[Structural mechanisms of interaction of cyanoacrylates with plant tubulin].

Structural mechanisms underlying the specific binding of cyanoacrylate compounds with tubulin of higher plants have been studied by example of interaction of ethyl-(2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA1) and isopropyl-(2Z)-3-amino-2-cyano-4-ethylhex-2-enoate (CA2) with Arabidopsis thaliana alpha-tubulin. It was revealed that the cyano group of cyanoacrylates is a functional analog of the nitrile group, which determines the processes of specific interaction with plant tubulin for dinitroaniline compounds. Based on the data of spatial structure fluctuations, dynamics of hydrogen bonds and interaction energy of the CA1 and CA2, the most probable binding mode for these compounds with plant alpha-tubulin was identified and appropriate site of interaction was characterized. Seven out of 10 residues composing this site (Gln-133, Asn-249, Val-250, Asp-251, Val-252, Asn-253 and Glu-254) are obligatory components of the dinitroanilines' binding site on the plant alpha-tubulin surface. Thus, the binding site on the alpha-tubulin surface characterized by us is able to recognize and specifically bind the substances which are cardinally different by their chemical nature and have no common pharmacophore groups, under the condition of a certain similarity of their electrostatic topology.

[Obtaining the transgenic lines of finger millet Eleusine coracana (L.) Gaertn. With dinitroaniline resistance].

The current data is dedicated to the study of bioballistic and Agrobacterium-mediated transformation of finger millet with the constructs carrying the mutant alpha-tubulin gene (TUAm 1), isolated from R-biotype goosegrass (Eleusine indica L.), for the decision of problem of dinitroaniline-resistance. It was found that 10 microM of trifluralin is optimal for the selection of transgene plants of finger millet. PCR analysis of transformed lines confirmed the transgene nature of plants. The analysis of seed of T1 oftransgene lines confirmed heterozygous character of inheritance of the resistance.

[cDNA library construction from panicle meristem of finger millet].

The protocol for production of full-size cDNA using SuperScript Full-Length cDNA Library Construction Kit II (Invitrogen) was tested and high quality cDNA library from meristematic tissue of finger millet panicle (Eleusine coracana (L.) Gaertn) was created. The titer of obtained cDNA library comprised 3.01 x 10(5) CFU/ml in avarage. In average the length of cDNA insertion consisted about 1070 base pairs, the effectivity of cDNA fragment insertions--99.5%. The selective sequencing of cDNA clones from created library was performed. The sequences of cDNA clones were identified with usage of BLAST-search. The results of cDNA library analysis and selective sequencing represents prove good functionality and full length character of inserted cDNA clones. Obtained cDNA library from meristematic tissue of finger millet panicle represents good and valuable source for isolation and identification of key genes regulating metabolism and meristematic development and for mining of new molecular markers to conduct out high quality genetic investigations and molecular breeding as well.

[Mapping a new secalin locus on the rye iRS arm].

A gene designated Sec-N encoding secalin was mapped in the introgressive winter common wheat line Hostianum 273/97 (H273) with the wheat-rye substitution (1B)1R from the octoploid triticale AD825. F2 seeds from crossing the line H273 with the line Hostianum 242/97-2 carrying the wheat-rye 1BL/1RS translocation were analysed. The studied component on the SDS-electrophoregram of total proteins was revealed to be a monomeric secalin which is encoded by the gene at the new locus Sec-N located distally with respect to the Sec-1 locus at a distance of 21.4 +/- 2.5% (22.9 +/- 3.1 cM). The arrangement of the secalin loci on the 1RS arm indicates that the Sec-N locus is to be homoeologous to the Gli-1 loci of common wheat.

[Creation of transgenic sugar beet lines expressing insect pest resistance genes cry1C and cry2A].

Impact of insect pests makes a significant limitation of the sugar beet crop yield. Integration of cry-genes of Bacillus thuringiensis into plant genome is one of the promising strategies to ensure plant resistance. The aim of this work was to obtain sugar beet lines (based on the MM 1/2 line) transformed with cry2A and cry1Cgenes. We have optimized transformation protocol and direct plant let regeneration protocol from leaf explants using 1 mg/l benzylaminopurine as well as 0,25 mg/l benzylaminopurine and 0,1 mg/l indole-butyric acid. Consequently, transgenic sugar beet lines transformed with vector constructs pRD400-cry1C and pRD400-cry2A have been obtained. PCR analysis revealed integration of cry2A and cry1C into genome of transgenic lines and expression of these genes in leaf tissues was shown by reverse transcription PCR.

[Alleles at storage protein loci in Triticum spelta L. accessions and their occurrence in related wheats].

Variation at eight storage protein loci was analyzed in the collection of T. spelta accessions from the National Centre of Plant Genetic Resources of Ukraine, most of which are European spelts. The analysis allowed identification of seven alleles at the Gli-B1 locus, five alleles at the Gli-A1 and Glu-B1 loci, three alleles at the Gli-A3 locus, two at the Gli-D1, Gli-B5, Glu-A1, and Glu-D1 loci. The majority of alleles are encountered among common wheat cultivars, only five alleles were specific for spelts. The high frequency of the alleles Gli-B1hs* and h encoding the 45-type gamma-gliadin in European spelts and durum wheat cultivars, as well as the occurrence of these alleles in T. dicoccum, in particular, in accessions from Switzerland and Germany, supports von Büren's hypothesis that European spelt resulted from hybridization between a tetraploid wheat with the 45-type y-gliadin and a hexaploid wheat. Analysis of genetic distances based on the genotypes at eight storage protein loci permitted differentiation of the Asian spelt accession from European spelts.

[Genetic transformation of the moss Ceratodon purpureus by novel polycationic carriers of DNA].

There is a big progress in application of genetic engineering for improving the biological properties of different organisms. Viral and non-viral carriers are used for delivery of genetic material into target cells. Nanoscale polymeric materials of natural and synthetic origin are the most promising gene delivery agents. These polymers have demonstrated high efficiency of DNA delivery into the mammalian cells, although they were not very effective in plant cells. Here, the procedure for genetic transformation of Ceratodon purpureus (Hedw.) Brid. moss protoplasts is described. Method is based on the application of novel surface-active polymeric carriers of the polyDMAEM structure and controlled length and charge. It allows obtaining more transient and stable moss transformants per microgram of plasmid DNA when compared with known protocol based on using polyethyleneglycol. It is easier, more convenient, and cheaper than the "gene gun" method. Perspectives for further improvement of structure and functional characteristics of novel polymeric carriers are considered for delivery of genetic material into plant cells.

[Biodiesel from microalgae: ways of increasing effectiveness of lipids accumulation by genetic engineering methods].

Microalgae are viewed as one of the most perspective producer of lipids for biodiesel production. The review shows the results of researches of genes' expression increase actually included in fatty acids biosynthesis. The increase of effectiveness of solar energy absorption and carbon dioxide fixation influences the microalgae productivity. Blocking expression of genes that are responsible for starch synthesis, changes the balance towards the quantity growth of lipids in the cell. The change of the length in fatty acids carbon backbone chain towards its shortening is important in the technology of biodiesel production. Operating processes of lipids' catabolism is another way of increasing their quantity. And at last using the methods of transcription analysis allows us to get deeper into the process of intensive accumulation of lipids in stressful conditions for the purpose of directing these processes.

[Alternative type of fuel--biobutanol].

Butanol--an alternative fuel that on amid dwindling global (accessible) oil reserves can serve as a source of energy. In the industrial-scale butanol is produced by chemical synthesis, although initially butanol production was due to microbiological synthesis. For cost-efficient production, a strain of microorganisms must have over production of butanol. In the review of butanol synthesis pathway with the help of microorganisms, their regulation, the principles and techniques of increasing the productivity of the most promising strains and producer strains for industrial production.

[Study of the effects of gamma-irradiation of common wheat F1 seeds using gliadins as genetic markers].

Effects of irradiation of dry F1 seeds with gamma rays in the dose of 200 Gy were studied. Hybrids between near-isogenic lines on the basis of the variety Bezostaya 1 served as the material of investigation. Irradiation markedly reduced productivity traits of F1 plants and did not affect the survival of F1 plants under the given growth conditions. A significant relative increase in the frequency of pollen grains with the 1BL/1RS translocation that formed F2 seeds in comparison with the control was one of the effects of irradiation of F1 seeds. Irradiation with gamma-rays induced mutations at gliadin loci with the frequency of 7,4 % (at 0,5 % in the control).

[Establishment of in vitro culture, plant regeneration and genetic transformation of Camelina sativa].

The results on in vitro culture establishment, plantlet regeneration and rooting of Camelina sativa cultivar sample Peremozhets and cultivar Mirazh are presented. Effective concentrations of sterilizing agents and duration of plant material treatment were estimated. Phytohormone ratio, sucrose concentration in nutrient medium that induce effective formation of C. sativa shoots and NAA concentration for plantlet rooting have been established. The method of Agrobacterium-mediated transformation of Camelina by using binary vector pGH217 carrying reporter beta-glucoronidase (gus) gene driven under 35S CaMV promoter and nos-terminator, and selective marker hpt gene conferring hygromycin-resistance in transgenic plant was elaborated.

[Allelic state of the molecular marker for the golden nematode (Globodera rostochiensis) resistance gene H1 among Ukrainian and world cultivars of potato (Solanum tuberosum ssp. tuberosum)].

The purpose of our investigation was determination of allelic state of the H1 resistance gene against the pathotypes Ro1 and Ro4 of golden potato cyst nematode (Globodera rostochiensis) among Ukrainian and world potato (Solanum tuberosum ssp. tuberosum) cultivars. The allelic condition of the TG689 marker was determined by PCR with DNA samples isolated from tubers of potato and primers, one pair of which flanks the allele-specific region and the other one was used for the control of DNA quality. Among analyzed 77 potato cultivars the allele of marker associated with the H1-type resistance was found in 74% of Ukrainian and 90% foreign ones although some of those cultivars proved to be susceptible to the golden potato nematode in field. The obtained data confirm the presence of H1-resistance against golden nematode pathotypes Ro1 and Ro4 among the Ukrainian potato cultivars and efficiency of the used marker within the accuracy that has been declared by its authors.

[Genetic transformation of flax (Linum usitatissimum L.) with chimeric GFP-TUA6 gene for visualisation of microtubules].

The data of Agrobacterium-mediated transformation of some Linum usitatissimum cultivars zoned on the territories of Belarus and Ukraine with the plasmid carrying chimeric GFP-TUA6 gene and nptII gene as selectable marker conferring resistance to kanamycin are presented in this study. Transformation was affected by a number of factors including optical density (OD600), time of inoculation of explants with Agrobacterium and co-culture conditions. Transgenic nature of obtained lines was confirmed by PCR analysis. Expression of GFP-TUA6 gene was detected with confocal laser scanning microscopy. The obtained transgenic lines can be used for further functional studies the role of microtubules in the processes of building the flax fibres and resistance to wind.

[Docking of low-molecular ligands on the plant FtsZ-protein with application of CUDA-accelerated calculations].

This article provides review and analysis of opportunities for application of the CUDA technology for acceleration of computations in structural biology and bioinformatics. On the example of work with the Hex 6.1 program, comparative analysis of increase in the speed and quality of results of hard-docking of a number of low-molecular compounds on the surface of the FtsZ protein from Arabidopsis thaliana was performed. Several potential benzimidazole--plant FtsZ protein binding sites were identified.

[Isolation and amplification cDNA of 8H07 gene conservative region of nematode Heterodera schachtii with high relationship to its rape homolog].

Original method of small regulatory si/miRNA isolation from plant cells was elaborated. PCR amplification of fragment cDNA 8H07 nematode Heterodera schachtii gene was carried out. Using Northern-blot method hybridization of plant si/miRNA with cDNA fragment of conservative region 8H07 gene the presence of their high homology is found out. The amplified cDNA fragment of nematode 8H07 gene in future will be used for creation recombinant gene with complementary antisense dsRNA sequence for increasing resistance of rape plants to parasitic nematodes.

[Genetically modified plants and the problems of plant protection: progress and estimation of potential risks].

The review deals with advances and prospects in development of transgenic plants. At present virtually all commercial GM crops are those created for solving plant protection problems--they carry transgenes conferring resistance to herbicides, pests, viruses. Approaches employed for development of commercial GM crops with herbicide, pest and virus resistance, as well as strategies and prospects of development of commercial GM plants with resistance to fungal and bacterial diseases and nematodes, are considered. Ecological (including agronomical) and social risks associated with commercial growing of transgenic plants are briefly discussed.

[Effect of inhibitors serine/threonine protein kinases and protein phosphatases on mitosis progression of synchronized tobacco by-2 cells].

In order to investigate the role of various serine/ threonine protein kinases and protein phosphatases in the regulation of mitosis progression in plant cells the influence of cyclin-dependent (olomoucine) and Ca2+ -calmodulin-dependent (W7) protein kinases inhibitors, as well as protein kinase C inhibitors (H7 and staurosporine) and protein phosphatases inhibitor (okadaic acid) on mitosis progression in synchronized tobacco BY-2 cells has been studied. It was found that BY-2 culture treatment with inhibitors of cyclin dependent protein kinases and protein kinase C causes prophase delay, reduces the mitotic index and displaces of mitotic peak as compare with control cells. Inhibition of Ca2+ -calmodulin dependent protein kinases enhances the cell entry into prophase and delays their exit from mitosis. Meanwhile inhibition of serine/threonine protein phosphatases insignificantly enhances of synchronized BY-2 cells entering into all phases of mitosis.

[Reconstruction of spatial structure of plant protein phosphatase type-1 and -2A in complex with okadaic acid].

The homology modeling, based on known temple structures of Homo sapiens protein phosphatase type-1 and -2A was implemented. The spatial structures of the human protein phosphatases and their plant homologs from Arabidopsis thaliana was predicted. The quality of models was confirmed by conformational analysis and root mean square deviations. The sites of okadaic acid binding in molecules of plant protein phosphatases (type-1 and -2A) were proved by the data of comparative analysis and molecular dynamics.

[Developing transgenic barley lines producing human lactoferrin using mutant alfa-tubulin gene as selective marker gene].

Method of biolistic transformation was used for genetic improvement of commercial barley cultivars (Oksamitoviy, Vodogray and Getman). The plasmid pHLFTuBA was used for particle bombardment that consists of the hLF gene under the control of the barley glutelin B-1 promoter and a selectable marker gene, alpha-tubulin conferring resistance to trifluralin (dinitroanilinr herbicide). Preliminary screening of different trifluralin concentration range from 0,1 to 30 microM was tested for determination of effective selective agent concentration. Two transgenic barley line of genotype Oksamitiviy and transgenic callus line of cultivar Getman were obtained after selection on 10 microM of trifluralin. To confirm the transgenic nature of regenerated plants, the PCR analysis was carried out. The 734bp length fragment of hLFgene was amplified from both regenerated plants.

[Morphological and histochemical analysis of mucous membrane transformation of the artificial urinary bladder].

The performed morphologic study of transformation mechanisms of the intestinal reservoir mucous membrane using modern immunochemical and histochemical methods revealed three morphologic phases of enteric epithelium adaptation; reactive-inflammatory, compensatory-protective and atrophic. Enteric urinary reservoir preserves the cambial apparatus typical for the enteric phenotype with an inherent apoptosis activity and proliferation which confirms a low level of tumor transformation risk.