Analysis of cellulose synthase gene expression strategies in higher plants using RNA-sequencing data.

The transcriptomes from different organs and tissues of western poplar, eucalyptus, soybean and common bean were studied. The expression level of cellulose synthase genes was notably different in different types of tissues and organs in studied plants. For common bean and eucalyptus transcriptome the domination of certain cellulose synthase genes was typical. These prevailing genes made up more than 50 % of the total expression pull of cellulose synthases. On the contrary, cellulose synthase expression pulls of wes-tern poplar and soybean were distributed between multiple genes. The different expression strategies of CesA-genes may reflect a phylogenetic processes that occurred in genomes studied.

Double helix of the journal «Ð¡ytology and genetics¼: fity years later.

This survey paper contains a brief analysis of publications included in current issue of scientific journal «Cytology and Genetics¼ dedicated to its 50th anniversary. These papers reflect scientific achievements of their authors in the field of genetics and cell biology and underine the potential of these two biological disciplines, forming «double helix¼ of the journal.

Diversity of ukrainian winter common wheat varieties with respect to storage protein loci and molecular markers for disease resistance genes.

Diversity of Ukrainian winter common wheat varieties was studied with respect to the storage protein loci Gli-A1, Gli-B1, Gli-D1, Glu-A1, Glu-B1, Glu-D1, Gli-A3, Gli-B5, and Gli-A6 (362 varieties) and markers for the Lr34/Yr18/Pm38/Sr57/Bdv1 gene conferring moderate resistance to a number of biotrophic pathogens, the Tsn1 gene for sensitivity to the toxins A of the necrotrophic fungi Pyrenophora triticirepentis and Stagonospora nodorum, the Tsc2 gene for sensitivity to the toxin B of P. triticirepentis, and the TDF_076_2D gene for moderate resistance to Fusarium head blight (181 varieties). Significant differences in frequencies of alleles at these marker loci between groups of varieties developed in different soil and climatic zones were revealed. The retention of a set of predominant alleles of a certain zone in different periods of breeding was confirmed. At the same time, the appearance of new allele associations in the groups of varieties of the Steppe (in particular Gli-A1g and Glu-B1al) and the Central Forest-Steppe (1AL/1RS and Glu-B1d) in the last two decades has been noted. Nonrandom associations between alleles of disease resistance genes as well as alleles of disease resistance genes and storage protein alleles were revealed

THE HOMOLOGOUS IDENTIFICATION OF THE STEM RUST RESISTANCE GENES RDg5, Adf3 AND RGA1 IN THE RELATIVES OF BARLEY.

The barley genes Rpg5, RGA1 and Adf3, which provide a strong resistance to many pathotypes of stem rust, were cloned a few years ago, but it was still unclear whether their homologues were represented in wheat and in related species. The paper describes the results of a bioinformatic research to determine the homologues of Rpg5, RGA1 and Adf3 in the genomes of Triticum aestivum and several wild grasses, which breeders usually use as sources of stem rust resistance, and which are available in the genome databases. It was found that the Th. elongatum sequence Q9FEC6 and T. aestivum sequence Q43655 were the high identical homologues of the Adf3 sequence. T. urartu M8A999 sequence and T. aestivum W5FCU1 sequence were found to be the closest homologues of Rpg5 complete protein sequence, but the identity of their kinase domains were not as clear as that of the other domains. The separate Rpg5 kinase part analysis did not provide the strong evidences that its orthologs were presented in our corn species. T urartu M7ZZX9 sequence and T. aestivum W5FFP0 and W5F133 sequences were showed to be the homologues of RGA1. The analysis of the predicted active sites allowed finding out the difference between sequences of Rpg5, RGA1, Adf3 protein and their homologues.

[ENHANCEMENT OF AGROBACTERIAL TRANSFORMATION OF PLANTS USING PROTEIN KINASE INHIBITORS TRIFLUOPERAZINE AND GENISTEIN].

The effect of different concentrations of protein tyrosine kinase inhibitor, genistein and serine/threonine protein kinase inhibitor, trifluoperazine, on the frequency of Agrobacterium-mediated transformation of leaf explants of N. tabacum was investigated. The influence of different concentrations of trifluoperazine in the range from 10 to 300 µM was investigated. It was found that 10 µM trifluoperazine provoked the increase of the frequency of agrobacterial transformation of tobacco leaf disks on 25%. In parallel, the influence of different concentrations of genistein in the range from 10 to 100 µM was investigated. It was found 100 µM genistein provoked the increase of the frequency of agrobacterial transformation of tobacco leaf disks on 12%.

[INVOLVEMENT OF PLANT CYTOSKELETON INTO CELLULAR MECHANISMS OF METALS TOXICITY].

This review summarizes published date and the results of the author's own researches cantering the participation of plant cells cytoskeleton. It is considered cytotoxic impact of metals on the cytoskeleton's components, including microtubules and actin filaments. Particular attention is paid to the cellular and molecular mechanisms of influence of metals on cytoskeleton. We discussed the most probable binding sites of heavy metals and alternative mechanisms of their impact on the cytoskeleton.

Genetic marking of glume colour in Triticum spelta L. var. caeruleum using gliadins.

Genetic control of dark color of glumes was studied in Triticum spelta L. var. caeruleum accessions using gliadins as genetic markers. F2 and BC1 plants from crosses between the spelt accessions and common wheat varieties with white glumes served as the material for the investigation. Analysis of segregation for glume colour fitted the monogenic control of the character. Using electrophoretical analysis of gliadin of seeds from the hybrid plants it was revealed that in the T. spelta var. caeruleum accessions the allele Gli-A1j* is linked to the allele for dark (black) glumes at the Rg-A1 locus.

Variation of microsatellite loci in Vincetoxicum Wolf. species in the south-east of Ukraine.

The genetic diversity of 13 Vincetoxicum Wolf species distributed in Ukraine was investigated using 4 of the 8 nuclear microsatellite markers, previously developed for Vincetoxicum atratum from Japan. The number of alleles ranged from 8 to 25. The expected heterozygosities were 0,690­0,938 and observed ones ranged from 0,205 to 0,806. In general, the level of genetic variation in studied representatives of the genus Vincetoxicum from Ukraine proved to be comparable with that of Vincetoxicum atratum. The microsatellite loci Vinc5, Vinc104, Vinc123, Vinc124 can be successfully used to assess intra- and interspecific polymorphisms of species of the genus Vincetoxicum Wolf in Ukraine.

Sr33 & Sr35 gene homolog indentification in genomes of cereals related with Aegilops tauschii & Triticum monococcum.

Using bioinformatics analysis, the homologues of the genes Sr33 and Sr35 were identifed in the genomes of Triticum aestivum, Hordeum vulgare and Triticum urartu. It is known that these genes provide resistance to hightly virulent wheat stem rust races (Ug99). To identify important for resistance amino acid sites, the comparison of the founded homologues with the Sr33 and Sr35 protein sequences was performed. It was found that the sequences S5DMA6 and E9P785 are the closest homologues of RGA1e protein ­ a product of the Sr33 gene, and the sequences M7YFA9 (CNL-C) and F2E9R2 are the homologues of CNL9 ­ a product of the gene Sr35. It is assumed that the homologues of the genes Sr33 and Sr35, which derived from the wild relatives of wheat and barley, can provide resistance to various forms of a stem rust and can be used in the future breeding programs for wheat improvement.

The influence of protein kinase KIN10 gene expression on root phenotype of Arabidopsis thaliana root system in condition of energy stress.

It was shown phenotypic changes in the root system of seedlings Arabidopsis thaliana in transgenic lines with overexpression and suppressed gene expression of serine-threonine protein kinase KIN10 in conditions of energy shortage and under normal conditions. The normal growth and development of KIN10 overexpressing plants with in energy deficiency conditions were detected. The significant inhibition of the plant development under normal conditions for these plant lines was obsereved. The levels of KIN10 gene expression under normal conditions in different organs A. thaliana, particularly in the roots, stems, leaves and flowers were analyzed. The highest level expression of the gene was fixed in the leaves.

Intra- and intertissular cytomictic interactions in mic-rosporogenesis of mono- and dicotyledonous plants.

A comparative cytological analysis of intra- and intertissular cytomictic interactions in early micro-sporogenesis of mono- and dicotyledonous plants was performed by the example of the two cellular systems - microsporocytes and tapetum. It is found that cytomixis is the component of intratissular interactions mainly. In the tapetum cells cytomixis is notable for structural and temporary taxon specific features. The nuclear migration in microsporocytes is confined mainly to zygotene-pachytene meiotic stages and characterized by a certain synchronism with cytomixis at the tapetum. Intertissular cytomictic interactions (tapetum - microsporocytes) were found in the monocot anthers only. Intertissular interactions are likely to reflect the intensification of competitive relations between the tapetum and microsporocytes for area in the process of anther tissue differentiation. Polyploid tapetum nucleus and syncytia being powerful acceptors are able to compete with microsporocytes and direct the chromatin translocation to their favor. The absence of intertissular interactions in dicots probably reflects a better balance between the processes of differentiation at somatic and generative tissues into microsporangium compared to monocots.

Influence of cold on organization of actin filament in various cell types root Arabidopsis thaliana.

The effect of the low temperature (4 ºC) on the orga-nization of actin filaments (microfilaments) of cells of different growth zones of the root of Arabidopsis thaliana (L.) have been studied. It was found that cold treatment inhibits growth of the main root and gives its morphology, causing a large number of deformed (ectopic) root hairs in the zone of differentiation. The temporal relationship of the disorientation and the organization of actin filaments and the detected changes of growth and morphology of roots under conditions of cold factor is shown. It has been found that the most sensitive to the cold are actin filaments of root hairs, meristematic cells, cells of elongation zone, and all epidermal cells of the root zones of A. thaliana.

Chornobyl thirty years later: evaluation of the consequence by biologists and medics Experience of thyroid gland status screening in postchernobyl period.

Summarizing paper contains short analysis of the publications resulting cell biological, molecular genetic and population genetic investigations devoted to studying of accute radiation exposure and chronic radiation contamination effects on biological systems after disaster on Chornobyl nuclear power station in 1986.

[BIOINFORMATIC SEARCH AND PHYLOGENETIC ANALYSIS OF THE CELLULOSE SYNTHASE GENES OF FLAX (LINUM USITATISSIMUM)].

A bioinformatic search of sequences encoding cellulose synthase genes in the flax genome, and their comparison to dicots orthologs was carried out. The analysis revealed 32 cellulose synthase gene candidates, 16 of which are highly likely to encode cellulose synthases, and the remaining 16--cellulose synthase-like proteins (Csl). Phylogenetic analysis of gene products of cellulose synthase genes allowed distinguishing 6 groups of cellulose synthase genes of different classes: CesA1/10, CesA3, CesA4, CesA5/6/2/9, CesA7 and CesA8. Paralogous sequences within classes CesA1/10 and CesA5/6/2/9 which are associated with the primary cell wall formation are characterized by a greater similarity within these classes than orthologous sequences. Whereas the genes controlling the biosynthesis of secondary cell wall cellulose form distinct clades: CesA4, CesA7, and CesA8. The analysis of 16 identified flax cellulose synthase gene candidates shows the presence of at least 12 different cellulose synthase gene variants in flax genome which are represented in all six clades of cellulose synthase genes. Thus, at this point genes of all ten known cellulose synthase classes are identify in flax genome, but their correct classification requires additional research.

[BIOINFORMATICS COMPARISON OF HUMAN AND PLANT PHOSPHATOMES].

The study presents the results of bioinformatic comparison of protein phosphatases from higher plants and human phosphatom (150 sequences). Based on sequence and profile comparison with known catalytic domains, 204 plant homologues from Physcomitrella patens and Arabidopsis thaliana where selected. Clustering of joint group of plant and mammalian protein phosphatases revealed fundamental differences in plant and human phosphatomes. At the same time, it was shown significant differences in the set of protein phosphatases in P. patens, A. thaliana, and such monocots as Orysa saliva and Zea mays.

[Role of transcription factors in transdifferentiation of the gastric mucosa].

The analysis of intestinal differentiation transcription factor CDX2 in the gastric mucosa biopsies has been carried out. It was established that CDX2 by itself promoter activation pathway can obtain intestinal phenotype for gastric mucosa cells. The loss of CDX2 expression in the nuclei of metaplastic epithelium may serve as a predictor of gastric mucosa malignization.

[PLANT GENETIC TRANSFORMATION USING CARBON NANOTUBES FOR DNA DELIVERY].

The possibility of exploiting carbon nanotubes (CNTs)-based nanocarriers to deliver genes into protoplasts, callus and mesophyll explants of plants was examined. Using single-walled CNTs (SWCNTs) at the concentration of 20 µg/ml and multi-walled CNTs (MWCNTs) at the concentration of 15 µg/ml genetic transformation of Nicotiana tabacum L. mesophyll protoplasts with plasmid pGreen 0029 was carried out and transient expression of reporter yfp gene in the protoplasts was observed. Using SWCNTs at the concentration of 40 µg/ml and MWCNTs at the concentration of 30 µg/ml genetic transformation of N. tabacum callus and leaf explants with nptII gene as a part of plasmid pGreen 0029 was carried out. As a result plant regeneration on selective medium containing 50 mg/lkanamycin was shown. SWCNTs-based nanocarriers de-onstrated their appli-ability to transform protoplasts as well as walled plant cells. Whereas, MWCNTs-based nano-arriers were suitable only for transformation of proto-lasts due to the limiting role of cellulose walls in cell penetration.

[DEVELOPMENT OF MARKER-FREE TRANSFORMANTS BY SITE-SPECIFIC RECOMBINASES].

To produce transgenic plants in current biotechnology selectable marker genes are used that lead to the selectivity.of transformants from non-transformed organisms. However, after the transgenic event has been occurred, the presence of these genes in transformed genome in general is uselless. Moreover, the continued presence of this kind of genes in transgenic plants with their further commercialization may raise certain public concern. Therefore, various techniques have been developed in recent years to obtain marker free transgenic plants. In the present review the main strategies for removal of selective marker DNA sequences that are used in genetic engineering are described. The most popular among them is site-specific recombination technology. The particular attention is paid to site-specific recombinase system Cre/loxP. The using of a new approach with site-specific recombinase system Cre/loxP under the control of 35S promoter to generate marker-free genetically modified plants is described.

Inhibitors of tyrosine kinases and phosphatases as a tool for the investigation of microtubule role in plant cold response.

Tyrosine phosphorylation plays a vital role in the variety of signal transduction pathways in eukaryotic cells, however its role and relevance in plants are still largely unknown. To investigate the functional role of tubulin tyrosine phosphorylation in plant cells the interplay between the effects of tyrosine kinases (herbimycin A) as well as tyrosine phosphatases (sodium orthovanadate) inhibitors on microtubules sensitivity to cold in A. thaliana root cells were studied. Since it was found that inhibition of tyrosine kinases significantly increased the microtubules sensitivity to cold, while inhibition of tyrosine phophatases enhanced their cold-resistance, we suggest an existence of certain functional interaction between the phosphorylation on tyrosine residues and sensitivity of cortical microtubules to low temperatures.

Effects of tyrosine kinase and phosphatase inhibitors on mitosis progression in synchronized tobacco BY-2 cells.

To test whether reversible tubulin phosphorylation plays any role in the process of plant mitosis the effects of inhibitors of tyrosine kinases, herbimycin A, genistein and tyrphostin AG 18, and of an inhibitor of tyrosine phosphatases, sodium orthovanadate, on microtubule organization and mitosis progression in a synchronized BY-2 culture has been investigated. It was found that treatment with inhibitors of tyrosine kinases of BY-2 cells at the G2/M transition did not lead to visible disturbances of mitotic microtubule structures, while it did reduce the frequency of their appearance. We assume that a decreased tyrosine phosphorylation level could alter the microtubule dynamic instability parameters during interphase/prophase transition. All types of tyrosine kinase inhibitors used caused a prophase delay: herbimycin A and genistein for 2 h, and tyrphostin AG18 for 1 h. Thereafter the peak of mitosis was displaced for 1 h by herbimycin A or genistein exposure, but after tyrphostin AG18 treatment the timing of the mitosis-peak was comparable to that in control cells. Enhancement of tyrosine phosphorylation induced by the tyrosine phosphatase inhibitor resulted in the opposite effect on BY-2 mitosis transition. Culture treatment with sodium orthovanadate during 1 h resulted in an accelerated start of the prophase and did not lead to the alteration in time of the mitotic index peak formation, as compared to control cells. We suppose that the reversible tyrosine phosphorylation can be involved in the regulation of interphase to M phase transition possibly through regulation of microtubule dynamics in plant cells.