In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bactéria
ABSTRACT 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 carr...
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Veröffentlicht in: | Pesquisa agropecuária tropical 2019-01, Vol.49 |
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Zusammenfassung: | ABSTRACT 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.
RESUMO 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 p |
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ISSN: | 1517-6398 1983-4063 1983-4063 |
DOI: | 10.1590/1983-40632019v4953673 |