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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.1982.tde-20240301-153647
Document
Author
Full name
Ilza Maria Sittolin
Institute/School/College
Date of Defense
Published
Piracicaba, 1982
Supervisor
Title in Portuguese
Interação fago-Xanthomonas campestris (Pammel) Dowson a nível de microscopia eletrônica, efeito do virazole e estudo radioautográfico da ação do fago sobre o hospedeiro
Keywords in Portuguese
ANTIVIRAIS
AUTORRADIOGRAFIA
BACTÉRIAS FITOPATOGÊNICAS
BACTERIÓFAGOS
MICROSCOPIA ELETRÔNICA
REPOLHO
Abstract in Portuguese
Na presente Dissertação foi isolado um bacteriófago de plantas de repolho infectadas com Xanthomonas campestris, provenientes da região de Botucatu-SP. O extrato purificado apresentou um título de 1010 partículas viáveis/ml e produziu placas de lise de 1 mm de diâmetro. O bacteriófago apresentou as seguintes características ao microscópio eletrônico: cabeça hexagonal com 82 nm de comprimento por 71 nm de largura, cauda com 112 nm de comprimento por 20 nm de largura e placa terminal com 45 nm de comprimento por 17 nm de altura. Devido a estas características o fago foi classificado no sistema LHT, como pertencente à Divisão Deoxyvira, Classe Deoxybinala, Ordem Urovirales e Família Phagoviridae e ao Grupo A na classificação de Bradley sendo denominado de Xac-CENA. O processo de infecção foi estudado através das técnicas de coloração negativa (PTA) e secções ultrafinas, permitindo distinguir as fases de adsorção, período latente, lise e liberação dos fagos. O exame das radioautografias mostrou que ocorreu um máximo de incorporação da timidina tritiada pelas bactérias infectadas com fago após 5 minutos de tratamento, indicando que o fago apresenta razoável domínio sobre o genoma da bactéria hospedeira desde o início do tratamento. O efeito do virazole sobre o processo de infecção do fago à bactéria, pode estar mostrando uma ação inibidora e competitiva sobre a enzima inosina 5’-fosfato desidrogenase no sistema fago-bactéria, tal como tem sido relatado em E. coli· As reações serológicas indicaram que ocorreu a indução de anticorpos específicos para o fago, como se pode ver através dos testes de dupla difusão em agar e de precipitação em tubos. O material obtido no teste de precipitação foi examinado ao microscópio eletrônico, observando-se agregados de partículas apenas entre as reações fago e anti-soro homólogo.
Title in English
Interaction of the phage-Xanthomonas campestris (Pammel) Dowson at the electronic microscopy level, virazole effect and radioautographic study of the phage action on the host
Abstract in English
A bacteriphage from the cabbage tissue infected with Xanthomonas campestris coming from the region of Botucatu-SP was isolated in the present work. The purified extract presented a title of 1010 viable particles/ml and produced clear plaques of 1 mm diameter. Electron microscopy revealed the following characteristics of the bacteriophage: hexagonal head with a length of 82 nm and a width of 71 nm, contractible tail with a length of 112 nm and a width of 20 nm. Terminal plaque with a length of 45 nm a height of 17 nm. Due to these characteristics the phage was classified, in the LHT system, as belonging to Deoxyvira Division, Deoxybinal Class, Urovirales Order and Phagoviridae Family and to the group A in the Bradley classification. We named it Xac-CENA. The infection process was studied through a negative staining technique (PTA) and ultrathin section, permitting to distinguish the phases of adsorption, latent period, lysis and release of the phages. Radioautography showed that a maximum incorporation of the tritiated thymidine in infected bacteria with the phage occurred after 5 minutes indicating that the phage presented a reasonable domain on the bacterial host genome since the beginning of the treatment. The effect of virazole on the infection process of the phage present in the bacteria, may show an inhibitive and competitive action of the inosine 5’-phosphate dehydrogenase enzyme of the phage-bacterium system, such as gas been reported in E. coli. Serological reactions indicated that induction of specific antibodies for the phage, occurred as shown by the diffusion tests in agar and precipitation tests in test tubes. The material obtained in the precipitation tests was examined by electronic microscopy. Aggregated virus particles were observed only in the reactions between the phage and homologous anti-serum.
 
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Publishing Date
2024-03-14
 
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