Fractionation of eukaryotic DNA in a pulsating electrical field. I. Detection and properties of discrete DNA fragments]

The fractionation of eukaryotic DNA by field inversion gel electrophoresis results in the appearance of discrete DNA-fragments. The set of these fragments is similar to that of different eukaryotic representatives and consists of various chromosomal DNAs, unified by size. The physical properties of DNA-fragments suggest that they can form multimeric structures due to the presence of sticky ends flanking discrete fragments. We suppose that the set of discrete DNA-fragments results in a specific cleavage of intact nuclear DNA and can reflect different levels of chromatin structural organization.     

Antimutagenesis as organism defense system against damaging factors of endogenous and exogenous origin

Antimutagenic system is represented as the system combining all enzymatic and metabolic processes of the organism at all levels. The problem of supporting of autoantimutagenic system with antimutagenic preparations is discussed. A number of possible mechanisms of antimutagenic action of different substances are considered.     

Bacterial test-system for a primary screening of the substances with potential anti-cancer activity

A principle possibility of antitumour activity test in bacterial system represented by Escherichia coil of the wild type and its MS2-induced mutant has been shown. The initial bacterial strain is an indicator of toxic properties of the tested substances and the mutant one is a specific test-culture modelling a tumour cell. The comparison of the data described for eucaryotes with the data obtained using the proposed bacterial test system confirms an adequate response of both strains to the substances with the proved antitumour properties. The data are considered as very promising for the further improvement of this test system towards its application for primary screening of antitumour substances.     

Dynamics of changes in the genome of callus tissues of <Emphasis Type="Italic">Rauwolfia serpentina</Emphasis> upon a switch to conditions of submerged cultivation

It is established that switching from surface to submerged cultivation in a liquid medium of a special composition does not induce noticeable changes in the callus tissues of Rauwolfia serpentina in the course of the first several passages. Polymorphism of the RAPD spectra is discovered following 4–6 growth passages of tissues in the submerged culture; this polymorphism may reflect both changes in the DNA sequence and in the genetic structure of the cell population forming the strain. Addition of an intermediate passage in a solid medium of simpler composition prior to a switch to the liquid medium does not exert a substantial influence on the level and nature of the changes in the genome.     

Mixoploidy in wild and cultivated species of Cruciferae capable of hybridizing with rapeseed Brassica napus

The number of chromosomes in cells of the root meristem of seedlings of wild and cultivated species of Cruciferae plants capable of hybridizing with rapeseed Brassica napus is studied. Only diploid metaphases are observed in seedlings of Brassica juncea, Diplotaxis tenuifolia, and Raphanus raphanistrum. In B. napus and B. cretica roughly 5% of the seedlings are mixoploid. Diploid cells are dominant in the mixoploids, though hypo-and hyperdiploids are also encountered. Nearly 20% of the seedlings of B. campestris and R. sativum are mixoploid, a significant fraction of which contains di-triploid chimeras. In B. nigra less than one-half of the seedlings are truly diploid, the majority of the plants being mixoploid. Seedlings containing preferentially tetraploid and triploid cells are dominant. The biological significance and possible causes of the newly discovered mixoploidy are discussed.     

Fractionation of eukaryotic DNA in a pulsed electrical field. II. Discrete DNA fragments and level of structural organization of chromatin]

The data presented confirm the possibility of enzymatic formation of discrete DNA-fragments appearing during fractionation of nuclear DNA by FIGE. Teniposide-dependent pattern of DNA-fragments as well as occurrence of protein-linked DNA breaks suggest that discrete cleavage of intact nuclear DNA is modulated by DNA topoisomerase II. The possible relationship between discrete DNA-fragments and the higher order chromatin folding are discussed.     

The influence of some environmental factors on cytological and biometric parameters and chlorophyll content of Deschampsia antarctica Desv. in the maritime Antarctic

Under the environmental conditions of the Point Thomas Oasis (King George Island, the South Shetland Islands), we studied the influence of month-long artificial treatment with fresh water, salt water, and guano solution on the biometric characteristics, chlorophyll content, as well as the nuclear area of leaf parenchymal cells and nuclear DNA content, in a maritime Antarctic aboriginal plant Deschampsia antarctica. The modeled factors induced an increase in the generative shoot height and the length of the largest leaf, but did not influence the number of flowers. Treatment with guano caused an increase in the chlorophyll a and b contents, while fresh water treatment only led to some increase in chlorophyll a. Fluctuations of physiologically significant traits, such as the nuclear area and DNA content in the leaf parenchyma cells of D. antarctica, have been traced under the influence of the studied factors. Understanding of the hierarchy of influence of these factors as well as and sensitivity of plants of this species to external agents require further investigation.     

Biosynthesis of poppy isoquinoline alkaloids in nature and in vitro culture. 2. Bracteum poppy (Papaver bracteatum Lindl.)

Literature data and the data of the author's investigations on production of isoquinoline alkaloids by Papaver bracteatum Lindl. have been analyzed. Information on the methods of regulation and cell localization of morphine and sanguinarine biosynthesis is presented. The works studying differentiation processes in tissue cultures of bracteum poppy and relationship thereof with thebaine biosynthesis have been analyzed. Possible mechanism determining the induction of somatic embryos development and thebaine biosynthesis in the culture in vitro are proposed.     

Efficiency of different PCR-based marker systems for assessment of Iris pumila genetic diversity

We investigated informativeness and effectiveness of different marker types (ISSR, IRAP, REMAP, RGAP and LP-PCR that employ primers based on the conservative sequences of abiotic stress response genes) to study genetic diversity of Iris pumila L. By the number of amplicons per primer, number of polymorphic amplicons per primer and resolving power index (Rp), ISSR-markers were the most efficient followed by LP-PCR-markers. In order of decreasing value of indicators of genetic diversity “the percentage of polymorphic bands”, and “the average Jaccard? genetic distance between plants”, marker systems may be arranged as follows: ISSR > RAPD > LP-PC > RGAP ≈ IRAP. For ISSR-markers, the percentage of polymorphic bands was 1.3–1.7 times higher than for the others, and the average genetic distance was 1.2–1.3 times higher. Different marker systems were ranked by the value of Nei? gene diversity and the Shannon? index as follows: ISSR > RAPD ≈ LP-PCR > RGAP ≈ IRAP, with the highest and the lowest values differing 1.4 times. Genetic population structure was investigated with program Structure 2.3. The data of all marker systems suggest that all genomes under study belonged to one population. The PCoA and cluster analyses based on genetic distances showed distinctions in clustering generated from different markers data and summarized data, as well as the lack of strong clusters. Mantel test revealed significant positive correlation between the matrices of genetic distances generated by the data of almost all marker systems. The strongest correlation was found between RGAP- and IRAP-markers (r = 0.452, p = 0.01) and between RGAP and ISSR (r = 0.430, p = 0.01). ISSR, RAPD and LP-PCR proved to be more effective for the study of I. pumila genetic diversity, nevertheless, joint use of different marker systems will provide a more comprehensive assessment of variation in different genomic regions.