The Bulletin of Vavilov Society of Geneticists and Breeders of Ukraine, Vol.10, N2

In this issue:

Original papers

Lipid peroxidation in Arabidopsis thaliana wild type and KO-Cat2 knock-out mutant upon heat stress

Doliba I.M., Rusnak T.O., Volkov R.A., Panchuk I.I. P.193-201

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Aim. Denaturation of proteins and increased generation of reactive oxygen species (ROS) results in plant cell injury upon heat stress. The aim of the study was to elucidate the functional role of different catalase (CAT) isoforms in plant protection upon increased temperatures. Methods. Leaves of Arabidopsis thaliana of wild type and -Cat2 knock-out mutant plants were subjected to heat treatment. Development of heat stress response was monitored comparing the intensities of membrane lipid peroxidation induced by ROS. Results. It was demonstrated that moderate (37°C) and, especially, severe (44°C) heat treatment activates lipid peroxidation in Arabidopsis leaves. Increased level of lipid peroxidation in -Cat2 knock-out mutant compared to wild type plants was revealed at early stage (1 hour) of moderate heat treatment and in the recovery phase after severe heat stress. However, no difference was found upon other stress regimes tested. Conclusions. The data indicate that CAT2 is involved in plant cell protection against thermooxidative stress; the loss of this isoform in the mutant can be at least partially compensated by activation of alternative protective mechanisms.
Key words: Arabidopsis thaliana, heat stress, lipid peroxidation, catalase, knock-out mutants.

Novel structural subclass of Lycaena tityrus 5S ribosomal DNA

Cherevatov O.V., Statna A.P., Volkov R.A. P.202-207

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Aims. 5S rDNA belongs to the class of moderately repeated sequences that are organized as tandem arrays. Each repeated unit consists of the conservative 5S rRNA coding region and of rapidly evolving intergenic spacer (IGS), which is widely used for molecular taxonomy. However, organisation and evolution of 5S rDNA of Lepidoptera still remains poorly understood. The aim of the study was to describe molecular organisation of 5S rDNA of Lycaena tityrus. Methods. 5S rDNA repeated units were amplified by PCR, cloned in bacterial vector and sequenced. Results. It was shown that the IGS from 5S rDNA of L. tityrus has the length of 60-61 bp to demonstrate only low level of sequence similarity compared to recently sequenced IGS of other butterfly species. Accordingly, the sequenced repeats of L. tityrus 5S rRNA belong to novel structural subclass Mc.
Key words: Lycena, Lepidoptera, 5S rDNA, molecular evolution and taxonomy.

An estimation of large white breed boars

Bodryashova K.V., Macovska N. N., Birukova O.D., Sidorenko O.V. P.208-213

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Aims. To estimate boars showing differing proportions of inheritance from large white breed pigs of English selection by the performance of the nests from sows inseminated by them; perform genotype analysis by the Methods. Nest performance was carried out individually for each boar by the results of reproduced qualities in sows inseminated by them. Gene polymorphism to be associated with economically valuable traits was evaluated by the estrogen genes (ESR), melanokartin-4 (MC4R) through PCR-PDRF. Stress resistance was determined by the eosinophil test. Results. Reproductive performances in boars of differing genotypes (inheritance proportion of English origin varied between ? and ?) in the herd of "Rus’” Ltd have been compared. The best results by the traits of plural pregnancy and piglet preservation were found to be peculiar to boars showing lesser inheritance proportion (?) of English selection. Estimation of genotype distribution by the loci of quantitative and qualitative traits (by ESR- and MC4R-markers) demonstrated that all sires of desired genotypes (ESR, MC4R) exhibited differing reiteration of own genotypes in their offspring. Being appraised as stress-resistant, hog Dougol 102 displayed better piglet preservation (97.3 %) during suction period. Conclusion. Use of several methods for genotype analysis allows singling out the best animals thus promoting improvement of herd gene pool.
Key words: Genotype, preservation, plural pregnancy, pedigree value, stress-resistance.

Changes in the content of flavonoids, carotenoids and anthocyaninsof Alhagi pseudalhagi (Bieb.) in conditions of chronic irradiation by natural radionuclides

Dzhafarov E.S., Gojayeva G.A., Babayev G.G., Orujova J.R. P.214-223

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Aim. Aim of this work was both to study the dynamics of seasonal accumulation and radiation-dependent changes in the content of flavonoids, anthocianins and carotenoids in the leaves and flowers of Alhagi pseudalhagi (Bieb.), vegetating on the area contaminated by various radionuclides. Methods. There were usedradiodozimetric, ?-spectrometric and spectrophotometric methods. Results. It was demonstrated that these pigments accumulation in various plant organs appears to be the dynamic process with their content being varied both during ontogenesis and upon effect of various environment stimuli. Conclusions. Stimulating effect of low doses of ionizing radiation can occur in different phases of development. In this case, it is also possible adaptation of plants to chronic exposure to radiation.
Key words: Alhagi pseudalhagi (Bieb.), high radiation background, pigments, seasonal and dose-dependent accumulation.

Influence of the raised radiation background on the content of photosynthetic pigments in leaves of the wild growing plants on the territory of Baku factory on iodine manufacture

Dzhafarov E.S., Jafarly A.K., Orujova J.R., Babayev H.G.P.224-234

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Aim.As a result of investigation into the dynamics of levels and proportions of photosynthetic pigments in leaves an attempt was made to uncover some details of low dosage ionizing radiation influence on photosynthesis in plants growing within the zone of increased radiation background. Pigment levels in leaves of the plants under study was determined in various phases of vegetation period. Methods. There were used radiodozimetric, ?-spectrometric and spectrophotometric methods. Results. In some cases densities of photosynthetic pigments in experimental and control plants somewhat differed. It was demonstrated that while ionizing radiation at particular phase of single plant development may result in activation of photosynthetic apparatus promoting accelerated chlorophyll synthesis at another phase it may unfavorably affect photosynthesis inhibiting pigment production. Conclusions. Ionizing radiation may affect favorably and unfavorably on plant photosynthesis with stimulating effect being manifested in various plants in different phases of their development.
Key words: increased radiation background, natural radionuclides, photosynthesis, chlorophyll, carotenoids.

Allelic polymorphism of genes associated with athletes physical performance in different sports

Drozdovska S.B., Dosenko V.E., Ilyin V.N.P.235-244

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Aim. In an effort to search for molecular and genetic markers of hereditary predisposition to various sports there was determined incidence of allele gene versions contributing to physical performance among Ukrainian population and athletes of various sports. Methods. Gene polymorphisms in groups of athletes from various sports and individuals having no experience of regular exercise was determined by PCR. Results. I/D polymorphism of ACE gene and Pro582 ? Ser polymorphism of HIF-1? were found to be associated with a predisposition to various sports; for the first time it was determined that T-786 ? C promoter genes polymorphism of eNOS influences on physical performance in sports. Conclusions. D-allele of the I/D polymorphism of ACE gene, 582Ser allele of Pro582 ? Ser HIF ? gene polymorphism, T allele of the T-786 ? C polymorphism in the promoter of eNOS gene are markers of predisposition to speed-strength sports.
Key words: gene polymorphisms, sports selection, genetic predisposition, molecular genetic markers, sports.

Genetic features by isozyme loci of scots pine plus trees selected in Ukraine and Belarus

Korshikov I.I., Padutov V.Ye., Ivanovskaya S.I., Demkovich A.Ye., Storozhenko V.I., Kalafat L.A., Pasternak G.A., Bagdasarova A.R. P.245-253

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Aim. To perform comparative analysis of genetic polymorphism in plus trees (43) sampled in Scots pine (Pinus sylvestris L.) plantations in the South-East of Ukraine, a random sample of trees (36) from the same plantations, and plus trees (51), sampled in the plantantions in Belarus.Methods. To determine plant genotype, we used electrophoretic analyses of five enzyme systems with the identification of 12 polymorphic loci. Results. Plus trees selected in Ukraine had insignificant differences from a random sample of trees in terms of genetic variation and differentiation. The observed heterozygosity (HO) of plus trees sampled in Ukrainian plantations was lower (0.203) than that of plus trees sampled in Belarus plantations (0.260) and the expected one of Ukrainian plus trees (HE) was higher (0.324 and 0.260, respectively). Conclusions. Geographical isolation of the southern Ukrainian and Belarus populations of P. sylvestris has an impact on the features of their gene pools and causes differences in genetic polymorphism of plus trees samples.
Key words: Pinus sylvestris L., plus-trees, genetic variation.

ISSR-analysis of amaranth (Amaranthus L.)

Lymanska S.V. P.254-261

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Aim. The aim was to study the genetic diversity of amaranth collection. Methods. It was used ISSR-analysis. Results. Amaranth collection displayed high polymorphism (81.9 %). There were detected 84 fragments among which 12 were polymorphic and 6 unique. There was carried out cluster analysis by the results of which amaranth samples were distributed in 3 classes in accord with their species belongings. Conclusions. It was ascertained high level of genetic variation for plant material involved. Monophyletic hypothesis of origin for corny amaranth species was confirmed to record their insignificant phylogenetic divergence. Unique loci were identified which may be used as genetic markers for particular amaranth samples.
Key words: amaranth, ISSR, polymorphism, phylogenetic divergence.

Generation of Tragopogon porrifolius and Althaea officinalis «hairy» roots using Agrobacterium rhizogenes

Matvieieva N.A. P.262-268

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Aim. Aim of this work was to generate cultures of Tragopogon porrifolius та Althaea officinalis “hairy” roots. Methods. Roots were generated by co-culturing of leaves with cell suspension of Agrobacterium rhizogenes carrying рСВ161 vector or with A. rhizogenes cells of А4 strain. Selection of transgenic roots was performed on Murashige and Skoog medium with twice reduced content of macrosalts. Results. In 10-14 days after transformation when wild strain of A. rhizogenes was used there was observed root formation with frequencies for A. officinalis and T. porrifolius 92.5 and 59.4 %, respectively. In case of using A. rhizogenes carrying рСВ161 vector root formation incidence for these plants was 100 and 37.5%, respectively. In number of cases there were observed changes in polyfructane content: both increase up to 2.6 times in "hairy" Althaea roots and decrease down to 4.6 times in T. porrifoliustissue culture. Conclusions. A. officinalis and T. porrifolius plants differed from each other by the rate of root formation and its incidence. Maximal incidence of "hairy" root generation made up 100 % upon transformation with wild strain of A. rhizogenes. Some of "hairy" root lines following transformation demonstrated changed polyfructane content.
Key words: Agrobacterium rhizogenes, Tragopogon porrifolius, Althaea officinalis.

Application of plant growth regulators for intensification of biomass growth and increase of polyfructan amount in the chicory «hairy» root cultures

Matvieieva N.A., Tsygankova V.А., Chapkevich S.O., Kuchuk N.V., Ponomarenko S.P.P.269-278

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Aim. Verification the possibility to intensify the biomass growth of chicory (Cichorium intybus L.) “hairy” roots, obtained by Аgrobacterium rhizogenes-mediated transformation and increase in them the synthesis of polyfructan (PF) using Ivin, Emistim, Biolan and Charkor plant growth regulators. Methods. Biomass growth increment for chicory roots from four transgenic lines to be maintained on nutrient media with growth regulators in concentrations 2,5 – 10 µl/l and carrying nptII selective and ifn - ?2b target genes has been studied. Results. For the first time it was found, that Ivin, Emistim, Biolan and Charkor growth regulators can increase biomass increment in cultures of chicory "hairy" roots and stimulate in them PF synthesis. It is shown that growth regulator of natural origin Biolan and growth regulator of synthetic origin Ivin considerably increase versus control the specific PF amount, up to 130 mg/g of dry root mass (regulator Biolan) and up to 130-216 mg/g of dry root mass (regulator Ivin), and general PF amount: 1.3-5.5 times (regulator Biolan) and 2.6 to 28.9 times (regulator Ivin) as well as elevate general root biomass increment, 7.6 times (regulator Biolan) and 2.4 to 20.25 times (regulator Ivin) depending on the line of chicory transgenic roots. Conclusions. Ivin, Emistim, Biolan and Charkor growth regulators were found to increase biomass growth increment of chicory “hairy” roots and general PF amount therefore it seems reasonable application of the above mentioned regulators for cultivation of transgenic chicory roots. At the same time, because of revealed for different lines of transgenic roots individual responsiveness there is a need for preliminary screening and selection of “hairy” roots most responsive to growth regulators.
Key words: “hairy” roots of chicory, plant growth regulators, polyfructans.

Assessment of gene pool of large black pig breed using immunogenetic study

Parasochka I.F. P.279-285

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Aim. To develop approaches for immunogenetic marking of breeding material for conserving the specific breed features in pig gene pool herds as exemplified by Large Black breed. Methods. Studies were conducted at the main farms for pig breeding of Large Black breed. Red blood cell antigens have been identified, immunogenetic parameters and quality of reproductive traits of experimental sows have been studied. Results. This article highlights the results of investigation on protection of genetic resources of Large Black breed. Immunogenetic marking of pedigree material in gene pool swine herds of Large Black breed pigs allows more accurately select sets of parents that promotes the preservation of specific features of the breed. Conclusions. Sows with homozygous genotypes have higher reproductive capacity, so it is appropriate to take into account this fact in forming the genetic structure of breeding stock herds.
Key words: Large Black, genetic markers, genetic resources, reproductive characteristic.

Phenotypic traits variability in Streptomyces globisporus 1912–б/п transfotmants

Polishchuk L.V., Kopeуko О.P., Lukyanchuk V.V. P.86-94

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Aim.The aim was to determine inheritance stability of some traits in 36 transformants carrying hybrid plasmid DNAs. Methods.Stability of morphological traits inheritance (sporulation and pigmentation) for transgenic cultures was examined visually. Antibiotical activity of cultures was determined by microbiological methods. Metabolite properties (antibiotics and carotenoids) were investigated by thin layer chromatography and spectrophotometry. Results. Recipient strain Streptomyces globisporus 1912-б/п is a stable sporulated unpigmented culture, which does synthesize neither antibiotic nor carotenoids. Simultaneous changes of some recipient phenotypic traits in 72% of examined transformants were revealed. Conclusions. These results suggest that the genes that determine the studied phenotypic traits are characterized by the increased mutability.
Keywords: Streptomyces, transformants, phenotypic trait, mutation, genetic instability.

Transgenic red clover (Trifolium pratense L.) plants obtaining by agrobacterial transformation method

Reshetnikov V. N., Fomenko T.I., Berdichevets L.G., Malyush M.K. P.295-302

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Aim.The obtaining of red clover transgenic lines carrying licB target gene. Methods. Methods of plant regeneration in vitro culture. Agrobacterial transformation in vitro culture with use of vector constructions containing gus and licB genes. Results. The method of red clover regeneration in vitro culture with selection of optimum type explants and phytohormones providing the greatest amount of plant regenerants was developed. Transformation of clover plants with use of vector construction containing gene gus is carried out that allowed to optimize transformation conditions with target gene licB. With use cotyledonous leaves agrobacterial transformation method of red clover cultivars Yantarny, Vitebchanin and Bjursele kanamycin resistant regenerants were obtained. Conclusions. The method of multiplex PCR allowed to choose12 lines of transgenic plants from primary red clover transformants free from agrobacterial contamination and carrying within genome sequences of selective and target genes.
Key words: red clover Trifolium pratense L., in vitro tissue culture, morphogenic potential, agrobacterial transformation.

Alanin as a somatic segregation inductor in apical meristem cells of Allium cepa L. roots

Savinski S.K., Vdovychenko Zh.V., Sytnyk K.S., Zimina O.V., Spyrydonov V.G., Parii M.F. P.303-310

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Aim. Aim of the study is to test the effectiveness of the amino acid alanine as an inductor of somatic reduction in seedlings of onion Allium cepa L. and identify the most optimal combination of alanine concentration and duration of seedlings treatment to induce the phenomenon. Methods. The roots of onion Allium cepa L were treated with alanin solutions of different concentrations and for different periods of time. Further study of the samples was carried out by light microscopy. Results. In the apical meristem cells of plants undergone to treatment with alanin the somatic segregation was observed with a medium frequency 2.1 %. Control plants failed to display this. Most effective for induction of the phenomenon was found the plant treatment with 0.10-0.15 M alanine solution for four hours. Conclusions. It is confirmed that alanine is an inductor of somatic segregation in onion cells. When the optimal conditions of induction were applied, the phenomenon was observed in 3.5% of dividing cells.
Key words: Allium cepa L., somatic segregation (somatic reduction), induction, inductor, alanin.

Screening of the winter wheat varieties for the presence of wheat-rye translocation by DNA markers

Stepanenko A.I., Morgun B.V., Chugunkova T.V., Adamenko N.I., Velykozhon L.G. P.311-318

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Aim. Using primers to loci Xrems 1303 and ?-secalin found in the short arm of rye 1R chromosome to identify the presence of wheat-rye translocations in the genome of soft winter wheat varieties by PCR analysis. Methods. DNA analysis by polymerase chain reaction and electrophoretic determination of amplification products. Results. Varieties of soft winter wheat with wheat-rye translocation were discovered. Conclusions. Specific primers to loci found in 1RS arm of rye chromosome can be used for screening of wheat varieties for the presence in them wheat-rye translocation.
Keywords: Triticum aestivum L., DNA markers, PCR-analysis, multiplex-PCR, rye-wheat translocation.

Transaminase activity in blood of donor-cows

Sheremeta V.I., Vergeles A.P., Bilanyuk L.M.P.319-323

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Aim. The aim of research was to study the dynamics of transaminase activities in blood donor cows upon stimulation of superovulation with gonadotropin FFA «Folligon ®», together with the biologically active agent "Stimulin-Vet." Methods. There were investigated the activity of aspartate aminotransferase, alanine aminotransferase, by enzymatic kinetic method, using semi-automatic biochemical analyzer Stat Fah 1904 (Awareness Technology, USA) in donor cows that were undergone superovulation according to PMSG scheme «Folligon ®» (3000 ME ) on day 10 of the sexual cycle, PGF2 ?«Estrofan ®» (2 ml) – on 12, 13 days. Experienced donors were injected intramuscularly on 8, 9, 10, 11 days of the sexual cycle drug "Stimulin-Vet" at a dose of 20 ml. Results. Gonadotropin injections to target donors altered physiological co-ordination of transaminases to result in lower de Ritis coefficient and may clearly indicate the change of the metabolic fluxes in the direction of intensified anabolic reactions. In the experimental group, de Ritis coefficient between the 8 and 12 days of cycle, didn’t changed, even considering increased activity of both enzymes after co-administration of gonadotropin and a biologically active drug "Stimulin-Vet." Perhaps this is due to the enzymes interrelationship via glutamic acid (component of the drug) thus forming substrate-enzymological regulation for the intensity of metabolic fluxes. Conclusions. Injection of the active drug "Stimulin-Vet" during foligonom stimulation of follicle growth in the ovaries of donor cows contributed to AST and ALT activities without changing the de Ritis coefficient.
Key words: transaminases, superovulation, cow-donor, "stimulin-vet" coeficient.

Correlation between phylloxera and mildew resistance features in the first family (F1) hybrids

Shikhlinski H.M., Akparov A.I., Mammadova N.Kh.P.324-330

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Aim. To investigate the correlation between phylloxera and mildew resistance features in the first family (F1) hybrids of grape. Method. Study was carried out on the first family (F?) hybrids of grape by using biometric methods. Results. Characterizing relation between phylloxera and mildew resistance features of the first family (F1) hybrids of grape upon hybridization of resistant components with tolerant ones it was determined that, relation between indicated features widely vary depending on combinations of crosses. Hybrids derived from various combinations showed different correlation coefficients. Conclusion. The findings obtained indicate the dependence of inheritance of the features involved that result in resistance to phylloxera and mildew diseases and hence the possibility of breeding grape varieties showing group resistance to fungal diseases and mildew, suitable for own-rooting cultivation and free of chemical protection from mildew, with good quality of yield.
Key words: grape, Vitis vinifera L., phylloxera, mildew, correlation.

Determination of tropane alkaloids in golden henbane (Hyoscyamus aureus) in vitro

Besher Sh., Al-Ammouri Y., Lawand S.P.331-336

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Aim. Aim of the work was to study tropane alkaloids in Hyoscyamus aureus plants. Methods. L-hyoscyamine and (?)-scopolamine were analyzed through GC-MS. Results. Density of the tropane alkaloids (hyoscyamine and scopolamine) in plants under study late in the vegetative phase of development to be maintained in vitro was shown to depend on ecotypes from which seeds were sampled. In general, hyoscyamine predominated in roots, while leaves demonstrated prevalence only one of the alkaloids with the other being in trace amounts. Conclusions. Amount of alkaloids in roots was found to depend on altitude of plant vegetation as determined by its estimation in offspring generated in vitro. The idea that environment-genotype interaction may affect the amount of biologically active compounds produced by plant organism was confirmed. Key words: Hyoscyamus aureus, tropane alkaloids, culture maintained in vitro, GC-MS, L-hyoscyamine, (?)-scopolamine

History

The V.M. Remeslo Myronivka Institute of Wheat of National academy of agrarian sciences of Ukraine (to the centenary of the founding)

Kochmarskyi V.S.

P.337-343

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Plant Breeding and Genetics Institute – history, directions and results of researches

Sokolov V.M. P.344-357

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Anniversaries

Nadezhda Grigor'evna Shestopalova (to 85th anniversary)

P.358-359

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Bogdan Pavlovych Matseliukh (to 80th anniversary)

P.360-361

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Academician Vadimir Nikolaevich Reshetnikov (to 75th anni¬versary)

P.362-364

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Academician Volodymyr Grygorovych Radchenko (to 60th anniversary)

P.365-368

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Information

IX congress of Vavilov Society of Geneticists and Breeders of Ukraine

P.369-371

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Report of the Presidium of Vavilov Society of Geneticists and Breeders of Ukraine on the activities between VIII and IX Congresses (2008-2012) (main statements)

P.372-389

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On the awarding of Diplomas from the Presidium of Vavilov Society of Geneticists and Breeders of Ukraine

P.380-382

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VIII International conference «Factors for experimental evolution of organisms». Information letter

P.383-384

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Language advisor

Language advisor

P.385-386

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