Estratégias para conservação e clonagem in vitro de mangabeira
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFS |
Texto Completo: | https://ri.ufs.br/jspui/handle/riufs/19202 |
Resumo: | Hancornia speciosa Gomes is a fruit tree, found in several regions of Brazil, but particularly important in the Northeast region. Due to the wide range of uses of its fruits, this tree species represents a source of income of families that extract the fruits from the natural populations. The genetic resources of mangabeira are mostly conserved by means of field collections, mainly due to the recalcitrant physiology of the seeds. Therefore, cryopreservation techniques and somatic embryogenesis are additionally explored, as alternatives to the conservation of the existing genetic variability and for the vegetative multiplication of promising genotypes. The objectives of this study were to: I – evaluate the efficiency of the droplet vitrification technique for long-term conservation of five mangabeira accessions (Água Boa; Japaratinga; Paratibe; Oiteiro; Terra Caída), and check if these accessions have any level of susceptibility to possible effects of cryopreservation; II – study maturation induction of somatic embryos from nodal and foliar explants of accession Terra Caída (TC) of the Germplasm Bank of Mangaba (BAGMangaba). For cryopreservation, shoot apices were subjected to different periods of exposure (30 and 50 min for test I; 25, 50 and 75 min for test II) to vitrification solution 2 (PVS2), prior to immersion in liquid nitrogen. To evaluate the effects of cryoprotective solutions and exposure times on shoot apices exposed or not to liquid nitrogen, regeneration percentages were recorded after 30 and 60 days of in vitro cultivation. For callus induction, leaf and nodal segments of the TC accession were cultivated in media with different putrescine concentrations (0, 250, 500, 750 and 1000 µM) in MS medium (Murashige & Skoog, 1962). After 120 days, the explants were transferred to a MS multiplication medium supplemented with 10 mg/L 2,4-D, 5mg/L benzylaminopurine (BAP), 1 g/L activated charcoal and 30 g/L sucrose and gelled with 3 g/ L Phytagel®. After 90 days in multiplication medium, embryogenic calli were identified and selected for the maturation phase of somatic embryos under water stress induced with polyethylene glycol (PEG, at 0 and 2%) and enriched with BAP (10 and 15mg/L). After 60 days, pro-embryos and maturing embryos were observed. The experimental results were analyzed by non-parametric tests using R software. It was possible to cryopreserve the studied accessions by droplet vitrification. Putrescine did not favor the development of embryogenic calli, and the presence of 2,4-D and BAP promoted differentiation into embryogenic calli. Nodal segments in combinations of 10 or 15 mg/L BAP with 2% PEG intensified the formation of pro-embryos and maturing embryos of the TC accession. The performance pattern of accessions from BAGMangaba could be evaluated by the cryopreservation technique. In addition, the concentrations of growth regulators and best types of explants for establishing a somatic embryogenesis protocol for mangabeira could be defined. |
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Santana, Fernanda VieiraLedo, Ana da SilvaSantos, Paulo Augusto Almeida2024-02-29T19:55:28Z2024-02-29T19:55:28Z2023-03-24SANTANA, Fernanda Vieira. Estratégias para conservação e clonagem in vitro de mangabeira. 2023. 57 f. Tese (Doutorado em Agricultura e Biodiversidade) - Universidade Federal de Sergipe, São Cristóvão, 2023.https://ri.ufs.br/jspui/handle/riufs/19202Hancornia speciosa Gomes is a fruit tree, found in several regions of Brazil, but particularly important in the Northeast region. Due to the wide range of uses of its fruits, this tree species represents a source of income of families that extract the fruits from the natural populations. The genetic resources of mangabeira are mostly conserved by means of field collections, mainly due to the recalcitrant physiology of the seeds. Therefore, cryopreservation techniques and somatic embryogenesis are additionally explored, as alternatives to the conservation of the existing genetic variability and for the vegetative multiplication of promising genotypes. The objectives of this study were to: I – evaluate the efficiency of the droplet vitrification technique for long-term conservation of five mangabeira accessions (Água Boa; Japaratinga; Paratibe; Oiteiro; Terra Caída), and check if these accessions have any level of susceptibility to possible effects of cryopreservation; II – study maturation induction of somatic embryos from nodal and foliar explants of accession Terra Caída (TC) of the Germplasm Bank of Mangaba (BAGMangaba). For cryopreservation, shoot apices were subjected to different periods of exposure (30 and 50 min for test I; 25, 50 and 75 min for test II) to vitrification solution 2 (PVS2), prior to immersion in liquid nitrogen. To evaluate the effects of cryoprotective solutions and exposure times on shoot apices exposed or not to liquid nitrogen, regeneration percentages were recorded after 30 and 60 days of in vitro cultivation. For callus induction, leaf and nodal segments of the TC accession were cultivated in media with different putrescine concentrations (0, 250, 500, 750 and 1000 µM) in MS medium (Murashige & Skoog, 1962). After 120 days, the explants were transferred to a MS multiplication medium supplemented with 10 mg/L 2,4-D, 5mg/L benzylaminopurine (BAP), 1 g/L activated charcoal and 30 g/L sucrose and gelled with 3 g/ L Phytagel®. After 90 days in multiplication medium, embryogenic calli were identified and selected for the maturation phase of somatic embryos under water stress induced with polyethylene glycol (PEG, at 0 and 2%) and enriched with BAP (10 and 15mg/L). After 60 days, pro-embryos and maturing embryos were observed. The experimental results were analyzed by non-parametric tests using R software. It was possible to cryopreserve the studied accessions by droplet vitrification. Putrescine did not favor the development of embryogenic calli, and the presence of 2,4-D and BAP promoted differentiation into embryogenic calli. Nodal segments in combinations of 10 or 15 mg/L BAP with 2% PEG intensified the formation of pro-embryos and maturing embryos of the TC accession. The performance pattern of accessions from BAGMangaba could be evaluated by the cryopreservation technique. In addition, the concentrations of growth regulators and best types of explants for establishing a somatic embryogenesis protocol for mangabeira could be defined.A Hancornia speciosa Gomes é uma frutífera que está distribuída em diversas regiões do País, mas que possui grande importância para a região Nordeste. Devido à ampla possibilidade de uso dos seus frutos, essa fruteira representa uma fonte de renda para famílias que praticam a atividade extrativista dessa espécie. A conservação de seus recursos genéticos é, sobretudo, baseada em coleções de campo, principalmente devido à fisiologia recalcitrante de suas sementes. Diante disso, a aplicação de técnicas de criopreservação e embriogênese somática inserem-se como alternativas complementares à conservação da variabilidade genética existente, e à multiplicação vegetativa de genótipos promissores. Os objetivos deste estudo foram: I - avaliar a eficiência da técnica vitrificação em gotas para a conservação a longo prazo de cinco acessos de mangabeira (Acessos Água Boa; Japaratinga; Paratibe; Oiteiro; Terra Caída), como também verificar se há algum tipo de suscetibilidade entre os acessos aos possíveis efeitos do processo de criopreservação; II - conduzir estudos de indução à maturação de embriões somáticos a partir de explantes nodais e foliares do acesso Terra Caída (TC) do BAGMangaba. Para a criopreservação, ápices caulinares foram submetidos a diferentes períodos (30 e 50 min para o ensaio I; 25, 50 e 75 min para o ensaio II) de exposição à solução de vitrificação 2 (PVS2) antes da imersão em nitrogênio líquido. Para avaliar os efeitos de soluções crioprotetoras e tempos de exposição em ápices caulinares expostos ou não ao nitrogênio líquido, foram observadas as porcentagens de regeneração aos 30 e 60 dias de cultivo in vitro. Já para a indução de calos segmentos foliares e nodais do acesso TC foram cultivados em meios com diferentes concentrações de putrescina (0, 250, 500, 750 e 1000 µM) em meio MS (Murashige & Skoog, 1962). Após 120 dias, os explantes foram transferidos para meio de multiplicação composto por MS suplementado com 10 mg/L 2,4-D, 5mg/L BAP, 1 g/L carvão ativado, 30 g/L sacarose e gelificado com 3 g/L de Phytagel®. Aos 90 dias de cultivo em meio de multiplicação, os calos embriogênicos foram identificados e selecionados para a fase de maturação dos embriões somáticos sob estresse hídrico induzido com PEG (0 e 2%) na presença de BAP (10 e 15mg/L), e após 60 dias a presença de pró-embriões e embriões em maturação foi observada. A análise dos experimentos foi realizada por meio dos testes não paramétricos utilizando o software R. Constatou-se que foi possível criopreservar os acessos estudados por meio da técnica de vitrificação em gotas. A putrescina não favoreceu o desenvolvimento de calos embriogênicos, e a presença de 2,4-D e BAP propocionou a diferenciação em calos embriogênicos. As combinações 10 ou 15 mg/L de BAP com 2% de PEG em segmentos nodais proporcionaram maior formação de pró-embriões e embriões em maturação no acesso TC. Foi possível avaliar o comportamento de acessos provenientes do BAGMangaba utilizando a técnica de criopreservação; como também definir concentrações de reguladores de crescimento e melhores tipos de explantes para o estabelecimento de um protocolo de embriogênese somática de mangabeira.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESSão CristóvãoporMangabeiraGermoplasma vegetalRecursos do germoplasma vegetalHancornia speciosa GomesVitrificação em gotasEmbriogênese somáticaDroplet vitrificationSomatic embryogenesisCIENCIAS AGRARIASEstratégias para conservação e clonagem in vitro de mangabeirainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisPós-Graduação em Agricultura e BiodiversidadeUniversidade Federal de Sergipe (UFS)reponame:Repositório Institucional da UFSinstname:Universidade Federal de Sergipe (UFS)instacron:UFSinfo:eu-repo/semantics/openAccessORIGINALFERNANDA_VIEIRA_SANTANA.pdfFERNANDA_VIEIRA_SANTANA.pdfapplication/pdf3410194https://ri.ufs.br/jspui/bitstream/riufs/19202/2/FERNANDA_VIEIRA_SANTANA.pdfc2f28b61d6e70b99ca506551073e197aMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81475https://ri.ufs.br/jspui/bitstream/riufs/19202/1/license.txt098cbbf65c2c15e1fb2e49c5d306a44cMD51riufs/192022024-02-29 16:55:34.144oai:ufs.br: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Repositório InstitucionalPUBhttps://ri.ufs.br/oai/requestrepositorio@academico.ufs.bropendoar:2024-02-29T19:55:34Repositório Institucional da UFS - Universidade Federal de Sergipe (UFS)false |
dc.title.pt_BR.fl_str_mv |
Estratégias para conservação e clonagem in vitro de mangabeira |
title |
Estratégias para conservação e clonagem in vitro de mangabeira |
spellingShingle |
Estratégias para conservação e clonagem in vitro de mangabeira Santana, Fernanda Vieira Mangabeira Germoplasma vegetal Recursos do germoplasma vegetal Hancornia speciosa Gomes Vitrificação em gotas Embriogênese somática Droplet vitrification Somatic embryogenesis CIENCIAS AGRARIAS |
title_short |
Estratégias para conservação e clonagem in vitro de mangabeira |
title_full |
Estratégias para conservação e clonagem in vitro de mangabeira |
title_fullStr |
Estratégias para conservação e clonagem in vitro de mangabeira |
title_full_unstemmed |
Estratégias para conservação e clonagem in vitro de mangabeira |
title_sort |
Estratégias para conservação e clonagem in vitro de mangabeira |
author |
Santana, Fernanda Vieira |
author_facet |
Santana, Fernanda Vieira |
author_role |
author |
dc.contributor.author.fl_str_mv |
Santana, Fernanda Vieira |
dc.contributor.advisor1.fl_str_mv |
Ledo, Ana da Silva |
dc.contributor.advisor-co1.fl_str_mv |
Santos, Paulo Augusto Almeida |
contributor_str_mv |
Ledo, Ana da Silva Santos, Paulo Augusto Almeida |
dc.subject.por.fl_str_mv |
Mangabeira Germoplasma vegetal Recursos do germoplasma vegetal Hancornia speciosa Gomes Vitrificação em gotas Embriogênese somática |
topic |
Mangabeira Germoplasma vegetal Recursos do germoplasma vegetal Hancornia speciosa Gomes Vitrificação em gotas Embriogênese somática Droplet vitrification Somatic embryogenesis CIENCIAS AGRARIAS |
dc.subject.eng.fl_str_mv |
Droplet vitrification Somatic embryogenesis |
dc.subject.cnpq.fl_str_mv |
CIENCIAS AGRARIAS |
description |
Hancornia speciosa Gomes is a fruit tree, found in several regions of Brazil, but particularly important in the Northeast region. Due to the wide range of uses of its fruits, this tree species represents a source of income of families that extract the fruits from the natural populations. The genetic resources of mangabeira are mostly conserved by means of field collections, mainly due to the recalcitrant physiology of the seeds. Therefore, cryopreservation techniques and somatic embryogenesis are additionally explored, as alternatives to the conservation of the existing genetic variability and for the vegetative multiplication of promising genotypes. The objectives of this study were to: I – evaluate the efficiency of the droplet vitrification technique for long-term conservation of five mangabeira accessions (Água Boa; Japaratinga; Paratibe; Oiteiro; Terra Caída), and check if these accessions have any level of susceptibility to possible effects of cryopreservation; II – study maturation induction of somatic embryos from nodal and foliar explants of accession Terra Caída (TC) of the Germplasm Bank of Mangaba (BAGMangaba). For cryopreservation, shoot apices were subjected to different periods of exposure (30 and 50 min for test I; 25, 50 and 75 min for test II) to vitrification solution 2 (PVS2), prior to immersion in liquid nitrogen. To evaluate the effects of cryoprotective solutions and exposure times on shoot apices exposed or not to liquid nitrogen, regeneration percentages were recorded after 30 and 60 days of in vitro cultivation. For callus induction, leaf and nodal segments of the TC accession were cultivated in media with different putrescine concentrations (0, 250, 500, 750 and 1000 µM) in MS medium (Murashige & Skoog, 1962). After 120 days, the explants were transferred to a MS multiplication medium supplemented with 10 mg/L 2,4-D, 5mg/L benzylaminopurine (BAP), 1 g/L activated charcoal and 30 g/L sucrose and gelled with 3 g/ L Phytagel®. After 90 days in multiplication medium, embryogenic calli were identified and selected for the maturation phase of somatic embryos under water stress induced with polyethylene glycol (PEG, at 0 and 2%) and enriched with BAP (10 and 15mg/L). After 60 days, pro-embryos and maturing embryos were observed. The experimental results were analyzed by non-parametric tests using R software. It was possible to cryopreserve the studied accessions by droplet vitrification. Putrescine did not favor the development of embryogenic calli, and the presence of 2,4-D and BAP promoted differentiation into embryogenic calli. Nodal segments in combinations of 10 or 15 mg/L BAP with 2% PEG intensified the formation of pro-embryos and maturing embryos of the TC accession. The performance pattern of accessions from BAGMangaba could be evaluated by the cryopreservation technique. In addition, the concentrations of growth regulators and best types of explants for establishing a somatic embryogenesis protocol for mangabeira could be defined. |
publishDate |
2023 |
dc.date.issued.fl_str_mv |
2023-03-24 |
dc.date.accessioned.fl_str_mv |
2024-02-29T19:55:28Z |
dc.date.available.fl_str_mv |
2024-02-29T19:55:28Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
SANTANA, Fernanda Vieira. Estratégias para conservação e clonagem in vitro de mangabeira. 2023. 57 f. Tese (Doutorado em Agricultura e Biodiversidade) - Universidade Federal de Sergipe, São Cristóvão, 2023. |
dc.identifier.uri.fl_str_mv |
https://ri.ufs.br/jspui/handle/riufs/19202 |
identifier_str_mv |
SANTANA, Fernanda Vieira. Estratégias para conservação e clonagem in vitro de mangabeira. 2023. 57 f. Tese (Doutorado em Agricultura e Biodiversidade) - Universidade Federal de Sergipe, São Cristóvão, 2023. |
url |
https://ri.ufs.br/jspui/handle/riufs/19202 |
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Pós-Graduação em Agricultura e Biodiversidade |
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Universidade Federal de Sergipe (UFS) |
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