In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bacteria
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , |
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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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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1799874820039507968 |