Characterization of Suspension Cultures of Arabidopsis thaliana (L.) Heynh Cells with Altered Expression of the Gene of Alternative Mitochondrial Oxidase Aox1a and Analysis of their Frost Resistance

The enzyme alternative cyanide-resistant oxidase (AOX) localized in mitochondria is involved in the processes of plant adaptation to various unfavorable biotic and abiotic factors. Transfer of electrons from ubiquinone to oxygen by alternative oxidase has a nonprotonmotive character and, by bypassing two sites of H+ pumping in complexes III and IV, lowers the energy efficiency of respiration and energy of electron flow through AOX is released as heat. In this work, we characterized heterotrophic suspension cultures of Arabidopsis thaliana (L.) Heynh cells obtained from seeds of plants with altered (reduced (AS-12 line) and increased (XX-2 line)) expression of the alternative oxidase gene AOX1a and studied their viability under subzero temperature (-10 °С for 3, 6, 9 hours). Cell viability and reactive oxygen species (ROS) production were assessed using fluorescence microscopy with fluorescein diacetate (FDA) and propidium iodide (PI) for cell viability measurement and H2DCF-DA for ROS measurement. The proportion of living cells was calculated as the proportion of FDApositive and PI-negative cells. Differences between the studied lines were determined in the content of mitochondrial proteins of the respiratory chain (AOX, COXII, NDB) and uncoupling protein (UCP), as well as in the intensity of formation of ROS and frost resistance. The obtained results confirmed the higher content of the AOX protein and its high contribution to mitochondrial respiration in line XX-2. Suspension culture cells of the AS-12 line showed a decrease in the AOX protein content and its contribution to mitochondrial respiration, compared to the wild type (Col-0) and line XX-2. Simultaneously with a decrease in the AOX protein content in the AS-12 cell culture, an increase in the content of the uncoupling protein UCP and subunit II of cytochrome oxidase (COXII) was observed. ROS generation was reduced in cell cultures of both XX-2 and AS-12. The obtained results indicate that the cells of the wildtype (Col-0) suspension culture were subjected to the most significant effect of subzero temperature. Long-term exposure (for 9 h) under -10 °С revealed significant differences in the viability of wild-type culture cells and lines with altered AOX1a gene expression. Cells of line XX-2 with an increased content of AOX turned out to be more resistant to subzero temperature compared to wild-type and AS-12 cells. However, while the proportion of living cells in the culture of the AS-12 line 48 h after exposure remained at the same level as immediately after it, in the suspension culture of the wild type cell death developed over time. The obtained results indicate the importance of alternative oxidase in the development of frost resistance in plant cell.

Diversity of Soils of Southern Transbaikalia (on the Example of the Daursky Reserve)

The paper presents the study of soil cover of the steppe areas on the territory of the Daursky State Nature Biosphere Reserve in Transbaikalia (East Siberia). We determined the types of soils in the area surrounding the currently dry Barun-Torey lake according to the modern classification: Chestnut typical and saline, Saline gleyed typical and sulfide (sor) typical. Typical chestnut soils are formed on the tops of lake terraces and form complexes with saline soils. Salted chestnut soils occupy transit landforms. Saline soils are confined to low relief forms of the shoreline and the bottom of the lake, where the accumulation of readily soluble salts occurs. Typical Chestnut and saline soils are characterized by a light granulometric composition and a low humus content. Sulphide and typical gleyed saline soils have a heavier granulometric composition, a highly alkaline reaction of the soil solution, a very low humus content and a high content of readily soluble salts. The natural factors of soil formation are: slightly dissected relief; arid and cryoarid climate; groundwater level; steppe and halophytic meadow vegetation. Chestnut soils that form at the tops of lake terraces are the most common. Solonchaks occupy low relief forms of the lake shoreline and lake bottom.

Chlorophyll Fluorescence in Assessing the Effect of Heavy Metal Compounds on Aquatic Organisms

Fluorescent methods allow to improve the bioassays because of time effectiveness. These methods give information on chlorophyll concentration and plant photosynthetic apparatus activity in a short period of time. The aim of this study was to estimate the sensitivity of aquatic organisms to heavy metals using chlorophyll fluorescent methods. The test-organisms of different taxonomic categories were used, including thermophilic strain Chlorella vulgaris Beijer, aquatic plants Elodea сanadensis Michx. and Lemna minor L., crustacean Daphnia magna Straus. Fluorimeter “Foton 10” developed in SibFU was employed to measure prompt and delayed fluorescence (PF and DF respectively). The experiments revealed that copper ions affected DF of chlorella in 30 minutes, resulting in a 50% decrease of relative indicator of DF (RIDF) in the range of concentrations 0.005-0.01 mg/l of Cu2+. Measuring the DF of duckweed was less time-consuming compared to registration the changing of morphological parameters when the plants were exposed to heavy metals. Copper, cadmium, and nickel exposures were found to decrease the RIDF of duckweed by more than 50% at the concentrations of 0.17, 0.31, and 1.89 mg/l respectively. However, zinc had no significant effect on the RIDF of Lemna minor in the range of 0.1-8 mg/l of Zn2+ within 24 h of exposure time. The analysis of fluorescent parameters of Canadian elodea showed the possibility of using the plant as a sorbent during the bioremediation of aquatic environments from heavy metals. Registration of chlorophyll fluorescence allowed revealing the toxic effects of negligible concentrations of heavy metals in experiments with the feeding rate of daphnids. The median effective concentrations (EC50) were 0.002, 0.02, 0.4, and 0.25 mg/l of Cd2+, Cu2+, Zn2+, and potassium dichromate respectively. This makes it possible to obtain information on the effects of pollution in the early stages of exposure and in a shorter time. Thus, the use of chlorophyll fluorescence in biomonitoring the state of the aquatic environment makes it possible to more quickly respond to changes in the ecological situation in water bodies.

Vital Dye Absorption by a Baikal Sponge as a Test Response to Surfactant Exposure

Baikal sponges account for a large, if not the largest proportion of zoobenthos of Lake Baikal, especially in the littoral. It is here that the largest number of pollutants comes. A fixed lifestyle, inertia, and the absence of visible manifestations of the life activity of the sponges create significant specific difficulties when working with them in a laboratory experiment. The lack of clear criteria, test reactions, and rapid biocontrol systems hinder the determination of the physiological state of the sponges, their activity, degree of infection, cure, and the influence of various stress factors on them. In this regard, there is relatively little information about the reactions of Baikal Porifera to toxicants. This report presents experimental materials on the change in dye absorption by Baicalospongia bacilifera sponge under the influence of one of the most common components of domestic wastewater – sodium dodecyl sulfate. A negative effect was noted with a 2-hour exposure to the sponge of this surfactant in a concentration of 25 mg/l and higher. The obtained experimental data suggest that, in further studies, bleaching of dye solutions with a sponge of B. bacilifera can serve as a test reaction for the presence of contaminants.

Characteristics of Serratula centauroides (Compositae) Coenopopulations in Transbaikalia (East Siberia)

The coenopopulations of Serratula centauroides in various plant communities of Transbaikalia have been studied. In the study areas 10 geobotanical descriptions were made in plant communities with the participation of S. centauroides. In the investigated area S. centauroides is found on dry rocky slopes, in herb and sandy steppes. Depending on the growing conditions, the density of individuals in coenopopulations ranges from 0.18 to 1.80 individuals per 1 m2. The highest occurrence of S. centauroides is noted in communities dominated by Gramineous and Artemisia: Gramineous-Artemisia-herb, Filifolium-Artemisia-Gramineous, Gramineous-Artemisia, Oxytropis-Gramineous and Allium-Artemisia. Studied coenopopulations S. centauroides are confined to the following communities: Filifolium-herb, Gramineous- Artemisia-herb, Sophora-herb, Filifolium-Artemisia-Gramineous, Gramineous-Artemisia, Agropyron- Carex, Carex-herb, Oxytropis-Gramineous and Allium-Artemisia. The phytomass, biological and exploitation resources, the possible volumes of annual harvesting of raw material of S. centauroides were determined. The phytomass of S. centauroides herb ranges from 8.10±3.46 to 75.93±19.55 g/m2 (air-dry raw material). The average phytomass of 1 individual of S. centauroides herb varies from 13.67±3.59 to 67.14±14.30 g (air-dry raw materials). The total biological resources of S. centauroides herb is 777.9 t (fresh raw materials) and 266.8 tons (air-dry raw materials). The total exploitation resources of S. centauroides herb is 224.7 t (fresh raw materials) and 79.8 t (air-dry raw materials). The possible volume of annual harvesting of raw materials of S. centauroides in the studied coenopopulations on the territory of the Trans-Baikal Territory and the Republic of Buryatia amounts to 37.5 t (fresh raw materials) and 13.3 t (air-dry raw materials).

First Findings of Sunbleak Leucaspius delineatus (Cyprinidae) in the Lake Baikal Basin

Since the early 1960s, the sunbleak Leucaspius delineatus (Heckel, 1843), along with carp acclimatized from European Russia, regularly entered the siberian carp ponds, first in the Ob’ basin, and then in the Yenisei and Angara basins. In the last decade, we have discovered the sunbleak in two water bodies of the Baikal basin: in May 2013 - in the water intake and discharge canals of the Gusinoozersk regional power plant situated on the large lake Gusinoe in the Selenga River basin (Republic of Buryatia), and in November 2016, the sunbleak was registered in the diet of pike inhabiting the reservoirs of the Talaya-Pokhabikha lake-bog complex in the immediate vicinity of the Baikal coastline at the southern end of the lake in Irkutsk Region. The age series in the samples of the sunbleak in the water intake canal of the power plant was represented by five age groups from 1+ (SL 34.2 mm and 0.59 g of weight) to 5+ (SL 73.9 mm and 6.73 g, respectively) with a predominance of individuals aged 1+ and 2+. In the discharge canal only group 1+ (SL 35.5 mm and 0.69 g) was noted. The sex ratio in catches at the age of 1+ was 1:1.2 with a predominance of males; by the age of 3+, the proportion of males increased significantly (1:8.6). Sexual maturity of the sunbleak from Lake Gusinoe occurs at the age of 3+. Spawning is portioned, the first portion is layed down not earlier than the second half of July. The total fecundity of the seven studied females at the age of 3–5 + varies from 632 to 1539 eggs, averaging 1312 eggs. The sunbleaks from Lake Gusinoe are predominantly benthophagous. In the first half of May 2013, near the water intake canal, the main food components were the nymphs of the Corixidae water bugs and larvae of Phryganea caddis flies, planktonic crustaceans and imago of aerial insects were also consumed. In July 2014, in the discharge channel, the main food item was the chironomid larvae. In the lake Sludyanskoye, three specimens were identified from the stomachs of the pike living there. All fish were partially digested, and therefore it was not possible to analyse it.

Biotopical Distribution of Orb-Weaver Spiders (Aranei: Araneidae) in the Ridge-top Zone of the South Sikhote-Alin Mountains

Until recently, the study of spiders in the mountainous conditions of the Primorsky Region of Russian Far East was carried out only with the example of ground dwelling spiders. The purpose of this work is to reveal the species composition and features of the biotopic distribution of orb-weaver spiders (Aranei: Araneidae) in the ridge-top zone of the Southern Sikhote-Alin mountains (Primorsky Region). The field material was collected from may to october in 2016 and 2017 at key sites located on different mountain ranges and their spurs in the altitude range from 1000 to 1560 m above sea level. Within each key sites, transect-sites with a size of 3 x 25 m were laid. The spider's collections were by the hands collecting and sweeping method. A total of 305 specimens of spiders belonging to 10 species from 7 genera were caught. Comparison of species lists of spiders was carried out by cluster analysis methods (unweighted pair group method with arithmetic mean, UPGMA) using the Jacquard coefficient. For automatic calculations data processing program Past v. 2.17 is used. The maximum diversity of spiders is observed among the herbage-reed meadows, which do not form a continuous belt of vegetation at altitudes of 1000–1400 m above sea level. seas. Seven species belonging to 6 genera were caught here. The minimal diversity is noted among the continuous open stony debris, devoid of vegetation. Here is the only species caught – Aculepeira matsudae. The species Plebs sachalinensis and Araniella yaginumai are most numerous and widespread in the ridge-top zone of the Southern Sikhote-Alin. The high abundance of Araniella yaginumai is achieved due to favorable conditions for this species in the herbage-reed grass lands. In all the studied biotopes in the ridge-top zone of the mountains the density of the orbweaver spiders is low and does not exceed 2.04 specimens per 10 m2.

Corticolous Lichens of Belarus. II. Obligate and Facultative Epiphytes

The lichen biota of Belarus lists 406 corticolous species. Of these, 213 (35.7%) species are obligate epiphytes. Crustose lichens make up the majority of obligate epiphytes (157 species, or 73.7%). Apparently, this can indicate the decisive role of the morphology and chemistry of the substrate for the closely contacted lichen thallus. Facultative epiphytes are represented by 193 species, which are quite widely represented by foliose and fruticose life forms (51.3%). Facultative epiphytes inhabit rotting and processed wood, stony substrates, soil (including forest litter), mosses, leaves (needles), root turnouts and metal objects. Wood is inhabited by 154 facultative epiphyte species, of which 80 lichens are strongly epiphyticlignicolous. The lichen diversity of tree bark and wood is relatively similar; the value of the Sørensen-Dice index is equal to 0.51. Soil is the second most important substrate after wood for facultative epiphytes. It is inhabited by 55 lichens, most of which are represented by Cladonia and Peltigera species. 46 species of facultative epiphytes were found on mosses. These are represented mainly by cyanobiont-containing lichens, broad-lobed species, as well as many by the representatives of the genus Cladonia. The stony substrate is suitable for 43 facultative epiphytes species and is characterized by a high specificity of lichen biota. Its Sørensen- Dice index is equal to 0.13. Most of these representatives are common in urban environments. Fungi, lichens, root inversions, leaves, and metal are predominantly inhabited by multisubstrate lichen species.

Assessment of the Effect of Low Glyphosate Concentrations in Forage Cereal Crops on the Biochemical Parameters of Blood and Organs of Laboratory Mise

Chemical methods for managing invasive plants in agriculture are dominant at present. Glyphosate-containing preparations are used as a desiccant and for drying of crops during pre-harvest period. Residual quantity of glyphosate founded in the grains of different crops was between 0.70 and 33.0 mg/kg. For example, the herbicide concentration in oats grown in the UK was about 0.9-14 mg/kg. As the result, herbicide is found in the environment, forage, food staff, animal tissues and human biomaterial. Chemical methods of analysis cannot conclude the toxic effects of herbicides on the environment, warm-blooded animals and humans. The purpose of the study was to examine the toxic effect of glyphosate on biochemical blood assays and organs of the laboratory rodents. The evaluation of influence of glyphosate was based on studies using 48 rodents. Chronic toxicity effects were determined by feeding rodents with grains containing 7, 14, 28 mg/kg of glyphosate. The exposure time was more than 4 months. The criteria for toxicity were based on biochemical changes of blood cells and tissues of rodents. Significant deviations of laboratory rodents’ physiological standards of blood indicators were determined during the study. The results of laboratory researches demonstrate quantitative and qualitative changes of blood cells. Glyphosate intoxication in concentration of 7, 14, 28 mg/kg caused leukopenia, anemia, by reducing the number of leukocytes and erythrocytes. As it was intimated, the average reduction of number of leukocytes in blood by 30% and 66 % was founded in the group of rodents fed with grains containing 7 mg/kg of glyphosate after 4 and 6 months of the experiment; in the group of rodents fed with grains containing 14 mg/kg of glyphosate after 4 and 6 months of the experiment the average reduction reached 30% and 62 %; in the group of rodents fed with grains containing 28 mg/kg of glyphosate after 4 and 6 months of the experiment the average reduction reached 38 % and 73 %. The decrease of erythrocytes in blood of laboratory rodents demonstrates a greater dose-effect relationship in comparison with leukocytes. The average reduction of number of erythrocytes in blood by 23% and 28 % was founded in the group of rodents fed with grains containing 7 mg/kg of glyphosate after 4 and 6 months of the experiment; in the group of rodents fed with grains containing 14 mg/kg of glyphosate after 4 and 6 months of the experiment the average reduction reached 36% and 47%; in the group of rodents fed with grains containing 28 mg/kg of glyphosate after 4 and 6 months of the experiment the average reduction reached 43% and 59%. Qualitative changes in erythrocytes were determined by changes in size and shape of cells. Postmortem examination of laboratory rodents founded changes in the liver, small and large intestine. The hepatotoxicity of glyphosate was expressed by changes in liver size, consistency and color. Negative influence of herbicide on the gut walls of small and large intestine was also expressed by the changes of intestinal mucosa color, consistency. A group of rodents fed with grains containing 28 mg/kg of glyphosate faced more abnormal changes of gut walls of small and large intestine, being reflected by the appearance of splinter hemorrhages under the intestinal mucosa of small intestine. The study also determined the negative effects of glyphosate on mammalian reproductive function and the viability of the posterity of laboratory rodents. The results demonstrate that chronic toxicity of herbicides contributes to the deviation of hematological and biochemical indices of blood, negatively affects differentiative state of liver, gut walls.

Supplements to the Flora of North of Irkutsk Region

During the floristic survey of certain sections of the Kirensky, Kazachinsko-Lensky and Ust-Kutsky districts of the Irkutsk region in 2016–2018, 79 species of vascular plants were identified, new or rare for the Prilensko-Katangsky (PR) floristic district in three areas: Lena- Angara plateau (PV-11), Cisbaikalian depression (PV-12) and Prilenskoe plateau (PS-14). For PR in general, 6 species were found for the first time, for PV-11 and PV-12 – one species each, for PS-14 – 30 species. It also provides information about the new locations of 9 plant species included in the red book of the Irkutsk region.