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Sorghum (Sorghum bicolor (L.) Moench) is one of the world’s leading cereal crops in agricultural production, which has a special importance in the arid regions. However, unlike other cereals, sorghum grain has a lower nutritional value, which is caused, inter alia, by the resistance of its seed storage proteins (kafirins) to protease digestion. One of the effective approaches to improve the nutritional value of sorghum grain is to obtain mutants with partially or completely suppressed synthesis or altered amino acid composition of kafirins. The employment of genome editing may allow to solve this problem by introducing mutations into the nucleotide sequences of the α- and γ-kafirin genes. In this study, genomic target motifs (23 bp sequences) were selected for the introduction of mutations into the α- and γ-KAFIRIN genes of sorghum. The design of the gRNAs was conducted using the online tools CRISPROR and CHOPCHOP. Two most suitable targets were chosen for α-KAFIRIN (k1C5) and two for γ-KAFIRIN (gKAF1) genes. The insertion of respective sequences in the generic vector pSH121 was performed at the BsaI (Eco31I) sites. Validation of the cloning procedure was performed by DNA sequencing. Subcloning of the resulting constructs was performed using the SfiI restriction sites into the compatible binary vector B479p7oUZm-LH. The correct assembly of binary vectors was confirmed by restriction analysis using the MluI and SfiI cleavage sites. The four vectors created (1C - 4C) were transferred by electroporation into the Agrobacterium tumefaciens strain AGL0. Currently, this vector series is used for stable transformation of sorghum using immature embryo explants.

Background. An analysis of the spatial distribution of some taxonomically and ecologically related legumes in the Ural showed a nontrivial spatial distribution of related species of the genus Oxytropis DC of the Orobia Bunge section within the Uchalinsky uplands. Despite the similarities in ecology, these species practically do not grow together. Explicit spatial segregation of closely related plants over a relatively small area allows this phenomenon to be used as a convenient model for studying the effect of segregation of closely related legume species on the genetic composition of their nodule bacteria. Materials and methods. The genetic diversity of nodule bacteria entering into symbiosis with O. kungurensi s, O. baschkiriensis , O. approximata and O. gmelinii plants was studied. In addition, the polymorphism of their symbiotic genes has also been analyzed. Results. Phylogenetic characteristics of nodule bacteria endemic for the Southern Ural belonging to 4 species of leguminous plants of the genus Oxytropis of the section Orobia: O. kungurensis , O. baschkiriensis , O. approximata , O. gmelinii which are characterized by spatial separation of the growth sites, also called plant segregation, are given. It was shown that all of them belong to the genus Mesorhizobium despite certain phylogenetic differences of bacteria. Analysis of the symbiotic genes of the analyzed strains revealed a lack of congruence of their phylogeny with the core part of the genome. It was found that the microsymbionts of O. baschkiriensis plants differ in the phylogeny of nod-genes from nodule bacteria of other plants of the Oxytropis genus and are close to microsymbionts of plants of the Lupinaster genus growing in the Southern Urals. Conclusion. Acquisition of the property to enter into symbiosis with nodule bacteria of plants of the genus Lupinaster may turn out to be an adaptive mechanism that arose as a result of segregation of O. baschkiriensis from other species of Oxytropis .

For the first time, summarized data on the production of nine species and ten hybrids of pondweed (Potamogeton, Stuckenia, Potamogetonaceae) are given for ecosystems of small and medium rivers in the north of European Russia. The pondweed is shown to be an important producer of primary organic matter. A significant contribution to primary production in rivers in the region is due to large-size pondweeds and their hybrids, which form dense monodominant stands: P. lucens, P. natans, P. perfoliatus, P. × angustifolius, P. × nitens, P. × salicifolius, P. × sparganiifolius. The production characteristics for the widespread species (P. lucens, P. natans, P. perfoliatus) in the rivers of the region are more than 1.5 times greater than those in reservoirs. Most pondweed species and hybrids show medium productivity and produce

This paper presents new data on the species of genus Vaucheria in Russia from analysis of original and museum collections. In total, 20 species were identified in 152 locations of 22 administrative regions of the European Russia, Western Siberia, and the Russian Far East. Vaucheria lii, a rare multiregional species new to Russia, has been found in 6 locations of the Vologda, Moscow, Nizhny Novgorod, Yaroslavl regions and North Ossetia. V. cruciata, V. schleicheri, V. taylorii, V. uncinata were recorded for the European Russia for the first time. V. birostris and V. pseudogeminata are new records for Western Siberia. Rank of V. terrestris var. nuoljae was raised up to separate species with V. terrestris var. major as a new synonym. The distributions of the frequently reported species, V. dichotoma and V. geminata, are critically analyzed.

Specific amplification of nucleic acids is a convenient and quick alternative to the culture-based method of detecting bacterial cells. However, conventional PCR and other amplification reactions can not differentiate between live bacteria and dead or dormant ones, and are also capable of amplifying DNA that persists for a long time and in a cell-free state. Several methods have been developed in order to establish the viability of microorganisms by amplification of specific sequences of nucleic acids, both those controlled by changing temperatures and isothermal ones. For some of them, DNA modified by phenanthridine dyes serves as a target, and simultaneous use of monoazides of ethidium and propidium was shown to be preferable for the purpose. For other methods, the targets are directly RNA molecules or their cDNA copies. Pre-rRNA detection seems to be the most preferable approach, due to the presence of these types of RNA exclusively in living cells.

We present current information on the conservation of aquatic vascular plants in West and East Siberia and the Russian Far East. This includes the region from the Ural Mountains in the west to the eastern seaboard of Russia from Kamchatka and Chukotka in the north to Vladivostok in the south, including Sakhalin, the Kuril and Commander Islands. A list of aquatic vascular plant species protected in the region is presented with an indication of their conservation status derived from regional Red Data Books, combined with population trend data from the literature, as well as field observations made by the authors. The region as a whole supports a total of 246 aquatic plant species, of which 96 are listed as threatened in regional Red Lists. The main threats to these species are habitat degradation, eutrophication and pollution of water bodies as a consequence of human activity and climate change. Many species which are specialized, relict or endemic, and others occurring here on the edge of their range, are vulnerable because of these factors. The main conservation action needed includes protection of natural habitats, establishment of monitoring protocols for the most severely threatened species and research into the autecology of threatened species. There is also a need to resolve taxonomic questions obstructing conservation.

The genetic diversity and phylogeny of root nodule bacteria, microsymbionts of plants of the genus Lupinaster Adans. (L. albus Link and L. pentaphyllus Moench), were studied, and also their symbiotic genes were analyzed. The bacterial strains studied were shown to be phylogenetically different; however, all of them are related to the Mezorhizobium genus with the exception of the single strain which is related to the Rhizobium genus. Analysis of the symbiotic genes nifH and nodC revealed their high homology among all strains independently of strain phylogeny, as well as the phylogenetic relation of these genes to those of Mezorhizobium. Mezorhizobium bacteria are likely specific microsymbionts of these plants, while the Rhizobium strain acquired its symbiotic genes and became capable of nodule formation in Lupinaster plants through horizontal gene transfer. Thus, the genetic composition of nodule bacteria inhabiting Lupinaster plants represents additional support for the idea that they do not belong to the Trifolium genus.

The dataset for charophytes from the Middle Volga Region, including Orenburg region, Penza region, Samara region, Ulyanovsk region, Chuvash Republic, Republic of Mariy El, Republic of Mordovia, Republic of Tatarstan, have been updated under study of Russian charophytes. To date, 18 species from four genera were known from the study area according to published records. The reliability of records has been checked with specimens stored in LE, IBIW, PVB and our new collections of 2009–2017 for the period from 1768 to 2017. Fourteen species of Chara, Nitella and Tolypella only have been confirmed with vouchers. The published records of six species and one genus need to be confirmed. The data are scarce nearly for all administrative regions except Samara region. Chara inconnexa and C. subspinosa have been found in the Middle Volga Region for the first time. The single locality of the latter species is remarkable from its distribution range perspective and based on a single specimen from the 18th century. Some species have been found in separate regions for the first time. The novel species record for Ulyanovsk region is the first data about charophytes in the region. The species richness of charophytes from the Middle Volga Region is evidently low in comparison with the Upper and the Lower Volga regions due to less diversity of some ecological groups and mainly insufficient knowledge of charophytes from the regions except for Samara region. The species composition of charophytes from the region studied could be tentatively recognised as impoverished compared to the Upper Volga Region. Its low similarity with the Lower Volga Region could be explained with the absence of many characteristic species of southern chorotype. Chara vulgaris and C. globularis are the most common species in the study area. They could be recognised as generalists. A correction of species lists for the regional Red Data Books has been suggested. The protection of Chara contraria, C. papillosa, C. subspinosa, C. tomentosa, C. virgata, Nitella gracilis, N. mucronata has been recommended according to the criteria of the Red Data Book of the Russian Federation. Chara subspinosa seems to be a regionally extinct species. Karst and oxbow lakes and to a lesser extent inundated quarries and ponds could be recognised as the most important for charophyte species protection. Three important plant areas have been identified as essential for long-term persistence of stable charophyte populations: the National Park «Mariy Chodra» in the Republic of Mariy El, the regional sanctuary «Golubye Ozyora» in the Republic of Tatarstan and the regional natural monument «Ozero Goluboe» in Samara region.

Plant infection with Agrobacterium rhizogenes leads to the development of a hairy root disease notable for the rapid agravitropic growth of roots on hormone-free nutrient media. In order to look into the interaction of A. rhizogenes with plants and assess opportunities of practical application of hairy root culture, new approaches to their production are elaborated. A method of bacterium-free and plasmid-free production of genetically modified roots (hairy roots) by means of biolistic transformation of leaf explants with a DNA fragment (size of 5461 bp) consisting of genes rolA, rolB, rolC, and rolD are proposed. In most cases, such transformation resulted in the emergence of only adventitious roots with transient expression of rol-genes, and the growth of such roots on hormone-free media ceased in 2–3 months in contrast to genuine hairy roots capable of unrestricted growth. Molecular analysis of different systems of target genes’ expression showed an important role of transgene rolC and host gene of cyclin-dependent protein kinase CDKB1-1 in the maintenance of rapid growth of hairy roots in vitro (in isolated cultures).

The peculiarities of root growth and stress tolerance of transgenic tobacco plants with constitutive expression of NtEXPA1 and NtEXPA5 genes, as well as plants with reduced expression of NtEXPA4 gene encoding α-expansins of Nicotiana tabacum, were studied during prolonged cultivation under conditions of drought, salinity, and low positive temperatures. Increased expression of expansin genes led to an increase in the growth rate and root length both under normal plant growth conditions and at 12°C and 50 mM NaCl. Increased expression of expansin genes influenced the changes in the fresh and dry mass of a shoot, leading to an increase in their exposure to hypothermia. Transgenic plants with a reduced level of NtEXPA4 expansin gene expression were characterized by a reduction in the fresh and dry weight of a shoot due to drought and low positive temperatures. The totality of the data obtained may indicate the involvement of NtEXPA1, NtEXPA4, and NtEXPA5 tobacco expansin genes in the regulation of growth under hypothermia, drought, and salinity.

Possible routes in which aquatic plants have spreaded across their vast ranges from ancestral areas and refugia still remain poorly known. We have chosen Eurasian yellow water-lilies (Nuphar Sm. sect. Nuphar, Nymphaeaceae) as a model group for a wide-scale phylogeographical study as they represent a common component of the continental freshwater flora. To elucidate the history of diversification of the section Nuphar, we sequenced nuclear ribosomal ITS and chloroplast intergenic spacers (trnH-psbA, trnL–trnF) from 117 localities from almost all the section area. Sequence data were analysed with a statistical parsimony approach using the network algorithm. We also incorporated data on morphological variation and pollen viability. The section Nuphar originated and diversified into several evolutionary lineages in Eastern Asia. Two lineages, represented by N. pumila and N. lutea, spread from the Russian Far East westwards to Siberia and further to Europe. Nuphar pumila also migrated from the Far East to North America (where it is referred to as N. microphylla). Nuphar lutea colonized deglaciated north-eastern Europe not only from the south, but also from the Siberian refugia. Nuphar lutea and N. pumila hybridize rarely in Eastern Europe and Asia and warrant species status. We provide the first evidence, based on a phylogeographical approach, for the origin and diversification of aquatic flowering plants in Eastern Asia. Our results indicate that phylogeographic analysis on the area-wide scale is one of the most efficient ways to deduce possible routes in which aquatic plants have spread across their total range from ancestral areas and refugia.

The expression level of the gene PtrXTH1 encoding xyloglucan endo-transglycosylase in the leaves of Populus tremula L. of wild-type and in response to exogenous phytohormones treatment was analyzed. The highest level of transcripts of PtrXTH1 was detected in young, intensively growing leaves of aspen. In young aspen leaves, the expression of PtrXTH1 was induced by cytokinins, auxins, and brassinosteroids. The content of PtrXTH1 transcripts increased under the constitutive expression of the PnARGOS-LIKE gene. Bioinformatic analysis of PtXTH1 putative promoter region in P. trichocarpa Torr. and A. Gray ex. Hook showed the presence of cis-regulatory elements associated with the regulation of growth and stress resistance. To determine the role of the gene under study, we also created transgenic tobacco plants with constitutive expression of the PnXTH1 gene (the ortholog of PtrXTH1 from the black poplar P. nigra). Transgenic tobacco plants were characterized by an increase in leaf size and fresh and dry weight of the aboveground part under normal growth conditions. When grown under conditions of salinization and drought, transgenic plants were distinguished by increased stress resistance due to the maintenance of cell expansion in roots and stems at a higher level and the ability to more effectively retain water in leaves compared with wild-type plants.

The genes of α-expansins of woody plants are of great interest for genetic engineering, since they can potentially be used to improve the tree growth parameters. In the flora of Russia, model woody plants for plant biotechnology are aspen (Populus tremula L.) and black poplar (Populus nigra L.). The objective of this study was to determine the role of α-expansin-encoding genes, aspen PtrEXPA3 and black poplar PnEXPA3, in the regulation and maintenance of woody plant growth. To achieve this goal, the PtrEXPA3 expression level were determined upon exogenous phytohormone treatment, the action of stress factors, and constitutive expression of the PnARGOS-LIKE gene. In addition, transgenic aspen plants with constitutive expression of the black poplar PnEXPA3 gene were generated, and their morphological analysis was carried out. The highest PtrEXPA3 mRNA level was detected in young intensely growing aspen leaves, and furthermore, expression of the gene was induced by exogenous cytokinins and auxins. In response to NaCl and constitutive expression of the PnARGOS-LIKE gene, the PtrEXPA3 mRNA level decreased. Transgenic aspen plants with constitutive PnEXPA3 expression were characterized by the decreased size of leaves, petioles, and internodes, as well as the increased size of leaf epidermal cells, while the stem size remained unchanged. Taken together, the data obtained enable the suggestion that the PtrEXPA3 and PnEXPA3 genes encode cytokinin- and auxin-regulated, leaf-specific expansins that are involved in the cell expansion.

Amaranth is a new and promising crop for the Russian climate, notable for its well-balanced amino acid composition. Yield increase using the methods of genetic engineering is a challenging task. We generated transgenic plants of amaranth with expression of the Arabidopsis thaliana ARGOS-LIKE gene under the control of the dahlia mosaic virus promoter. We achieved 1.4% transformation effectiveness. In comparison with wild-type amaranth, we observed a 21% increase in stem length, 79% increase in leaf length, and 190% increase in fresh weight of transgenic plants. It was shown that ARGOS-LIKE gene of A. thaliana along with the dahlia mosaic virus promoter can be used to increase primarily the green weight of shoot and leaf size of amaranth.

The ability to synthesize milk-clotting (rennet) proteinases was studied in eight strains of Irpex Lacteus basidiomycetes. It was found that the studied I. lacteus strains are characterized by different enzyme activities in their culture liquid. For I. lacteus strains 2425, 2426, and 2427, the maximum milk-clotting activity was detected during the exponential growth phase on the 15th day of cultivation on glucose–peptone medium. These I. lacteus strains are the most prospective milk-clotting enzyme producers for further research and practical application. I. lacteus strains 2421, 2422, 2423, 2424, and 2428 showed lower values of the enzyme activity and require additional research to determine the optimal culture conditions.

We identified a Populus nigra auxin-regulated gene involved in organ size (PnARGOS)-LIKE, encoding one organ size related protein in black poplar. It is homologous to AtARGOS and AtARGOS-LIKE genes of Arabidopsis thaliana. ABRE-like, G-box, GATA and I-box motifs were discovered in the promoter region of the poplar ARGOS-LIKE gene. In wild type aspen (Populus tremula) plants, an ortholog of the PnARGOS-LIKE gene (PtrARGOS-LIKE) was noticeably expressed in actively dividing and expanding young leaves and calli, whereas its mRNA content increased in response to exogenous 6-benzylaminopurine, 1-naphthaleneacetic acid, and 24-epibrassinolide. Expression of the PtrARGOS-LIKE gene was reduced under a salinity treatment. In addition, we generated transgenic tobacco and aspen plants with an up-regulated expression of the PnARGOS-LIKE gene. A constitutive expression of the gene contributed to an increase in size of stems and leaves of the transgenic tobacco plants. In the transgenic aspen, a constitutive expression of the PnARGOS-LIKE gene promoted an increase in the frequency of leaf initiations and in leaf length and area. The size of transgenic tobacco and aspen leaves increased due to the enlargement of individual cells. The results show the significance of the PnARGOS-LIKE gene for control of leaf initiation and organ growth by cell expansion in poplar.

Eight species of charophytes have been found in the vast area of the ubiquitous permafrost in Far North-East Asia (i.e. the Republic of Sakha (Yakutia), the Magadan Region, and the Chukotka Autonomous Area, Russia): Chara contraria, C. globularis, C. strigosa, C. virgata, Nitella flexilis, N. opaca, N. wahlbergiana and Tolypella canadensis. Charophytes are found in a variety of water bodies and watercourses in areas with various thicknesses of permafrost, within the boreal and subarctic climate zones. Charophyte habitats are mostly associated with the distribution of carbonate rocks. The number of species and their records decrease towards north and east mainly due to shortening of the vegetation period. The highest density of records of charophytes and their most northern records are in river valleys, where the topography and heating effect of the river mitigate the influence of climate and permafrost. This association can be used to model and predict charophyte distribution in periglacial areas during glaciation. Widespread eurytopic species with a flexible life cycle, Chara contraria and C. globularis, are found in valley water bodies due to their ability to withstand environmental variation associated with marked changes in water temperature and flood disturbance. Rare perennial species such as C. strigosa and Tolypella canadensis are represented in deep lakes with less variation in environmental conditions. This paper discusses the distribution, habitat and life history strategies of charophytes that exist in the severe conditions of the permafrost area in Far North-East Asia based on previously published and new data.

The Nicotiana tabacum AINTEGUMENTA-like gene (NtANTL), encoding one of AP2/ERF transcription factors, is a putative ortholog of the AtANT gene from Arabidopsis thaliana. In wild-type tobacco plants, the NtANTL gene was expressed in the actively dividing young flowers, shoot apices, and calluses, while the level of its mRNA increased considerably after treatment with exogenous 6-benzylaminopurine, indoleacetic acid and 24-epibrassinolide. We found a positive correlation among the expression levels of NtANTL, cyclin NtCYCD3;1 and cyclin-dependent kinase NtCDKB1-1 genes, suggesting possible molecular links between AINTEGUMENTA and cell cycle regulators in tobacco plants. However, no correlation was observed between NtANTL, NtCYCD3;1 and NtCDKB1-1 expression levels in response to NaCl and ABA. These observations indicate that the transcription factor NtANTL was not involved in the regulation of the cellular response to salinity nor did it affect the expression of NtCYCD3;1 and NtCDKB1-1 when tobacco plants were exposed to salt stress and ABA. In addition, we generated transgenic tobacco plants with both up-regulated and down-regulated expression of the NtANTL gene. Constitutive expression of the NtANTL gene contributed to an increase in the size of leaves and corolla of transgenic plants. Transgenic plants with reduced expression of the NtANTL gene had smaller leaves, flowers and stems, but showed a compensatory increase in the cell size of leaves and flowers. The results show the significance of the NtANTL gene for the control of organ growth by both cell division and expansion in tobacco plants.

Screening of nodule bacteria (rhizobia) forming symbiotic relationships with legumes has been performed in order to isolate strains resistant to cadmium ions in a wide range of concentrations (6-132 mg/kg). The effect ofcadmium salts (6, 12, 24 mg/kg) on the legume-rhizobium symbiosis ofthe pea Pisum sativum L. with Rhizobium leguminosarum and of the fodder galega Galega orientalis Lam. with Rhizobium galegae has been studied under experimental laboratory conditions. No statistically significant differences have been revealed in the growth and biomass of plants with regard to the control in the range of concentrations given above. However, it was found that cadmium inhibited nodulation in P. sativum and stimulated it in G. orientalis.

Transgenic tobacco plants expressing Arabidopsis thaliana ARGOS and ARGOS-LIKE genes under the control of the chalcone synthase promoter of Petunia hybrida L., as well as the estradiol inducible XVE system, have been obtained. The part of transgenic plants with flower-specific expression of the target genes was characterized by increased flower size, caused by an increase in cell size and quantity in the case of the ARGOS gene and by a stimulation of cell growth via stretching in the case of the ARGOS-LIKE gene. An enhanced expression level of the NtEXPA1, NtEXPA4 genes encoding expansins, NtEXGT gene encoding endo-xyloglucan transferase, and the AINTEGUMENTA-like gene was detected in the flowers of transgenic tobacco plants. In the case of inducible expression of ARGOS and ARGOS-LIKE genes, an increase in leaf, stem and flower size was revealed in several lines of transgenic plants as compared to control. Expression of the ARGOS gene also affected cell number and size in this case, while the ARGOS-LIKE gene mainly influenced cell size via stretching. Inducible expression of the ARGOS gene in flowers mainly provided an enhanced containment of AINTEGUMENTA-like mRNA, while ARGOS-LIKE gene expression resulted in the activation of NtEXPA1 and NtEXGT genes.

Aegagropila linnaei (Chlorophyta, Cladophoraceae) is reported for the first time for the territory of the Udmurt Republic from Pazelinskiy Bay of Izhevsk Reservoir. The habitat description and morphological characteristics of the alga are provided. It is suggested that the species is threatened in the habitat. A. linnaei is recommended to include in the Red Data Book of the Udmurt Republic.

The effect of cadmium acetate (200 and 400 μM Cd(CH3COO)2) on the transcriptional promoter activity of rice (Oryza sativa L.) phytochelatin synthase gene 900 bp in size was studied. The reporter gene of glucuronidase was placed under the control of this promoter in transgenic tobacco (Nicotiana tabacum L.) plants. For determining boundaries of this gene promoter responsive to cadmium, deletion constructs were created and their transcriptional activity was measured under the influence of different cadmium concentrations. Deletion of 189 bp from the 5′-terminus of this gene promoter resulted in some transcription activation, whereas deletion of successive 95 bp from the 5′-terminus resulted in cadmium-independent activation of the promoter.

Hybrid plants were obtained by pollination of a Nicotiana rustica and Nicotiana tabacum Black Cuban cultivar with pollen from transgenic Nicotiana tabacum plants of a Petit Havana SR1 cultivar bearing the ARGOS-LIKE and AtEXPA10 genes. RT-PCR analysis showed high levels of ARGOS-LIKE and AtEXPA10 expression in the hybrids, which resulted in increased sizes of their leaves and stems. The organs of both transgenic and hybrid plants were enlarged due to the enlargement of individual cells. The inheritance was stable, while their phenotypic effects varied from insignificant to levels exceeding those in the parental transgenic plants.

Transgenic tobacco plants overexpressing the PnEXPA3 gene of black poplar (Populus nigra), which encodes α-expansin, were obtained. The transgenic plants were characterized by increased size of epidermic and mesophyll cells of leaves. However, the size of leaves remained normal. Overexpression of the PnEXPA3 gene provided stimulatory effect only on the stem length. Other morphological traits of the transgenic plants remained unchanged.

We obtained transgenic tobacco plants demonstrating overexpression of NtEXPA5 gene that encodes alpha-expansin of Nicotiana tabacum. The transgenic plants were characterized by increased size of leaves and stems. However, size of flowers remained almost unchanged. The increase of organ sizes was induced by cell stretching only. Moreover, the number of cell divisions was even decreased. The obtained data suggest tight interaction between cell stretching regulation and cell division, which together provide the basic mechanism aimed at the controlling of plant organ sizes.

Changes in the expression levels of tobacco expansin genes NtEXPA1, NtEXPA4, NtEXPA5, and NtEXPA6 were studied in different organs of tobacco (Nicotiana tabacum L.) as well as in response to phytohormone and stress treatments. It was shown that NtEXPA1, NtEXPA4 and NtEXPA5 transcripts were predominantly expressed in the shoot apices and young leaves, but almost absent in mature leaves and roots. The NtEXPA6 mRNA was found at high levels in calluses containing a large number of undifferentiated cells, but hardly detectable in the leaves of different ages and roots. In young leaves, expression levels of NtEXPA1, NtEXPA4 and NtEXPA5 genes were induced by cytokinins, auxins and gibberellins. Cytokinins and auxins were also found to increase NtEXPA6 transcripts in young leaves but to the much lower levels than the other expansin mRNAs. Expression analysis demonstrated that brassinosteroid phytohormones were able either to up-regulate or to down-regulate expression of different expansins in leaves of different ages. Furthermore, transcript levels of NtEXPA1, NtEXPA4, and NtEXPA5 genes were increased in response to NaCl, drought, cold, heat, and 10μM abscisic acid (ABA) treatments but reduced in response to more severe stresses, i.e. cadmium, freezing, and 100μM ABA. In contrast, no substantial changes were found in NtEXPA6 transcript level after all stress treatments. In addition, we examined the involvement of tobacco expansins in the regulation of abiotic stress tolerance by transgenic approaches. Transgenic tobacco plants with constitutive expression of NtEXPA1 and NtEXPA5 exhibited improved tolerance to salt stress: these plants showed higher growth indices after NaCl treatment and minimized water loss by reducing stomatal density. In contrast, NtEXPA4-silenced plants were characterized by a considerable growth reduction under salinity and enhanced water loss. Our findings indicate that expression levels of all studied tobacco expansins genes are modulated by plant hormones whereas NtEXPA1, NtEXPA4, and NtEXPA5 expansins may be involved in the regulation of stress tolerance in tobacco plants.

The AINTEGUMENTA gene encodes a transcription factor involved in the regulation of growth of both reproductive and vegetative plant organs. Two conserved regions of this gene were amplified and cloned in antisense orientation. The produced genetic constructs were used to obtain transgenic tobacco plants with the AINTEGUMENTA gene expression downregulated. Over half of the transgenic plants were characterized by smaller leaves, stems, and flowers with their shapes and symmetry not changed. The transgenic plants ceased to grow earlier, flowered later, and had fewer flowers than controls. The main reason for smaller organs was shown to be the lower cell number in an organ with cell sizes unchanged. The genetic constructs can be used to obtain transgenic plants of various species with smaller organs.

In the process of obtaining transgenic plants suitable for phytoremediation of heavy metal-contaminated soils, a synthetic pseudophytochelatin gene coding for a phytochelatin analog Met(GluCys)6Gly was de novo designed and cloned. Contrary to natural enzymatically synthesized phytochelatins, this peptide can be made by a template synthesis. A construct carrying the gene in question under the control of the viral constitutive 35S promoter was made on the basis of a binary vector pCAMBIA 1305.1. This construct was used to express the pseudophytochelatin gene in the model transgenic tobacco plants (Nicotiana tabacum L.), and the above plants acquired additional resistance to various cadmium concentrations.

Rhizobia have the ability to increase growth of non-legume plants due to the production of phytohormones and protection of plant from diseases and pathogens. However, the practical use of these beneficial bacteria sometimes fails because of their inability to effectively colonize rhizoplane and rhizosphere of inoculated plants. We chose the legume lectins as a factor that allows plants to form associative symbiosis with rhizobia. To test the fact that transgenic tobacco, tomato and rape roots with pea lectin gene may affect specific interaction with rhizobia, transgenic roots have been artificially inoculated by fluorescently-labeled pea rhizobia R. leguminosarum and east galega rhizobia Rhizobium galega. Microscopic and microbiological tests have shown that the number of adhered R. leguminosarum onto tobacco, rape and tomato roots which transformed with pea lectin gene is higher in comparison with the control, but no such effect through inoculation of these plants with R. galegae has been found. This confirms the interaction of R. leguminosarum with pea lectin at the surface of transformed roots. Undoubtedly, the improvement of recognition and attachment processes by using lectins can lead to the achievement of a stable associative relationship between non-symbiotic plants and rhizobia.

Communities of macroscopic red algae in rivers of the Upper Volga region and adjacent territories are considered. Their classification based on the Braun-Blanquet approach is proposed. Phytocoenoses belong to the class Lemaneetea fluviatilis and the order Lemaneetalia fluviatilis. Two alliances Lemaneion fluviatilis and Batrachospermion gelatinosi separate the plant communities of fast and slow flowing waters. The first mentioned alliance includes the association Lemaneetum fluviatilis only, the second one unites the associations Batrachospermetum gelatinosi, Batrachospermetum turfosi and the community Sirodotia suecica. Almost all syntaxa are published validly for the first time. The characteristics of syntaxonomical units are given. Data on species composition, structure, ecology and distribution of the revealed phytocoenoses are presented. The communities of red algae are the most diverse in the rivers of the western part of the Vologda region, where all studied syntaxa are recorded. The plant communities of the association Batrachospermeum gelatinosi are the most common in watercourses of the region. Investigated rhodophyte phytocoenoses are rather sensitive to any changes of environmental conditions indicating the purity of river waters. They can be recommended for conservation purposes.

Expansins are non-enzymatic plant proteins breaking hydrogen bonds between cellulose microfibrils and hemicellulose polymer matrix. Each plant has many expansin genes, whose protein products participate in the regulation of plant growth and development mainly by regulating cell expansion. To analyze the effects of elevated expansin expression on the plant organ sizes, we cloned the AtEXPA10 gene from Arabidopsis thaliana and PnEXPA1 gene from Populus nigra. Transgenic tobacco plants expressing the target genes were obtained. The obtained transgenic tobacco plants were shown to have significantly larger leaves and longer stems compared to control plants. The flowers were quite insignificantly larger, but at the same time transgenic plants had more flowers. The microscopic studies showed that the organs of AtEXPA10-carrying plants were larger mainly due to stimulated cell proliferation, whereas the overexpression of the PnEXPA1 gene activated cell expansion.

The auxin-inducible gene ARGOS from Arabidopsis thaliana is expressed in growing tissues and controls the plant organ size by regulating cell proliferation and meristematic competence. The promoter of the dahlia (Dahlia pinnata Cav.) mosaic virus (DMV) resembles the well-known cauliflower mosaic virus 35S promoter but shows a higher activity in transgenic tobacco plants (Nicotiana tabacum L.). We obtained transgenic tobacco plants expressing the Arabidopsis ARGOS gene under the control of the DMV promoter. Several of the T0 generation plants exhibited an accelerated transition to flowering, a slight increase in flower size, and a significant increase in the leaf size. The T1 transgenic plants were characterized by faster growth, the increased leaf size, and somewhat enlarged flowers as compared with control plants. These phenotypic traits, as well as stability and inheritance of the transgene were demonstrated also in T2 transgenic plants.

"Barbate roots" in tobacco and colza transgenic on lectin gene were obtained with the use of a wild strain of Agrobacterium rhizogenes 15834 transformed with pCAMBIA1305.1 plasmid containing the full-size lectin gene (psl) from the Pisum sativum. Influence of expression oflectin gene on colonization oftransgenic roots with symbiont of pea (Rhizobium leguminosarum) was investigated. The number of adhered bacteria onto the roots transformed with lectin gene was 14-fold and 37-fold higher in comparison with the control; this confirms the interaction of R. leguminosarum with pea lectin at the surface of the transformed roots of tobacco and colza. The developed experimental approach, based on the simulation of recognition processes and early symbiotic interactions with lectins of pea plants, may, in perspective, be used for obtaining stable associations of economically valuable, nonsymbiotrophic plant species with rhizobia.

Nitellopsis obtusa (Charophyta) is reported for the Bering Island (Commander Islands) for the first time. It is the most north-eastern record in Russia and Eurasia. The species is found in atypical habitat, in a shallow lakelet of coastal tundra. Possibility of distribution of the species by migratory water birds is discussed.

By the techniques of DNA shuffling, PCR, and restriction-ligation, chimeric forms of cauliflower (Brassica oleracea) mosaic virus (CaMV), dahlia (Dahlia pinnata) mosaic virus (DMV), and carnation (Dianthus caryophillus) etching ring virus (CERV) promoters were obtained at various combinations. Twelve chimeric promoters were cloned into pCambia binary vectors comprising the reporter GUS gene, and their activities in transgenic tobacco (Nicotiana tabacum) plants were determined fluorimetrically. 35S promoter and those of DMV (442 bp) and CERV (371 and 501 bp) were used as controls. Seven of analyzed promoters displayed higher and seven promoters lower activity in transgenic tobacco plants than 35S promoter. The highest activity was characteristic of natural DMV promoter, and the least one — natural CERV promoter 501 bp in size. The CERV promoter 371 bp in size was approximately similar in strength to 35S promoter.

The effects of hybrid lectins—full-sized pea Pisum sativum lectin (PSL) with the carbohydrate-binding region of white melilot Melilotus albus lectin or wild licorice Astragalus glycyphyllos lectin substituted for the corresponding PSL region (PSL/MAL and PSL/AGL, correspondingly)—on the legume-rhizobium symbiosis were studied. The treatment of the Rhizobium leguminosarum bv. viciae in the alfalfa (Medicago sativa) rhizosphere with PSL induced formation of uninfected pseudonodules on its roots, whereas the treatment of the bacteria from Astragalus cicer nodules with PSL/AGL rendered these bacteria able to form infective nodules on alfalfa roots. This ability is associated with expanded and unusual carbohydrate-binding properties (combined specificity for Gal and Glc) of this hybrid protein as compared with the natural legume lectins.

Promoters of carnation etched ring virus (CERV) and dahlia mosaic virus (DMV) were cloned into binary vectors pCambia 1304, pCambia 1281Z, and pCambia 1291Z with reporter GFP and GUS genes. Activities of these promoters in tobacco protoplasts and transgenic plants were determined using these constructs. Histochemical GUS analysis demonstrated the absence of tissue-specificity in transgenic plants transformed with these promoters. The quantitative analysis of these promoter activities in transgenic tobacco plants, using 4-methylumbelliferone as a substrate, showed that 35S CaMV, CERV, and DMV promoters displayed approximately similar activities in transgenic tobacco plants.

Competitiveness and genetic variation of the Rhizobium galegae strains from the collection of the All-Russia Institute of Agricultural Microbiology, Russian Academy of Agricultural Sciences, causing nodulation of oriental goat's rue under conditions of Bashkortostan soils (lacking this rhizobial species) were studied. It was demonstrated that of all the tested strains, the strains CIAM 0702 and CIAM 0704, each carrying two megaplasmids of 1500 and 2000 MDa, were the most competitive. RAPD (random amplified polymorphic DNA) analysis showed that R. galegae strains were able to intensively exchange the genetic material in the host plant rhizosphere. We did not succeed in detecting the local root nodule bacteria that were either initially able to infect oriental goat's rue or had adapted to infecting this species due to various genetic rearrangements.

The carbohydrate-binding sequences (CBS) in the lectin genes of Trifolium repens, T. pratense, and T. trichocephalum were sequenced. The gene regions encoding lectin CBS of T. pratense and T. repens displayed a considerable similarity; however, the CBS of these species differed essentially. Moreover, T. repens formed a compact cluster with Melilotus albus and M. officinalis in the phylogenetic trees constructed according to the nucleotide sequences and the corresponding CBS of legume lectins. T. trichocephalum does not fall into the group of the tribe Trifolieae members according to both the amino acid sequence of lectin carbohydrate-binding region and the nucleotide sequence of lectin gene.

The process of accumulation of knowledge on wheat and related wild species during the 20th century is briefly reviewed with special reference to the evidence of the recent years on evolution of polyploid wheats and the role of diploid species. The latter serve as potential donors of the genomes, detection of which is particularly important because of the continuing speciation in the tribe Triticeae and artificial development of synthetic forms. The arguments in favor of the donor role for various diploid wheat and aegilops species from the section Sitopsis are compared. It is stated that in the formation of the both lines of polyploid wheats turgidum–aestivumand timopheevi,diploid Aegilops speltoides acted as a maternal form. In addition to cytoplasmic genomes, this aegilops species introduced into them also the B and G nuclear subgenomes. A comparison of nucleotide sequences in the variable part of the promoter of evolutionary conserved rRNA genes in polyploid wheats with their counterparts in diploid wheats and aegilops species confirmed the accepted wheat phylogenies.

We have found a taxonomically unclear linear-leaved Potamogeton (section Graminifolii , Potamogetonaceae) in Central Yakutia (Asian Russia). This form resembles P . friesii and P . pusillus , two common species in the republic, and the mainly European species P . rutilus and North American P . strictifolius . These species, except P . strictifolius , did not differ by variation of the nuclear ITS region. The chloroplast markers petL-psbE and rpl32-trnL clearly separated the unclear Yakutian plants from P . friesii and P . pusillus , and the latter marker revealed similarity between the European P . rutilus and the Yakutian samples. The latter fit well within the morphological circumscription of P . rutilus . We confirmed the presence of P . rutilus in five localities in Central Yakutia and 13 more in other regions of North Asia. These records greatly extends to the east the distribution range of this species known before mainly west of the Middle Volga. Potamogeton rutilus prefers clean, mainly mesotrophic lakes with alkaline waters, and occurs in coastal areas or in limestone and sandstone catchments. The species usually grow in deep waters on silt-sandy bottom, in species-rich communities. It is sensitive to pollution and competition with more robust aquatic plants. Due to local distribution and specific habitat requerments P . rutilus can be regarded as threatened species and deserve the category Vulnerable in regional and global Red Lists.

The nucleotide sequences of 280–360-bp domains of lectin genes from 20 legume species belonging to 17 genera have been determined. A computer analysis of the sequences has been performed with the LASERGENE package. Based on this analysis, we constructed the phylogenetic tree of the lectins, which reflects their phylogenetic and evolutionary relationships, and predicted the amino-acid sequences of the corresponding protein domains. Features of the structure of the hydrocarbon-binding lectin domains were elucidated in some species of legume genera from the temperate climatic zone. The domains were highly variable and contained the consensus sequence AspTrePheXxxAsxXxxXxxTrpAspProXxxXxxIns/DelArgHis bearing the bulk of amino acid replacements, insertions, and deletions. An association between legume groups (including species from different genera and tribes) symbiotic with the same rhizobium species and the similarity between the hydrocarbon-binding domains of lectins from these plants was found.