A new subarctic strain of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis>—part I: identification and fatty acid profiling

We isolated a new subarctic strain Tetradesmus obliquus IPPAS S-2023 (Scenedesmaceae, Chlorophyceae) from rock baths in the White Sea. To verify its taxonomic assignment, internal transcribed spacer 2 (ITS2) of the strain was sequenced and its secondary structure was compared with predicted ITS2 secondary structures of Scenedesmaceae. The analysis of the ITS2 made it possible to assign the new strain IPPAS S-2023 to the species T. obliquus. The ultrastructural studies and energy-dispersive X-ray spectroscopy (EDX) analysis revealed a marked accumulation of vacuolar inclusions enriched in phosphorus and nitrogen (N) as well as cytoplasmiс oil bodies. Most of predicted properties of biodiesel derived from the fatty acids profile of the strain grown in the N-free medium complied with the requirements of European and American standards. The results suggest that the new subarctic strain T. obliquus IPPAS S-2023 is a promising candidate for nutrients biosequestration and for biodiesel production. In a companion paper, we assess its biomass production capability and suitability and demonstrated suitability of IPPAS S-2023 as a reference strain for studies on elevated CO₂ stress effects selection of carbon dioxide-tolerant microalgae by comparison with a CO₂-tolerant strain IPPAS S-2014.     

Dialysis cultivation of microorganisms as the appropriate model of population control in ecosystem research

Opportunities for application of methods of ecological control of anthropogenic impact on ecosystems have been considered. The biotic approach based on dialysis cultivation of microorganism populations has been suggested in order to evaluate the ecological state of living systems. Dialysis cultures are characterized with a high level of biomass accumulation and considerable extension of exponential and stationary growth phases. The cell state at dialysis cultivation corresponds to the state of the organisms in native nature conditions. The method for analysis of responses from microbe cells on external factors’ influence has been suggested. The method is based on the registration of alterations of time-spatial parameters in cell populations by layer-by-ayer noninvasive analysis using the spectroscopy of internal reflection.     

Methodology for determination of the state of microbial culture by degree of spatial organization of whole cells and their external structures

The proposed methodology for determination of biosystems state is based on registration of changes in the space-time parameters of cells and their external structures in a population of microorganisms from different taxonomic groups by the method of noninvasive layer-by-layer analysis using internal reflection spectroscopy. The dependence between the state of an organism and the changes in spatial organization of whole cells and their surface layers have been revealed. The proposed method allows estimation of the character and degree of dynamics of biosystem variations irrespective of the kind of influence. This method can be used for assessing the state of other organisms by the differences between values A for the surface structures and cells of these organisms.     

Possibilities of bioconversion of agricultural waste with the use of microalgae

A new methodology of biological treatment and conversion of farm waste (manure and wash water) with the use of intensively cultivated phototrophic microorganisms (microalgae) is reviewed. Criteria for selection of microalgae and peculiarities of their intensive cultivation for efficient removal of biogenic elements from and destruction of the organic components of the wastes as well as the possibilities of cost-effective utilization of the resulting microalgal biomass are considered. Advantages and drawbacks of the new methodology are compared with those of conventional anaerobic techniques. Special attention is paid to the integrated technologies combining the aerobic conversion methods with microalgal post-treatment.     

Formation of Phenolic Compounds in Apogeotropic Roots of Cycad Plants

Formation and location of phenolic compounds in apogeotropic roots (coralloid roots) were studied in six cycad species, which belong to the genera Cycas, Encephalartos, and Ceratozamia. Total contents of soluble phenolic compounds in coralloid roots in all species studied varied insignificantly, with except for Ceratozamia mexicana that accumulated three times higher amounts of phenolic compounds. Phenolic compounds were accumulated in cell walls of cortical parenchyma of coralloid roots, in intercellular spaces, and in specialized storage cells, found in all zones of apogeotropic roots. The greatest number of phenol-storing cells was situated in the cortical parenchyma of the central part of coralloid roots, adjacent to a zone where active symbiotic cyanobacteria were localized, and in the coralloid root basal region lacking viable forms of cyanobionts. It was suggested that phenolic compounds affect the formation of symbiosis between cyanobacteria and apogeotropic roots of cycad plants, as well as their metabolism.     

Ion-exchange characteristics of the cell walls isolated from the thallus of the lichen <Emphasis Type="Italic">Peltigera aphthosa</Emphasis> (L.) Willd

Ion-exchange characteristics of the cell walls isolated from different zones of the foliose lichen Peltigera aphthosa (L.) Willd were determined. Four types of ionogenic groups were revealed in the thallus cell walls of P. aphthosa, namely amino groups, carboxylic groups of uronic acids, carboxylic groups of phenolic acids, and phenolic OH groups. They may participate in the ion-exchange reactions with the ions of the environment. The amount of ionogenic groups in P. aphthosa cell walls was found to depend on the zone and age of the thallus.     

pH and CO<Subscript>2</Subscript> effects on <Emphasis Type="Italic">Coelastrella</Emphasis> (<Emphasis Type="Italic">Scotiellopsis</Emphasis>) <Emphasis Type="Italic">rubescens</Emphasis> growth and metabolism

We studied effects of рН and СО₂ enrichment on the physiological condition and biochemical composition of a carotenogenic microalga Coelastrella (Scotiellopsis Vinatzer) rubescens Kaufnerová et Eliás (Scenedesmaceae, Sphaeropleales, Chlorophyceae), a promising source of natural astaxanthin. The microalga was grown at a constant pH (5, 6, 7 or 8) maintained by direct СО₂ injection. The air-sparged culture served as the control. Cell division rate and size, dry biomass productivity, the rates of nitrogen and phosphorus uptake as well as photosynthetic pigment and total lipid content and fatty acid composition were followed. С. rubescens possessed a narrow-range рН tolerance (the optimum рН 6–7). Under these conditions, the highest values of the maximum (1.0–1.1 1/day) and average (0.3–0.35 1/day) specific growth rate, chlorophyll а (4.8–4.9%) and total carotenoid dry weight percentages (1.7–1.8%) were recorded. Cell lipid fatty acid unsaturation index (1.851) and polyunsaturated fatty acid percentage (36–39%) and С18:3 ω3/С18:1 ω9 ratio (3.8–4.5) were also the highest under these conditions. A decline of рН to 5 brought about severe stress manifesting itself as a cell division cessation, photosynthetic apparatus reduction, two-fold increase in cell volume, accumulation of dry weight and lipids and a considerable decline in fatty acid unsaturation. Cultivation of С. rubescens without СО₂ enrichment resulted in a rapid alkalization of the medium to рН 9.5–10.5 impairing the physiological condition of the cells. Reasons of the deteriorative effects of suboptimal pH values on the physiological condition of C. rubescens are discussed.     

A new subarctic strain of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis>. Part II: comparative studies of CO<Subscript>2</Subscript>-stress tolerance

A huge interest in CO₂-tolerant microalgae is fueled by development of CO₂-biomitigation methods based on intensive cultivation of microalgae. Still, the mechanisms of CO₂-tolerance are scarcely investigated. Previously, we described a symbiotic Desmodesmus sp. IPPAS S-2014 from a White Sea hydroid tolerant to extremely high (20–100%) CO₂ levels. In the present work, we compared its ultrastructural and physiological responses to those of a novel free-living White Sea strain of Tetradesmus obliquus IPPAS S-2023 characterized in the companion paper. The strain S-2023 is closely related to Desmodesmus sp. IPPAS S-2014 but lacks its tolerance to extremely high CO₂ (it is unable to survive at 100% CO₂ and exhibits a reduced-growth phenotype when sparged with 20% CO₂: air mixture). We compared the responses of the cell organization and photosynthetic activity to 20% CO₂ in the tolerant and the intolerant White Sea chlorophytes using chlorophyll fluorescence measurements and ultrastructural analysis (transmission electron microscopy). The features peculiar to the CO₂-intolerant chlorophyte include (i) inability to maintain pH homeostasis, (ii) a steady decline in the photosynthetic activity of the cells, (iii) a reduction of the photosynthetic membranes, and (iv) delayed accumulation of starch (starch grains) and its subsequent conversion to reserve lipids (oil bodies). Nitrogen starvation enhances the effects of high-CO₂ stress in the CO₂-intolerant microalga. The results of this work are discussed in the context of selection of tolerant algal strains for CO₂ biomitigation applications.     

Microbial complexes from apogeotropic roots and from rhizosphere of cycad plants

The microbial complexes of soil, the rhizosphere, and the rhizoplane of the apogeotropic (coralloid) roots of cycad plants were comparatively studied. The aseptically prepared homogenates of the surface-sterilized coralloid roots did not contain bacterial microsymbiont, indicating that it was absent in the root tissues. At the same time, associated bacteria belonging to different taxonomic groups were detected in increasing amounts in the cycad rhizoplane, rhizosphere, and the surrounding soil. The bacterial communities found in the cycad rhizoplane and the surrounding soil were dominated by bacteria from the genus Bacillus. The saprotrophic bacteria and fungi colonizing the cycad rhizosphere and rhizoplane were dominated by microorganisms capable of degrading the plant cell walls. The local degradation of the cell wall was actually observed on the micrographs of the thin sections of cycad roots in the form of channels, through which symbiotic cyanobacterial filaments can penetrate into the cortical parenchyma.