Microbial Community Composition and Rates of the Methane Cycle Microbial Processes in the Upper Sediments of the Yamal Sector of the Southwestern Kara Sea

Geochemical, biogeochemical, and molecular genetic investigation of the upper (0–5 cm) bottom sediments of the Yamal sector of the Kara Sea was carried out. The Yamal sector is well-protected from the massive inflow of river water. The sediments were oxidized at the surface and weakly reduced in the 3?5-cm layer. C_org content varied from 0.1 to 1.3%, while the level of dissolved СН₄ was 1.9 to 20.3 μmol L^–1. The isotopic composition of organic matter (OM) carbon, δ¹³Corg, varied from–27.5 to–22.2‰ (–25.4‰ on average). The share of terrigenous OM was 13.3 to 72.2% (48.9% on average). The rate of methane production, methane oxidation, and sulfate reduction varied from 0.8 to 9.0 (2.7 on average) nmol СН₄ dm^–3 day^–1, from 9.9 to 103 (31.6 on average) nmol СН₄ dm^–3 day^–1, and from 0.49 to 2.2 (1.1 on average) μmol S dm–3 day–1, respectively. High-throughput sequencing of the amplicons of the 16S rRNA genes was used to reveal the physiological groups of microorganisms responsible for the processes of methane production and oxidation, sulfate reduction, and oxidation of reduced sulfur compounds. Members of the phylum Woesearchaeota were predominant among archaea. Methanogenic archaea belonged to the families Methanobacteriaceae, Methanococcaceae, and Methanosarcinaceae (Euryarchaeota). Methanotrophs of the family Methylococcaceae were revealed among the Gammaproteobacteria, with their share in the sediments ~1%. In the class Deltaproteobacteria (15.4%), three orders of sulfate reducers were predominant: Desulfobacterales, Desulfovibrionales, and Desulfuromonadales. Oxidation of reduced sulfur compounds was carried out by chemolithoautotrophic bacteria of the genera Sulfurovum, Sulfurimonas, and Arcobacter of the class Epsilonproteobacteria (1.1% of the total microbial number).     

Microbiological and biogeochemical processes in a pockmark of the Gdansk depression,Baltic Sea

Comprehensive microbiological and biogeochemical investigation of a pockmark within one of the sites of gas-saturated sediments in the Gdansk depression, Baltic Sea was carried out during the 87th voyage of the Professor Shtokman research vessel. Methane content in the near-bottom water and in the underlying sediments indicates stable methane flow from the sediment into the water. In the 10-m water layer above the pockmark, apart from methane anomalies, elevated numbers of microorganisms and enhanced rates of dark CO₂ fixation (up to 1.15 µmol C/(l day)) and methane oxidation (up to 2.14 nmol CH₄/(l day)) were revealed. Lightened isotopic composition of suspended organic matter also indicates high activity of the near-bottom microbial community. Compared to the background stations, methane content in pockmark sediments increased sharply from the surface to 40–60 ml/dm³ in the 20–30 cm horizon. High rates of bacterial sulfate reduction (SR) were detected throughout the core (0–40 cm); the maximum of 74 µmol S/(dm³ day) was located in subsurface horizons (15–20 cm). The highest rates of anaerobic methane oxidation (AMO), up to 80 µmol/dm³ day), were detected in the same horizon. Good coincidence of the AMO and SR profiles with stoichiometry close to 1: 1 is evidence in favor of a close relation between these processes performed by a consortium of methanotrophic archaea and sulfate-reducing bacteria. Methane isotopic composition in subsurface sediments of the pockmark (from ?53.0 to ?56.5‰) does not rule out the presence of methane other than the biogenic methane from the deep horizons of the sedimentary cover.     

Successional Changes in the Microbial Community of the Alkaline Lake Khilganta during the Dry Season

Microbial processes in a shallow, saline, alkaline Lake Khilganta (Southern Siberia) were studied during the dry season. During the drought, a crust was formed on the lake surface, where low rates of production processes were observed, with predominance of anoxygenic photosynthesis at 2.3 mg C/(dm³ day). The rates of microbial processes increased after short-term rains. During this period, a thin cyanobacterial mat was formed on the bottom, in which filamentous cyanbacteria Geitlerinema spp. predominated and the rate of oxygenic photosynthesis was up to 18 mg C/(dm³ day). Subsequent water evaporation and salinity increase resulted in altered community types and their activity. Red spots emerged on the mat surface, where anoxygenic prototrophic members of the genus Ectothiorhodospira predominated. Anoxygenic photosynthesis became the main production process in microbial mats, with the rate of 60 mg C/(dm³ day). At salinity increase to 200 g/L, the water remained in small depressions on the bottom, where extremophilic green algae Dunaliella sp. predominated, and the rate of oxygenic photosynthesis was 0.877 mg C/(dm³ day). These changes in the type and activity of microbial communities is an example of succession of microbial communities in Southern Siberia saline lakes during drought.     

Microbial processes at the Lost City vent field,Mid-Atlantic Ridge

Microbiological and biogeochemical measurements showed that the intensities of CO₂ assimilation, methane oxidation, and sulfate reduction at the Lost City vent field (3° N) reach 3.8 µg C/(1 day), 0.06 µg C/(1 day), and 117 µg S/(1 day), respectively. On the surface of the carbonate structures occurring at this field, two varieties of bacterial mats were found. The first variety, which is specific to the Lost City alkaline vent field, represents jellylike bacterial mats dominated by slime-producing bacteria of several morphotypes. This mat variety also contains chemolithotrophic and heterotrophic microorganisms, either microaerobic or anaerobic. The intensities of CO₂ assimilation, methane oxidation, and sulfate reduction in this variety reach 747 µg C/(dm³ day), 0.02 µg C/(dm³ day), and 28000 µg S/(dm³ day), respectively. Bacterial mats of the second variety are formed by nonpigmented filamentous sulfur bacteria, which are close morphologically to Thiothrix. The intensities of CO₂ assimilation, methane oxidation, and sulfate reduction in the second mat variety reach 8.2 µg C/(dm³ day), 5.8 µg C/(dm³ day), and 17000 µg S/(dm³ day), respectively. These data suggest the existence of subsurface microflora at the Lost City vent field.Translated from Mikrobiologiya, Vol. 74, No. 1, 2005, pp. 111–118.Original Russian Text Copyright © 2005 by Dulov, Lein, Dubinina, Pimenov.     

Substrate Specificity of Methanogenic Communities from Lake Baikal Bottom Sediments Associated with Hydrocarbon Gas Discharge

Methane production by microbial communities from Lake Baikal bottom sediments with different chemical composition of pore water was studied. Methane production was more active in the media supplemented with H₂: CO₂ and H₂ + CH₃COONa, rather than on media with acetate as the sole source of carbon and energy. Addition of methanol stimulated methane production only in the case of microbial communities from upper silts. Ability of the communities to produce methane correlated reliably with the concentrations of the NO^3–, SO₄^2?, Cl^–, and CH₃COO^– ions in the pore water of the relevant sediments. Cultivation of communities from the mud volcano sediments resulted in development of methanogenic archaea of the family Methanocellaсеае in the media supplemented with H₂: CO₂ and H₂ + CH₃COONa, while methanogenic archaea in the communities cultivated without additional substrates belonged to the genera Methanoregula, Methanobacterium, and Methanosaeta.     

The role of <Emphasis Type="Italic">Viviparus contectus</Emphasis> (Millet) (Mollusca,Viviparidae) in the sedimentation of suspension and transformation of organic matter in the Tnya River (Ukraine)

As active filterers and sedimentators, mollusks Viviparus contectus (Millet) clear water from suspensions in the Tnya River. The sedimentation ability of mollusks is assessed on the basis of experimental studies at three stations using the funneling technique. Mollusks at the age of 3 and 5 years are used. The rate of sedimentation of V. contectus is 7.3 to 13.7 mg/(spec. day) in July 2014. The efficiency of suspended sedimentation is higher in younger individuals. The rate of sedimentation is determined by the total body weight of mollusks; the ratio between these parameters is described by the exponential function. V. contectus contributes to the decrease in the content of organic matter in the water with a rate of 0.055 to 0.069 mg of O₂/(dm³ spec. h), as well as to the enrichment of the near-bottom water layer with organic matter.     

Growth and photosynthetic response of two larches exposed to O<Subscript>3</Subscript> mixing ratios ranging from preindustrial to near future

In this study, we questioned whether ground-level ozone (O₃) induces hormesis in Japanese larch (Larix kaempferi) and its hybrid F₁ (L. gmelinii var. japonica × L. kaempferi). In order to answer the question, we exposed seedlings of both taxa to four O₃ treatments [ranging from ≈10 to 60 nmol(O₃) mol^–1] in open-top chambers for two consecutive growing seasons. We found a hormetic response in maximum photosynthetic rate (P_Nmax) at 1700 μmol(CO₂) mol^–1 and maximum rates of carboxylation (V_cmax) and electron transport (J_max) in both larches. Stimulation of P_Nmax, V_cmax, and J_max did not lead to suppressed plant productivity in Japanese larch, which followed a stress-tolerant strategy, but it did lead to suppressed plant productivity in hybrid larch which followed a competitive strategy. These findings are the first to suggest that stimulation of physiological functions by low O₃ exposures may have negative consequences for larch reproduction.     

Phylogenetic and functional prokaryotic diversity in the Hoito-Gol mesothermal mineral spring (Eastern Sayan Mountains,Buryat Republic)

High-throughput sequencing was used for comparative analysis of microbial communities of the water and mat from the Hoito-Gol mesothermal mineral sulfide spring (Eastern Sayan Mountains, Buryat Republic). Activity of microbial communities was determined. While both spring biotopes were dominated by members of three bacterial phyla—Proteobacteria, Bacteroidetes, and Firmicutes—they differed drastically in the composition of predominant phylotypes (at the genus level). In the water, the organisms widespread in aquatic environments were predominant, mostly aerobic chemoorganotrophs of the genera Acinetobacter, Pedobacter, and Flavobacterium. In the microbial mat, the organisms actively involved in the sulfur cycle predominated, including sulfur-reducing bacteria Sulfurospirillum, sulfate-reducing deltaproteobacteria, sulfuroxidizing chemoautotrophic bacteria, anoxygenic phototrophic bacteria of the phyla Chloroflexi and Chlorobi, as well as purple bacteria belonging to the α-, ß-, and γ-Proteobacteria. Microbial mats of the spring exhibited higher phylogenetic diversity compared to high-temperature mats containing photosynthetic microorganisms.     

Gas exchange,carbon balance and stomatal traits in wild and cultivated rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) genotypes

Carbon balancing within the plant species is an important feature for climatic adaptability. Photosynthesis and respiration traits are directly linked with carbon balance. These features were studied in 20 wild rice accessions Oryza spp., and cultivars. Wide variation was observed within the wild rice accessions for photosynthetic oxygen evolution or photosynthetic rate (A), dark (R _d), and light induced respiration (LIR) rates, as well as stomatal density and number. The mean rate of A varied from 10.49 μmol O₂ m^?2 s^?1 in cultivated species and 13.09 μmol O₂ m^?2 s^?1 in wild spp., The mean R _d is 2.09 μmol O₂ m^?2 s^?1 and 2.31 μmol O₂ m^?2 s^?1 in cultivated and wild spp., respectively. Light induced Respiration (LIR) was found to be almost twice in wild rice spp., (16.75 μmol O₂ m^?2 s^?1) compared to cultivated Oryza spp., Among the various parameters, this study reveals LIR and A as the key factors for positive carbon balance. Stomatal contribution towards carbon balance appears to be more dependent on abaxial surface where several number of stomata are situated. Correlation analysis indicates that R _d and LIR increase with the increase in A. In this study, O. nivara (CR 100100, CR 100097), O. rufipogon (IR 103404) and O. glumaepatula (IR104387) were identified as potential donors which could be used in rice breeding program. Co-ordination between gas exchange and patchiness in stomatal behaviour appears to be important for carbon balance and environmental adaptation of wild rice accessions, therefore, survival under harsh environment.     

Non-native liana, <Emphasis Type="Italic">Euonymus fortunei</Emphasis>, associated with increased soil nutrients,unique bacterial communities,and faster decomposition rate

Invasive plants have wide-ranging impacts on native systems including reducing native plant richness and altering soil chemistry, microbes, and nutrient cycling. Increasingly, these effects are found to linger long after removal of the invader. We examined how soil chemistry, bacterial communities, and litter decomposition varied with cover of Euonymus fortunei, an invasive evergreen liana, in two central Kentucky deciduous forests. In one forest, E. fortunei invaded in the late 1990s but invasion remained patchy and we paired invaded and uninvaded plots to examine the associations between E. fortunei cover and our response variables. In the second forest, E. fortunei had completely invaded the forest by 2005; areas where it had been selectively removed by 2010 were paired with an adjacent invaded plot. Where E. fortunei had patchily invaded, E. fortunei patches had up to 3.5× nitrogen, 2.7× carbon, and 1.9× more labile glomalin in soils than uninvaded plots, whereas there were no differences in soil characteristics between invaded and removal plots. In the patchily invaded forest, bacterial community composition varied among invaded and non-invaded plots, whereas bacterial communities did not vary among invaded and removal plots. Finally, E. fortunei leaf litter decomposed faster (k = 4.91 year^?1) than the native liana (k = 3.77 year^?1), Vitis vulpina; decomposition of both E. fortunei and V. vulpina was faster in invaded (k = 7.10 year^?1) than removal plots (k = 4.77 year^?1). Our findings suggest that E. fortunei invasion increases the rate of leaf litter decomposition via high-quality litter, alters the decomposition environment, and shifts in the soil biotic communities associated with a dense mat of wintercreeper. Land managers with limited resources should target the densest mats for the greatest restoration potential and remove wintercreeper patches before they establish dense mats.     

Microbial processes of the carbon and sulfur cycles in the Chukchi Sea

The research performed in August 2004 within the framework of the Russian-American Long-term Census of the Arctic (RUSALCA) resulted in the first data concerning the rates of the key microbial processes in the water column and bottom sediments of the Bering strait and the Chukchi Sea. The total bacterial counts in the water column varied from 30 × 10³ cells ml^?1 in the northern and eastern parts to 245 × 10³ cells ml^?1 in the southern part. The methane content in the water column of the Chukchi sea varied from 8 nmol CH₄l^?1 in the eastern part of the sea to 31 nmol CH₄l^?1 in the northern part of the Herald Canyon. Microbial activity occurred in the upper 0–3 cm of the bottom sediments; the methane formation rate varied from 0.25 to 16 nmol CH₄dm^?3 day^?1. The rates of methane oxidation varied from 1.61 to 14.7 nmol CH₄dm^?3 day^?1. The rates of sulfate reduction varied from 1.35 to 16.2 μmol SO ₄ ^2? dm^?1 day^?1. The rate of methane formation in the sediments increased with depth, while sulfate reduction rates decreased (less than 1 μmol SO ₄ ^2? dm^?3 day^?1). These high concentrations of biogenic elements and high rates of microbial processes in the upper sediment layers suggest a specific type of trophic chain in the Chukchi Sea. The approximate calculated balance of methane emission from the water column into the atmosphere is from 5.4 to 57.3 μmol CH₄m^?2 day^?1.     

Analysis of applicability of triplet-state emission of molecular hydrogen for spectral diagnostics of a DC discharge

The applicability of emission of the N ³Λ_σ triplet states of molecular hydrogen for spectral diagnostics of the positive column of a dc glow discharge in hydrogen at translational gas temperatures of 360–600 K, specific absorbed powers of 0.8–4.25 W/cm, gas pressures of p = 0.3–15.0 Torr, reduced fields of E/N = 30–130 Td, and electron densities of n _e = 4.0 × 10⁹–6.5 × 10¹⁰ cm^–3 is analyzed by using an advanced level-based semi-empirical collisional?radiative model. It is found that secondary processes make the main contribution to the population and decay of the N ³Λ_σ = a ³Σ⁺ _g , c ³Π _u , g ³Σ⁺ _g , h ³Σ⁺ _g , and i ³Π _g triplet states. The dipole-allowed transitions e ³Σ⁺ _g → a ³Σ⁺ _g , f ³Σ⁺ _g → a ³Σ⁺ _g , g ³Σ⁺ _g and k ³Π _u → a ³Σ⁺ _g can be used for spectral diagnostics of a dc discharge within a simplified coronal model.     

Modelling photosynthesis in shallow algal production ponds

Shallow ponds with rapidly photosynthesising cyanobacteria or eukaryotic algae are used for growing biotechnology feedstock and have been proposed for biofuel production but a credible model to predict the productivity of a column of phytoplankton in such ponds is lacking. Oxygen electrodes and Pulse Amplitude Modulation (PAM) fluorometer technology were used to measure gross photosynthesis (P _G) vs. irradiance (E) curves (P _G vs. E curves) in Chlorella (chlorophyta), Dunaliella salina (chlorophyta) and Phaeodactylum (bacillariophyta). P _G vs. E curves were fitted to the waiting-in-line function [P _G = (P _Gmax × E/E_opt) × exp(1 — E/E_opt)]. Attenuation of incident light with depth could then be used to model P _G vs. E curves to describe P _G vs. depth in pond cultures of uniformly distributed planktonic algae. Respiratory data (by O2-electrode) allowed net photosynthesis (P _N) of algal ponds to be modelled with depth. Photoinhibition of photosynthesis at the pond surface reduced P _N of the water column. Calculated optimum depths for the algal ponds were: Phaeodactylum, 63 mm; Dunaliella, 71 mm and Chlorella, 87 mm. Irradiance at this depth is ≈ 5 to 10 μmol m^?2 s^?1 photosynthetic photon flux density (PPFD). This knowledge can then be used to optimise the pond depth. The total net P _N [μmol(O2) m^?2 s^?1] were: Chlorella, ≈ 12.6 ± 0.76; Dunaliella, ≈ 6.5 ± 0.41; Phaeodactylum ≈ 6.1 ± 0.35. Snell’s and Fresnel’s laws were used to correct irradiance for reflection and refraction and thus estimate the time course of P _N over the course of a day taking into account respiration during the day and at night. The optimum P _N of a pond adjusted to be of optimal depth (0.1–0.5 m) should be approximately constant because increasing the cell density will proportionally reduce the optimum depth of the pond and vice versa. Net photosynthesis for an optimised pond located at the tropic of Cancer would be [in t(C) ha^?1 y^?1]: Chlorella, ≈ 14.1 ± 0.66; Dunaliella, ≈ 5.48 ± 0.39; Phaeodactylum, ≈ 6.58 ± 0.42 but such calculations do not take weather, such as cloud cover, and temperature, into account.     

Assessment of photosynthetic potential of indoor plants under cold stress

Photosynthetic parameters including net photosynthetic rate (P_N), transpiration rate (E), water-use efficiency (WUE), and stomatal conductance (g_s) were studied in indoor C₃ plants Philodendron domesticum (Pd), Dracaena fragans (Df), Peperomia obtussifolia (Po), Chlorophytum comosum (Cc), and in a CAM plant, Sansevieria trifasciata (St), exposed to various low temperatures (0, 5, 10, 15, 20, and 25°C). All studied plants survived up to 0°C, but only St and Cc endured, while other plants wilted, when the temperature increased back to room temperature (25°C). The P_N declined rapidly with the decrease of temperature in all studied plants. St showed the maximum P_N of 11.9 μmol m^?2 s^?1 at 25°C followed by Cc, Po, Pd, and Df. E also followed a trend almost similar to that of P_N. St showed minimum E (0.1 mmol m^?2 s^?1) as compared to other studied C₃ plants at 25°C. The E decreased up to ≈4-fold at 5 and 0°C. Furthermore, a considerable decline in WUE was observed under cold stress in all C₃ plants, while St showed maximum WUE. Similarly, the g_s also declined gradually with the decrease in the temperature in all plants. Among C₃ plants, Pd and Po showed the maximum g_s of 0.07 mol m^?2 s^?1 at 25°C followed by Df and Cc. However, St showed the minimum g_s that further decreased up to ～4-fold at 0°C. In addition, the content of photosynthetic pigments [chlorophyll a, b, (a+b), and carotenoids] was varying in all studied plants at 0°C. Our findings clearly indicated the best photosynthetic potential of St compared to other studied plants. This species might be recommended for improving air quality in high-altitude closed environments.     

Internally illuminated photobioreactor using a novel type of light-emitting diode (LED) bar for cultivation of <Emphasis Type="Italic">Arthrospira platensis</Emphasis>

The biochemical properties of Spirulina platensis in an internally illuminated photobioreactor (IlPBR) were investigated under different light-emitted diode (LED) wavelengths; blue (λ_max= 450 and 460 nm), green (λ_max= 525 nm), red (λ_max = 630 and 660 nm), and white (6,500K), with various light intensities (200, 500, 1,000, and 2,000 μmol/m²/sec) were examined. The highest specific growth rate, maximum biomass, and phycocyanin productivity occurred under the red LEDs (0.39/day, 0.10 g/L/day, and 0.14 g/g-cell/day, respectively) at 1,000 μmol/m²/sec; the lowest growth rate was obtained under blue LEDs. Indeed, the size of trichomes was changed into short form under blue LEDs at all light intensities or all LEDs at 2,000 μmol/m²/sec for the first 2 days after inoculation, and S. platensis did not grow in the IlPBR under the dark condition. These results provide a base for different approaches for designing the pilot scale photobioreactor and developing cost-effective light sources.     

Selenate reduction rates and kinetics across depth in littoral sediment of the Salton Sea,California

To predict selenium cycling in sediments, it is crucial to identify and quantify the processes leading to selenium sequestration in sediments. More specifically, it is essential to obtain environmentally-relevant kinetic parameters for selenium reduction and information on how they spatially vary in sediments. The Salton Sea (California, USA) is an ideal model system to examine selenium processes in sediments due to its semi-enclosed conditions and increasing selenium concentration over the last century. Selenium enters the Salton Sea mainly as selenate and might be sequestered in the sediment through microbial reduction. To determine the potential selenium sequestration of Salton Sea littoral sediments and which sediment properties are controlling selenate reduction kinetics, we determined the centimeter-scale vertical distribution of potential selenate reduction rates and apparent kinetic parameters (maximum selenate reduction rates, V_max, and selenate half-saturation concentration, K_m) using flow-through reactor (FTR) experiments. We compared sediments from two littoral sites (South and North) and four depth intervals (0–2, 2–4, 4–6 and 6–8 cm). Furthermore, we characterized the selenium fractions in the sediment recovered from the FTR experiments to identify the processes leading to the sequestration of selenium. Our results reveal higher potential for selenium reduction and sequestration in the topmost sediment (0–2 cm) suggesting that microorganisms inhabiting surface sediment are well adapted to reduce selenate entering the Salton Sea. As apparent K_m values (103–2144 µM) exceed the average selenium concentration in the overlying water (6–25 nM), in situ selenate reduction is limited by the low availability of selenate and the resident selenate-reducing microorganisms operate well below their V_max (11 and 43 nmol cm^?3 h^?1). Selenium speciation after FTR experiments confirms the primary sequestration of reduced biomass-associated and elemental selenium (68–99% of total selenium) in the sediment. Further, the absence of correlation between the tested sediment physical (porosity, bulk density, clay content), chemical (C_org, N_tot, total selenium content) and biological characteristics (abundance of culturable selenate-reducers) with the kinetic parameters of selenate reduction indicates that these sediment characteristics cannot be used as predictors of apparent V_max or K_m. Conclusively, microbial selenate reduction is an important, if not the primary process, leading to the sequestration of reduced selenium in the Salton Sea sediments and making the surficial Salton Sea sediments an important selenium sink.     

Use of specific PCR primers for the study of sulfate-reducing bacteria diversity in microbial mats of Ebro Delta,Spain

Microbial mats are prokaryotic communities that provide model systems to analyze microbial diversity and ecophysiological interactions. Sulfate-reducing bacteria (SRB) play a key role in sulfur and nutrient recycling in these ecosystems. In this work, specific primers for 16S rRNA encoding gene, previously described, were used to study the diversity of SRB in microbial mats of the Ebro Delta. We confirm that this method is reliable to identify the diversity of SRB in these ecosystems. However, some mismatches in obtained sequences had been observed in our system and must be taken under consideration. Various genera of SRB in Ebro Delta microbial mats were identified, such as Desulfonema, Desulfatitalea, Desulfosalsimonas, Desulfoccocus, and Desulfovibrio. The diversity observed in our samples is very similar to previously reported in other microbial mats communities.     

Construction of flavocytochrome <Emphasis Type="Italic">b</Emphasis><Subscript>2</Subscript>-overproducing strains of the thermotolerant methylotrophic yeast <Emphasis Type="Italic">Hansenula polymorpha</Emphasis> (<Emphasis Type="Italic">Pichia angusta</Emphasis>)

L-Lactate cytochrome c oxidoreductase (flavocytochrome b ₂, FC b ₂) from the thermotolerant methylotrophic yeast Hansenula polymorpha (Pichia angusta) is, unlike the enzyme form baker’s yeast, a thermostable enzyme potentially important for bioanalytical technologies for highly selective assays of L-lactate in biological fluids and foods. This paper describes the construction of flavocytochrome b ₂ producers with over-expression of the H. polymorpha CYB2 gene, encoding FC b ₂. The HpCYB2 gene under the control of the strong H. polymorpha alcohol oxidase promoter in a plasmid for multicopy integration was transformed into the recipient strain H. polymorpha C-105 (grc1 catX), impaired in glucose repression and devoid of catalase activity. A method was developed for preliminary screening of the transformants with increased FC b ₂ activity in permeabilized yeast cells. The optimal cultivation conditions providing for the maximal yield of the target enzyme were found. The constructed strain is a promising FC b ₂ producer characterized by a sixfold increased (to 3 μmol min^?1 mg^?1 protein in cell-free extract) activity of the enzyme.     

Effect of red and blue light on acclimation of <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> to CO<Subscript>2</Subscript>-limiting conditions

Effects of red (RL) and blue (BL) light on acclimation of the unicellular green alga Chlamydomonas reinhardtii to the low level of ambient CO₂ were studied. C. reinhardtii cells grown at 5% CO₂ and under white light (170 μmol/(m²s)) had a relatively low activity of extracellular carbonic anhydrase (CA), a low affinity for dissolved inorganic carbon, and a low rate of photosynthesis under CO₂-limiting conditions. These cells readily started acclimation to the low CO₂ concentration when they were exposed to atmospheric air (～ 0.03% CO₂) under RL or BL (150 μmol/(m² s) each). The acclimation was manifested in a significant increase in the CO₂-limited rate of photosynthesis, the affinity for dissolved inorganic carbon, and the extracellular CA activity with no difference between RL-and BL-cells. Independently of light quality, the acclimation was completed for 5–7 h after cell exposure to air. As is evident from RL-and BL-dependent changes in the sum of chlorophylls and chlorophyll a/b ratio, transfer of C. reinhardtii cells to air and RL or BL triggered also the process of algal photosynthetic adaptation to light quality. However, this process did not interfere with acclimation to low CO₂ because started 4 h later. On the basis of similarity in the low CO₂-induced changes under RL and BL, it is concluded that acclimation of C. reinhardtii to CO₂-limiting conditions does not depend on light quality.     

Plasma diagnostics from intensities of resonance line series of He-like ions

The possibility of using the relative intensities of the 1snp ¹P₁–1s ^{2 1}S0 transitions with n = 3–6 in He-like multicharged ions to diagnose plasma in a nonstationary ionization state is considered. The calculations performed for F VIII ions show that, at electron temperatures of T _e = 10–100 eV, the intensity ratios are sensitive to the plasma electron density in the range of N _e = 10¹⁶–10²⁰ cm^–3. The universal calculated dependences can be used to diagnose various kinds of recombining or ionizing plasmas containing such ions.