Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia

Detalhes bibliográficos
Autor(a) principal: Timoteo, Caroline de Oliveira
Data de Publicação: 2022
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFLA
Texto Completo: http://repositorio.ufla.br/jspui/handle/1/49854
Resumo: Coffee is one of the most popular beverages in the world, comprising the basis of economy for several tropical countries, including Brazil. Coffee production is mainly based on Coffea arabica species, preferred by consumers due to the quality of the drink. Given the importance of coffee to the world economy, it is essential that genetic resources are conserved in the long term. Considering that arabica coffee is propagated mainly by seeds and reproduction occurs predominantly by self-pollination, seeds or embryonic axes are ideal for conserving the genetic resources. In this context, the objective of this study was to establish a protocol for the cryopreservation of zygotic embryos of cultivars of Coffea arabica (IAC 62 and IAC 144) through the techniques of encapsulation-dehydration and encapsulation-vitrification. To start this process, the zygotic embryos were extracted from the seeds and encapsulated in a sodium alginate matrix and culture medium. In the encapsulation-dehydration technique, after pre-culture, the encapsulated zygotic embryos were dehydrated and immediately immersed in liquid nitrogen (-196 °C) or inoculated in a culture medium. For the encapsulation-vitrification technique, after pre-culture, the encapsulated zygotic embryos were immersed in Loading Solution and subsequently immersed in Plant vitrification solution 2 (PVS2) and immersed in liquid nitrogen or inoculated in culture medium. Evaluations were carried out regarding seedling length, weight, and number of leaves, anatomical and biochemical analyzes regarding the activities of antioxidant metabolism enzymes (SOD, POD, CAT and APX), quantification of hydrogen peroxide, lipid peroxidation and proline content. After the regeneration of the seedlings formed by the development of cryopreserved zygotic embryos, acclimatization was performed in a hydroponic system with substrate. The results indicated that the zygotic embryos were able to survive and regenerate after the cryopreservation process when dehydrated after 8 hours. Seedlings regenerated from cryopreserved zygotic embryos showed statistically similar growth to seedlings whose zygotic embryos were not cryopreserved. The anatomical evaluations showed a retraction in the volume of the fundamental meristem cells because of the dehydration process and the biochemical analyzes indicated a higher content of proline and activity of SOD and POD enzymes. Acclimatized plants originated from zygotic embryos that were cryopreserved reached the same phenotypics characteristics of plants whose embryos were not cryopreserved. Regarding the encapsulation-vitrification technique, the zygotic embryos were not able to survive the freezing process. Thus, it is concluded that it is possible to cryopreserve zygotic embryos of arabica coffee cultivars through the encapsulation-dehydration technique, considering that the moisture content is a key factor for the survival of the explants and that the acclimatization can be carried out through of the hydroponic system with substrate.
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spelling Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponiaCoffea arabica L. zygotic embryos cryopreserved by encapsulation-dehydration and acclimatized by hydroponicsCafé - SementesSementes - CriopreservaçãoSistemas hidropônicosSemi-hidroponiaSistema antioxidanteProlinaCultivo in vitroCoffee - SeedsSeeds - CryopreservationHydroponic systemsSemi-hydroponicsAntioxidant systemProlineIn vitro cultivationFisiologia VegetalCoffee is one of the most popular beverages in the world, comprising the basis of economy for several tropical countries, including Brazil. Coffee production is mainly based on Coffea arabica species, preferred by consumers due to the quality of the drink. Given the importance of coffee to the world economy, it is essential that genetic resources are conserved in the long term. Considering that arabica coffee is propagated mainly by seeds and reproduction occurs predominantly by self-pollination, seeds or embryonic axes are ideal for conserving the genetic resources. In this context, the objective of this study was to establish a protocol for the cryopreservation of zygotic embryos of cultivars of Coffea arabica (IAC 62 and IAC 144) through the techniques of encapsulation-dehydration and encapsulation-vitrification. To start this process, the zygotic embryos were extracted from the seeds and encapsulated in a sodium alginate matrix and culture medium. In the encapsulation-dehydration technique, after pre-culture, the encapsulated zygotic embryos were dehydrated and immediately immersed in liquid nitrogen (-196 °C) or inoculated in a culture medium. For the encapsulation-vitrification technique, after pre-culture, the encapsulated zygotic embryos were immersed in Loading Solution and subsequently immersed in Plant vitrification solution 2 (PVS2) and immersed in liquid nitrogen or inoculated in culture medium. Evaluations were carried out regarding seedling length, weight, and number of leaves, anatomical and biochemical analyzes regarding the activities of antioxidant metabolism enzymes (SOD, POD, CAT and APX), quantification of hydrogen peroxide, lipid peroxidation and proline content. After the regeneration of the seedlings formed by the development of cryopreserved zygotic embryos, acclimatization was performed in a hydroponic system with substrate. The results indicated that the zygotic embryos were able to survive and regenerate after the cryopreservation process when dehydrated after 8 hours. Seedlings regenerated from cryopreserved zygotic embryos showed statistically similar growth to seedlings whose zygotic embryos were not cryopreserved. The anatomical evaluations showed a retraction in the volume of the fundamental meristem cells because of the dehydration process and the biochemical analyzes indicated a higher content of proline and activity of SOD and POD enzymes. Acclimatized plants originated from zygotic embryos that were cryopreserved reached the same phenotypics characteristics of plants whose embryos were not cryopreserved. Regarding the encapsulation-vitrification technique, the zygotic embryos were not able to survive the freezing process. Thus, it is concluded that it is possible to cryopreserve zygotic embryos of arabica coffee cultivars through the encapsulation-dehydration technique, considering that the moisture content is a key factor for the survival of the explants and that the acclimatization can be carried out through of the hydroponic system with substrate.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)O café é uma das bebidas mais populares do mundo, compreendendo a base da economia de vários países tropicais, incluindo o Brasil. A produção cafeeira é baseada principalmente na espécie Coffea arabica, preferida pelos consumidores devido a qualidade da bebida. Dada a importância do café para a economia mundial, é indispensável que os recursos genéticos dessa cultura sejam conservados em longo prazo. Tendo em vista que o café arábica é propagado principalmente por sementes e que a reprodução ocorre predominantemente por autopolinização, as sementes ou eixos embrionários são ideais para conservação dos recursos genéticos dessa espécie. Nesse contexto o objetivo desse estudo foi estabelecer um protocolo para a criopreservação de embriões zigóticos de cultivares de Coffea arabica, (IAC 62 e IAC 144) por meio das técnicas de encapsulation-dehydration e encapsulation-vitrification. Para iniciar esse processo os embriões zigóticos foram extraídos das sementes e encapsulados por uma matriz de alginato de sódio e meio de cultivo. Na técnica de encapsulation-dehydration, após o pré-cultivo, os embriões zigóticos encapsulados foram desidratados e imersos imediatamente em nitrogênio líquido (-196 °C) ou inoculados em meio de cultivo. Para a técnica de encapsulation-vitrification, após o pré-cultivo, os embriões zigóticos encapsulados foram imersos na Loading Solution e posteriormente foram imersos em solução de Plant vitrification solution 2 (PVS2) e imersos em nitrogênio líquido ou inoculados em meio de cultivo. Foram realizadas avalições quanto ao comprimento das plântulas, peso e número de folhas, análises anatômicas e bioquímicas, quanto as atividades de enzimas do metabolismo antioxidante (SOD, POD, CAT e APX), quantificação de peróxido de hidrogênio, peroxidação lipídica e teor de prolina. Após a regeneração das plântulas formadas pelo desenvolvimento dos embriões zigóticos criopreservados foi realizada a aclimatização em sistema hidropônico com substrato. Os resultados indicaram que os embriões zigóticos foram capazes de sobreviver e regenerar após o processo de criopreservação quando desidratados a partir de 8 horas. As plântulas regeneradas de embriões zigóticos criopreservados apresentaram crescimento estatisticamente semelhante ao de plântulas cujos embriões zigóticos não foram criopreservados. As avaliações anatômicas demonstraram uma retração do volume das células do meristema fundamental em decorrência do processo de desidratação e as análises bioquímicas indicaram maior teor de prolina e atividade das enzimas SOD e POD. As plantas aclimatizadas originadas de embriões zigóticos que foram criopreservados atingiram as mesmas características fenotípicas de plantas cujos embriões não foram criopreservados. Em relação a técnica de encapsulation-vitrification, os embriões zigóticos não foram capazes de sobreviver ao processo de congelamento. Dessa forma conclui-se que é possível criopreservar embriões zigóticos de cultivares de café arábica por meio da técnica de encapsulation-dehydration, tendo em vista que o teor de umidade é fator chave para a sobrevivência dos explantes e que a aclimatização pode ser realizada por meio do sistema hidropônico com substrato.Universidade Federal de LavrasPrograma de Pós-Graduação em Agronomia/Fisiologia VegetalUFLAbrasilDepartamento de AgriculturaPaiva, RenatoReis, Michele Valquíria dosSilva, Diogo Pedrosa Corrêa daReis, Michele Valquíria dosPrudente, Débora de OliveiraHerrera, Raírys CravoRosa, Sttela Dellyzete Veiga Franco daTimoteo, Caroline de Oliveira2022-05-03T19:45:41Z2022-05-03T19:45:41Z2022-05-032022-02-18info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfTIMOTEO, C. de O. Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia. 2022. 87 p. Tese (Doutorado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2022.http://repositorio.ufla.br/jspui/handle/1/49854porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFLAinstname:Universidade Federal de Lavras (UFLA)instacron:UFLA2023-05-09T13:31:29Zoai:localhost:1/49854Repositório InstitucionalPUBhttp://repositorio.ufla.br/oai/requestnivaldo@ufla.br || repositorio.biblioteca@ufla.bropendoar:2023-05-09T13:31:29Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)false
dc.title.none.fl_str_mv Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
Coffea arabica L. zygotic embryos cryopreserved by encapsulation-dehydration and acclimatized by hydroponics
title Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
spellingShingle Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
Timoteo, Caroline de Oliveira
Café - Sementes
Sementes - Criopreservação
Sistemas hidropônicos
Semi-hidroponia
Sistema antioxidante
Prolina
Cultivo in vitro
Coffee - Seeds
Seeds - Cryopreservation
Hydroponic systems
Semi-hydroponics
Antioxidant system
Proline
In vitro cultivation
Fisiologia Vegetal
title_short Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
title_full Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
title_fullStr Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
title_full_unstemmed Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
title_sort Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia
author Timoteo, Caroline de Oliveira
author_facet Timoteo, Caroline de Oliveira
author_role author
dc.contributor.none.fl_str_mv Paiva, Renato
Reis, Michele Valquíria dos
Silva, Diogo Pedrosa Corrêa da
Reis, Michele Valquíria dos
Prudente, Débora de Oliveira
Herrera, Raírys Cravo
Rosa, Sttela Dellyzete Veiga Franco da
dc.contributor.author.fl_str_mv Timoteo, Caroline de Oliveira
dc.subject.por.fl_str_mv Café - Sementes
Sementes - Criopreservação
Sistemas hidropônicos
Semi-hidroponia
Sistema antioxidante
Prolina
Cultivo in vitro
Coffee - Seeds
Seeds - Cryopreservation
Hydroponic systems
Semi-hydroponics
Antioxidant system
Proline
In vitro cultivation
Fisiologia Vegetal
topic Café - Sementes
Sementes - Criopreservação
Sistemas hidropônicos
Semi-hidroponia
Sistema antioxidante
Prolina
Cultivo in vitro
Coffee - Seeds
Seeds - Cryopreservation
Hydroponic systems
Semi-hydroponics
Antioxidant system
Proline
In vitro cultivation
Fisiologia Vegetal
description Coffee is one of the most popular beverages in the world, comprising the basis of economy for several tropical countries, including Brazil. Coffee production is mainly based on Coffea arabica species, preferred by consumers due to the quality of the drink. Given the importance of coffee to the world economy, it is essential that genetic resources are conserved in the long term. Considering that arabica coffee is propagated mainly by seeds and reproduction occurs predominantly by self-pollination, seeds or embryonic axes are ideal for conserving the genetic resources. In this context, the objective of this study was to establish a protocol for the cryopreservation of zygotic embryos of cultivars of Coffea arabica (IAC 62 and IAC 144) through the techniques of encapsulation-dehydration and encapsulation-vitrification. To start this process, the zygotic embryos were extracted from the seeds and encapsulated in a sodium alginate matrix and culture medium. In the encapsulation-dehydration technique, after pre-culture, the encapsulated zygotic embryos were dehydrated and immediately immersed in liquid nitrogen (-196 °C) or inoculated in a culture medium. For the encapsulation-vitrification technique, after pre-culture, the encapsulated zygotic embryos were immersed in Loading Solution and subsequently immersed in Plant vitrification solution 2 (PVS2) and immersed in liquid nitrogen or inoculated in culture medium. Evaluations were carried out regarding seedling length, weight, and number of leaves, anatomical and biochemical analyzes regarding the activities of antioxidant metabolism enzymes (SOD, POD, CAT and APX), quantification of hydrogen peroxide, lipid peroxidation and proline content. After the regeneration of the seedlings formed by the development of cryopreserved zygotic embryos, acclimatization was performed in a hydroponic system with substrate. The results indicated that the zygotic embryos were able to survive and regenerate after the cryopreservation process when dehydrated after 8 hours. Seedlings regenerated from cryopreserved zygotic embryos showed statistically similar growth to seedlings whose zygotic embryos were not cryopreserved. The anatomical evaluations showed a retraction in the volume of the fundamental meristem cells because of the dehydration process and the biochemical analyzes indicated a higher content of proline and activity of SOD and POD enzymes. Acclimatized plants originated from zygotic embryos that were cryopreserved reached the same phenotypics characteristics of plants whose embryos were not cryopreserved. Regarding the encapsulation-vitrification technique, the zygotic embryos were not able to survive the freezing process. Thus, it is concluded that it is possible to cryopreserve zygotic embryos of arabica coffee cultivars through the encapsulation-dehydration technique, considering that the moisture content is a key factor for the survival of the explants and that the acclimatization can be carried out through of the hydroponic system with substrate.
publishDate 2022
dc.date.none.fl_str_mv 2022-05-03T19:45:41Z
2022-05-03T19:45:41Z
2022-05-03
2022-02-18
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.uri.fl_str_mv TIMOTEO, C. de O. Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia. 2022. 87 p. Tese (Doutorado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2022.
http://repositorio.ufla.br/jspui/handle/1/49854
identifier_str_mv TIMOTEO, C. de O. Embriões zigóticos de Coffea arabica L. criopreservados por encapsulation-dehydration e aclimatizados por hidroponia. 2022. 87 p. Tese (Doutorado em Agronomia/Fisiologia Vegetal) – Universidade Federal de Lavras, Lavras, 2022.
url http://repositorio.ufla.br/jspui/handle/1/49854
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dc.publisher.none.fl_str_mv Universidade Federal de Lavras
Programa de Pós-Graduação em Agronomia/Fisiologia Vegetal
UFLA
brasil
Departamento de Agricultura
publisher.none.fl_str_mv Universidade Federal de Lavras
Programa de Pós-Graduação em Agronomia/Fisiologia Vegetal
UFLA
brasil
Departamento de Agricultura
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collection Repositório Institucional da UFLA
repository.name.fl_str_mv Repositório Institucional da UFLA - Universidade Federal de Lavras (UFLA)
repository.mail.fl_str_mv nivaldo@ufla.br || repositorio.biblioteca@ufla.br
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