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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2005.tde-18072005-152305
Document
Author
Full name
Flávia Mandolesi Pereira de Melo
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2005
Supervisor
Committee
Fiore, Marli de Fatima (President)
Kleiner, Aline Aparecida Pizzirani
Moraes, Luiz Alberto Beraldo de
Title in Portuguese
Atividade antifúngica de metabólitos secundários produzidos pelo endófito de mandioca Bacillus pumilus MAIIIM4a.
Keywords in Portuguese
antifúngico
bactéria endofítica
controle biológico (fitossanidade)
fungo fitopatogênico
mandioca
microbiologia
microscopia eletronônica de varredura
Abstract in Portuguese
Microrganismos endofíticos são definidos como organismos que habitam em pelo menos durante um período de seu ciclo vital, o interior de um vegetal, sem causar aparentemente nenhum dano a este. Na busca de novos organismos e novos metabólitos secundários, um estudo foi conduzido visando avaliar a diversidade química de bactérias endofíticas por meio do isolamento e identificação de metabólitos secundários produzido por bactérias endofíticas de etnovariedades de mandioca, mantidas por tribos indígenas da Amazônia brasileira. Sessenta e sete bactérias endofíticas de mandioca foram selecionadas e submetidas à uma seleção através de testes de antagonismo in vitro. A bactéria endofítica, Bacillus pumilus foi a que apresentou forte ação inibitória contra os fitopatógenos Rhizoctonia solani, Pythium e Sclerotium rolfsii. Essa bactéria foi identificada através do sequenciamento de um fragmento do gene de 16S rRNA e por meio da análise de ácidos graxos (FAME). A obtenção dos metabólitos da bactéria selecionada foi realizada através da extração dos meios de cultura com os solventes hexano, acetato de etila e diclorometano. Após a concentração dos extratos, seus constituintes químicos foram realizados através de cromatografia em camada delgada, cromatografia em coluna, cromatografia gasosa acoplada ao espectrômetro de massas CG-EM) e cromatografia líquida acoplada ao espectrômetro de massas (CL-EM). Os extratos obtidos por meio de diclorometano e acetato de etila continham diferentes componentes químicos que mostraram atividade inibitória contra os três fitopatógenos testados. O método cromatográfico CL-EM permitiu a identificação de uma substância antifúngica produzida pela bactéria endofítica, Bacillus pumilus, conhecida como pumilacidina.
Title in English
Antifungal activity of secondary metabolites produced by Bacillus pumilus MAIIIM4a, an endophytic from cassava.
Keywords in English
antifungal
biological control (phytosanity)
cassava
endophitic bacterium
microbiology
phytopathogenic fungi
scanning electron microscope
Abstract in English
Endophytic microorganisms are defined as organisms that inhabit the interior of a vegetable at least during a period of the vital cycle, without causing any apparent damage. In the search for new organisms and new secondary metabolites, a study was conducted to evaluate the chemical diversity of endophytic bacteria through the isolation and identification of secondary metabolites produced by endophytic bacteria of cassava cultivated by Brazilian Amazon Indian tribes. Sixty seven endophytic bacteria isolated from cassava were screened using in vitro antagonisms tests. An endophytic bacterium, the Bacillus pumilus, which showed a strong inhibitory activity against Rhizoctonia solani, Pythium aphanidermatum and Sclerotium rolfsii was selected. This bacterium was identified by sequencing of 16S rRNA genes and by FAME. The bacterial endophytic localization was confirmed by cassava cell tissue examination using scanning electron microscopy. The bacterial metabolites were extracted from the culture media using the solvents hexan, ethyl acetate and dichloromethane. After concentrate the extracts their chemical constituents were analyzed using thin layer chromatography (TLC), chromatography column, gas chromatography coupled with mass spectrometry (GC/MS) and liquid chromatography coupled with mass spectrometry (LC/MS). The extracts obtained with dichloromethane and ethyl acetate contained different chemical compounds that showed inhibitory activity against the three plant-pathogenic fungi tested. The LC/MS method allowed the identification of an antifungal compound produced by the B. pumilus, which is known as pumilacidin.
 
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FlaviaMelo.pdf (3.29 Mbytes)
Publishing Date
2005-08-08
 
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