Caracterização isoenzimática de clones e somaclones de cana-de-açúcar

Sawazaki, Haiko Enok; Silvarolla, Maria Bernadete; Alvarez, Raphael

doi:10.1590/S0006-87051989000100001

Resumos

Foram caracterizados os padrões de isoenzimas de peroxidase e esterase de extrato de folha e raiz de cana-de-açúcar de variedade e somaclones de NA56-79 aos 7 meses de idade e de variedades e somaclones de NA56-79, IAC68-12 e IAC68-144 aos 9 meses. Avaliaram-se também as atividades específicas de peroxidase com relação ao tempo e perfil isoenzimático das três variedades e respectivos somaclones. Os zimogramas de peroxidase e esterase de folhas mostraram diferenças distintas entre as três variedades; a esterase apresentou menor número de bandas em relação à peroxidase, porém de mais fácil caracterização. Os zimogramas de raízes, devido à grande variação de atividade enzimática, em conseqüência de raízes velhas, não foram conclusivos quanto à identificação das variedades. Não ocorreram diferenças nos zimogramas de peroxidase ou esterase entre somaclones da mesma variedade. NA56-79 apresentou menor número de isoenzimas e menor atividade especifica de peroxidase em relação à IAC68-144, sugerindo que a diminuição das isoenzimas seja associada ao decréscimo de atividade enzimática. A atividade especifica de peroxidase variou com a maturação, sendo a 9 meses maior do que a 7.

cana-de-açúcar; isoenzimas; variedades; somaclones

Extracts from leaves and roots of three sugarcane varieties and their somaclones: NA56-79 (at seven and nine month old plants), IAC68-12 and IAC68-144 (both at nine month old plants) were obtained. The peroxidase and esterase isoenzyme contents of the extracts were determined to verify the genetic variability of tissue culture somaclones. The peroxidase specific activity was studied in relation to the time and isoenzymatic pattern. The peroxidase and esterase zymograms showed distinct differences among varieties, and the esterase presented less bands but easier to characterize than the peroxidase bands. The root zymograms exhibited great enzymatic variation due to the presence of old roots thus not allowing to characterize the varieties. No differences in peroxidase or esterase zymograms among somaclones of the same variety were noticed. The NA56-79 presented lower specific activity of peroxidase and number of isoenzymes than IAC68-144, suggesting that the decrease of isoenzyme bands is associated to the reduction of enzymatic activity. The specific activity of peroxidase changed with maturity and was higher at nine than at seven month old plants.

sugarcane; isoenzymes; varieties; somaclones

I. BIOTECNOLOGIA E FITOQUÍMICA

Caracterização isoenzimática de clones e somaclones de cana-de-açúcar

Isoenzymatic characterization of sugarcane clones and somaclones

Haiko Enok SawazakiI, ^* * Com bolsa de pesquisa do CNPq. ; Maria Bernadete Silvarolla^II; Raphael Alvarez^III

^ISeção de Fitoquímica, Instituto Agronômico (IAC), Caixa Postal 28, 13001 Campinas, SP

^IISeção de Genética, IAC

IIISeção de Cana-de-Açúcar, IAC

RESUMO

Foram caracterizados os padrões de isoenzimas de peroxidase e esterase de extrato de folha e raiz de cana-de-açúcar de variedade e somaclones de NA56-79 aos 7 meses de idade e de variedades e somaclones de NA56-79, IAC68-12 e IAC68-144 aos 9 meses. Avaliaram-se também as atividades específicas de peroxidase com relação ao tempo e perfil isoenzimático das três variedades e respectivos somaclones. Os zimogramas de peroxidase e esterase de folhas mostraram diferenças distintas entre as três variedades; a esterase apresentou menor número de bandas em relação à peroxidase, porém de mais fácil caracterização. Os zimogramas de raízes, devido à grande variação de atividade enzimática, em conseqüência de raízes velhas, não foram conclusivos quanto à identificação das variedades. Não ocorreram diferenças nos zimogramas de peroxidase ou esterase entre somaclones da mesma variedade. NA56-79 apresentou menor número de isoenzimas e menor atividade especifica de peroxidase em relação à IAC68-144, sugerindo que a diminuição das isoenzimas seja associada ao decréscimo de atividade enzimática. A atividade especifica de peroxidase variou com a maturação, sendo a 9 meses maior do que a 7.

Termos de indexação: cana-de-açúcar, isoenzimas, variedades, somaclones.

SUMMARY

Extracts from leaves and roots of three sugarcane varieties and their somaclones: NA56-79 (at seven and nine month old plants), IAC68-12 and IAC68-144 (both at nine month old plants) were obtained. The peroxidase and esterase isoenzyme contents of the extracts were determined to verify the genetic variability of tissue culture somaclones. The peroxidase specific activity was studied in relation to the time and isoenzymatic pattern. The peroxidase and esterase zymograms showed distinct differences among varieties, and the esterase presented less bands but easier to characterize than the peroxidase bands. The root zymograms exhibited great enzymatic variation due to the presence of old roots thus not allowing to characterize the varieties. No differences in peroxidase or esterase zymograms among somaclones of the same variety were noticed. The NA56-79 presented lower specific activity of peroxidase and number of isoenzymes than IAC68-144, suggesting that the decrease of isoenzyme bands is associated to the reduction of enzymatic activity. The specific activity of peroxidase changed with maturity and was higher at nine than at seven month old plants.

Index terms: sugarcane, isoenzymes, varieties, somaclones.

Texto completo disponível apenas em PDF.

Full text available only in PDF format.

Recebido para publicação em 23 de junho de 1988 e aceito em 6 de abril de 1989.

DILLEWIJN, C. van. Botany of sugarcane. Walthan, Mass., Chronica Botanica, 1952.371 p.
FERNANDEZ, A.M.F.; ANTONI, H.J. & CANELADA, M.E.L. Estudios sobre variabilidad genetica de isoperoxidasas y caracteres morfológicos en subclones de cana de azucar obtenidos mediante cultivos "in vitro". Revista Agronomica del Noroeste Argentino, 12:79-91,1975.
GONÇALVES, C.H.R.P.; CURY, J.A. & CROCOMO, O.J. Isoenzimas na identificação e seleção de variedades de cana-de-açúcar (Saccharum spp). In: CONGRESSO NACIONAL DA SOCIEDADE DE TÉCNICOS AÇUCAREIROS DO BRASIL, 2., Rio de Janeiro, 1981. Anais. Rio de Janeiro, STAB, 1981. p.329-340.
HEINZ, D.J. & MEE, G.W.P. Morphologic, cytogenetic and enzymatic variation in Saccharum species hybrid clones derived from callus tissue. American Journal Botany, 58(3):237-262,1971.
LIU, Y.T.; CHEN, S.S.; LEE, S. & LEE, H.C. Detection of isoenzymes in sugarcane leaves by disc electrophoresis. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 15., Durban, 1974. Proceedings, p.1048-1057.
LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the Folin-Phenol reagent. Journal Biological of the Chemistry, 193:265, 1951.
MAEHLY, A.C. & CHANCE, B. The assay of catalase and peroxidases: methods of biochemical analysis. New York, Interscience Publishers, 1954. 357p.
MARIBONA, R.H.; KORNEVA, S.B.; RUIZ, A. & GONZALEZ, S. Obtention of sugarcane plants by tissue culture from different plant organs. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 18., Havana, 1983.Proceedings. Havana, Executive Commitee of the ISSCT, 1983. p.610-621.
MARKERT, C.L. & MOLLER, F. Multiple forms of enzymes: tissue ontogenetic, and species specific patterns. Proceedings of the National Academy of Sciences, 45:753-763, 1959.
RUIZ, A. & MARIBONA, R.H. Peroxidase isoenzymes analysis: a massive method for the identification of sugarcane varieties. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 18., Havana, 1983. Proceedings. Havana, Executive Commitee of the ISSCT, 1983. p.437-442.
RUDOLPH, K. & STAHMANN, M.A. Multiple hydrolases in bean leaves (Phaseolus vulgaris L.) and the effect of the halo blight disease caused by Pseudomonas phaseolicola (Burkl) Dowson. Plant Physiology, 44:389-394, 1966.
SCANDALIOS, J.G. Isozymes in development and differentiation. Annual Review of Plant Physiology, 25:225-258, 1974.
SCHWARTZ, D. Genetic studies on mutant enzymes in maize. V. In vitro interconversion of allelic isozymes. Proceedings of the National Academy of Science, 52:222-226, 1964.
SONDAHL, M.R. & CARLUCCI, M.V. Variabilidade somaclonal em cultura de tecidos de cana-de-açúcar. Ciência e Cultura, 35(7):689, 1983. (Suplemento)
WALDRON, J.C. & GLASZIOU, K.T. Isoenzymes as a method of varietal identification in sugarcane. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 14., New Orleans, 1971. Proceedings. Baton Rouge, Executive Commitee of the ISSCT, 1972. p.249-256.

*

Com bolsa de pesquisa do CNPq.

Datas de Publicação

Publicação nesta coleção
29 Nov 2007
Data do Fascículo
1989

Histórico

Aceito
06 Abr 1989
Recebido
23 Jun 1988

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] DILLEWIJN, C. van. Botany of sugarcane. Walthan, Mass., Chronica Botanica, 1952.371 p.

[2] FERNANDEZ, A.M.F.; ANTONI, H.J. & CANELADA, M.E.L. Estudios sobre variabilidad genetica de isoperoxidasas y caracteres morfológicos en subclones de cana de azucar obtenidos mediante cultivos "in vitro". Revista Agronomica del Noroeste Argentino, 12:79-91,1975.

[3] GONÇALVES, C.H.R.P.; CURY, J.A. & CROCOMO, O.J. Isoenzimas na identificação e seleção de variedades de cana-de-açúcar (Saccharum spp). In: CONGRESSO NACIONAL DA SOCIEDADE DE TÉCNICOS AÇUCAREIROS DO BRASIL, 2., Rio de Janeiro, 1981. Anais. Rio de Janeiro, STAB, 1981. p.329-340.

[4] HEINZ, D.J. & MEE, G.W.P. Morphologic, cytogenetic and enzymatic variation in Saccharum species hybrid clones derived from callus tissue. American Journal Botany, 58(3):237-262,1971.

[5] LIU, Y.T.; CHEN, S.S.; LEE, S. & LEE, H.C. Detection of isoenzymes in sugarcane leaves by disc electrophoresis. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 15., Durban, 1974. Proceedings, p.1048-1057.

[6] LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the Folin-Phenol reagent. Journal Biological of the Chemistry, 193:265, 1951.

[7] MAEHLY, A.C. & CHANCE, B. The assay of catalase and peroxidases: methods of biochemical analysis. New York, Interscience Publishers, 1954. 357p.

[8] MARIBONA, R.H.; KORNEVA, S.B.; RUIZ, A. & GONZALEZ, S. Obtention of sugarcane plants by tissue culture from different plant organs. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 18., Havana, 1983.Proceedings. Havana, Executive Commitee of the ISSCT, 1983. p.610-621.

[9] MARKERT, C.L. & MOLLER, F. Multiple forms of enzymes: tissue ontogenetic, and species specific patterns. Proceedings of the National Academy of Sciences, 45:753-763, 1959.

[10] RUIZ, A. & MARIBONA, R.H. Peroxidase isoenzymes analysis: a massive method for the identification of sugarcane varieties. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 18., Havana, 1983. Proceedings. Havana, Executive Commitee of the ISSCT, 1983. p.437-442.

[11] RUDOLPH, K. & STAHMANN, M.A. Multiple hydrolases in bean leaves (Phaseolus vulgaris L.) and the effect of the halo blight disease caused by Pseudomonas phaseolicola (Burkl) Dowson. Plant Physiology, 44:389-394, 1966.

[12] SCANDALIOS, J.G. Isozymes in development and differentiation. Annual Review of Plant Physiology, 25:225-258, 1974.

[13] SCHWARTZ, D. Genetic studies on mutant enzymes in maize. V. In vitro interconversion of allelic isozymes. Proceedings of the National Academy of Science, 52:222-226, 1964.

[14] SONDAHL, M.R. & CARLUCCI, M.V. Variabilidade somaclonal em cultura de tecidos de cana-de-açúcar. Ciência e Cultura, 35(7):689, 1983. (Suplemento)

[15] WALDRON, J.C. & GLASZIOU, K.T. Isoenzymes as a method of varietal identification in sugarcane. In: CONGRESS OF THE INTERNATIONAL SOCIETY OF SUGARCANE TECHNOLOGISTS, 14., New Orleans, 1971. Proceedings. Baton Rouge, Executive Commitee of the ISSCT, 1972. p.249-256.

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Caracterização isoenzimática de clones e somaclones de cana-de-açúcar

Resumos

Datas de Publicação

Histórico