RESUMO
Identification of nucleotide sequence of the gene 16S rRNA of cyanobacteria Anabaena variabilis str.21 (Uzb1) exhibits that the strain from root sphere of cotton plants is 99% homologous to a known strain of Anabaena variabilis (EF488831.1). This data confirms the phylogenetic relationship of the strain of cyanobacteria A. variabilis Uzb1 to other described representatives of genus Anabaena as addition to morphological characteristics (presence of akinets (spores) and heterocysts).
Assuntos
Anabaena variabilis/genética , Anabaena variabilis/isolamento & purificação , Genes Bacterianos , RNA Ribossômico 16S/genética , Esporos Bacterianos/genética , Anabaena variabilis/classificação , Sequência de Bases , Gossypium/microbiologia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/classificação , RNA Ribossômico 16S/isolamento & purificação , Rizosfera , Esporos Bacterianos/classificação , Esporos Bacterianos/isolamento & purificaçãoRESUMO
The effect of Acholeplasma laidlawii var. granulum 118 on activity of phenylalanine-ammonia-lyase (PAL) in callus cultures of sugar beat was researched. The optimal conditions of enzyme reaction were: using the L-phenilalanine as a substrate, pH 8.4-8.8, the temperature optimum 38-40 degrees C. It was established that at the infecting of sugar beat callus culture by phytopathogenic mollicute the PAL activity was temporarily increased and reached its maximum after 2 h of infecting. Then it gradually decreased and in 24 h reached its initial level. An increase of PAL activity of plant is considered as protective reaction in response to the action of pathogen.
Assuntos
Acholeplasma laidlawii/crescimento & desenvolvimento , Beta vulgaris/enzimologia , Fenilalanina Amônia-Liase/metabolismo , Fenilalanina/metabolismo , Proteínas de Plantas/metabolismo , Beta vulgaris/imunologia , Beta vulgaris/microbiologia , Concentração de Íons de Hidrogênio , Imunidade Vegetal/fisiologia , Estereoisomerismo , Especificidade por Substrato , Temperatura , Técnicas de Cultura de TecidosRESUMO
The dynamics of lectin activity of sugar beet calluses infected by mollicute Acholeplasma laidlawii var granulum str. 118 was studied. Activity of acid-soluble lectins of sugar beet cell cultures increased 4.5 times during some first hours after infecting by acholeplasma. At early stages of infecting by mollicute qualitative changes were also revealed in the protein spectra of acid-soluble lectins of sugar beet calluses.
Assuntos
Acholeplasma laidlawii/crescimento & desenvolvimento , Beta vulgaris/metabolismo , Beta vulgaris/microbiologia , Lectinas de Plantas/metabolismo , Beta vulgaris/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controleRESUMO
Effect of oligodesoxyribonucleotide, complementary to 3'-end sequence (3'-UCUUUCCUCCAC) of 16S-rRNA of mollicutes, and its phenasine derivatives on the process of translation of mollicute has been studied in the system of in vitro translation, created on the basis of ribosomes of Acholeplasma laidlawii var. granulum str. 118 and germ extracts of the wheat and optimized in respect of the temperature (23 degrees C), translation time (70 min), concentration of potassium and magnesium ions (150 and 5 mM, respectively). It is shown that acholeplasma ribosomes are most efficiently inhibited (60%) under their interaction with oligonucleotide containing one phenasine insert on the 3'-end, nonmodified oligonucleotide exerted a bit less inhibiting effect (58%). Oligonucleotide containing intercalating inserts in 3'- and 5'-positions manifested the least inhibiting effect (35%). It is noted that the efficiency translation inhibition by synthetic oligonucleotides is conditioned by nuclease activity of the system and by the length of the section of active binding of oligonucleotide with the sequence target on rRNA. It is supposed that oligonucleotides complementary to certain unique rRNA sequences can become promising highly specific drugs for prophylaxis and treatment of mycoplasmoses.
Assuntos
Oligodesoxirribonucleotídeos/metabolismo , Biossíntese de Proteínas/fisiologia , RNA Ribossômico 16S/metabolismo , Ribossomos/metabolismo , Tenericutes/metabolismo , Meios de Cultura , Relação Dose-Resposta a Droga , Magnésio/farmacologia , Potássio/farmacologia , Biossíntese de Proteínas/efeitos dos fármacos , RNA Ribossômico 16S/efeitos dos fármacos , Ribossomos/efeitos dos fármacos , Temperatura , Tenericutes/efeitos dos fármacos , Fatores de Tempo , TriticumRESUMO
Active cell-free protein-synthesizing systems are created on the basis of ribosomes of Acholeplasma laidlawii var. granulum str. 118, A. laidlawii PG-8 and active lysates of eucaryotic cells (lysate of rabbit reticulocytes and wheat germ extract) in which activity of 80S-ribosomes was suppressed by addition of actidione in concentrations of 150 and 200 mkg/ml, respectively. It is shown that preincubation of active lysates of eucaryotic cells with actidione does not influence their ribosome inhibition level. It was found that the system of in vitro translation on the basis of wheat germ extract was more specific for ribosomes of phytopathogenic acholeplasmas (str. 118), while ribosomes of the saprophytic strain (PG-8) revealed no such specificity with respect to cell-free translation systems.
Assuntos
Acholeplasma laidlawii/metabolismo , Biossíntese de Proteínas , Ribossomos/metabolismo , Acholeplasma laidlawii/efeitos dos fármacos , Fracionamento Celular , Cicloeximida/farmacologia , Depressão Química , Relação Dose-Resposta a Droga , Biossíntese de Proteínas/efeitos dos fármacos , Ribossomos/efeitos dos fármacosRESUMO
6-Azacytidine (6-AC) is shown to have an inhibitory effect on the Mollicutes of the different systematic position. The growth of type strains of Mollicutes (Acholeplasma laidlawii PG-8, Mycoplasma pneumoniae FH and M. fermentans PG-18) completely ceased in the nutrient medium at concentration of the above substance in it within the range of 125-250 micrograms/ml. 50% inhibiting concentration of 6-AC equaled: for M. fermentans PG-8: 23.43 micrograms/ml; M. pneumoniae FN: 46.8 micrograms/ml; Acholeplasma laidlawii PG-8: 62.5 micrograms/ml. 6-AC concentration 5 micrograms/ml decreased the process of DNA-dependent DNA synthesis in the in vitro system more than by 60%. 6-AC exerted less effect on the DNA-dependent RNA synthesis in the in vitro system: at different concentrations of 6-AC (up to 400 micrograms/ml) RNA synthesis decreased only by 20%. Translation on ribosomes of Mollicutes in the in vitro system completely ceased at 6-AC concentration 100 micrograms/ml. The results obtained indicate that for 6-AC in cells of Mollicutes and, possibly, for other microorganisms there are two targets: ribosomes and DNA-dependent DNA-polymerase. Total effect of blocking of the translation and replication processes by 6-azacytidine causes death of Mollicutes. Since 6-AC has no harmful effect on the human cells, it can be used as an efficient method for treatment of respiratory and urogenital diseases induced by Mollicutes.
Assuntos
Azacitidina/análogos & derivados , Tenericutes/efeitos dos fármacos , Acholeplasma laidlawii/efeitos dos fármacos , Acholeplasma laidlawii/enzimologia , Acholeplasma laidlawii/genética , Azacitidina/farmacologia , DNA Polimerase Dirigida por DNA/efeitos dos fármacos , Depressão Química , Relação Dose-Resposta a Droga , Mycoplasma fermentans/efeitos dos fármacos , Mycoplasma fermentans/enzimologia , Mycoplasma fermentans/genética , Mycoplasma pneumoniae/efeitos dos fármacos , Mycoplasma pneumoniae/enzimologia , Mycoplasma pneumoniae/genética , Biossíntese de Proteínas/efeitos dos fármacos , Tenericutes/enzimologia , Tenericutes/genética , Transcrição Gênica/efeitos dos fármacosRESUMO
Ribosomes are produced from Acholeplasma laidlawii PG-8 and A. laidlawii var. granulum str. 118. It is proved as possible to use the standard method of differential centrifugation for isolation of mollicute ribosomes. The physico-chemical properties of acholesplasma ribosomes are studied. The protein content for A. laidlawii PG-8 amounted to 39.6%, rRNA content--60.4% and for A. laidlawii var. granulum--39.1 and 60.8%, respectively. The RNA: protein ratio equals 1.5:1, the ratio of optic density indicates at 260 and 280 nm--2, that is peculiar to treated preparations of ribosomes. Sedimentation coefficient of acholeplasma ribosomes is 70S. The produced preparations of acholeplasma ribosomes can be used subsequently for creating the system of translation in vitro to study the biosynthesis processes of mollicute protein.