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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2019.tde-20191108-122826
Document
Author
Full name
Cantídio Fernando Gouvêa
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2001
Supervisor
Title in Portuguese
Avaliação enzimática do envelhecimento durante o crescimento de brotações de Eucalyptus spp. "in vitro"
Keywords in Portuguese
ATIVIDADE ENZIMÁTICA
BROTAÇÃO
CRESCIMENTO
ENVELHECIMENTO
EUCALIPTO
MICROPROPAGAÇÃO VEGETAL
Abstract in Portuguese
O gênero Eucalyptus é de grande importância para a silvicultura brasileira, principalmente as espécies Eucalyptus grandis e Eucalyptus urophylla e seus híbridos, que são plantados em extensas áreas e sob diferentes condições edafoclimáticas. Programas de melhoramento genético têm selecionado materiais superiores que são propagados vegetativamente, sendo a cultura de tecidos uma das técnicas de propagação utilizadas. O cultivo "in vitro" do eucalipto permite promover o seu rejuvenescimento, maximizar a sua propagação, e possibilita o estudo dos fenômenos fisiológicos relacionados à ontogenia, heterocronia e necessidades nutricionais. Assim, pesquisas sobre regulação gênica e sobre as atividades de enzimas ligadas a passos metabólicos essenciais são importantes para o processo produtivo. O presente trabalho teve por objetivo monitorar a variação do número de formas alélicas e das atividades de enzimas ligadas ao envelhecimento durante um ciclo de crescimento de brotações de Eucalyptus spp "in vitro". Dez clones foram estudados por um período de 55 dias, numa única subcultura, com amostras coletadas a cada 7 dias, utilizando-se um delineamento experimental inteiramente casualizado. A eletroforese de isoenzimas utilizada no estudo foi realizada em gel de amido a 13% (p/v), sendo avaliadas as seguintes isoenzimas: fosfatase ácida, peroxidase, glucose-6-fosfato desidrogenase, 6-fosfogluconato desidrogenase, leucina aminopeptidase, malato desidrogenase, isocitrato desidrogenase, xiquimato desidrogenase e fosfoglucose isomerase. Os géis foram digitalizados após coloração e analisados posteriormente, sendo avaliado o número e densidade das bandas em cada amostra. As curvas de crescimento dos clones e os tempos de máximo acúmulo de matéria seca foram determinados para o monitoramento do crescimento dos clones. Estes resultados mostraram um padrão diferenciado de crescimento entre os clones e uma variação no acúmulo de matéria seca. As atividades enzimáticas medidas pela densitometria de bandas, mostrou grande amplitude durante o crescimento, mas não foi observada variação quanto ao número de bandas, exceto para a enzima peroxidase
Title in English
Enzymatic evaluation of the aging of Eucalyptus spp. clone shoot growth in vitro
Abstract in English
The genus Eucalyptus is of major importance to Brazilian commercial forestry, specially Eucalyptus grandis, E. urophylla and their hybrids. Large plantations are found in a range of soil and climate conditions. Breeding programs have generated superior genotypes that are propagated vegetatively. Tissue culture is one of the techniques of choice. In vitro culture of Eucalyptus allows juvenile characteristics recovery,maximization of propagation rates and a better understanding of physiological phenomena related to ontogeny, heterochrony and nutritional demands. Under this approach, studies on gene regulation and on the activity of enzymes related to essential metabolic pathways are important to the productive process. This work aimed the accessment of the number of allelic forms and the activity of enzymes related to different metabolic steps, during an in vitro growth cycle. Ten Eucalyptus clones were studied over a period of 55 days of culture. Samples were collected every 7 days. Growth curves were determined for all clones. The following isoenzyme systems were analysed: acid phosphatase, peroxidase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, leucyl amino peptidase, malate dehydrogenase, isocitrate dehydrogenase, shikimic acid dehydrogenase and phosphoglucoisomerase. Electrophoresis were undertaken using 13% (w/v) starch gels. A positive control was used in all gels. Gel images were digitalyzed after staining and than analysed. Each sample was evaluated concerning the number and density of bands. The results showed a differentiated growth pattern among the clones, specially the time of maximum dry matter accumulation. The enzymatic activity measured by band densitometry has shown a large amplitude of variation during the culture period analysed, but there was no variation conceming the number of bands, except to peroxidase enzyme
 
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Publishing Date
2019-11-08
 
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