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Doctoral Thesis
DOI
10.11606/T.46.1993.tde-15102012-160938
Document
Author
Full name
Isaac Stringueta Machado
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 1993
Supervisor
Committee
Crocomo, Otto Jesu (President)
Gonçalves, Antonio Natal
Maia, José Carlos da Costa
Rodrigues, Joao Domingos
Rossi, Celso
Title in Portuguese
Atividade de enzimas do metabolismo de compostos secundários comprometidos com o enraizamento "in vitro" de Eucalyptus grandis HILL ex MAIDEN
Keywords in Portuguese
Bioquímica
Biotecnologia
Compostos secundários
Enraizamento \"in vitro\"
Enzimas
Eucalyptus grandis
Micropropagação
Abstract in Portuguese
O trabalho realizado constitui uma estratégia em Biotecnologia de Plantas para a investigação dos esquemas bioquímicos controladores da diferenciação celular no processo de enraizamento. Através da técnica da cultura de tecidos "in vitro" foram micropropagados clones de Eucalyptus grandis HILL ex MAIDEN (GO 250, GO 682 e GO 669) selecionadas no CEBTEC/USP. Em uma pesquisa preliminar foram estabelecidas e multiplicadas gemas, seguida de testes da composição de meios de cultura para elongação e enraizamento. Foram estudadas diversas possibilidades de balanceamento das substâncias reguladoras de crescimento - ácido indolilbutírico (AIB)/ benzilaminopurina (BAP). Os clones apresentaram comportamentos anatômico-fisiológicos (massa fresca e seca; teor protéico; taxas de multiplicação das brotações epicórmicas, elongação e enraizamento) diferenciados entre si. As frequências de enraizamento mostraram-se superiores às já observadas por outros autores. Foram estabelecidas, ainda, algumas alterações na composição dos meios de cultura empregados. Através de experimentos cinéticos foram determinados os padrões de indução e/ou inibição das enzimas fenilalanina amônia-liase (PAL) e glucose 6-fosfato-desidrogenase (G-6P-DH) importantes para a biossíntese de compostos flavonóides, considerados como os principais cofatores das auxinas endógenas no complexo iniciador do enraizamento. Verificou-se que embora o suprimento exógeno crescente de AIB tenha resultado em maiores taxas de enraizamento, as atividades da PAL e G-6P-DH não se mostraram significativamente alteradas. Contudo, foi observado também que as gemas que enraizaram apresentaram atividade da PAL significativamente superior, evidenciando maior fluxo do Metabolismo do Fenilpropano durante o enraizamento.
Title in English
Enzyme activity of metabolism of secondary compounds related to in vitro rooting of Eucalyptus grandis HILL ex MAIDEN
Keywords in English
Biochemistry
Biotechnology
Enzyme
Eucalyptus grandis
in vitro rooting
Micropropagation
Secondary compounds
Abstract in English
This study constitutes a strategy in Plant Biotechnology with the purpose to investigate the regulating biochemical development of the cellular differentiation in the rooting process. Through plant tissue culture "in vitro" technique, clones of Eucalyptus grandis Hill ex Maiden (GO 250, GO 682 e GO 669) selected at CEBTEC/USP, were micropropagated. In a preliminary study shoots were multiplied, followed by tests of media culture composition for shoot elongation and root initiation. Several possibilities and rates of plant growth regulators, indolilbutyric acid (IBA)/benzylaminopurine (BAP) were studied. The clones showed different anatomic-physiological behaviours (fresh and oven-dry weight; proteic content; epicormic shoots multiplication rates, elongation and rooting). The frequences of rooting were higher than those described by other authors. Some modifications in the composition of used cuIture media were also defined. Through kinetic experiments, induction and inhibition patterns of phenylalanine ammonia-lyase (PAL) and glucose 6- phosphate-dehydrogenase (G-6P-DH) important to the biossyntesis of flavonoid compounds were determined. They were considered as the main cofactors of endogenous auxins in the root stimulating complex. Although, the increasing exogenous supply of AIB resulted in higher rooting rates, the activities of PAl and G-6P-DH enzyme did not present alterations. However, it was also observed that shoots wich rooted, in the treatments, showed higher PAL activity. resulting in higher flux of phenylpropanoid metabolism during the rooting.
 
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Publishing Date
2012-10-16
 
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