Tomsk State University Journal of Biology

7-19

Authors: Nina S. Evseeva, Galina E. Pashneva, Zoia N. Kvasnikova

The results of long-term researches (1985–2012) of deluvial process in agrolandscapes of the southeast of the Western Siberia are presented. Deluvial process is washing away of soils by thawed and rain snow (a water erosion of soils); it is one of the main forms of migration of substance in the agrolandscapes, leading to a decrease in fertility of soils and efficiency of a soil cover, etc. The work purpose is an assessment of intensity and dynamics of development of diluvial process on an arable land of the southern part of Tomsk region and identification of ecological aspects of this process. It is established that: 1) Natural factors such as relief, lithology and climate are favorable for development of diluvial process. The steepness of slopes of an arable land changes from 0–1° to 10–12°, sometimes more. On the surface there are deposited loess rocks that soak overnight in water (for 2–120 minutes); soils have low resistance to erosion and runoff. For the territory, intensive storm activity is characteristic, during one heavy rain there are up to 80.5 mm of precipitations. Average annual water reserves in snow vary from 100 to 140 mm, in some years up to 180 mm. 2) The intensity of soil erosion, heavy rains in the area between Tom-Yaya is 40–100 m3/ha in the event of loss of rainfall over 50 mm/day, more runoff from the slopes of arable land is 0.5–8 m3/ha. 3) The intensity of soil erosion by melted snow water varies from 0.5–1.0 m3/ha to 50–80 m3/ha, depending on weather conditions of snowmelt, slope, length, exposure, slope, soil fertility status, etc. The main contribution to the development of soil erosion is made by thawed snow water. 4) The analysis of literature and our data show that during snowmelt from 0.6 to 15 kg/ha of humus, from 0.13 to 1.8 kg/ha of nitrogen, 0.11–0.90 kg/ha of phosphorus, etc. are annually carried away by melting waters from 1 ha of arable land.

36-46

Authors: Olga A. Nekrasova, Maria I. Dergacheva

The growth of rare elements used in modern industrial production requires research activation with a view to their monitoring and regulation. The comparison of trace elements content in modern soils with ancient analogues withdrawn from active functioning, can help to assess the accumulation of elements in soils of the date hereof. Modern background soil, as well as paleosols and cultural layers of the archaeological site Stepnoe 7 of Bronze epoch, located in the Southern Urals in Chelyabinsk region are the objects of the study. Profile of modern soil background in morphology corresponds to leached chernozems, has a slightly alkaline environment and the magnetic susceptibility of organic-horizon 2.3–3.5×10–6 CGSE per/g of sediment. The humus horizon contains on average 3.2% of organic carbon, about 50% of it is humic acids, which exceeds the number of fulvic acids in more than 2 times and causes humate type of humus. Humic acids of the A horizons upper thickness of the soil profile contains 50.5± 3.4% C, 4.2 ± 0.3% H, 41.5 ± 2.87% O and 3.3 ± 0.3% N from mass. Buried soils and cultural layers have similar characteristics: the humus content is about 1%, it is characterized by humate composition, and the elemental composition of humic acids corresponds to steppe conditions of their functioning. Humic acids were allocated from 0.1n NaOH extracts after preliminary decalcification by their sedimentation 2n HCl, with the subsequent dissolution in alkali and resedimentation. The total content of rare earth elements in humic acids and soils was determined by multi-element neutron activation analysis in the laboratory of Tomsk Polytechnic University. A comparison of the content of rare earth elements in modern soils, paleosols and cultural layers with Clark in the lithosphere allows us to consider them non-contaminated lanthanides. Waning row of average element content is the same for all objects under study and looks as follows: Ce> Nd> La> Sm> Yb> Eu> Tb> Lu. However, for almost all of the elements the average content in the modern background soil is much higher than in the buried soils and cultural layers, although significant differences using the Student’s t-test for the objects being compared are not identified for any metal. In order to establish the contribution of HA in the binding of rare earth element content of soils, the contents of elements in the preparations were recalculated to their number in the soil, per humic acid. The content of total organic carbon in the soil, the share of humic acids in the humus composition and carbon content in HA preparations were taken into account. The total percentage of all studied rare-earth metals in humic acids in relation to their content is 4/03% in the modern soil, 1.56% in the cultural layers and 1.28% in the buried paleosols. Thus, it clearly shows a tendency of increasing the content of rare earth elements in modern southern forest-steppe soil of Zauralye in the last four thousand years. The share of trace elements associated with humic acids of modern soils in the total pool of rare earth elements of more than 3 times higher than in paleosols and cultural layers.

67-76

Authors: Tatjyna M. Pankina, Vladimir N. Romanenko, Svetlana I. Istratkina, Aleksandr V. Shichin, Tatjyna M. Poltorackaj

There are some centers of natural diseases, trans-missed by ixodes ticks оn the territory of Tomsk region. The most dangerous of this are ixodes ticks encephalitis (ITE) and ixodes ticks borreliosus (ITB); but some other diseases have been registered (erlihiosus, West Nile fever). Between 16000 to 25000 suction attacks of ticks and up to 1000 morbid events are registered on the territory of Tomsk region annually. Ixodes pavlovskyi Pom 1946 dominated on town and suburbs territories, but as far as from town and the Tom river valley Ixodes persulcatus P.Sch 1930 became dominant species (up to 100%). Seasonal activity of ticks in Tomsk district characterized by a cleary shown one pike. Maximal density of ticks is registered in the first part of June, but in August most part of ticks disappears. On the town’s territory the pike of activity is registered early – in the 2–3 decades of May. On the town’s territory ticks disappear at the end of June and on the suburbs territories up to mid-July respectively (2–4 weeks earlier). The whole period of ticks activity on the south part of Tomsk region is 120–200 days in case of absence of human impact. Ticks virus infectiousness collected from the grass on the territory of Tomsk district was specified. The part of ticks infected by ITE was 1.9–7.7% during 2002–2011. The lowest part of infected ticks was nearby Kolarovo (0–6.3%) and Kurlek (0–0.5%) villages. There were no viruses in ticks collected nearby Semiluzhki and kuzovlevo villages, The high part of infected ticks were collected in the forests nearby Bogashevo (up to 11.1%), Haldeevo (up to 12.7%) and Nekrasovo (up to 13.3%) villages. Virus of ITE was specified more often (by the IFA method) in ticks suckinghuman blood. The virus of ITE was found in the ticks sucking human blood as often as: in Kolarovo 21.2–29.7% in Bogashevo – 15.7–33.3%, in Haldeevo – 16.0–22.2%. In the ticks sucked to a human in Tomsk and in suburbs the virus of ITE was found in Ixodes pavlovskyi (from 34.76%) and in Ixodes persulcatus (49.78%). Among ticks inhabited in the natural ecosystems of Tomsk the percent of ticks infected by ITB was up to 63,8 in Academgorodok and up to 36,4% nearby Irkutsky road. The virus of ITB was found in ticks in Kolarovo (62%), in Bogashevo – up to 36.6%, in Haldeevo – up to 40% , in Nekrasovo – 15.4% and in Kuzovlevo – 13.3%. The middle percent of ITB infected ticks in Tomsk region was between 21.3 and 49.8% during last ten years. There were maximal infectiousness among ticks collected from the grass in spring period (3.1–7.1%); in the mid-July there were few infectious ticks only. The percent of infectiousness among over-wintering ticks were at the minimum in spring (19–25%), but increasing during summer up to 32–35%. The most effective measure against ticks was on the territories with clear borders and with good preliminary preparation for the action. For example, on the territories of summer child camps the effectiveness of actions was 95–100% and the number of sucking ticks were as less as ten during the season.

98-108

Authors: Gleb N. Artemov, Gleb A. Sapunov, Vladimir N. Stegniy

Chromosome attachment to the nuclear lamina is one of the main mechanisms of chromosome positioning in the nuclear interior. Previously, contacts of chromosomes with nuclear envelope in semi-squashed malaria mosquitoes nurse cells nuclei were discovered and its species-specificity was showed. We localized nuclear debris on polytene chromosomes of nurse cells of sibling species Anopheles atroparvus Thiel., An. messeae Fall. and An. beklemishevi Steg. et Kab. by chromosome immunostaining with using antibodies to fruit fly lamin Dm0. Distribution of lamin on the shortest chromosomes of the set – XL chromosome was analyzed. Lamin was detected in distinct chromosome regions, mainly in interbands. We used only signals, which were found more than in four nuclei. These regions looked like bright bands and were detected in all analyzed specimens. If intra-specific polymorphism by lamin distribution take s place, it cannot be shown by using our methodical approach. However, we found some single nuclei, which had different signals in each homolog chromosomes. The brightest lamin signals quantity, width and their distribution along chromosomes are species-specific features. In An. messeae and An. beklemishevi lamin revealed in chromosome attachment regions, which was mapped during semi-squashed nurse cells nuclei investigation. The lack of lamin signals in XL chromosome attachment region of An. atroparvus is connected with specific morphology of pericentric regions and with limitation of preparation making. We did not observe significant concordance in lamin localization on XL chromosome in analyzed species. However, we found one similarity between An. messeae and An. atroparvus in lamin localization in 2b-2c region. In spite of similar localization, these regions differ from each other by intensity signals, which can reflect differentiation in the strength of chromosomes contacts. The obtained results do not prove data about evolution conservatism of lamin-associated domains localization in distinct cell types. Interspecific differences in lamin distribution along the XL chromosome suggest chromosome region structure transformation, which results in acquisition or loss of the attachment function or modification of the strength of chromosome contact with nuclear envelope.

117-123

Authors: Konstantin E. Usov, Alina A. Kokhanenko, Vladimir N. Stegniy

It is known that DNA sequences associated with heterochromatin are difficult for sequencing and subsequent analysis because of repeats enrichment. So, even finished and published genome sequencing projects of different organisms only contain information about the DNA sequences associated with euchromatin. Now, however, it is clear that understanding the principles of the eukaryotic genome functioning requires detailed information on the composition of its sequences. It should be noted that a comparative analysis of DNA in heterochromatin both phylogenetically close and distant taxa is relevant from the point of view of the problem of the evolution of genomes. Previously, we obtained region-specific DNA library chromocenter of polytene chromosomes of D. orena by microdissection. Next, on its basis we got a set of DNA clones and carried out their sequencing. The analysis of the clones using various software packages allowed to find out a wide variety of repetitive sequences among them (mobile genetic elements; tandem repeats) and the identified sequences homologous to the protein-coding genes. In addition, using microdissection method we obtained a region-specific DNA library chromocenter of polytene chromosomes of D. virilis. As part of this work, we carried out dot blot hybridization region-specific DNA libraries from the chromocenter D. virilis with characterized and annotated 42 DNA clones chromocenter D. orena. That allowed us to obtain new data on the molecular composition of DNA chromocenter D. virilis and estimate, generally, conservative composition of pericentromeric heterochromatin DNA in these relatively distant in the phylogenetic relation to species like D. orena and D. virilis. It should be noted that of 42 examined clones, only 8 failed to detect clones in the polytene chromosome DNA chromocenter D. virilis. Thus, it is established that, despite the evolutionary distance D. orena and D. virilis, DNA pericentromeric heterochromatin of polytene chromosomes in these species contains a significant amount of common sequences such as dispersed repeats (mobile genetic elements) − LTR-retrotransposons, LINE-elements, DNA transposons, tandem repeats (minisatellites), and sequences homologous to the protein-encoding genes (the genes, apparently, belong to the so-called orthologs).

133-144

Authors: Irina F. Golovatskaya, Vyacheslav Yu. Dorofeev, Yulia V. Medvedeva, Pavel E. Nikiforov, Raisa A. Karnachuk

The method of isolating apical meristems and obtaining from them virus-free regenerants is the main way to make potato plants free from pathogens. Further cultivation of the improved potato in hydroponics allows obtaining virus-free tubers, thereby solving the problems of modern seed-growing of this crop. The in vitro cultivation of micropropagated plants is carried out at artificial light. In this connection it is necessary to choose a light regime to optimize the growth processes of microcuttings. The effect of additional irradiated with red, white and blue light on the morphogenesis of potato cv. Lugovskoy in vitro was investigated. Different growth reactions of micropropagated plantlets during subcultivation were revealed. The influence of the correction of white light on the formation of root, shoot and the photosynthetic apparatus of Solanum tuberosum L. cv. Lugovskoy plantlets in the process of cultivation in vitro was studied. The supplementary lighting with blue, red and white light was used. The greatest increase in dry weight of shoots was observed in plants under red light. At the same time, the blue light inhibited or changed the direction of synthetic processes in the shoot, causing a decrease in its dry biomass. Red light increased the root volume and the total area of leaves, compared with the action of the blue light. Irradiating with blue light increased the area of leaf surface 3–5 tiers, whereas irradiating with a white and red light – the area of 6–9 tiers, while the smallest number of tiers was formed under blue light conditions. The levels of chlorophyll a, chlorophyll b and carotenoids were increased at red and blue light compared with a content of pigments under white light. Red and blue light played an important photoregulation role in the growth and morphogenic responses of potato plants, both at the shoot and root system. Specific effect of light of different spectral composition on morphogenesis of microcuttings potato cv. Lugovskoy should be considered for optimizing cultivation regime in vitro. The practical significance of the research results is to use the method of irradiating with selective light at work to develop virus-free potato micropropagated plants with a view to accelerating the growth in vitro, the successful adaptation of plants in vivo, the reduction of the vegetation period and the increase in productive process for obtaining the maximum output of the improved minitubers in the conditions of hydroponics.

152-161

Authors: Igor V. Volkov, Irina I. Volkova

The analysis of long-term dynamics of two high-mountain phytocoenoses of Aktru valley (Altai Republic) growing in different landscapes shows that the species compositions, horizontal and vertical structure as well as ecological features of communities have undergone various modifications. The most perceptible changes for the period of 2000–2011 have been revealed for the community occupying the lateral side of moraine near the glacier body; this fact is connected to the intensifying glacier melting at the ablation zone and decreasing environmental influence of the glacier to the adjacent territory. The changes in periglacial zone were reflected at considerable increase in substrate humidification; that was ascertained by the method of ecological scales. Microclimate change caused disappearance of ultra-oreophytes and mountainsteppe plant species. Biomorphological transformation of the community is proceeding from predominance of small cushion life forms, to prevalence of prostrate life forms under essential growth of the ground coverage. Pinus sibirica turned from procumbent (elfin wood) to arboraceous life form. Transformation of the alpine meadow situated out of the zone of influence of glacier body for the period of 1993–2010 has extended over the gradual modification of the plant community structure, increase in species quantity and herbaceous layer height, with relatively constant optimums of plants on such ecological factors as moisture, nutrient richness and salinity of substrate. These changes reflect the gradual shift of the mountain belts as a result of annual temperature growth. The results of our research have revealed that the most significant transformation of the vegetation connected with climate features change can be observed in the vegetation of the moraines at the ablation zone of glaciers.

7-18

Authors: Matrena V. Okoneshnikova

The data describing the state of soil and soil cover prior the anthropogenic impact were obtained during geotechnical and ecological investigations which were carried out for the assessment of potential impact on the environment caused by gas pipeline and bridge construction across the Lena River. The studied area is located in the central part of the Lena River valley and is adjacent to the city of Yakutsk (61°5’ N). It covers all geomorphological levels of the floodplain complex and includes lower fluvial terraces (I and II) on the both banks of the river. The soils of the floodplain complex are represented by typical gley, gleyic, chernozemic and muck subtypes of alluvial sod soils. On the basis of morphological, physical and chemical properties, typical alluvial sod soils with thick humus horizon (up to 20–30 cm), which are optimally moistened and packed, are the most resistant to mechanical treatment and chemical pollution of all floodplain soils. On the lower fluvial terraces, meadow-chernozemic and chernozemic-meadow soils with different salinity are widely represented. The type of salinity is predominantly sulphatic and chloridic that is accounted for the main role of sulphates and chlorides in saline composition of the ascending solutions. Sod-meadow soils with clearly marked sod horizon and neutral pH are less saline and more fertile. Almost all soil types in the upper part consist of packed sod-humus horizon of sandy-loam and light-loam granulometric composition. Mechanical disruption of humus horizon may cause deterioration of physical properties of soil and an increase or initiate gleization and thixotropic processes; it may also lead to thawing of permafrost and activation of water erosion. Within the right of the way of earthmoving machines, the disruption of the entire soil profile was observed, particularly mixing and transforming horizons, thermal erosion and unwanted cryogenic processes (thermokarst, mudflow and subsidence) increased as well. In over-moistened areas of alluvial sod-gley and sod-muck soils the disruption of soil and vegetation cover may be accompanied by temporal or permanent swamping of the territory. It was also determined, that the average concentration of heavy metals and oil products in all the studied soils is significantly lower than their permissible concentrations recommended in a series of Russian regulatory documents for sandy loam soils. However, the poor ability of soils of central Yakutia to self-purification predetermines a high risk of stable accumulation of pollutants in the studied soils.

38-45

Authors: Anna N. Vorobyeva, Dementii D. Basargin

The leaf epidermis of taxonomically difficult Far East species of Saussurea pulchella and S. neopulchella were studied. The results of the morphometric analysis of the epidermal cells and stoma are presented. A comparative research of the epidermal cells and stoma of the leaf of S. pulchella and S. neopulchella allowed revealing constant parameters at the level of the genus: they are the form of around stomataand epidermal cells, the type and the form of stoma and trichomes. For both species of plants, the stomata of anomocytic type are characteristic. The stomata are of elongated and roundish form and they are surrounded by 3–7 cells. The size, the amplitude and the frequency of waves of around stomata cells correspond to the parameters of the epidermal cells. The form and the size of cells adjoining the guard cells do not differ from the epidermal cells. A certain orientation of around stomata cells is not observed. The epidermal cells are of a polygonal or wrong form with rectilinearly-roundish or twisting outlines. The projection of epidermal cells is polygonal, roundish or spread. Both species of plants have single-row multicellular simple trichomes on the upper and the lower surface of the leaf. They are formed by cells of a cubical form with the extended apical cell. In the basis of the trichome, there is a cell of a roundish form which is surrounded by polygonal epidermal cells dispersing in radial directions. The parameters identifying species are revealed; they are the type of the leaf, the size of the epidermal cells on the longitudinal (long) axis and the stomatal index. The leaf of S. neopulchella is hypostomatic (the stomata on the upper surface of the leaf are absent), whereas the leaf of S. pulchella is amfistomatic (the stomata are present on the upper and the lower surfaces of the leaf). The sizes of the epidermal cells on the longitudinal axis of S. neopulchella on the epidermis of the upper surface of the leaf change within the limits from 51.24 to 78.84 microns and on the epidermis of the lower surface of the leaf – from 27.93 to 56.4 microns. For S. pulchella, these indicators are within the limits from 48,89 to 84,28 microns for the epidermis of the upper surface of the leaf and from 42.52 to 82.97 microns for the epidermis of the lower surface of the leaf. The stomatal index of the species also differs: for the upper epidermis of S. neopulchella it is 0% and for the lower epidermis – 19.93%; for the upper epidermis of S. pulchella the index is 8.48% and for the lower epidermis – 22.79%. The researched parameters

56-72

Authors: Galina Ya. Stepanyuk, Lyubov V. Khotskova

The phenorythmic features of seven species of Dendrobium Sw. during cultivation in hothouses of the Siberian botanical garden of Tomsk state university were studied. The studied Dendrobium species are divided into four phenorhythmic groups on terms of flowering in garden greenhouses: blossoming during winter period, blossoming during winter and spring periods, blossoming during summer period and blossoming during autumn and winter periods. Artificial pollination of Dendrobium flowers was carried out. Biometric characteristics of fruits of the studied Dendrobium species and terms of their maturing during cultivation under conditions of greenhouses of the Siberian botanical garden of TSU were submitted. When cultivating the species of Dendrobium in greenhouses of the Siberian botanical garden, the duration of maturing of D. phalaenopsis fruits makes 155 days, D. nobile fruits – 247 days and D. kingianum fruits – 67 days. The technique of sterilization of fruits and seed crops on artificial nutrient mediums was fulfilled. The stages of the development of Dendrobium seedlings on nutrient mediums in vitro were studied. It is established that plants are ready to disembarkation in an intermediate substratum on the average in 330 days after crops of seeds. The period of adaptation of Dendrobium seedlings in an intermediate substratum made 10–15 months, then the seedlings were replaced in constant substrata. Average survival of Dendrobium seedlings in an inter mediate substratum made 72.5 ± 6.5%. It is established that D. phalaenopsis is characterized by slow growth. An increase in D. phalaenopsis shoot makes 2.25 cm a year, and the average growth rate of a leaf – 2.34 cm a year. For the sixth year intensification of growth of D. phalaenopsis shoots is observed; that is, connected with transition of plants to the generative period of development. Young plants of D. phalaenopsis blossomed in greenhouses of the Siberian botanical garden on the sixth year of their life. The influence of mineral and organic nutrition on the intensity of growth and development of young plants of D. phalaenopsis and D. nobile was investigated. It is established that optimum nutrition is solution of mineral salts on Murashige&Skoog’s copy-book in concentration of 1 ml of the MS solution on 1 l of water. A way of reproduction of four Dendrobium species by «live children» was developed. It is established that from one shoot of D. phalaenopsis and D. nobile it is possible to receive up to 5 top shoots-suckers a year, and from one shoot of D. kingianum and D. delicatum – 3 top shoots-suckers a year. Survival of «children» of these Dendrobium species averaged 81.3%. It is established that Dendrobium plants multiplied by this method and blossomed for the third year of their life in hothouses of Siberian botanical garden.

83-90

Authors: Vadim V. Yartsev, Valentina N. Kuranova

Genus Siberian Salamanders Salamandrella Dybowski, 1870 includes two species which are considered cryptic: S. keyserlingii Dybowski, 1870 and Salamandrella tri dactyla Nikolsky, 1905. The data on reproductive isolation between species are absent. We have conducted experiments of the hybridization between females of S. keyserlingii (n = 3) and males of S. tridactyla (n = 24) in the laboratory. Females of S. keyserlingii were captured in the vicinities of Tomsk (Western Siberia), and males of S. tridactyla – in the vicinities of Khabarovsk (Far East) during the breeding season. The animals were observed in aquariums. Males performed tail and body vibrations to attract females. Females may also swing their tails to attract males. In general, the reproductive behavior of the S. tridactyla and the S. keyserlingii is similar. Two of three clutches of females of S. keyserlingii were fertilized by males of S. tridactyla. Embryos developed, and then larvae appeared. The similarity of behavior and the development of hybrid embryos indicate the absence of behavioral and gametal barriers between species. The results show the possibility of hybridization between species. Further studies of evaluation of the viability of embryos, larvae and postmetamorphic stages, as well as hybrids’ fertility are necessary.

100-110

Authors: Vladimir А. Ternovoy, Viktor А. Svyatchenko, Мargarita V. Таrаsоvа, Nikolay N. Кiselev, Еlena V. Chub, Тamara P. Mikryukova, Еlena V. Protopopova, Valeryi B. Loktev, Petr М. Chumakov, Sergey V. Netesov

The use of oncolytic viruses is one of the promising areas of malignant diseases modern treatment methods. In numerous studies of the adenoviruses strains oncolytic properties, their efficacy and safety in the treatment of various cancers were shown. The vast majority of the studies were carried out with the use of adenovirus serotype 5. To improve these anticancer agents means to increase their specificity for cancer cells and their lytic activity. To this end, adenovirus serotype 5 recombinant variants were constructed expressing apoptin (VP3 protein of chicken anemia virus) and causing p53 and bcl-2 independent apoptosis, increasing the lytic properties of viruses. Furthermore, to increase the specificity of adenovirus replication towards tumor cells, telomerase reverse transcriptase promoter (hTERT) was used. According to the data indicating that the apoptin expression in the cells is accompanied by induction of apoptosis in tumor cells only, the adenoviruses variants were constructed in which apoptin expression is regulated by cytomegalovirus promoter (CMV), which is stronger than hTERT. As a result, the design of apoptin gene sequences, CMV and hTERT promoters and the construction of recombinant plasmids bearing them, which are necessary for constructing recombinant adenoviruses, was created. Using the methodology of homologous recombination and co-transfection of cells, two replications of defective recombinant adenovirus AdΔE1a-hT-Apo and AdΔE1a-CM-Apo with insertions of apoptin gene under the control of the hTERT-and CMV-promoter, respectively, were obtained. In these recombinants, E1A and E1B genes, necessary for viral replication, are deleted, which makes such agents safer. The third constructed recombinant represents replication competent adenovirus AdhTE1a-CM-Apo with deletion of E1B-55K protein gene and insertion of tumor-specific promoter in this recombinant strain allows the virus to replicate only in the cells with an active telomerase. Furthermore, the specificity of this adenoviral variant to tumor cells is enhanced by deletion of the gene coding E1B-55K protein, limiting its replication in normal cells, which are not able to complement the function of this protein. The sequencing of recombinant adenoviruses genomic DNA demonstrated the presence of engineered gene cassettes in their genomes, the validity of the introduced transgenes and regulatory sequences and their compliance with the chosen design scheme. The obtained adenovirus vectors can serve as a basis for creating perspective oncolytic agents with high specificity due to the control of viral replication by hTERT promoter and high lytic activity due to the presence of the apoptin gene as a transgene causing p53 and bcl-2-independent apoptosis in tumor cells.

120-126

Authors: Alexander A. Ermoshin, Marianna N. Tsybizova, Irina S. Kiselyova

Environmental degradation caused by anthropogenic impact affects the living activity of organisms, including plants. Especially a lot of stresses are caused by heavy metal ions, p.e. copper. Investigation of the effects of copper ions on living organisms is important because this element is a very common pollutant, as metallic copper is one of the most wide-spread constructing materials. However, copper is an important trace element involved in the transfer of electrons in the ETC in mitochondria and chloroplasts, pigment synthesis and activation of many enzymes. The excess of copper in the environment lowers the level of chlorophyll biosynthesis, changes the protein composition of the chloroplast and inhibits photosynthetic electron transport. Aluminum at high concentrations is also a toxic element for plants. It is one of the most abundant elements in the lithosphere. Due to its light weight, high thermal and electrical conductivity, it is widely used as a structural material. Despite the fact that aluminum is not a heavy metal, it is quite toxic to plants, especially in acidic soils. Aluminum toxicity is significantly caused by its antagonism to calcium and magnesium. All these facts cause the interest to creating forms of plants tolerable to these ions and to the study of the mechanisms of their toxicity and detoxification. The aim of our study was to investigate the influence of copper and aluminum ions on germination and seedling growth in white clover. Seed germination, root and hypocotyls sizes as well as the content of photosynthetic pigments were studied under the influence of copper sulphate and aluminum sulphate solutions at 50, 100, 200 and 1 000 μM on metal. Seeds were germinated in Petri dishes on filter paper moistened with test solutions. The germination of seeds was tested on the third day after sowing, on the fifth day the linear size of seedlings and concentration of photosynthetic pigment were determined after ethanol extraction spectrophotometrically. It was shown that the most sensitive indicator of copper and aluminum stress is the root growth compared to hypocotyl growth. Copper ions were significantly more toxic to clover sprouts than aluminum ones. 50 μM copper sulfate caused a depression in the development of plants, and aluminum ions at the same concentration had no effect on plant development. 100–200 μM CuSO4 caused a twofold reduction in all the studied parameters while the Al2(SO4)3 at 200 and 1 000 μM reduced them to no more than one third. Maximum concentration of aluminum ions (1 mM) had less effect on plant development than copper ions at concentration of 100 and 200 μM. The obtained data could be used for clover breeding to get more heavymetal resistant forms.

137-151

Authors: Evgeniya A. Golovatskaya, Liliya G. Nikonova (Abzalimova)

The study of the transformation of organic matter is of great importance at present time, due to climate change and the increasing level of human impact on natural ecosystems. In peatlands there is a constant exchange of minerals between plants and peat soils. The rate of decomposition of plant residues and release of these nutrients depend on individual characteristics of the chemical composition of plants and the environment in which these processes occur. Peatlands have a lower, compared with the net primary production, speed of transformation of organic matter of plants, due to which there is a constant accumulation of organic matter in the form of peat. To estimate the rate of peat accumulation and the amount of carbon stored in the form of peat from decomposition of crop residues, we carried out an investigation of the destruction of peat-forming plants (13 species), growing on oligotrophic peatlands of the southern taiga subzone of Western Siberia. The studies revealed that the most intense transformation processes are at an early stage of decomposition, and then the degradation rate decreases. The most resistant to decomposition are sphagnum mosses, except Sph. angustifolium. The least resistant – herbaceous plants: Menyanthes trifoliata, Rubus chamaemorus. Shrubs are characterized by an average rate of decomposition. Thus, the main peatforming plants, at present, in the studied oligotrophic bogs are sphagnum moss, as also evidenced by botanical composition of peat deposits. The dynamics of the removal of carbon from residues peat-forming plants have the same trends as the weight loss during organic matter destruction. Removal of carbon during decomposition of peatforming plants during the year depending on the type of plant is from 3% (Sphagnum fuscum) to 72% (Menyanthes trifoliata), and the average for phytocenosis is 33%. immobilization of nitrogen in many of the studied plants (S. fuscum, S. magellanicum, Vaccinium vitis-idea, Andromeda polifolia, Ledum palustre, Chamaedaphne calyculata, Vaccinium oxycoccus, Rubus chamaemorus). The maximum accumulation of nitrogen is characteristic for S. fuscum (187% of the initial quantity). A comparison of net primary production of the studied plant communities and the rate of destruction showed that the amount of carbon stored in the process of photosynthesis is much higher than the loss of carbon during decomposition of plant residues both for certain species of plants (2–25 times depending on the plant species) and for phytocenosis overall (4–7 times depending on phytocenosis).

7-22

Authors: Anatoly G. Dyukarev, Natalia N. Pologova

Pinus sibirica forests near settlements are formed on the rich habitats in the absence of competition from other forest species. Tree stands here have fast rate of biological processes and high productivity. These forests ecosystems inherit the background conditions of the environment and are formed in habitats with a wide variety of moisture conditions and soils. Most of studied P. sibirica forests at the south of Tomsk region are dedicated to landscapes subtaiga forest zone with zonal gray soils. In the north of forest-steppe zone P. sibirica forests are rarer and confined to the river valleys due to high standards of P. sibirica to moisture conditions. Complex combinations of soil cover in each P. sibirica forest patch is determined by proximity to river valleys. There are several levels of heterogeneity of soil. Zonal level is defined by climatic conditions. Regional level is defined by position in the macro-relief. Local level of inhomogeneity is related with the microrelief and soil position in the system of tree stand edificator. Zonal automorphic soils are typical of upland habitats. Accumulation – transitive or eluvial soils are typical of sloping positions. Semi-hydromorphic and hydromorphic soils are typical of accumulative positions. The patterns of distribution of soil are found in each patch of P. sibirica forest, but in general, the soils of slope positions are found for a large part of their area. Tree stand structure in biogeocenosis affects through redistribution of precipitation by crowns and litter inflow. Edificator influence of forests determines capacity of forest litter, sod horizon, bulk density of humus and humus content in the upper part of soil. The main factor of differentiation of the soil cover in South taiga is lithological. It is heterogeneity of the underlying rocks. The variety of lithological basis was analyzed by using combined indicator – the average diameter of fractions. The soilforming processes and patterns of the soil profile were analyzed by distribution of silt fraction. The peculiarity of soils of Pinus sibirica forests is loose constitution and high aggregation of humus horizons. These facts create an optimal water-air and thermal conditions in the root zone. Basic soil characteristics determine the type and shape of Pinus sibirica trees and their productivity. Pure P. sibirica forests of park type with low resistance to pests and diseases are formed in rich soils, but stable P. sibirica forests of taiga type with complex structure are formed on podzolic soils. Modern degradation of P. sibirica forests near settlements occurs due to their accelerated development, high levels of anthropogenic pressure and the lack of renewal.

43-51

Authors: Tatyana P. Alekseeva, Tatyana I. Burmistrova, Larisa D. Stakhina, Natalia N. Tereshchenko

For intensification microbiological destruction of oil in in oil contaminated soils may use ecological ameliorant based on peat, it has good sorption properties and it is enriched active hydrocarbon oxidiring microorganisms. Rate of biodegradation in soil may defined by form and dose nitrogen fertilizer in consist of peat ameliorant. Laboratory trials revealed that the ameliorants which contain the peat and nitrogen and phosphorus fertilizers are able due to active stimulation of microflora to increase the rate of decomposition of oil in soil by 1.5–2.0 times. The influence of carbamide and ammonia nitrate contents and their ratios to others components on the rate of destruction of oil pollutants in soil have been studied. The most optimal kind of nitrogen fertilizer in composition of peat ameliorant is cаrbamide. Application of peat ameliorant with carbamide in comparison with ameliorant containing ammonium nitrate promote more active flow of microbiological and enzymatic process during the whole period of experiment. This factor serve higher ratio of destruction oil hydrocarbon. These ratios are 32.4 and 54.3% vs. 21.6 and 42.5% for 6 month of experiment. Changing of carboxylic group contains for 6 month of experiment argue about more significant oxidizing process, that it is passing in oil in case of using peat ameliorant. In these versions in comparison with version where ammonia nitrate are used, is marked lower content of carboxylic group. It is connected to further transformation of carboxylic acid into another oxygen-containing compound. In this case it has been reached maximum degree of paraffin- naphthenic hydrocarbon’s destruction – 2.6 vs. 1.9. The high initial dosage in content peat ameliorant of nitrogen fertilizer added to oil-contaminated soil don’t lead to maximal rate of oil carbon destruction. This rate is higher with more wide ratio C:N:P (863:1:1 in this case) and for 6 month experiment is 42.5 and 54.3% vs. 21.6 and 32.4% with C:N:P ratio equal to 20:1:0.15.

57-69

Authors: Marina N. Dirks

Colonization peculiarities and ontogenetic structure of coenopopulations of two shrubs Juniperus sibirica and Betula rotundifolia were investigated at different stages of vegetation development on new moraines of the Maly Aktru glacier (2200–2290 m a.s.l., Aktru headwater basin, Severo-Chuiskiy range). Colonization of J. sibirica can occur in early years after glacier retreating. The first individuals on some sites probably don’t survive dying when they are young because of unfavorable conditions. B. rotundifolia colonizes new moraines not until 40 years after glacier retreating at the herb-moss-willow stage of vegetation development and depends on the presence of plane sites of fine fragments of rocks protected from cooling by the glacier and with a considerable snow cover. The time period of new moraine forming is not enough for passing overall ontogenesis by individuals of this species. Seemingly, the absence of the youngest individuals of J. sibirica is connected with unfavorable weather conditions in the years of investigations and/or low germinating ability. A settlement of the youngest individuals of B. rotundifolia at the herb-moss-willow-birch stage is hampered by dense moss cover and leaf litter. These are reasons why the ontogenetic spectra of all investigated coenopopulations of this species are incomplete. The series of J. sibirica cenopopulations from invasional cenopopulation at the pioneer stage to the young and normal ones at the herb-moss-willow-birch stage and on the end moraine of the middle of the 19th century were observed. The series of B. rotundifolia cenopopulations from the young and normal cenopopulation at the herb-moss-willow stage to the mature and normal one at the herb-moss-willow-birch stage were revealed. Coenopopulation of J. sibirica at the pioneer stage isn’t capable yet of self-sustaining and completely depends on seed influx from without because of the absence of generative individuals in its structure. Self-sustaining of its other coenopopulations can occur by their own seeds as there are generative individuals. All coenopopulations of B. rotundifolia are capable of self-sustaining by seed reproduction due to the presence of generative individuals and also by partial formations at the herb-moss-willow-birch stage. The character of ontogenetic spectra and demographic characteristics of species cenopopulations depend on the age of moraine deposits, substrate features, micro- and mesorelief peculiarities, ecotopes protected from cooling by the glacier and a sufficient snow cover.

84-100

Authors: Natalia I. Makunina

The Saksary hill massif is situated in the western part of the South-Minusinskaya basin in rain shadow of the Range Abakansky. It is limited by the Abakan river and its affluents the Uybat and the Kamyshta rivers. It extends from west to east by 22 km, from north to south - by 20 km. The climate is ultracontinental. The average annual temperature is about 0 ºC, a minimal one - -48º…-52ºC and a maximal one +34º…39 ºC. The sum of temperatures above 10 ºC is 1600 ºC, the annual amount of precipitation is 250-300 mm. In the western part of Mynusinskaya basins the upper limit of steppe belt is 600 m above s.l. Only the marginal parts of the massif are characterized by heights below 600 m. The basic central part has the heights from 600 to 900 m above s.l. Nevertheless, it is covered with steppe vegetation. At the top part of the northern steep slopes of several highest hills we can only see some trees. The steppe vegetation of the Saksary hill massif in outline was characterized by V.V. Reverdatto (1928), A.V. Polozhyi and A.T. Maltseva (1974). The principal aims of this work are to reveal the diversity of steppe communities of the Saksary hill massif, to classify them according to the principals of Braun-Blanquet classification and to describe the structure of vegetation cover. The diversity of the Saksary hill massif steppe communities is presented with 5 variants and 5 subassociations of 7 associations belonging to Central Asian class Cleistogenetea squarrosa and European Siberian class Festuco-Brometea. The altitude above the sea level, steepness and stoniness of hill slopes determine the pattern of steppe associations. The Saksary hill massif has three altitudinal steps: 1) the pediment of hill massif (350-450 m above s.l.); the foot, marginal part of hill massif (450-600 m above s.l.); 3) the top, central part of hill massif (600.900 m above s.l.). The first hill massif step is characterized by predominance of flat surfaces, the second one - with gentle slopes (3-7º) and the third - with mean (7-15º) slopes. The steep slopes (15.25º) are present on the third hill massif step and more rarely on the second one. The altitudinal-belt steppe types cover the gentle (3-7º) and mean (7-15º) slopes. They succeed one another as the altitude increases. On the hill massif pediment bunchgrass steppes of subass. Artemisio frigidae-Stipetum krylovii artemisietosum scopariae predominate. On the foot part of hill massif (450-600 m above s.l.) they give place to bunchgrass steppes of ass. Thalictro foetidi-Festucetum valesiacae, above 600 m gentle slopes and sun exposed mean slopes are covered with tussock steppes of ass. Artemisio glaucae-Caricetum pediformis, on shadowed slopes the meadow steppes of ass. Bupleuro multinervi-Helictotrichetum desertorum predominate. The steep slopes (15-25 ºC) are covered with crushed stones and have the shape of stairs, their pattern of steppe communities is the same on its all altitudinal range. The crushed stone “steps” are covered with short grass petrophytic steppes of ass. Androsaco dasyphyllae-Caricetum pediformis, on the steep parts of stony slopes tussock petrophytic steppes of ass. Youngio tenuifoliae-Helictotrichetum desertorum predominate. The steppe communities of the first and the second hill massif step belong to the class Cleistogenetea squarrosae and the third one - to the class Festuco Brometea.

115-126

Authors: Sergey A. Abramov1, Vladislav V. Vinogradov2, Alexander S. Zolotyuh3

Characterising of habitat associations of species is fundamental for understanding of basic mechanism of animal communities organization. Differences in species diversity between different localities can be attributed to ecological, biogeographic and evolutionary processes. The main indicator which shows the relation of the species to the environmental conditions is its abundance. Population diversity of rodent species with different ecological specialisation is a marker of different habitats. Mouse-like rodents form the most abundant communities among vertebrate animals inhabiting the forest belt of the mountain region. Based on the long-term research and literature data the authors discuss the distribution regularities of rodent species in the mountain forest ecological space of the Altai-Sayan Mountains. The forest belt of the Altai-Sayan Mountains is inhabited by 15 mouse-like rodents species. A comprehensive analysis of distribution of rodent species within the forest zone allowed us to determine the attitude of species to the range of environmental factors and to estimate the collocation of rodent species in the multidimensional ecological space. Three main axes of environmental space were allocated with a multidimensional scaling method. To identify the ecological niches of the studied rodent species, each axis was regressed to the abundance of rodent species. Regression analysis was also used to assess the association between the environmental parameters and the axes of the environmental space. The analysis showed that the differences in the sensitivity of species to environmental factors cause a divergence of their ecological niches, which provides the independence of these species within their community. The most significant ecological factors which provide the distribution of rodents in the forest belt are density of canopy, altitude elevation of the territory, humidity and the understory layer (shrubs and herbs) structure.

141-149

Authors: Petr A. Popov

The information in this article about the characteristics of ichthyofauna composition in a number of lakes and rivers of Gorny Altay shows an adaptive nature of fish distribution in ponds of this mountain country. In rivers and lakes located at relatively high altitudes above sea level, ichthyocenoses are poor according to the number of their constituent species, and the number of fishe is low. The main reason for this is a relatively small value of radiation balance and the part of the solar energy that is absorbed by high mountain reservoirs. A significant limiting role in the formation of species diversity of ichthyocenoses is played by small sizes and the modules of rivers, their high velocity and strong bias, the mobility of bottom substrate, the lack of floodplain system, localized icings freezing, a short period of open water and low temperatures as well as forage supply of fish. A significant role in limiting the spread of fish upstream is played by waterfalls and areas with great steepness of the stream. The presence of insurmountable physical barriers to fish is the main cause of total absence of fish in a large number of lakes and river systems in Gorny Altay. In alpine lakes, the taxonomic diversity of ichthyofauna composition is also limited by large depths, a short vegetation period, low water temperatures, a low quantity of nutrients in the water, a low degree of diversity and productive efficiency of biocenoses plankton and benthos. The factor that significantly restricts species diversity of ichthyocenoses in some rivers and lakes in Altai is a great number of mineral suspensions in water as a result of snowfields and glaciers melting. The adaptive nature of fish distribution in the conditions of Gorny Altay is manifested not only in a particular lake-river system as a whole, but within a lake or a short length of a river. We often meet Thymallus arcticus Pallas, 1776, Barbatulus toni Dybowski, 1869 or Сottus altaicus Kaschenko, 1899 and С. sibiricus Warpachowski, 1889 in a rough river stretch in the shade of large boulders or in pits. In lakes, fish, during summer feeding, as a rule, prefer the most productive area of littoral zone on a forage basis, and during winter – the deepest parts of a water body. In general, the adaptive nature of fish distribution in the waters of Gorny Altay should be taken into account while developing a strategy for protection and sport fishing in the region of Western Siberia.

160-171

Authors: Irina S. Kovtun, MarinaV. Efimova

Present-day science must base on molecular biology methods to conduct detailed and multifaceted researches. Biochemical, morphological, biophysical and other methods are not enough for description and interpretation of phenomena and mechanisms that occur in a plant cell. The polymerase chain reaction (PCR) is an integral part of many molecular investigations. The PCR is the basis of many research methods, in particular, of reverse transcription polymerase chain reaction (RT-PCR). RT-PCR is characterized by high sensitivity and is used for relative quantitative assessment of gene expression on mRNA level. The initial phase while preparing to carry out the polymerase chain reaction is the primer design for specific gene. For successful realization of this process it is necessary to have some experience in work with information databases, nucleotide sequences and primer design. Also, it is necessary to find primers not only for specific genes but also for control gene. As a means of control, it is recommended to use the «housekeeping» gene or genes (constitutive gene), the expression of which is relatively constant regardless of external conditions. This fact will help to assess the change of expression of the target genes. This scope of work doesn’t take a researcher a lot of time, but for a young explorer this search of nucleic acid sequences and primers canpresent a significant challenge. Wherewith, especially this phase of work is the initial stage of the initiation of molecular biological studies. All mistakes and inexperience could bring a great time and financial costs. In the present article a step-wise sequence of actions when searching for nucleotide sequences of genes and primer design to them are described, which may contribute to a general idea of the versatility, value and scope of bioinformatics approach. The description is made on a specific example using gene sequences of the constitutive protein tubulin of different plant objects. Described basic skills are quite enough for a young researcher to make a successful primer design, who has encountered the need for molecular studies for the first time.