Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods

Detalhes bibliográficos
Autor(a) principal: Frizzo, Caroline
Data de Publicação: 2018
Outros Autores: Quoirin, Marguerite
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Pesquisa Florestal Brasileira (Online)
Texto Completo: https://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/1498
Resumo: Araucaria angustifolia (Bertol.) O. Kuntze is one of the most important native species in Southern Brazil. However, its naturally recalcitrant seeds represent obstacles for long-term conservation and thus cryopreservation is a viable alternative for germplasm storing. Embryonic axes (EA) excised from araucaria seeds were encapsulated, dehydrated, and submitted to two cryopreservation methods: flash-cooling by freezing in liquid nitrogen (LN) for 2 h and pre-cooling at – 40 °C, followed by freezing in LN for 2 h. Subsequently, the EA were quickly thawed and assessed for DNA integrity, tetrazolium test, in vitro germination and oxidation occurrence. The DNA of both not cryopreserved and cryopreserved EA maintained their integrity. The tetrazolium test results indicated that the majority of flash-cooled EA were viable. After 15 days of in vitro culture, the EA did not germinate and presented signs of oxidation. Dehydration method by direct plunge in LN is promising for cryopreservation of araucaria EA, as demonstrated through the results of tetrazolium test and the maintenance of total DNA integrity.
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spelling Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methodsViabilidade de eixos embrionários de Araucaria angustifolia após o uso de dois métodos de criopreservaçãoDehydrationEncapsulationLiquid nitrogenDesidrataçãoEncapsulamentoNitrogênio líquidoAraucaria angustifolia (Bertol.) O. Kuntze is one of the most important native species in Southern Brazil. However, its naturally recalcitrant seeds represent obstacles for long-term conservation and thus cryopreservation is a viable alternative for germplasm storing. Embryonic axes (EA) excised from araucaria seeds were encapsulated, dehydrated, and submitted to two cryopreservation methods: flash-cooling by freezing in liquid nitrogen (LN) for 2 h and pre-cooling at – 40 °C, followed by freezing in LN for 2 h. Subsequently, the EA were quickly thawed and assessed for DNA integrity, tetrazolium test, in vitro germination and oxidation occurrence. The DNA of both not cryopreserved and cryopreserved EA maintained their integrity. The tetrazolium test results indicated that the majority of flash-cooled EA were viable. After 15 days of in vitro culture, the EA did not germinate and presented signs of oxidation. Dehydration method by direct plunge in LN is promising for cryopreservation of araucaria EA, as demonstrated through the results of tetrazolium test and the maintenance of total DNA integrity.Araucaria angustifolia (Bertol.) O. Kuntze é uma das espécies nativas do Brasil com maior importância na Região Sul. No entanto, suas sementes são recalcitrantes, constituindo um obstáculo para a sua conservação em longo prazo, sendo a criopreservação uma alternativa viável para o armazenamento de seu germoplasma. Os eixos embrionários (EE) excisados das sementes de araucária foram encapsulados, desidratados e submetidos a dois métodos de criopreservação: o resfriamento rápido, congelando em nitrogênio líquido (NL) durante 2 h, e o pré-resfriamento a - 40 °C, seguido do congelamento em nitrogênio líquido durante 2 h. Posteriormente, os EE foram descongelados rapidamente e avaliados quanto à integridade do DNA pelo teste bioquímico do tetrazólio, germinação in vitro e ocorrência de oxidação. Tanto o DNA dos EE que não foram criopreservados quanto o daqueles que foram criopreservados mantiveram sua integridade. Os resultados do teste de tetrazólio indicaram que a maioria dos EE desidratados pelo congelamento rápido permaneceu viável. Após 15 dias de cultivo in vitro, os EE não germinaram e apresentaram sinais de oxidação. Os métodos de encapsulamento-desidratação seguidos de congelamento rápido são promissores para a criopreservação de EE de araucária, como demonstrado pelo teste de tetrazólio e da manutenção da integridade total do DNA.Embrapa Florestas2018-12-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/149810.4336/2018.pfb.38e201701498Pesquisa Florestal Brasileira; v. 38 (2018)Pesquisa Florestal Brasileira; Vol. 38 (2018)1983-26051809-3647reponame:Pesquisa Florestal Brasileira (Online)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPAenghttps://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/1498/748https://creativecommons.org/licenses/by-nc-nd/4.0info:eu-repo/semantics/openAccessFrizzo, CarolineQuoirin, Marguerite2019-05-10T19:43:45Zoai:pfb.cnpf.embrapa.br/pfb:article/1498Revistahttps://pfb.cnpf.embrapa.br/pfb/index.php/pfb/PUBhttps://pfb.cnpf.embrapa.br/pfb/index.php/pfb/oaipfb@embrapa.br || revista.pfb@gmail.com || patricia.mattos@embrapa.br1983-26051809-3647opendoar:2019-05-10T19:43:45Pesquisa Florestal Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false
dc.title.none.fl_str_mv Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
Viabilidade de eixos embrionários de Araucaria angustifolia após o uso de dois métodos de criopreservação
title Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
spellingShingle Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
Frizzo, Caroline
Dehydration
Encapsulation
Liquid nitrogen
Desidratação
Encapsulamento
Nitrogênio líquido
title_short Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
title_full Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
title_fullStr Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
title_full_unstemmed Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
title_sort Viability of embryonic axes of Araucaria angustifolia after freezing using two cryopreservation methods
author Frizzo, Caroline
author_facet Frizzo, Caroline
Quoirin, Marguerite
author_role author
author2 Quoirin, Marguerite
author2_role author
dc.contributor.author.fl_str_mv Frizzo, Caroline
Quoirin, Marguerite
dc.subject.por.fl_str_mv Dehydration
Encapsulation
Liquid nitrogen
Desidratação
Encapsulamento
Nitrogênio líquido
topic Dehydration
Encapsulation
Liquid nitrogen
Desidratação
Encapsulamento
Nitrogênio líquido
description Araucaria angustifolia (Bertol.) O. Kuntze is one of the most important native species in Southern Brazil. However, its naturally recalcitrant seeds represent obstacles for long-term conservation and thus cryopreservation is a viable alternative for germplasm storing. Embryonic axes (EA) excised from araucaria seeds were encapsulated, dehydrated, and submitted to two cryopreservation methods: flash-cooling by freezing in liquid nitrogen (LN) for 2 h and pre-cooling at – 40 °C, followed by freezing in LN for 2 h. Subsequently, the EA were quickly thawed and assessed for DNA integrity, tetrazolium test, in vitro germination and oxidation occurrence. The DNA of both not cryopreserved and cryopreserved EA maintained their integrity. The tetrazolium test results indicated that the majority of flash-cooled EA were viable. After 15 days of in vitro culture, the EA did not germinate and presented signs of oxidation. Dehydration method by direct plunge in LN is promising for cryopreservation of araucaria EA, as demonstrated through the results of tetrazolium test and the maintenance of total DNA integrity.
publishDate 2018
dc.date.none.fl_str_mv 2018-12-07
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://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/1498
10.4336/2018.pfb.38e201701498
url https://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/1498
identifier_str_mv 10.4336/2018.pfb.38e201701498
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://pfb.cnpf.embrapa.br/pfb/index.php/pfb/article/view/1498/748
dc.rights.driver.fl_str_mv https://creativecommons.org/licenses/by-nc-nd/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Embrapa Florestas
publisher.none.fl_str_mv Embrapa Florestas
dc.source.none.fl_str_mv Pesquisa Florestal Brasileira; v. 38 (2018)
Pesquisa Florestal Brasileira; Vol. 38 (2018)
1983-2605
1809-3647
reponame:Pesquisa Florestal 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 Florestal Brasileira (Online)
collection Pesquisa Florestal Brasileira (Online)
repository.name.fl_str_mv Pesquisa Florestal Brasileira (Online) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
repository.mail.fl_str_mv pfb@embrapa.br || revista.pfb@gmail.com || patricia.mattos@embrapa.br
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