex are poorly studied. In particular, the genus Prochlorothrix is represented by a small environmental DNA database and reports of only two cultured species from Northern Europe. In this work, marine and freshwater habitats of Northwestern Russia were investigated. PCR with Prochlorothrix 16S rRNA gene specific primers, FISH analysis with a Prochlorothrix 16S rRNA-targeted probe, Prochlorothrix culture isolation, and phylogenetic analysis of Prochlorothrix diversity were carried out. We identified Prochlorothrix 16S rDNA in samples from the St. Petersburg region and corroborated this finding by FISH. Attempts to isolate PCR- and FISH-detected Prochlorothrix strains were unsuccessful. Phylogenetic analysis revealed that the Prochlorothrix 16S rDNA sequences identified were very similar and formed a single cluster with high bootstrap support. Some of these sequences represent environmental strains of the species Prochlorothrix hollandica and P. scandica, while the others belong to new Prochlorothrix species or even to a new Prochlorothrix-related genus. Our results suggest a broader distribution and greater diversity in Prochlorothrix than previously thought.

Polyphasic emended description of the filamentous prochlorophyte Prochlorothrix scandica Skulberg 2008

Abstract: Chlorophyll b -containing, phycobiliprotein-less cyanobacterium (“prochlorophyte”) Prochlorothrix scandica Skulberg 2008 represents the second species of the genus Prochlo-rothrix . Distinction between the strains P. scandica NIVA-8/90 and Prochlorothrix hollandica PCC 9006 (CCAP 1490/1 T ) is based predominantly on genomic DNA properties. The degree of DNA reassociation of the two strains is 5 %. Compared to P. hollandica, P. scandica possesses an additional rrn operon. In addition, there is a distinct polymorphism of the highly iterated short palindromes (Hip1). Among the pheno-typic features different in the two species are cell morphology, growth temperature optimum and fatty acids content. Keywords: polyphasic taxonomy, prochlorophytes, cyanobacteria, ITS, Hip1, Prochlorothrix, Prochlorothrix scandica The GenBank accession number for the single 16S−23S ITS sequence of Prochlo-rothrix hollandica PCC 9006 T is HQ316172 and the accession numbers for the two 16S−23S ITS sequences of

Taxonomic Attribution of “Oscillatoriales” Strains within the Bacteriological System of Cyanobacteria: Identification Algorithm for Operational Genera

In “Oscillatoriales” cyanobacteria (Cyanophyceae), relatively simple and uniform morphology superimposes on high genetic diversity that impedes reliable identification. The system of Cyanobacteria set forth in Bergey’s Manual of Systematic Bacteriology-2001/Systematics of Archaea and Bacteria-2015 deals with operational taxa—form-genera (“larger” genera represented by strains) unlike true cyanophycean genera represented by species. Form-genera were established on morphological criteria shared with Cyanophyceae, although they were typified by Pasteur Culture Collection (PCC) strains. Despite being important in determinative cyanobacteriology, old diagnoses of form-genera should be reappraised because, in them: (i) vague and/or ephemeral morphological characters are considered taxonomically significant; (ii) phylogenetic character, such as 16S rRNA gene sequence (16S) is missing. We identified 32 “Oscillatoriales” strains from CALU collection (St. Petersburg University, Russia) basing on core morphology traits, 16S of PCC type strains, and 16S from GenBank database. We proposed that, in experimentally oriented and ecology oriented studies, unequivocal identification can be attained via triple match: streamlined form-genus diagnosis— 16S of PCC reference strain—GenBank most similar 16S. Additionally, we traced the phylogeny of “Oscillatoriales” form-genera via 16S clustering and HIP1 fingerprinting, and suggested that these operational taxa should be replaced with monophyletic assemblages. Nucleotide sequence data reported are available in the GenBank database under the accession numbers KX263921−KX263950.

Testing culture purity in prokaryotes: criteria and challenges

Reliance on pure cultures was introduced at the beginning of microbiology as a discipline and has remained significant although their adaptive properties are essentially dissimilar from those of mixed cultures and environmental populations. They are needed for (i) taxonomic identification; (ii) diagnostics of pathogens; (iii) virulence and pathogenicity studies; (iv) elucidation of metabolic properties; (v) testing sensitivity to antibiotics; (vi) full-length genome assembly; (vii) strain deposition in microbial collections; and (viii) description of new species with name validation. Depending on the specific task there are alternative claims for culture purity, i.e., when conventional criteria are satisfied or when looking deeper is necessary. Conventional proof (microscopic and plating controls) has a low resolution and depends on the observer’s personal judgement. Phenotypic criteria alone cannot prove culture purity and should be complemented with genomic criteria. We consider the possible use of DNA high-throughput culture sequencing data to define criteria for only one genospecies, axenic state detection panel and only one genome. The second and third of these are preferable, although their resolving capacity (depth) is limited. Because minor contaminants may go undetected, even with deep sequencing, the reliably pure culture would be a clonal culture launched from a single cell or trichome (multicellular bacterium). Although this type of culture is associated with technical difficulties and cannot be employed on a large scale (the corresponding inoculums may have low chances of growth when transferred to solid media), it is hoped that the high-throughput culturing methods introduced by ‘culturomics’ will overcome this obstacle.

Molecular analysis of cell division genes and adjacent genes in the cyanobacterium Pleurocapsa sp. CALU 1126

The nucleotide sequences of the cell division gene ftsZ and adjacent genomic regions have been determined for the first time for cyanobacteria of the Pleurocapsa group (Pleurocapsa sp. CALU 1126). The comparison of this locus with the respective genomic regions of other cyanobacteria shows the differences in phylogeny retraced by different genes.