Isoenzimas na identificação precoce de híbridos e clones nucelares no melhoramento de citros

Ballve, Rosa Maria Lizana; Bordignon, Rita; Medina Filho, Herculano Penna; Siqueira, Walter Jose; Teófilo Sobrinho, Joaquim; Rompeu Júnior, Jorgino

doi:10.1590/S0006-87051991000100007

Resumos

Um dos problemas relacionados ao melhoramento de citros é a poliembrionia e a conseqüente dificuldade, após cruzamentos controlados, de distinguir clones nucelares dos híbridos. Estudou-se um método para eletroforese de isoenzimas em gel de amido para doze sistemas isoenzímicos, desenvolvendo-se procedimento apropriado para a resolução simultânea dos sistemas fosfatase ácida, peroxidase anódica e catódica, glutamato oxaloacético transaminase, fosfoglucoisomerase, fosfoglucomutase e enzima málica. Caracterizaram-se por esses sistemas 31 clones de citros com possível interesse para o programa de melhoramento do IAC. Estão envolvidos 7 locos e 25 alelos, permitindo uma identificação precoce de híbridos antes do plantio no campo para avaliações agronômicas. Implicações desses estudos básicos no melhoramento e na determinação da taxa de seedlings nucelares de porta-enxertos comerciais são discutidas.

citros; genética; melhoramento; isoenzimas; híbridos; clones nucelares; embriões zigóticos

One of the problems related to citrus breeding is the polyembryony and the ensuing difficulty in distinguishing nucellar clones from hybrids obtained from controlled crosses. Twelve isozyme systems using starch gel electrophoresis were studied. A procedure for simultaneous resolution of acid phosphatase, anodic and cathodic peroxidase, glutamate oxaloacetate transaminase, phosphoglucoisomerase, phosphoglucomutase and malic enzyme was developed. Thirty one clones with possible use in the Instituto Agronômico de Campinas, State of São Paulo, Brazil breeding program were characterized. Seven loci and twenty five alleles are involved allowing for an early identification of hybrids before transplanting them into the field for agronomic evaluations. Implications in breeding strategies and in the determination of the degree of nucellar embryony of commercial rootstocks are discussed.

citrus; genetics; breeding; isozymes; hybrids; nucellar clones; zygotic embryos

II. GENÉTICA E MELHORAMENTO DE PLANTAS

Isoenzimas na identificação precoce de híbridos e clones nucelares no melhoramento de citros

Isozymes in the early identification of hybrids and nucellar clones in citrus breeding

Rosa Maria Lizana BallveI, ^** ** Com bolsa CAPES/UNICAMP. ; Rita Bordignon^I; Herculano Penna Medina FilhoI, ^* * Com bolsa de pesquisa do CNPq. ; Walter Jose SiqueiraI, ^* * Com bolsa de pesquisa do CNPq. ; Joaquim Teófilo Sobrinho^II; Jorgino Rompeu Júnior^III

^ISeção de Genética, Instituto Agronômico (I AC), Caixa Postal 28, 13001 Campinas (SP)

^IIEstação Experimental Dr. Sylvio Moreira, IAC

^IIISeção de Citricultura, IAC

RESUMO

Um dos problemas relacionados ao melhoramento de citros é a poliembrionia e a conseqüente dificuldade, após cruzamentos controlados, de distinguir clones nucelares dos híbridos. Estudou-se um método para eletroforese de isoenzimas em gel de amido para doze sistemas isoenzímicos, desenvolvendo-se procedimento apropriado para a resolução simultânea dos sistemas fosfatase ácida, peroxidase anódica e catódica, glutamato oxaloacético transaminase, fosfoglucoisomerase, fosfoglucomutase e enzima málica. Caracterizaram-se por esses sistemas 31 clones de citros com possível interesse para o programa de melhoramento do IAC. Estão envolvidos 7 locos e 25 alelos, permitindo uma identificação precoce de híbridos antes do plantio no campo para avaliações agronômicas. Implicações desses estudos básicos no melhoramento e na determinação da taxa de seedlings nucelares de porta-enxertos comerciais são discutidas.

Termos de indexação: citros, genética, melhoramento, isoenzimas, híbridos, clones nucelares, embriões zigóticos.

ABSTRACTS

One of the problems related to citrus breeding is the polyembryony and the ensuing difficulty in distinguishing nucellar clones from hybrids obtained from controlled crosses. Twelve isozyme systems using starch gel electrophoresis were studied. A procedure for simultaneous resolution of acid phosphatase, anodic and cathodic peroxidase, glutamate oxaloacetate transaminase, phosphoglucoisomerase, phosphoglucomutase and malic enzyme was developed. Thirty one clones with possible use in the Instituto Agronômico de Campinas, State of São Paulo, Brazil breeding program were characterized. Seven loci and twenty five alleles are involved allowing for an early identification of hybrids before transplanting them into the field for agronomic evaluations. Implications in breeding strategies and in the determination of the degree of nucellar embryony of commercial rootstocks are discussed.

Index terms: citrus, genetics, breeding, isozymes, hybrids, nucellar clones, zygotic embryos.

Texto completo disponível apenas em PDF.

Full text available only in PDF format.

Recebido para publicação em 6 de agosto de 1990 e aceito em 30 de marco de 1991.

ARULSEKAR, S.; PARFITT, D.E.; BERES, W. & HANSCHE, P.E. Genetics of malate dehydrogenase isozymes in peach. Journal of Heredity, Washington, 77(1):49-51, 1986.
^__________;^__________& McGRANAHAN, G.H. Isozyme gene markers in Juglans species: inheritance of GPl and AAT in J. regia and J. hindsii. Journal of Heredity, Washington, 76(2):103-106, 1985,
BRINGHURST, R.S.; ARULSEKAR, S.; HANCOCK JUNIOR, J.F. & VOTH, V. Electrophoretic characterization of strawberry cultivars. Journal of the American Society ofr Horticultural Science, Alexandria, 106(5):684-687, 1981.
BUTTON, J.; VARDI, A & SPIEGEL-ROY, P. Root Peroxidase isoenzymes as an aid in Citrus breeding and taxonomy. Theoretical and Applied Genetics, Berlin, 47:119-123, 1976.
BYRNE, D.H. & LITTLETON, T.G. Characterization of isozyme variability in apricots. Journal of the American Society for Horticultural Science, Alexandria 114(4):674-678, 1989.]
^__________&^__________ Electrophoretic characterization of diploid plums of the southeastern United States. Journal of the American Society for Horticultural Science, Alexandria, 113(6):919-924, 1988.
CAMERON, J.W. & FROST, H.B. Genetics, breeding, and nucellar embryony. In: REUTHER, W., BATCHELOR, L.D. & WEBBER, H.J., eds. The citrus industry, v.ll. Anatomy, physiology, genetics, and reproduction. Riverside, University of California-Division of Agricultural Science, 1968. cap.5, p.325-370.
ESEN, A. & SOOST, R.K. Peroxidase polymorphism in Citrus. Journal of Heredity, Washington, 67(4):199-203, 1976.
FAO TRADE YEARBOOK. Rome, FAO, v.42, 1988.
GERACI, G.; MANZOCCHI, L.A.; TUSA, N.; OCCORSO, G.; RADOGNA, L & DE PASQUALE, F. Comparison of different methods for identifying zygotic and nucellar seedlings in Citrus. In: INTERNATIONAL SOCIETY OF CITRICULTURE, 5. TOKYO, 1981. Proceedings. Tokyo, 1982. v.1, p.1-4.
HAUAGGE, R.; KESTER, D.E.; ARULSEKAR, S.; PARFITT, D.E. & LIU, L. Isozyme variation among California almond cuftivars: II. Cultivar characterization and origins. Journal of the American Society for Horticultural Science, Alexandria, 112(4):693-698, 1987.
IGLESIAS, L; LIMA, H. & SIMON, J.P. Isoenzyme identification of zygotic and nucellar seedlings in Citrus. Journal of Heredity, Washington, 65(2):81-84, 1974.
LLANOS, J.L & H. LIMA. Peroxidase in healthy grapefruit trees and those affected by blight. ln:INTERNATIONAL SOCIETY OF CITRICULTURE,5.,Tokyo, 1981. Proceedings. Tokyo, 1982. v.1, p.476-478.
LOUKAS,M.; STAVRAKAKIS,M.N. & KIMBRAS, C.B. Inheritance of polymorphic isoenzymes in grape cultivars. Journal of Heredity, Washington, 74(3):181-183, 1983.
MANSHARDT, R.M. & WENSLAFF, T.F. Zygotic polyembryony in interspecific hybrids of Carica papaya and C. cauliflora. Journal of the American Society for Horticultural Science, Alexandria, 114(4):684-689, 1989.
MEDINA FILHO, H.P. Eletroforese em gel de amido: aplicações em genética e melhoramento de plantas. Campinas, Instituto Agronômico. 1983. 15p. (Circular, 121)
^__________& BALLVE, R.M.L. Protocolo laboratorial para preparação e corrida de géis de amido e revelação para is o enzimas vegetais. In: CURSO TEÓRICO-PRÁTICO DE ELETROFORESE, 5., Nova Odessa, 1989. Nova Odessa, Instituto de Zootecnia, 1989. 59p.
MOORE, G. A. & CASTLE, W.S. Morphological and isozymic analysis of open-pollinated Citrus rootstock population. Journal of Heredity, Washington, 79(1):59-63, 1988.
PARFITT, D.E.; ARULSEKAR, S. & RAMMING, D. W. Identification of plumx peach hybrids by isoenzyme analysis. HortScience, Alexandria, 20(2):246-248, 1985.
ROOSE, M.L Isozymes and DNA restriction fragment length polymorphisms in citrus breeding and systematics. In: INTERNATIONAL CITRUS CONGRESS, 6., Tel Aviv, 1988. Proceedings. Philadelphia Rehovot, Balaban, 1988. p. 155-165.
^__________& TRAUGH, S.N. Identification and performance of citrus trees on nucellar and zygotic rootstocks. Journal of the American Society for Horticultural Science, Alexandria, 113(1):100-105, 1988.
SHAW, C.R. & PRASAD, R. Starch gel eletrophoresis of enzymes: a compilation of recipes. Biochemical Genetics, New York, 4:297-320, 1970.
SOOST, R.K. & CAMERON, J.W. Citrus. In: JANICK, J. & MOORE, J.N., eds. Advances in fruit breeding. West Lafayette, Purdue University Press, 1975. p.507-540.
^__________& TORRES, A.M. Leaf isozymes as genetic markers in Citrus. In: INTERNATIONAL SOCIETY OF CITRICULTURE, 5., Tokyo, 1981. Proceedings. Tokyo, 1982. v.1, p. 7-10.
SPIEGEL-ROY, P.; VARDI, A. & SHANI, A. Peroxidase isozymes as a tool for early separation of nucellar and zygotic citrus seedlings. In: INTERNATIONAL SOCIETY OF CITRICUTURE, 3., Orlando, 1977. Proceedings. Orlando, 1977. v.2, p.619-624.
TANAKA, T. Species problem in citrus: a critical study of wild and cultivated units of Citrus,based upon field studies in their native homes. Tokyo, Japanese Society for the Promotion of Science, 1954. 152p.
TANKSLEY, S.D. An efficient and economical design for starch gel electrophoresis. Report of the Tomato Genetics Cooperative, Davis (29):37-38, 1979.
^__________; MEDINA FILHO, H.P. & RICK, C.M. The effect of isozyme selection on metric characters in an interspecific backcross of tomato: basis of an early screening procedure. Theoretical and Applied Genetics, Berlin, 60:291 -296, 1981.
^__________;^__________&^__________ Use of naturally-occurring enzyme variation to detect and map genes controlling quantitative traits in an interspecific backcross of tomato. Heredity, London, 49(1):11-25, 1982.
TORRES,A.M.; DIEDENHOFEN,U.; BERGH,B.O. & KNIGHT, R.J. Enzyme polymorphisms as genetic markers in the avocado. American Journal of Botany, Baltimore, 65(2):134-139, 1978a.
^__________;MAU-LASTOVICKA,T.;WILLIAMS,T.E.&SOOST,R.K. Segregation distortion and linkage of Citrus and Poncirus isozyme genes. Journal of Heredity, Washington, 76(4):289-294, 1985.
^__________; SOOST, R.K. & DIEDENHOFEN, U. Leaf isozymes as genetic markers in Citrus. American Journal of Botany, Baltimore, 65(8):869-881, 1978b.
^__________;^__________& MAU-LASTOVICKA, T. Citrus isozymes: genetics and distinguishing nucellar from zygotic seedlings. Journal of Heredity, Washington, 73(5):335-339, 1982.
VALLEJOS, C.E. Enzyme activity staining. In: TANKSLEY, S.D. & ORTON, T.J., eds. Isozymes in plant genetics and breeding: part A. Amsterdam, Elsevier, 1963. p. 469-516.
WEEDEN, N.F. & LAMB, R.C. Identification of apple cultivars by isozyme phenotypes. Journal of the American Society for Horticultural Science, Alexandria, 110(4):509-515, 1985.

*

Com bolsa de pesquisa do CNPq.

**

Com bolsa CAPES/UNICAMP.

Datas de Publicação

Publicação nesta coleção
19 Nov 2007
Data do Fascículo
1991

Histórico

Aceito
30 Mar 1991
Recebido
06 Ago 1990

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

[1] ARULSEKAR, S.; PARFITT, D.E.; BERES, W. & HANSCHE, P.E. Genetics of malate dehydrogenase isozymes in peach. Journal of Heredity, Washington, 77(1):49-51, 1986.

[2] ^__________;^__________& McGRANAHAN, G.H. Isozyme gene markers in Juglans species: inheritance of GPl and AAT in J. regia and J. hindsii. Journal of Heredity, Washington, 76(2):103-106, 1985,

[3] BRINGHURST, R.S.; ARULSEKAR, S.; HANCOCK JUNIOR, J.F. & VOTH, V. Electrophoretic characterization of strawberry cultivars. Journal of the American Society ofr Horticultural Science, Alexandria, 106(5):684-687, 1981.

[4] BUTTON, J.; VARDI, A & SPIEGEL-ROY, P. Root Peroxidase isoenzymes as an aid in Citrus breeding and taxonomy. Theoretical and Applied Genetics, Berlin, 47:119-123, 1976.

[5] BYRNE, D.H. & LITTLETON, T.G. Characterization of isozyme variability in apricots. Journal of the American Society for Horticultural Science, Alexandria 114(4):674-678, 1989.]

[6] ^__________&^__________ Electrophoretic characterization of diploid plums of the southeastern United States. Journal of the American Society for Horticultural Science, Alexandria, 113(6):919-924, 1988.

[7] CAMERON, J.W. & FROST, H.B. Genetics, breeding, and nucellar embryony. In: REUTHER, W., BATCHELOR, L.D. & WEBBER, H.J., eds. The citrus industry, v.ll. Anatomy, physiology, genetics, and reproduction. Riverside, University of California-Division of Agricultural Science, 1968. cap.5, p.325-370.

[8] ESEN, A. & SOOST, R.K. Peroxidase polymorphism in Citrus. Journal of Heredity, Washington, 67(4):199-203, 1976.

[9] FAO TRADE YEARBOOK. Rome, FAO, v.42, 1988.

[10] GERACI, G.; MANZOCCHI, L.A.; TUSA, N.; OCCORSO, G.; RADOGNA, L & DE PASQUALE, F. Comparison of different methods for identifying zygotic and nucellar seedlings in Citrus. In: INTERNATIONAL SOCIETY OF CITRICULTURE, 5. TOKYO, 1981. Proceedings. Tokyo, 1982. v.1, p.1-4.

[11] HAUAGGE, R.; KESTER, D.E.; ARULSEKAR, S.; PARFITT, D.E. & LIU, L. Isozyme variation among California almond cuftivars: II. Cultivar characterization and origins. Journal of the American Society for Horticultural Science, Alexandria, 112(4):693-698, 1987.

[12] IGLESIAS, L; LIMA, H. & SIMON, J.P. Isoenzyme identification of zygotic and nucellar seedlings in Citrus. Journal of Heredity, Washington, 65(2):81-84, 1974.

[13] LLANOS, J.L & H. LIMA. Peroxidase in healthy grapefruit trees and those affected by blight. ln:INTERNATIONAL SOCIETY OF CITRICULTURE,5.,Tokyo, 1981. Proceedings. Tokyo, 1982. v.1, p.476-478.

[14] LOUKAS,M.; STAVRAKAKIS,M.N. & KIMBRAS, C.B. Inheritance of polymorphic isoenzymes in grape cultivars. Journal of Heredity, Washington, 74(3):181-183, 1983.

[15] MANSHARDT, R.M. & WENSLAFF, T.F. Zygotic polyembryony in interspecific hybrids of Carica papaya and C. cauliflora. Journal of the American Society for Horticultural Science, Alexandria, 114(4):684-689, 1989.

[16] MEDINA FILHO, H.P. Eletroforese em gel de amido: aplicações em genética e melhoramento de plantas. Campinas, Instituto Agronômico. 1983. 15p. (Circular, 121)

[17] ^__________& BALLVE, R.M.L. Protocolo laboratorial para preparação e corrida de géis de amido e revelação para is o enzimas vegetais. In: CURSO TEÓRICO-PRÁTICO DE ELETROFORESE, 5., Nova Odessa, 1989. Nova Odessa, Instituto de Zootecnia, 1989. 59p.

[18] MOORE, G. A. & CASTLE, W.S. Morphological and isozymic analysis of open-pollinated Citrus rootstock population. Journal of Heredity, Washington, 79(1):59-63, 1988.

[19] PARFITT, D.E.; ARULSEKAR, S. & RAMMING, D. W. Identification of plumx peach hybrids by isoenzyme analysis. HortScience, Alexandria, 20(2):246-248, 1985.

[20] ROOSE, M.L Isozymes and DNA restriction fragment length polymorphisms in citrus breeding and systematics. In: INTERNATIONAL CITRUS CONGRESS, 6., Tel Aviv, 1988. Proceedings. Philadelphia Rehovot, Balaban, 1988. p. 155-165.

[21] ^__________& TRAUGH, S.N. Identification and performance of citrus trees on nucellar and zygotic rootstocks. Journal of the American Society for Horticultural Science, Alexandria, 113(1):100-105, 1988.

[22] SHAW, C.R. & PRASAD, R. Starch gel eletrophoresis of enzymes: a compilation of recipes. Biochemical Genetics, New York, 4:297-320, 1970.

[23] SOOST, R.K. & CAMERON, J.W. Citrus. In: JANICK, J. & MOORE, J.N., eds. Advances in fruit breeding. West Lafayette, Purdue University Press, 1975. p.507-540.

[24] ^__________& TORRES, A.M. Leaf isozymes as genetic markers in Citrus. In: INTERNATIONAL SOCIETY OF CITRICULTURE, 5., Tokyo, 1981. Proceedings. Tokyo, 1982. v.1, p. 7-10.

[25] SPIEGEL-ROY, P.; VARDI, A. & SHANI, A. Peroxidase isozymes as a tool for early separation of nucellar and zygotic citrus seedlings. In: INTERNATIONAL SOCIETY OF CITRICUTURE, 3., Orlando, 1977. Proceedings. Orlando, 1977. v.2, p.619-624.

[26] TANAKA, T. Species problem in citrus: a critical study of wild and cultivated units of Citrus,based upon field studies in their native homes. Tokyo, Japanese Society for the Promotion of Science, 1954. 152p.

[27] TANKSLEY, S.D. An efficient and economical design for starch gel electrophoresis. Report of the Tomato Genetics Cooperative, Davis (29):37-38, 1979.

[28] ^__________; MEDINA FILHO, H.P. & RICK, C.M. The effect of isozyme selection on metric characters in an interspecific backcross of tomato: basis of an early screening procedure. Theoretical and Applied Genetics, Berlin, 60:291 -296, 1981.

[29] ^__________;^__________&^__________ Use of naturally-occurring enzyme variation to detect and map genes controlling quantitative traits in an interspecific backcross of tomato. Heredity, London, 49(1):11-25, 1982.

[30] TORRES,A.M.; DIEDENHOFEN,U.; BERGH,B.O. & KNIGHT, R.J. Enzyme polymorphisms as genetic markers in the avocado. American Journal of Botany, Baltimore, 65(2):134-139, 1978a.

[31] ^__________;MAU-LASTOVICKA,T.;WILLIAMS,T.E.&SOOST,R.K. Segregation distortion and linkage of Citrus and Poncirus isozyme genes. Journal of Heredity, Washington, 76(4):289-294, 1985.

[32] ^__________; SOOST, R.K. & DIEDENHOFEN, U. Leaf isozymes as genetic markers in Citrus. American Journal of Botany, Baltimore, 65(8):869-881, 1978b.

[33] ^__________;^__________& MAU-LASTOVICKA, T. Citrus isozymes: genetics and distinguishing nucellar from zygotic seedlings. Journal of Heredity, Washington, 73(5):335-339, 1982.

[34] VALLEJOS, C.E. Enzyme activity staining. In: TANKSLEY, S.D. & ORTON, T.J., eds. Isozymes in plant genetics and breeding: part A. Amsterdam, Elsevier, 1963. p. 469-516.

[35] WEEDEN, N.F. & LAMB, R.C. Identification of apple cultivars by isozyme phenotypes. Journal of the American Society for Horticultural Science, Alexandria, 110(4):509-515, 1985.

Brasil

Brasil

Isoenzimas na identificação precoce de híbridos e clones nucelares no melhoramento de citros

Isozymes in the early identification of hybrids and nucellar clones in citrus breeding

Resumos

Datas de Publicação

Histórico