Scientific Journal of KubSAU

Polythematic online scientific journal
of Kuban State Agrarian University
ISSN 1990-4665
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288 kb

NUTRIENT MEDIUM FOR INDUCTION GEMMOGENESIS TO CREATE SOMATIC CLONES OF TEA PLANTS IN VITRO CULTURE

abstract 1321708100 issue 132 pp. 1258 – 1267 31.10.2017 ru 427
The article presents the first results of studies of somaclonal variability, which take place during cultivation of tea plants tissues and organs in vitro culture. As a starting material, there was used morphogenic callus, isolated from the basal part of tea microshoots, because callus increases somaclonal variability. An optimized protocol of the nutrient medium for induction gemmogenesis to product somatic clones of tea plants in vitro culture was developed. Morphogenesis calluses, initiated from the basal part of tea microshoots and subcultured on the nutrient medium with 6 – BAP – 2,5 ml + gibberellic acid – 1,0 ml + Tidiazuron – 4,0 ml + Tryptophan – 1000 mg, distinguished by high rates of induction of gemmogenesis – 63,3 %. The article studies the effect of exogenous growth regulators on morphological and growth indices of tea callus culture in vitro
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MORPHOLOGICAL FEATURES OF CONIDIOGENESIS OF VENTURIA INAEQUALIS (COOKE) WINTER IN LABORATORY CONDITIONS

abstract 1321708106 issue 132 pp. 1310 – 1322 31.10.2017 ru 526
The pathogenic agent of apple scab disease, Venturia inaequalis (Cooke) Winter, causes significant damage to the industrial production of apples. Studying the pathogen in pure culture is important for solving the theoretical and practical issues of its biology. The sporulation ability of the Venturia inaequalis laboratory culture is an important diagnostic feature in morphological culture analysis, in the test for sensitivity to fungicides, when creating a collection inoculum for an artificial infection. We have studied the morphological features of conidiogenesis of pure V. inaequalis culture, such as the location of conidiogenic structures and their form. For the first time in the laboratory culture of the pathogen, sporulation in the thickness of agar on the substrate mycelium was demonstrated. Conidiogenesis proceeded according to the blastic-annellidic type. Depending on the location of the conidiogenic structures on the aerial or substrate mycelium, their morphology was different. Conidiogenic areas in the substrate mycelium could be observed with the unaided eye, in the form of hyphal grit, while they were conidia conglomerations in each annelid. Annellide had a curved shape. On the aerial mycelium, annelids were straight and always had only one conidium. Differences in the morphology of conidiogenic structures are supposed to be related to the physical conditions of the environment in which sporulation takes place. In the agar, each mature conidium remaining at the apex of the annelid interferes with the formation of the next one, which results in its bending. The fixed arrangement of conidiogenic structures and forming conidia in the thickness of agar allows the use of substrate sporification for model studies of the conidiogenesis process
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DEVELOPMENT OF OPTIMAL WAY OF OBTAINING A HYDROLYZATE OF LACTIC ACID BACTERIA

abstract 1321708121 issue 132 pp. 1465 – 1477 31.10.2017 ru 561
The work was carried out in the scientific-research laboratory of the chair of Biotechnology, Biochemistry and Biophysics of Kuban SAU, the aim of which is the search of optimal conditions of obtaining the hydrolyzates of lactic acid bacteria by thermoacid and enzymatic ways. The pure cultures of microflora of gastrointestinal tract of quails − Lactobacillus agilis, Lactobacillus intermedius and Lactobacillus salivarius were the object of researches. Under conducting the thermal acid hydrolysis there were used the same correlations of bacterial mass and water as well as procedures of clearing of hydrolyzates. The variable conditions were рН, temperature and time of hydrolysis. When carrying out the enzymatic hydrolysis in this study there was used the enzyme from the group of muramidase lysozyme. The amount of enzymes and time of carried out hydrolysis were the variable conditions under obtaining the enzymatic hydrolyzates. In the process of researches there were studied: qualitative content of total protein, peptides with М.м. < 1500, protein and peptides with M.m. > 1500, aminoacids and as well as GMDP (glucosaminilmuramilpentapeptide). On the basis of carried out experiments there was stated that according to all components of biologically active substances the thermal acid way is more effective than enzymatic. The maximum accumulation of BAA in hydrolyzates by different ways was revealed at destruction of cells Lb. intermedius. Content of GMDP in thermal acid hydrolyzate was higher than in enzymatic on 0,028 g/100 ml (51,8 %)
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ICONOGRAPHY OF WHEAT

abstract 1331709021 issue 133 pp. 240 – 262 30.11.2017 ru 630
The study of agronomy plants history, its distribution and use through image analysis and iconography have been presented. As an object of study, the image of wheat has been taken as the ancient culture of the world among agricultural plants. After many years, the illustrative series takes on a completely different meaning, namely, it becomes an independent resource of information. The analysis includes images of wheat in painting, sculpture, mosaic, tapestries, murals, botanical illustrations, stamps, porcelain and scientific drawings. In the work, we have made an attempt to analyze the images of wheat in the works of art in order to search for species diversity, directions of selection and genetic research, and also to present the morphological characteristics of culture. The method of analysis is a method of visual notes or sketches, which consists in comparing and searching information by image. The analysis is carried out on grounds: tall wheat, multiflorous forms of wheat, nonspreading, thickness of straw. The visual analysis of the images of wheat in works of art by the method of sketches made it possible to reveal the species diversity of culture along the ear, to see the ancient forms that were cultivated in the world before the "green revolution". The analysis included works of art from the 14th to the 21st century, the masters of painting in Italy, Germany, Russia, the Netherlands, France, the United States, and others
263 kb

DESIGN OF THERAPEUTIC PHAGE COCKTAILS BASED ON T4-TYPE BACTERIOPHAGES: ADVANTAGES AND DISADVANTAGES

abstract 1331709063 issue 133 pp. 823 – 850 30.11.2017 ru 827
In the review, the stages of designing therapeutic cocktails of T4 type bacteriophages based on works by Harald Brüssow from Nestlé S.A. (Switzerland) are considered. The main stages of this process are identified: analysis of existing cocktails, selection of phages; creating a collection; cultivation of the host bacterium, multiplication of viruses; purification of the preparation; contamination testing; preservation, stabilization and storage; preclinical and clinical trials. H. Brüssow first studied the Russian drug " Coli-Proteus bacteriophage " of "Microgen" with the help of metagenomic analysis, electron microscopy and conducted its clinical studies. Prof. Brüssow considered the advantages of T4 bacteriophages for the treatment of Escherichia coli infections. Researchers studied methods of cultivation in Erlenmeyer flasks, in a bioreactor, in disposable sack cultivators for the propagation of viruses. For its purification the chromatography, centrifugation, filtration and polyethylene glycol precipitation were studied. To quickly check the contamination of phage cocktails, a mass spectrometry method is proposed. Researchers considered basic strategies, such as lyophilization, spray drying, the formation of microcrystals and microspheres to stabilize the preparations. They also reviewed the results of clinical trials of phage cocktails. We have listed the problems of selecting T4 bacteriophages from the point of view of modern knowledge. H. Brüssow and his colleagues carried out an interesting work on the construction of phage cocktails based on T4 type bacteriophages, and also revealed the problems of the current state of phage therapy
749 kb

APPROBATION OF ISSR DNA-MARKERS FOR GENOTYPING OF GALÁNTHUS WORONOWII LOSINSK.. AND ANALYSIS OF GENETIC STABILITY OF PLANTS, OBTAINED BY IN VITRO CULTURE

abstract 1331709088 issue 133 pp. 1166 – 1178 30.11.2017 ru 675
In the course of the work, 33 ISSR markers were evaluated for efficacy in the detection of genetic changes in regenerants of Galanthus woronowii Losinsk.. Ten markers were found suitable for genotyping according to the species under study. Five samples from the selected ten were analyzed for a sample of 20 plants of regenerants and a mother plant. The obtained data testify to genetic stability of plant material in the process of microclonal propagation
493 kb

STRUCTURAL AND PHYLOGENETIC ANALYSIS OF VVMYBA1, VVMYBA2 AND VVMYBA3 GRAPEVINE GENES

abstract 1341710026 issue 134 pp. 303 – 327 29.12.2017 ru 1122
The article presents the structural and phylogenetic analysis of VIT_02s0033g00410, VIT_02s0033g00390 and VIT_02s0033g00450 genes of the grapevine genome and the closely related orthologous gene MYB114 of the Arabidopsis genome. These genes are responsible for the biosynthesis of anthocyanin in the organs of model plants and are of interest not only for practical production and breeding, but also for fundamental research. These genes were analyzed for GCcomposition of nucleotides, the presence of cisregulatory elements and promoter regions. DNA and protein sequences were aligned to look for similar elements, which allowed further analysis of the ultraconservative domains of four genes. Based on the results of search and identification of the conservative regions, a cluster tree was constructed, which made it possible to identify the separation of gene sidelines from, presumably, the main one. At the same time, the construction of consensus trees based on DNA and protein sequences revealed their absolute similarity. "The Minimal Evolution Tree" allowed calculating the approximate dates of the appearance of the mutations and the divergence times of the gene branches between each other. At the same time, the appearance of the Vitis genus and its separation from the Rosales was taken as the time first divergence point. In the end, homologous metabolic pathways were searched between grapevine and Arabidopsis, which revealed the presence of homologous proteins in the grape proteome. In this turn, it already confirms the existence of similar biosynthetic pathways and, as a consequence, interactions such as "DNA-protein" and "protein-protein"
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CONSTRUCTION OF MUTANTS OF BACTERIOPHAGE T4 WITH REDUCED ANTIGENICITY

abstract 1341710034 issue 134 pp. 404 – 426 29.12.2017 ru 1475
A method of obtaining insertion mutants for the hoc gene, which encodes for the main phage antigen, was developed on the model of bacteriophage T4. This gene was cloned in the plasmid pBSL0+ and was disrupted by insertion of foreign DNA. The phage mutants were obtained by in vivo phage-plasmid recombination. The construction of insertion bacteriophage mutants was carried out in two stages. The resulting mutants on this procedure could be grown on wild-type E. coli strains, which is convenient for the production and use of these phages in therapy. The mutants obtained had reduced antigenicity. At the same time, the yield of the mutant strains was high when they were grown on the non-suppressor E. coli laboratory strains. A number of stages of purification of the bacteriophage mutants obtained were performed. Preparations have been studied by transmission electron microscopy and mass spectrometry. By several periodic cultivations of the mutant bacteriophages, it was shown that mutations of this type are stably maintained during more than 50 generations. T4 related bacteriophages of the family Myoviridae, for example, T-even, have the significant homology amongst their genomes, which makes possible to produce similar mutants. Thus, our method was developed to obtain mutants with reduced antigenicity which can be used for both the treatment of systemic infections, and diarrhea in the case, when, bacteriophages penetrate into the bloodstream. Such phages can be used in medicine and veterinary. The reported study was partially supported by RFBR, research projects No. 13-04-00991, 16-44-230855
223 kb

SPECIFICITY OF HEAVY METALS ACCUMULATION IN STEVIA REBAUDIANA

abstract 1341710043 issue 134 pp. 525 – 533 29.12.2017 ru 1044
The analysis of the accumulation and distribution of heavy metals in the stevia raw material grown in different agroecological regions is carried out. The elements present in the soils always have a complex effect on the plant and at the same time enhance or weaken each other's action. Plants of stevia are able to regulate the flow of the heavy metals under study, assimilating organs accumulate the optimal number of elements necessary for their growth and development, regardless of the growing conditions
296 kb

USING OF RETROTRANSPOSONE CASSANDRA BASED DNA-MARKERS FOR EVALUATION OF GENETIC DIVERSITY OF PRUNUS SPINOSA SPECIES

abstract 1341710094 issue 134 pp. 1166 – 1176 29.12.2017 ru 362
This article presents the results of testing IRAP DNA markers Cass1 and Cass2 applied to Prunus spinosa. The findings suggest the high perspectiveness of their using for the study of genetic diversity of the gene pool of this species. According to the results of the analysis of the sample 12 genotypes were identified from 6 to 13 fragments in the spectrum of Cass1 and from 5 to 11 fragments for Cass2. As a result of cluster analysis in the sample formed three groups of samples. In one of the groups, which is most distant from the other two, includes samples taken in Ukraine, while the remaining two groups included samples from Armenia, the Krasnodar region region, the Republic of Adygea, Ukraine and Moldova, and three cultural large-fruited form. The distribution of samples in clusters corresponded to their geographical origin that favors the objective assessment of genetic distances between the samples using Cass1 and Cass2 markers. Thus, it was concluded that the prospects of using DNA markers to study the genetic diversity within a species of Prunus spinosa
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