Molecular identification and ultrastructural and phylogenetic studies of cyanobacteria from association with the white sea hydroid Dynamena pumila (L., 1758).

Three new cyanobacterial strains, that have been previously purified from the hydroid Dynamena pumila (L., 1758), isolated from the White Sea, were studied using scanning and transmission electron microscopy methods and were characterized by using almost complete sequence of the 16S rRNA gene, internal transcribed spacer 16S-23S rRNA, and part of the gene for 23S rRNA. The full nucleotide sequences of the rRNA gene clusters were deposited to GenBank (HM064496.1, GU265558.1, JQ259187.1). Comparison of rRNA gene cluster sequences of Synechococcus cyanobacterium 1Dp66E-1, Oscillatoriales cyanobacterium 2Dp86E, and Nostoc sp. 10Dp66E with all sequences present at the GenBank shows that these cyanobacterial strains do not have 100% identity with any organisms investigated previously. Furthermore, for the first time heterotrophic bacterium, associated with Nostoc sp. 10Dp66E, was identified as a member of the new phylum Gemmatimonadetes, genus of Gemmatimonas (GenBank accession number is JX437625.1). Phylogenetic analysis showed that cyanobacterium Synechococcus sp. 1Dp66E-1 forms the unique branch and belongs to a cluster of Synechococcus, including freshwater and sea strains. Oscillatoriales cyanobacterium 2Dp86E belongs to a cluster of Leptolyngbya strains. Isolate Nostoc sp. 10Dp66E forms unique branch and belongs to a cluster of the genus Nostoc, with the closest relative of Nostoc commune isolates.

The pleiotropic effects of ftn2 and ftn6 mutations in cyanobacterium Synechococcus sp. PCC 7942: an ultrastructural study.

Two cell division mutants (Ftn2 and Ftn6) of the cyanobacterium Synechococcus sp. PCC 7942 were studied using scanning electron microscopy and transmission electron microscopy methods. This included negative staining and ultrathin section analysis. Different morphological and ultrastructural features of mutant cells were identified. Ftn2 and Ftn6 mutants exhibited particularly elongated cells characterized by significantly changed shape in comparison with the wild type. There was irregular bending, curving, spiralization, and bulges as well as cell branching. Elongated mutant cells were able to initiate cytokinesis simultaneously in several division sites which were localized irregularly along the cell. Damaged rigidity of the cell wall was typical of many cells for both mutants. Thylakoids of mutants showed modified arrangement and ultrastructural organization. Carboxysome-like structures without a shell and/or without accurate polyhedral packing protein particles were often detected in the mutants. However, in the case of Ftn2 and Ftn6, the average number of carboxysomes per section was less than in the wild type by a factor of 4 and 2, respectively. These multiple morphological and ultrastructural changes in mutant cells evinced pleiotropic responses which were induced by mutations in cell division genes ftn2 and ftn6. Ultrastructural abnormalities of Ftn2 and Ftn6 mutants were consistent with differences in their proteomes. These results could support the significance of FTN2 and FTN6 proteins for both cyanobacterial cell division and cellular physiology.

[Permeability of cyanobacterial mucous surface structures for macromolecules].

The space of diffusive distribution of neutral hydrophilic macromolecules (dextrans with molecular sizes of 1.5 to 9 nm in the Stokes radius values) in the mucous surface structures (MSS) of intact bacterial cells has been studied for the first time on cyanobacteria. Cyanobacterial species and strains under study belong to different taxonomic groups, the members of which form MSS of various morphology and ultrastructure and can grow in association with plants and animals, inter alia as mucous microcolonies. The range of permeability has been determined by the fractionation of polydisperse dextrans method, previously applied for plants, in combination with electron microscopy. Dextrans are supposedly distributed in the MSS polysaccharide matrix in accordance with their sizes, in much the same way as in a macroporous unitary gel. The similarity of the chemical composition and macromolecular organization of cyanobacterial MSS with pectins of plant cell walls and the role of MSS and the intercellular matrix as permeability barriers in associative interactions of microorganisms are under consideration.

[Communication of cyanobacteria with plant partners during association formation].

Data are presented on the physiological diagnostics of cyanobacterial communication with higher plants in natural symbioses (plant syncyanoses) and in model associations, as well as on the interaction of the partners without spatial integration. Emphasis is placed on changes in cyanobacterial features important for symbiogenesis. The multicomponent structure and the possible nature of the factors that enable partner communications are discussed with hormogonia formation and taxis as an example.

[The accumulation and degradation dynamics of cyanophycin in cyanobacteria grown in symbiotic associations with plant tissues and cells]. / Dinamika nakopleniia i destruktsii tsianofitsina v kletkakh tsianobakterii pri vzaimodeistvii s rastitel'nymi tkaniami.

Five different artificial associations of cyanobacterial cells with the cells or tissues of nightshade and rauwolfia were studied. The associations grown on nitrogen-containing media produced heterocysts. Cyanobacterial cells in the associations retained their ability to take up bound nitrogen from the medium, to store it in the form of cyanophycin granules, and to use them in the process of symbiotic growth. The synthesis and degradation of cyanophycin granules in cyanobacterial cells were more active in the associations than in monocultures. In the symbiotic associations of Chlorogloeopsis fritschii ATCC 27193 with Solanum laciniatum cells and of Nostoc muscorum CALU 304 with the Rauwolfia serpentina callus, heterocysts were produced at 3- to 30-fold higher cyanophycin contents than in cyanobacterial monocultures. In contrast, in the association of N. muscorum CALU 304 with the Solanum dulcamara callus, heterocysts were produced at lower cyanophycin contents than in the N. muscorum CALU 304 monoculture. The degradation of cyanophycin granules in N. muscorum CALU 304 cells grown in associations with plant tissues or cells was subjected to mathematical analysis. The activation of cyanophycin degradation and heterocyst production in the associations N. muscorum CALU 304-R. serpentina and C. fritschii-S. laciniatum was accompanied by an enhanced synthesis of the nitrogen-containing alkaloids in plant cells. The data obtained suggest that an integrated system of nitrogen homeostasis can be formed in symbiotic associations. Depending on the growth stage of an association, its plant member can either stimulate the accumulation of bound nitrogen in vegetative cyanobacterial cells in the form of cyanophycin granules, or activate their degradation, or initiate the formation of heterocysts independently of the cyanobacterial sensory-signalling system.

[Formation of giant and ultramicroscopic forms of Nostoc muscorum CALU 304 during cocultivation with Rauwolfia tissues]. / Obrazovanie gigantskikh i ultramikroform Nostoc muscorum CALU 304 pri vzaimodeistvii s kul'tiviruemymi tkaniami rauvol'fii.

The study of heteromorphic Nostoc muscorum CALU 304 cells, whose formation was induced by 6- to 7-week cocultivation with the Rauwolfia callus tissues under unfavorable conditions, revealed the occurrence of giant cell forms (GCFs) with a volume which was 35-210 times greater than that of standard cyanobacterial cells. Some GCFs had an impaired structure of the murein layer of the cell wall, which resulted in the degree of impairment of the cell wall ranging from the mere loss of its rigidity to its profound degeneration with the retention of only small peptidoglycan fragments. An analysis of thin sections showed that all GCFs had enlarged nucleoids. The photosynthetic membranes of spheroplast-like GCFs formed vesicles with the contents analogous to that of nucleoids (DNA strands and ribosomes). About 60% of the vesicles had a size exceeding 300 nm. With the degradation of GCFs, the vesicles appeared in the intercellular slimy matrix. It is suggested that the vesicles are analogous to elementary bodies, which are the minimal and likely primary reproductive elements of L-forms. The data obtained in this study indicate that such L-forms may be produced in the populations of the cyanobionts of natural and model syncyanoses. Along with the other known cyanobacterial forms induced by macrosymbionts, L-forms may represent specific adaptive cell forms generated in response to the action of plant symbionts.

[Ultrastructure of the cell surface of heteromorphic Nostoc muscorum CALU 304 cocultured with Rauwolfia tissue]. / Ul'trastruktura kletochnoi poverkhnosti geteromorfnykh kletok Nostoc muscurum CALU 304 v smeshannoi kul'ture s tkan'iu Rauvol'fia.

The ultrastructure of the heteromorphic cells (HMCs) of the cyanobacterium Nostoc muscorum CALU 304 grown in pure culture, monoculture, and a mixed culture with the Rauwolfia callus tissue was studied. The comparative analysis of the cell surface of HMCs, the frequency of the generation of cell forms with defective cell walls (DCWFs), including protoplasts and spheroplasts, and the peculiarities of the cell surface ultrastructure under different growth conditions showed that, in the early terms of mixed incubation, the callus tissue acts to preserve the existing cyanobacterial DCWFs, but begins to promote their formation in the later incubation terms. DCWFs exhibited an integrity of their protoplasm and were metabolically active. It is suggested that structural alterations in the rigid layer of the cell wall may be due to the activation of the murolytic enzymes of cyanobacteria and the profound rearrangement of their peptidoglycan metabolism caused by the Rauwolfia metabolites diffused through the medium. These metabolites may also interfere with the functioning of the universal cell division protein of bacteria, FtsZ. In general, the Rauwolfia callus tissue promoted the unbalanced growth of the cyanobacterium N. muscorum CALU 304 and favored its viability in the mixed culture. The long-term incubation of the Rauwolfia tissue with the N. muscorum CALU 304 cells led to their transformation to L-forms.

[Spatial integration of partners and heteromorphism of cyanobacterium Nostoc muscoum CALU 304 in mixed culture with Rauwolfia]. / Prostranstvennaia integratsiia partnerov i geteromorfizm tsianobakteriiNostoc muscorum CALU 304 v smeshanoi kul'ture s tkan'iu rauvol'fii.

A comparative morphological study was conducted of Nostoc muscorum CALU 304 grown either as a pure culture on standard media or as a mixed culture with Rauwolfia callus tissue on a medium for plant tissue cultivation. The interaction of the cyanobacterial and plant partners results in their spatial integration into aggregates of specific anatomy, which arise periodically during the mixed culture growth. The morphology of the cyanobacterial cells varies depending on their localization in the combined aggregate. The degree of cyanobacterial heteromorphism increases with time of growth of the association. Evidence of the plant origin of the factors inducing heteromorphic changes in N. muscorum was obtained, as well as evidence indicating that these factors can rapidly diffuse in agarized medium. A conclusion is inferred that the heteromorphic cells correspond to bacterial forms that appear during unbalanced growth as an adaptation to altered environmental conditions.