In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria

Detalhes bibliográficos
Autor(a) principal: Pasqualini, Ana Paula de Azevedo
Data de Publicação: 2019
Outros Autores: Schneider, Gabriela Xavier, Fraga, Hugo Pacheco de Freitas, Biasi, Luiz Antonio, Quoirin, Marguerite
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Pesquisa Agropecuária Tropical (Online)
Texto Completo: https://revistas.ufg.br/pat/article/view/53673
Resumo: In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test. KEYWORDS: Ralstonia, bamboo, micropropagation.
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spelling In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteriaEstabelecimento in vitro de Bambusa oldhamii Munro a partir de matrizes cultivadas a campo e identificação molecular de bactéria endofíticaIn plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test. KEYWORDS: Ralstonia, bamboo, micropropagation.Na micropropagação de plantas, a etapa de estabelecimento é dificultada pela presença de micro-organismos em tecidos oriundos de matrizes do campo, especialmente para bambu. Objetivou-se estabelecer um protocolo eficiente de assepsia de explantes de Bambusa oldhamii oriundos de plantas em campo e efetuar a identificação molecular de um possível isolado bacteriano endofítico. Os explantes foram expostos subsequentemente a álcool 70 %, hipoclorito de sódio 1 % (NaOCl), cloreto de mercúrio 0,1 % (HgCl2), tiofanato-metílico (Cercobin®) e digliconato de clorexidina (Riohex® 2 %), em diferentes combinações, e introduzidos em meio Murashige and Skoog (sólido ou líquido), acrescido ou não de 4 mL L-1 de Plant Preservative Mixture (PPMTM), em sete tratamentos. A assepsia e imersão dos explantes no meio de cultura líquido contendo 4 mL L-1 de PPMTM inibiram visualmente o crescimento de bactérias e fungos, permitiram o desenvolvimento de brotações com comprimento médio de 2,2 cm e posteriores subcultivos, sendo o melhor tratamento usado para o estabelecimento in vitro de B. oldhamii. A identificação molecular da bactéria por meio do sequenciamento do 16S rDNA permitiu identificar o isolado bacteriano como Ralstonia sp., com 100 % de similaridade, e as análises filogenéticas o agruparam com Ralstonia pickettii. Além disso, a bactéria se mostrou sensível a 4 mL L-1 de PPMTM pelo teste de concentração mínima inibitória. PALAVRAS-CHAVE: Ralstonia, bambu, micropropagação.Escola de Agronomia - Universidade Federal de Goiás2019-03-28info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistas.ufg.br/pat/article/view/53673Pesquisa Agropecuária Tropical [Agricultural Research in the Tropics]; Vol. 49 (2019); e53673Pesquisa Agropecuária Tropical (Agricultural Research in the Tropics); Vol. 49 (2019); e53673Pesquisa Agropecuária Tropical; v. 49 (2019); e536731983-4063reponame:Pesquisa Agropecuária Tropical (Online)instname:Universidade Federal de Goiás (UFG)instacron:UFGenghttps://revistas.ufg.br/pat/article/view/53673/32881Pasqualini, Ana Paula de AzevedoSchneider, Gabriela XavierFraga, Hugo Pacheco de FreitasBiasi, Luiz AntonioQuoirin, Margueriteinfo:eu-repo/semantics/openAccess2020-03-02T15:42:43Zoai:ojs.revistas.ufg.br:article/53673Revistahttps://revistas.ufg.br/patPUBhttps://revistas.ufg.br/pat/oaiaseleguini.pat@gmail.com||mgoes@agro.ufg.br1983-40631517-6398opendoar:2024-05-21T19:56:22.965499Pesquisa Agropecuária Tropical (Online) - Universidade Federal de Goiás (UFG)true
dc.title.none.fl_str_mv In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
Estabelecimento in vitro de Bambusa oldhamii Munro a partir de matrizes cultivadas a campo e identificação molecular de bactéria endofítica
title In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
spellingShingle In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
Pasqualini, Ana Paula de Azevedo
title_short In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
title_full In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
title_fullStr In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
title_full_unstemmed In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
title_sort In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
author Pasqualini, Ana Paula de Azevedo
author_facet Pasqualini, Ana Paula de Azevedo
Schneider, Gabriela Xavier
Fraga, Hugo Pacheco de Freitas
Biasi, Luiz Antonio
Quoirin, Marguerite
author_role author
author2 Schneider, Gabriela Xavier
Fraga, Hugo Pacheco de Freitas
Biasi, Luiz Antonio
Quoirin, Marguerite
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Pasqualini, Ana Paula de Azevedo
Schneider, Gabriela Xavier
Fraga, Hugo Pacheco de Freitas
Biasi, Luiz Antonio
Quoirin, Marguerite
description In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test. KEYWORDS: Ralstonia, bamboo, micropropagation.
publishDate 2019
dc.date.none.fl_str_mv 2019-03-28
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://revistas.ufg.br/pat/article/view/53673
url https://revistas.ufg.br/pat/article/view/53673
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://revistas.ufg.br/pat/article/view/53673/32881
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 Escola de Agronomia - Universidade Federal de Goiás
publisher.none.fl_str_mv Escola de Agronomia - Universidade Federal de Goiás
dc.source.none.fl_str_mv Pesquisa Agropecuária Tropical [Agricultural Research in the Tropics]; Vol. 49 (2019); e53673
Pesquisa Agropecuária Tropical (Agricultural Research in the Tropics); Vol. 49 (2019); e53673
Pesquisa Agropecuária Tropical; v. 49 (2019); e53673
1983-4063
reponame:Pesquisa Agropecuária Tropical (Online)
instname:Universidade Federal de Goiás (UFG)
instacron:UFG
instname_str Universidade Federal de Goiás (UFG)
instacron_str UFG
institution UFG
reponame_str Pesquisa Agropecuária Tropical (Online)
collection Pesquisa Agropecuária Tropical (Online)
repository.name.fl_str_mv Pesquisa Agropecuária Tropical (Online) - Universidade Federal de Goiás (UFG)
repository.mail.fl_str_mv aseleguini.pat@gmail.com||mgoes@agro.ufg.br
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