Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L.
Ano de defesa: | 2007 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , , |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Viçosa
|
Programa de Pós-Graduação: |
Doutorado em Botânica
|
Departamento: |
Botânica estrutural; Ecologia e Sistemática
|
País: |
BR
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://locus.ufv.br/handle/123456789/383 |
Resumo: | Bixa Orellana L. and Ginkgo biloba L. are woody species that synthesize exclusive medicinal components of economic importance. The objectives of this study were: (1) to evaluate and select genotypes of B. orellana with high bixin content; (2) to rescue vegetative genotypes of B. orellana aiming the production of explants for the in vitro culture; (3) to establish a system of in vitro propagation of B. orellana from axillaries shoots in nodal segments of adult plants, and adventitious shoots from juvenile explants; (4) to induce the formation of calli from leaf explants of G. biloba and to evaluate the in vitro production of secondary metabolites; (5) to establish a system of in vitro propagation of G. biloba from axillaries shoots in nodal segments of adult plants. Genotypes of B. orellana with high bixin content had been rescued by air layering technique and propagated in vitro by the induction and growth of axillaries shoots from nodal segments. The explants were disinfested with of sodium hipochloride solution (10 minutes 1.25%) and PPMTM (20 minutes 20%), and cultivated on MS medium culture. The addition of PPMTM 1%, Timentin® 600 mg.L-1 and 300 Agrimicin® mg.L-1 to the culture medium partially controlled the development of bacteria on the explants. The development of axillaries shoots was observed in 28% of the nodal segments. Adventitious shoots were induced in cotyledon discs, hypocotyl and root segments cultivated on MS and WPM media with ZEA 4.56 µM. It was identified a gradient of morphogenic potential that increase from the apex in direction to the base of the hypocotyls and roots explants. High frequencies of regeneration of shoots were induced in cultivated hypocotyl segments on WPM medium culture with 4.56 µM TDZ (85%) and with 4.56 µM ZEA (80%). The AIB 4.9 µM induced greater frequency of rooted shoots (25%) and roots number (2). Complete plants in vitro produced were transferred to substract and successfully acclimatized in a greenhouse. The histological analysis had proven the adventitious origin of the shoots from the organization of meristematic centers in cortical cells of hypocotyl segments, characterizing direct organogenesis in B. orellana. In G. biloba leaf explants callogenesis is dependent of an exogenous source of auxin. Greater callus frequencies had been induced from leaf discs cultivated on MS medium with ANA 4.4 µM combined with 0.4 BAP µM (100%) or 4.4 µM (92%). The maximum production of callus fresh mass was observed with carbohydrate sucrose (394,57 mg) and the subcultivation in the dark (678,3 mg). The BAP modified the pattern of the primary callus from white yellowish friable to green compact one when used by three subcultivation period. The terpene trilactones ginkgolides A and B, and the flavonol glycosides quercetin, kaempferol and rutin have not been observed in calli cultivated in vitro and in leaves of G. biloba through TLC and HPLC methods. The in vitro propagation of G. biloba was carried out by means of axillary shooting in nodal segments excised from herbaceous branches, harvested from adult plants maintained in greenhouse. The hydrolyzed casein (500 mg.L-1) in MS medium, was essential for the induction of higher frequencies of axillary shoots in nodal segments (85%), for multiplication of these shoots (66.6%), and also for formation of multiple shoots in the explants. The kinetin and the activated charcoal were inhibitory to this process. |
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Mantovani, Nilton Césarhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706084U9Grando, Magali Ferrarihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782193J1Xavier, Aloisiohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782565D0Otoni, Wagner Camposhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6Calvete, Eunice Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783766P5Nienow, Alexandre Augustohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785438H9Carneiro, Cerci Mariahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4776102A02015-03-26T12:19:27Z2007-12-132015-03-26T12:19:27Z2007-09-06MANTOVANI, Nilton César. Vegetative propagation and in vitro culture of Bixa orellana L. and Ginkgo biloba L.. 2007. 149 f. Tese (Doutorado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2007.http://locus.ufv.br/handle/123456789/383Bixa Orellana L. and Ginkgo biloba L. are woody species that synthesize exclusive medicinal components of economic importance. The objectives of this study were: (1) to evaluate and select genotypes of B. orellana with high bixin content; (2) to rescue vegetative genotypes of B. orellana aiming the production of explants for the in vitro culture; (3) to establish a system of in vitro propagation of B. orellana from axillaries shoots in nodal segments of adult plants, and adventitious shoots from juvenile explants; (4) to induce the formation of calli from leaf explants of G. biloba and to evaluate the in vitro production of secondary metabolites; (5) to establish a system of in vitro propagation of G. biloba from axillaries shoots in nodal segments of adult plants. Genotypes of B. orellana with high bixin content had been rescued by air layering technique and propagated in vitro by the induction and growth of axillaries shoots from nodal segments. The explants were disinfested with of sodium hipochloride solution (10 minutes 1.25%) and PPMTM (20 minutes 20%), and cultivated on MS medium culture. The addition of PPMTM 1%, Timentin® 600 mg.L-1 and 300 Agrimicin® mg.L-1 to the culture medium partially controlled the development of bacteria on the explants. The development of axillaries shoots was observed in 28% of the nodal segments. Adventitious shoots were induced in cotyledon discs, hypocotyl and root segments cultivated on MS and WPM media with ZEA 4.56 µM. It was identified a gradient of morphogenic potential that increase from the apex in direction to the base of the hypocotyls and roots explants. High frequencies of regeneration of shoots were induced in cultivated hypocotyl segments on WPM medium culture with 4.56 µM TDZ (85%) and with 4.56 µM ZEA (80%). The AIB 4.9 µM induced greater frequency of rooted shoots (25%) and roots number (2). Complete plants in vitro produced were transferred to substract and successfully acclimatized in a greenhouse. The histological analysis had proven the adventitious origin of the shoots from the organization of meristematic centers in cortical cells of hypocotyl segments, characterizing direct organogenesis in B. orellana. In G. biloba leaf explants callogenesis is dependent of an exogenous source of auxin. Greater callus frequencies had been induced from leaf discs cultivated on MS medium with ANA 4.4 µM combined with 0.4 BAP µM (100%) or 4.4 µM (92%). The maximum production of callus fresh mass was observed with carbohydrate sucrose (394,57 mg) and the subcultivation in the dark (678,3 mg). The BAP modified the pattern of the primary callus from white yellowish friable to green compact one when used by three subcultivation period. The terpene trilactones ginkgolides A and B, and the flavonol glycosides quercetin, kaempferol and rutin have not been observed in calli cultivated in vitro and in leaves of G. biloba through TLC and HPLC methods. The in vitro propagation of G. biloba was carried out by means of axillary shooting in nodal segments excised from herbaceous branches, harvested from adult plants maintained in greenhouse. The hydrolyzed casein (500 mg.L-1) in MS medium, was essential for the induction of higher frequencies of axillary shoots in nodal segments (85%), for multiplication of these shoots (66.6%), and also for formation of multiple shoots in the explants. The kinetin and the activated charcoal were inhibitory to this process.Bixa orellana L. e Ginkgo biloba L. são espécies lenhosas que sintetizam compostos medicinais exclusivos e de relevante importância econômica. Esta pesquisa teve como objetivos: (1) avaliar e selecionar genótipos de B. orellana com altos teores de bixina nas sementes; (2) resgatar vegetativamente genótipos de B. orellana, visando à produção de explantes para o cultivo in vitro; (3) estabelecer um sistema de propagação in vitro de B. orellana, a partir de brotos axilares em segmentos nodais de plantas adultas, e de brotos adventícios em explantes juvenis; (4) induzir a formação de calos em explantes foliares de G. biloba e avaliar a produção de metabólitos secundários in vitro e, (5) estabelecer um sistema de propagação in vitro de G. biloba, a partir de brotos axilares em segmentos nodais de plantas adultas. Genótipos de B. orellana, com alto teor de bixina, foram resgatados por alporquia e propagados in vitro a partir da indução de brotos axilares em segmentos nodais. Os explantes foram desinfestados com soluções de hipoclorito de sódio (10 minutos a 1,25%) e PPMTM (20 minutos a 20%), e cultivados em meio MS. Nestas condições foram eliminados os contaminantes fúngicos, porém, em concentrações mais altas, as soluções desinfestantes provocaram necrose e morte dos explantes. A adição de PPMTM 1%, Timentin® 600 mg.L-1 e Agrimicina® 300 mg.L-1 ao meio de cultura, controlou parcialmente o desenvolvimento de bactérias nos explantes. O desenvolvimento de brotos axilares ocorreu em 28% dos segmentos nodais. Brotos adventícios foram induzidos em discos cotiledonares, segmentos hipocotiledonares e radiculares, cultivados nos meios MS e WPM, com 4,56 µM de ZEA. Identificou-se um gradiente de potencial morfogênico, que cresce do ápice em direção à base, em hipocótilos e raízes. Altas freqüências de regeneração de brotos foram induzidas em segmentos hipocotiledonares, cultivados em meio WPM, com 4,56 µM de TDZ (85%) e com 4,56 µM de ZEA (80%). O AIB a 4,9 µM induziu maior freqüência de brotos enraizados (25%) e número de raízes (2). Plantas completas produzidas in vitro foram transferidas para substrato e aclimatizadas com sucesso em casa de vegetação. As análises histológicas comprovaram a origem adventícia dos brotos, a partir da organização de centros meristemáticos em células corticais de segmentos de hipocótilo, caracterizando a organogênese direta em B. orellana. Em G. biloba, a calogênese em explantes foliares, é dependente de uma fonte exógena de auxina. Altas freqüências calogênicas foram induzidas em discos foliares, cultivados no meio MS, com ANA 4,4 µM combinada com BAP a 0,4 µM (100%) ou 4,4 µM (92%). A máxima produção de massa fresca de calos foi obtida com o carboidrato sacarose (394,57 mg) e o subcultivo na ausência de luz (678,3 mg). O BAP alterou o padrão dos calos primários, branco amarelados e friáveis, para verdes e compactos, quando utilizado por três subcultivos. As lactonas terpênicas ginkgolídeos A e B, e os glicosídeos flavonóides quercetina, camferol e rutina, não foram detectados nos calos ou folhas de G. biloba, através dos métodos de TLC e HPLC. A propagação in vitro de G. biloba foi obtida a partir da indução e multiplicação de brotos axilares em segmentos nodais, excisados de ramos herbáceos de plantas matrizes adultas cultivadas em casa de vegetação. A caseína hidrolisada (500 mg.L-1), no meio MS, foi essencial para a indução de altas taxas de emissão de brotos axilares nos segmentos nodais (85%), de multiplicação (66,6%), e de formação de múltiplos brotos nos explantes. A cinetina e o carvão ativado apresentaram efeitos inibitórios neste processo.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaDoutorado em BotânicaUFVBRBotânica estrutural; Ecologia e SistemáticaCultura de tecidosPropagaçãoMorfogênese in vitroBixa orellanaGinkgo bilobaTissue culturePropagationIn vitro morfogenesisBixa orellanaGinkgo bilobaCNPQ::CIENCIAS BIOLOGICAS::BOTANICA::BOTANICA APLICADAPropagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L.Vegetative propagation and in vitro culture of Bixa orellana L. and Ginkgo biloba L.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf3264136https://locus.ufv.br//bitstream/123456789/383/1/texto%20completo.pdf44bb3af2b74068e8a5d47ed5a51cebecMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain261563https://locus.ufv.br//bitstream/123456789/383/2/texto%20completo.pdf.txt65fcd9bee70d0836321172847b6c4f6dMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3592https://locus.ufv.br//bitstream/123456789/383/3/texto%20completo.pdf.jpg41d9abf2d2023589a8f583edc883d9ffMD53123456789/3832016-04-06 23:05:47.889oai:locus.ufv.br:123456789/383Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-07T02:05:47LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
dc.title.alternative.eng.fl_str_mv |
Vegetative propagation and in vitro culture of Bixa orellana L. and Ginkgo biloba L. |
title |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
spellingShingle |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. Mantovani, Nilton César Cultura de tecidos Propagação Morfogênese in vitro Bixa orellana Ginkgo biloba Tissue culture Propagation In vitro morfogenesis Bixa orellana Ginkgo biloba CNPQ::CIENCIAS BIOLOGICAS::BOTANICA::BOTANICA APLICADA |
title_short |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
title_full |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
title_fullStr |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
title_full_unstemmed |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
title_sort |
Propagação vegetativa e cultivo in vitro de Bixa orellana L. e Ginkgo biloba L. |
author |
Mantovani, Nilton César |
author_facet |
Mantovani, Nilton César |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706084U9 |
dc.contributor.author.fl_str_mv |
Mantovani, Nilton César |
dc.contributor.advisor-co1.fl_str_mv |
Grando, Magali Ferrari |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782193J1 |
dc.contributor.advisor-co2.fl_str_mv |
Xavier, Aloisio |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782565D0 |
dc.contributor.advisor1.fl_str_mv |
Otoni, Wagner Campos |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6 |
dc.contributor.referee1.fl_str_mv |
Calvete, Eunice Oliveira |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783766P5 |
dc.contributor.referee2.fl_str_mv |
Nienow, Alexandre Augusto |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785438H9 |
dc.contributor.referee3.fl_str_mv |
Carneiro, Cerci Maria |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4776102A0 |
contributor_str_mv |
Grando, Magali Ferrari Xavier, Aloisio Otoni, Wagner Campos Calvete, Eunice Oliveira Nienow, Alexandre Augusto Carneiro, Cerci Maria |
dc.subject.por.fl_str_mv |
Cultura de tecidos Propagação Morfogênese in vitro Bixa orellana Ginkgo biloba |
topic |
Cultura de tecidos Propagação Morfogênese in vitro Bixa orellana Ginkgo biloba Tissue culture Propagation In vitro morfogenesis Bixa orellana Ginkgo biloba CNPQ::CIENCIAS BIOLOGICAS::BOTANICA::BOTANICA APLICADA |
dc.subject.eng.fl_str_mv |
Tissue culture Propagation In vitro morfogenesis Bixa orellana Ginkgo biloba |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA::BOTANICA APLICADA |
description |
Bixa Orellana L. and Ginkgo biloba L. are woody species that synthesize exclusive medicinal components of economic importance. The objectives of this study were: (1) to evaluate and select genotypes of B. orellana with high bixin content; (2) to rescue vegetative genotypes of B. orellana aiming the production of explants for the in vitro culture; (3) to establish a system of in vitro propagation of B. orellana from axillaries shoots in nodal segments of adult plants, and adventitious shoots from juvenile explants; (4) to induce the formation of calli from leaf explants of G. biloba and to evaluate the in vitro production of secondary metabolites; (5) to establish a system of in vitro propagation of G. biloba from axillaries shoots in nodal segments of adult plants. Genotypes of B. orellana with high bixin content had been rescued by air layering technique and propagated in vitro by the induction and growth of axillaries shoots from nodal segments. The explants were disinfested with of sodium hipochloride solution (10 minutes 1.25%) and PPMTM (20 minutes 20%), and cultivated on MS medium culture. The addition of PPMTM 1%, Timentin® 600 mg.L-1 and 300 Agrimicin® mg.L-1 to the culture medium partially controlled the development of bacteria on the explants. The development of axillaries shoots was observed in 28% of the nodal segments. Adventitious shoots were induced in cotyledon discs, hypocotyl and root segments cultivated on MS and WPM media with ZEA 4.56 µM. It was identified a gradient of morphogenic potential that increase from the apex in direction to the base of the hypocotyls and roots explants. High frequencies of regeneration of shoots were induced in cultivated hypocotyl segments on WPM medium culture with 4.56 µM TDZ (85%) and with 4.56 µM ZEA (80%). The AIB 4.9 µM induced greater frequency of rooted shoots (25%) and roots number (2). Complete plants in vitro produced were transferred to substract and successfully acclimatized in a greenhouse. The histological analysis had proven the adventitious origin of the shoots from the organization of meristematic centers in cortical cells of hypocotyl segments, characterizing direct organogenesis in B. orellana. In G. biloba leaf explants callogenesis is dependent of an exogenous source of auxin. Greater callus frequencies had been induced from leaf discs cultivated on MS medium with ANA 4.4 µM combined with 0.4 BAP µM (100%) or 4.4 µM (92%). The maximum production of callus fresh mass was observed with carbohydrate sucrose (394,57 mg) and the subcultivation in the dark (678,3 mg). The BAP modified the pattern of the primary callus from white yellowish friable to green compact one when used by three subcultivation period. The terpene trilactones ginkgolides A and B, and the flavonol glycosides quercetin, kaempferol and rutin have not been observed in calli cultivated in vitro and in leaves of G. biloba through TLC and HPLC methods. The in vitro propagation of G. biloba was carried out by means of axillary shooting in nodal segments excised from herbaceous branches, harvested from adult plants maintained in greenhouse. The hydrolyzed casein (500 mg.L-1) in MS medium, was essential for the induction of higher frequencies of axillary shoots in nodal segments (85%), for multiplication of these shoots (66.6%), and also for formation of multiple shoots in the explants. The kinetin and the activated charcoal were inhibitory to this process. |
publishDate |
2007 |
dc.date.available.fl_str_mv |
2007-12-13 2015-03-26T12:19:27Z |
dc.date.issued.fl_str_mv |
2007-09-06 |
dc.date.accessioned.fl_str_mv |
2015-03-26T12:19:27Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
MANTOVANI, Nilton César. Vegetative propagation and in vitro culture of Bixa orellana L. and Ginkgo biloba L.. 2007. 149 f. Tese (Doutorado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2007. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/383 |
identifier_str_mv |
MANTOVANI, Nilton César. Vegetative propagation and in vitro culture of Bixa orellana L. and Ginkgo biloba L.. 2007. 149 f. Tese (Doutorado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2007. |
url |
http://locus.ufv.br/handle/123456789/383 |
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por |
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por |
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Universidade Federal de Viçosa |
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Doutorado em Botânica |
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UFV |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Botânica estrutural; Ecologia e Sistemática |
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Universidade Federal de Viçosa |
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