Method of clonal micropropagation of different willow species and hybrids

Conditions of cultivation and micropropagation of selected biotypes of five willow species (Salyx dasyclados Wimm., S. caspica Pall., S. triandra L., S. purpurea L., and S. viminalis L.) and two hybrids (×S. acuminata S. and ×S. palustris Host.) were optimized. Data on in vitro propagation of S. caspica, S. triandra, S. purpurea together with hybrids S. acuminata and S. palustris were obtained for the first time. It has been demonstrated that the outcome of cultivation and propagation of willows strongly depends on genotypic peculiarities of initial plants. The optimal terms of isolation and sterilization of single-node segments for obtaining 50–75% of aseptic viable developing cultures were estimated. The nutritive media were selected providing induction of stem development (to 67%), their rooting (to 91%), elongation (to 3–6 cm), and multiplication (propagation coefficient of 4). The designed method (adopted to different genotypes) can be applied for obtaining aseptic in vitro cultures serving as initial plant material for genetic transformation and mass propagation of plants with new agriculturally valuable characteristics which are of interest for construction of bioenergetic plantations and for needs of the paper industry.     

Association of gene polymorphisms of matrix metalloproteinases with reproductive losses in the first trimester of pregnancy

In the present study, the frequencies of genotypes and alleles of candidate genes with respect to polymorphisms associated with increased pregnancy loss in the first trimester of pregnancy, including MMP1–1607insG, MMP9 A–8202G, and TIMP1 С536T, were reported. The frequency of homozygotes for allele MMP9 A–8202 was increased by a factor of two among women with miscarriage in the first trimester compared to the control. Significant models of interaction of genes MMPs and TIMP1 were revealed. The genotypes of genes MMP1 (rs1799750), MMP9 (rs11697325), and TIMP1 (rs11551797) increasing the risk of pregnancy loss in the first trimester were determined.     

Antimutagenic activity of mitochondria-targeted plastoquinone derivative

The ability of cationic plastoquinone derivative 10-(6′-plastoquinonyl) decyltriphenylphosphonium (SkQ1) to modify processes of spontaneous and induced mutagenesis was studied. It is shown that daily introduction of this compound into male Wistar rats in doses of 25 and 250 nmol/kg during two weeks decreases spontaneous level of chromosome aberrations in anaphase in the eye cornea from 0.39 ± 0.09 to 0.13 ± 0.08 and 0.14 ± 0.05, respectively. The level of 8-hydroxy-2′-deoxyguanosine in blood serum of the investigated animals decreases from 32.12 ± 1.55 to 25.90 ± 2.26 and 25.76 ± 1.50 ng/ml, respectively. These facts indicate that the decrease in spontaneous clastogenesis is caused by decreased level of DNA damage by endogenous reactive oxygen species. A higher dose of SkQ1 also decreases to control level chromosome aberrations caused by oxygen under pressure of 0.5 MPa for 60 min. It is also shown in experiments with bacterial biosensors that SkQ1 is able to efficiently protect cells against genotoxic effect of UV radiation at 300–400 nm.     

Responses of sunflower chlorophyll mutants to increased temperature and oxidative burst

A tolerance of nuclear, plastome, and mitochondrial mutants of sunflower (Helianthus annuus L.) to extreme factors, increased temperature and oxidative stress, which was simulated by hyperbaric oxygenation was studied. In model experiments, activities of superoxide dismutase and catalase were assayed; cytogenetic analysis was used for the assessments of proliferative activity of cells in the root meristem; in field experiments, seedling emergence and growth at the stage of 3–4 leaf pair were evaluated. Most tolerant to increased temperature and oxygen pressure were a plastome en:chlorina-5 mutant and a partial revertant pr6-en:chlorina-7 with a changed structure of mitochondrial DNA.     

Rodent lymphocytes express functionally active glutamate receptors

RT-PCR demonstrated that ionotropic (iGluR NR1) and metabotropic (mGluR Group III) glutamate receptors are expressed in rodent lymphocytes. Flow cytometry showed that activation of iGluR NR1 by N-methyl-D-aspartate (NMDA) increased intracellular free calcium and reactive oxygen species (ROS) levels and activated caspase-3. The latter effect was attenuated by the NMDA antagonist, 5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801), by the antioxidant N-acetylcysteine and by cyclosporin A. Treatment with L-2-amino-4-phosphonobutyric acid (L-AP4), an mGluR Group III agonist, increased lymphocyte ROS levels but to a lower extent than did NMDA. Activation of lymphocytes with both NMDA and L-AP4 caused a synergistic increase in ROS levels and induced necrotic cellular death without elevating the caspase-3 activation observed in the presence of NMDA alone. These results show that lymphocyte iGluR NR1 and mGluR Group III receptors may be involved in controlling rodent lymphocyte functions and longevity as they regulate events in cell proliferation, maturation, and death.     

Mutagenic effect of nitrosomethylurea, modified by heat shock at early stages of sunflower seedling development]

A comparative study of chlorophyll mutations induced in sunflower seedlings of the initial line 3629 and its nuclear and plastom mutant derivatives by nitrosomethylurea (NMU) (0.015%), heat shock (HS, 40 degrees C), and their combination was performed. The spectrum and frequency of chlorophyll mutations depended on the treatment conditions and on the genotype of the lines tested. Nuclear mutant displayed highest sensitivity to NMU, HS, and their combination. The line 3629 plants displayed highest tolerance to NMU and HS. However, plastom mutants were more tolerant to the combination of the mutagens compared to the plants of the initial line.     

The Use of the abundance ratio of 13C and 12C isotopes for characteristic of the origin of ethyl alcohol

During alcohol fermentation, the carbon isotope composition of ethyl alcohol produced depended on the substrate used and was characterized by the value of delta 13C equal to -24.7 +/- 0.8/1000 (wheat grain), -22 +/- 0.1/1000 (rye grain), -22 +/- 0.5/1000 (products of wood hydrolysis), -15.3 +/- 0.3/1000 (maize grain) and -10 +/- 0.1/1000 (sugar cane). The isotope composition of carbon of ethyl alcohol obtained during catalytic hydroxylation of ethylene has a delta 13C of -30.6 +/- 0.3/1000. The possibility of quantitative determination of specific components in mixtures of ethanol samples with various isotope compositions (chemical synthesis and alcohol fermentation of raw material from C3- or C4-plants) was shown.     

Mutagenic Effect of Nitrosomethylurea Modified by Heat Shock at Early Stages of the Sunflower Seedlings Development

A comparative study of chlorophyll mutations induced in sunflower seedlings of the initial line 3629 and its nuclear and plastom mutant derivatives by nitrosomethylurea (NMU), heat shock (HS, 40°C), and their combination was performed. The spectrum and frequency of chlorophyll mutations depended on the treatment conditions and on the genotype of the lines tested. Nuclear mutant displayed highest sensitivity to NMU, HS, and their combination. The line 3629 plants displayed highest tolerance to NMU and HS. However, plastom mutants were more tolerant to the combination of the mutagens compared to the plants of the initial line.