RESUMEN
BOX, ERIC and REP--genomic fingerprints of 12 isolated and 10 typical pathogenic for rape bacterial strains Pseudomonas, Xanthomonas and Pectobacterium genera have been analyzed. The affinity of isolated strains with representatives of P. marginalis pv. marginalis, Pseudomonas fluorescens and Xanthomonas campestris pv. campestris species has been determined.
Asunto(s)
Brassica rapa/microbiología , ADN Bacteriano/genética , Pectobacterium/aislamiento & purificación , Enfermedades de las Plantas/microbiología , Pseudomonas/aislamiento & purificación , Xanthomonas/aislamiento & purificación , Electroforesis en Gel de Agar/métodos , Pectobacterium/genética , Reacción en Cadena de la Polimerasa/métodos , Pseudomonas/genética , Xanthomonas/genéticaRESUMEN
Analysis of molecular-genetic polymorphism of barley varieties was performed using AFLP-method. A system for identification and differentiation of barley varieties based on AFLP-markers was developed. Results of testing for 19 varieties indicate a high differential ability of the developed system. Identification of varieties can be conducted using one of two proposed discriminatory sets of AFLP-markers. Based on the calculated genetic distances a dendrogram of phylogenetic relations between the varieties was constructed. The dendrogram revealed a separated origin of varieties of malting and feed directions.
Asunto(s)
ADN de Plantas/análisis , Hordeum/genética , Filogenia , Polimorfismo Genético , Selección Genética , Alelos , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Alimentación Animal , Cerveza , Cartilla de ADN/biosíntesis , ADN de Plantas/clasificación , ADN de Plantas/genética , Frecuencia de los Genes , Marcadores Genéticos , Hordeum/clasificación , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
The fatty acid composition of cell lipids of 15 strains isolated from the affected plants of rape and five collection strains has been studied. According to the results of chemotaxonomic analysis it has been found that 9 isolated strains are similar to representatives of species Pseudomonas marginalis and Pseudomonas fluorescens, and 6 - to those of Xanthomonas campestris pv. campestris. The authors have established the efficiency of certain methods for the extraction of fatty acids used for the identification of bacteria pathogenic for rape which belong to the genera Pseudomonas and Xanthomonas.
Asunto(s)
Brassica rapa/microbiología , Ácidos Grasos/aislamiento & purificación , Enfermedades de las Plantas/microbiología , Pseudomonas fluorescens/química , Pseudomonas/química , Xanthomonas campestris/química , Cromatografía Liquida , Ácidos Grasos/química , Espectrometría de Masas , Metilación , Filogenia , Pseudomonas/clasificación , Pseudomonas/aislamiento & purificación , Pseudomonas fluorescens/clasificación , Pseudomonas fluorescens/aislamiento & purificación , Xanthomonas campestris/clasificación , Xanthomonas campestris/aislamiento & purificaciónRESUMEN
Bacterial destruction of the culture was described and its agents identified in the spring and winter rape crops. Typical symptoms are the following: browning of stem tissue and its mucilagization, chlorosis of leaves, yellowing and beginning of soft rot in the place of leaf stalks affixion to stems, loss of pigmentation (violet). Pathogenic properties of the collection strains and morphological, cultural, physiological, and biochemical properties of the agents of rape's bacterial diseases isolated by the authors have been investigated. It was found that all the isolates selected by the authors are highly or moderately aggressive towards different varieties of rape. According to the complex of phenotypic properties 44% of the total number of isolates selected by the authors are related to representatives of the genus Pseudomonas, 37% - to Xanthomonas and 19% - to Pectobacterium.
Asunto(s)
Brassica napus/microbiología , Pectobacterium/patogenicidad , Pseudomonas/patogenicidad , Xanthomonas/patogenicidad , Enfermedades de las Plantas/microbiología , Hojas de la Planta/microbiología , Tallos de la Planta/microbiologíaRESUMEN
In this work we have demonstrated two independent real-time PCR methods for detection single nucleotide polymorphism (SNP) of genes csn and acyl-CoA: diacylglycerol acyltransferase 1 (dgat) in cattle. We have analyzed 296 samples of milk production cattle of Ukrainian breeding. The genotype frequencies were AA--0.58, AB--0.34, BB--0.08 for csn gene and for dgat gene--AA--0.7, AK--0.26, KK--0.04. High efficiency of so called "anti-primer" method was shown. Duration of anti-primer PCR reaction was about 2-2.5 hours only and provided full investigation of unknown gene allele.
Asunto(s)
Caseínas/genética , Cartilla de ADN , Diacilglicerol O-Acetiltransferasa/genética , Lactancia/genética , Polimorfismo de Nucleótido Simple , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Alelos , Animales , Bovinos , Industria Lechera , Femenino , Frecuencia de los Genes , UcraniaRESUMEN
The WHO approved protocol of pandemic influenza virus A/H1N1 identification by the method of one-step RT-PCR was adopted. The cost of research was decreased due to adoption of RNA purification method and utilizing of the common enzyme systems for RT-PCR.
Asunto(s)
Brotes de Enfermedades , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Diagnóstico Diferencial , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Gripe Humana/epidemiología , Mucosa Bucal/virología , Sensibilidad y Especificidad , UcraniaRESUMEN
Investigations of the genetic structure of internal noncoding transcribed spacer 2 (ITS2) of ribosomal DNA of five Trichogramma species - T. pintoi Voeg., T. evanescens Westw., T. dendrolimi Mats., T. cacoeciae Meyer. and T. sembidis Auriv. were carried out. The performed PCR-analyses with the following nucleotide sequence determi- nation of ITS2 regions allowed to reveal essential interspecific distinctions. The data can be used for T. pintoi, T. dendrolimi and T. sembidis species identification.
Asunto(s)
ADN Espaciador Ribosómico , Avispas/genética , Animales , Cartilla de ADN , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Ácido Nucleico , Especificidad de la EspecieRESUMEN
The influence of simulated microgravitation (clinostating) on the proceeding of mixed virus infection in Zhuravushka variety potato plants, infected with potato viruses X and M (PVX, PVM) was investigated. It was shown, that sensitivity of "virus-plant" system to microgravitation depends both on pathogene species and clinostating mode. Vertical clinostating was found to be more effective than horizontal one. By means of biotest, PCR and ELISA it was detected that potato plants, infected with PVX and PVM are released from PVX under the influence of clinostating during 19-47 days. Assume, that microgravitation is able to induce some protective reactions in plants that influence virus reproduction.
Asunto(s)
Enfermedades de las Plantas/virología , Virus de Plantas/aislamiento & purificación , Solanum tuberosum/crecimiento & desarrollo , Solanum tuberosum/virología , Simulación de Ingravidez , Electroforesis en Gel de Agar , Genes Virales/genética , Microscopía Electrónica , Enfermedades de las Plantas/etiología , Virus de Plantas/crecimiento & desarrollo , Virus de Plantas/ultraestructura , Reacción en Cadena de la Polimerasa , RotaciónRESUMEN
The chemical synthesis of a modified gene fragment that codes IgG-binding domain of protein G of Streptococcus sp. (SPG) was done. Two copies of gene fragmentes SPG were cloned into plasmid vector pET-24(a), for expression of recombinant truncated protein G (rSPG) in the culture of Escherichia coli. IgG-binding capacity and specificity for rSPG was confirmed by model assay such interaction in vitro. Basing on the obtained data we affirm that truncated recombinant protein G is able to bind immunoglobulin G in mammalians and can be used for purification of IgG from sera by affinity chromatography or for synthesis of immunoassay conjugates with broad range specificity.
Asunto(s)
Proteínas Bacterianas , Técnicas para Inmunoenzimas/métodos , Inmunoglobulina G/metabolismo , Proteínas Recombinantes , Streptococcus/metabolismo , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Streptococcus/genéticaRESUMEN
We have carried out comparative studies of double-stranded RNA (dsRNA) of viral nature isolated from sugar beet leaves and from mycelium of microscopic fungi using different methods such as PAAG electrophoresis and by polymerase chain reaction (PCR). It was shown that the fragments of dsRNA from sugar beet leaves and from mycelium microscopic fungi had the identical electrophoretic pattern and the same size (1.8 and 2.0 kbp). Using PCR technique it was shown, that isolated dsRNA have a common template for amplification. Electron microscopy of PCR-positive mycelium allows us to detect the virus particles of the spherical form with diameter 30-40 nm. The obtained data confirm our previous suppositions, concerning the belonging of isolated dsRNAs (size 1.8 and 2.0 kbp) to new mycovirus targeted a microscopic fungus, instead of beet cryptic viruses.
Asunto(s)
Basidiomycota/virología , Beta vulgaris/microbiología , Virus de Plantas/aislamiento & purificación , ARN Bicatenario/análisis , ARN Viral/análisis , Basidiomycota/ultraestructura , Beta vulgaris/virología , Electroforesis en Gel de Poliacrilamida , Microscopía Electrónica , Micelio/ultraestructura , Micelio/virología , Reacción en Cadena de la PolimerasaRESUMEN
Species composition of fungi, isolated from the sugar beet leaves, roots and rhizosphere, collected in Poltava and Kyiv regions has been studied. Species Alternaria, Trichoderma, Cladosporium, Acremonium, Penicillium, Fusarium, Mycelia sterilia were found in all investigated samples. Basidiomycetes were isolated from the leaf samples in separate cases.
Asunto(s)
Beta vulgaris/microbiología , Hongos Mitospóricos/aislamiento & purificación , Hojas de la Planta/microbiología , Raíces de Plantas/microbiología , Acremonium/aislamiento & purificación , Alternaria/aislamiento & purificación , Basidiomycota/aislamiento & purificación , Cladosporium/aislamiento & purificación , Fusarium/aislamiento & purificación , Penicillium/aislamiento & purificación , Trichoderma/aislamiento & purificación , UcraniaRESUMEN
We have developed diagnostic system for revealing 35S promoter CaMV and transgene CryIIIA in transgenic potato NL Russet Burbank and NL Superior companies Monsanto. Using the data about nucleotide sequences of 35S promoter (AF234316 GenBank) and gene CryIIIA (X70979 GenBank) oligonucleotides primers were synthesized for polymerase chain reaction (PCR). The technique of genomic DNA isolation from plant material, suitable for PCR has been optimized. Using Touchdown PCR method presence of specific transgene in a Bt-potato was shown. This method can be used in routine laboratory practice for revealing 35S promoter in genetically modified plants and for identification of specific transgene CryIIIA in the transformed Bt-plants.
Asunto(s)
Toxinas Bacterianas , Genes de Plantas , Plantas Modificadas Genéticamente/genética , Solanum tuberosum/genética , Toxinas de Bacillus thuringiensis , Proteínas Bacterianas/genética , Cartilla de ADN/biosíntesis , Endotoxinas/genética , Proteínas Hemolisinas , Virus del Mosaico/genética , Reacción en Cadena de la Polimerasa/métodos , Regiones Promotoras Genéticas , TransgenesRESUMEN
The virus gene CP AlMV cloned to the vector pBI121 (Clontech) was analyzed using the method of restriction analysis and polymerase chain reaction. The restriction analysis with the use of ApaI enzyme has shown presence of the specific fragment with a part of CP gene about 2.5 kb. The same procedure with EcoRI and HindIII enzymes has shown difference in electrophoretic migration of the formed fragments. Polymerase chain reaction analysis has shown the existence of the specific amplification product of the CP gene 666 bp long. Such investigations have proved successful cloning and presence of the CP AlMV gene in vector pBI121 and could afford to make following transformation of the higher plants with attaching genetic tolerance to the virus from Bromoviridae family.
Asunto(s)
Virus del Mosaico de la Alfalfa/genética , Cápside/genética , Genes Virales , Virus del Mosaico de la Alfalfa/enzimología , Clonación Molecular , Amplificación de Genes , Vectores Genéticos , Plantas Modificadas Genéticamente/genética , Mapeo RestrictivoRESUMEN
Virus tolerance of the three varieties of Bt-potato New Leaf--Atlantic, Russet Burbank and Superior from "Monsanto company" has been studied. Using serology, biochemistry methods and biotest the mixed virus infecting of experimental plants has been shown. Virus transmission by infected potato root crops under reproduction and accelerated degradation processes during storage have been registered.