Plant regeneration from protoplast of Brazilian citrus cultivars
Autor(a) principal: | |
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Data de Publicação: | 2000 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Pesquisa Agropecuária Brasileira (Online) |
Texto Completo: | https://seer.sct.embrapa.br/index.php/pab/article/view/5848 |
Resumo: | A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 mM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 mM), NAA (1.07 mM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development. |
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Plant regeneration from protoplast of Brazilian citrus cultivarsRegeneração de plantas a partir de protoplastos de cultivares Brasileiros de citroscultivar improvement; polyethylene glycol; rootstock; tissue culturecultura de tecidos; melhoramento; polietileno glicol; porta-enxerto A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 mM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 mM), NAA (1.07 mM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development.O objetivo deste trabalho foi regenerar plantas a partir de protoplastos de cultivares brasileiras de citros, após isolamento, fusão e cultura. Protoplastos foram isolados a partir de células embriogênicas em suspensão e de mesófilo foliar de plântulas germinadas in vitro. A solução enzimática para o isolamento de protoplastos foi composta de manitol (0,7 M), CaCl2 (24,5 mM), NaH2PO4 (0,92 mM), MES (6,15 mM), celulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) e pectolyase Y-23 (Seishin, 0,2%). A cultura dos protoplastos em meio líquido após fusão química levou ao desenvolvimento de microcolônias e calos, posteriormente adaptados para meio semi-sólido. A embriogênese somática ocorreu espontaneamente, após dois subcultivos em meio de cultura MT suplementado com 500 mg/L de extrato de malte. Embriões bem desenvolvidos germinaram em meio MT modificado, com a adição de GA3 (2,0 mM) e extrato de malte (500 mg/L). A regeneração de plantas também foi obtida por organogênese direta, com a formação de ápices caulinares a partir de embriões menores, em meio MT modificado com a adição de extrato de malte (500 mg/L), BAP (1,32 mM), NAA (1,07 mM) e água de coco (10 mL/L). As plântulas foram transferidas para meio de enraizamento e, posteriormente, para casa de vegetação, para fins de adaptação e desenvolvimento. Pesquisa Agropecuaria BrasileiraPesquisa Agropecuária BrasileiraGlória, Fernanda Januzzi Mendes daFilho, Francisco de Assis Alves MourãoMendes, Beatriz Madalena Januzzi2000-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.sct.embrapa.br/index.php/pab/article/view/5848Pesquisa Agropecuaria Brasileira; v.35, n.4, abr. 2000; 727-732Pesquisa Agropecuária Brasileira; v.35, n.4, abr. 2000; 727-7321678-39210100-104xreponame:Pesquisa Agropecuária Brasileira (Online)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPAenghttps://seer.sct.embrapa.br/index.php/pab/article/view/5848/2952info:eu-repo/semantics/openAccess2010-07-30T12:43:46Zoai:ojs.seer.sct.embrapa.br:article/5848Revistahttp://seer.sct.embrapa.br/index.php/pabPRIhttps://old.scielo.br/oai/scielo-oai.phppab@sct.embrapa.br || sct.pab@embrapa.br1678-39210100-204Xopendoar:2010-07-30T12:43:46Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false |
dc.title.none.fl_str_mv |
Plant regeneration from protoplast of Brazilian citrus cultivars Regeneração de plantas a partir de protoplastos de cultivares Brasileiros de citros |
title |
Plant regeneration from protoplast of Brazilian citrus cultivars |
spellingShingle |
Plant regeneration from protoplast of Brazilian citrus cultivars Glória, Fernanda Januzzi Mendes da cultivar improvement; polyethylene glycol; rootstock; tissue culture cultura de tecidos; melhoramento; polietileno glicol; porta-enxerto |
title_short |
Plant regeneration from protoplast of Brazilian citrus cultivars |
title_full |
Plant regeneration from protoplast of Brazilian citrus cultivars |
title_fullStr |
Plant regeneration from protoplast of Brazilian citrus cultivars |
title_full_unstemmed |
Plant regeneration from protoplast of Brazilian citrus cultivars |
title_sort |
Plant regeneration from protoplast of Brazilian citrus cultivars |
author |
Glória, Fernanda Januzzi Mendes da |
author_facet |
Glória, Fernanda Januzzi Mendes da Filho, Francisco de Assis Alves Mourão Mendes, Beatriz Madalena Januzzi |
author_role |
author |
author2 |
Filho, Francisco de Assis Alves Mourão Mendes, Beatriz Madalena Januzzi |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
|
dc.contributor.author.fl_str_mv |
Glória, Fernanda Januzzi Mendes da Filho, Francisco de Assis Alves Mourão Mendes, Beatriz Madalena Januzzi |
dc.subject.por.fl_str_mv |
cultivar improvement; polyethylene glycol; rootstock; tissue culture cultura de tecidos; melhoramento; polietileno glicol; porta-enxerto |
topic |
cultivar improvement; polyethylene glycol; rootstock; tissue culture cultura de tecidos; melhoramento; polietileno glicol; porta-enxerto |
description |
A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M), CaCl2 (24.5 mM), NaH2PO4 (0.92 mM), MES (6.15 mM), cellulase (Onozuka RS - Yakult, 1%), macerase (Onozuka R10 - Yakult, 1%) and pectolyase Y-23 (Seishin, 0.2%). Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 mM) and malt extract (500 mg/L). Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L), BAP (1.32 mM), NAA (1.07 mM) and coconut water (10 mL/L). Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development. |
publishDate |
2000 |
dc.date.none.fl_str_mv |
2000-04-01 |
dc.type.none.fl_str_mv |
|
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://seer.sct.embrapa.br/index.php/pab/article/view/5848 |
url |
https://seer.sct.embrapa.br/index.php/pab/article/view/5848 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://seer.sct.embrapa.br/index.php/pab/article/view/5848/2952 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Pesquisa Agropecuaria Brasileira Pesquisa Agropecuária Brasileira |
publisher.none.fl_str_mv |
Pesquisa Agropecuaria Brasileira Pesquisa Agropecuária Brasileira |
dc.source.none.fl_str_mv |
Pesquisa Agropecuaria Brasileira; v.35, n.4, abr. 2000; 727-732 Pesquisa Agropecuária Brasileira; v.35, n.4, abr. 2000; 727-732 1678-3921 0100-104x reponame:Pesquisa Agropecuária Brasileira (Online) instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa) instacron:EMBRAPA |
instname_str |
Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Pesquisa Agropecuária Brasileira (Online) |
collection |
Pesquisa Agropecuária Brasileira (Online) |
repository.name.fl_str_mv |
Pesquisa Agropecuária Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa) |
repository.mail.fl_str_mv |
pab@sct.embrapa.br || sct.pab@embrapa.br |
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1793416658397691904 |