A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry.
Braz J Med Biol Res
; 50(1): e5492, 2017 Jan 16.
Article
em En
| MEDLINE
| ID: mdl-28099582
ABSTRACT
The conventional method for quantification of polyhydroxyalkanoates based on whole-cell methanolysis and gas chromatography (GC) is laborious and time-consuming. In this work, a method based on flow cytometry of Nile red stained bacterial cells was established to quantify poly-3-hydroxybutyrate (PHB) production by the diazotrophic and plant-associated bacteria, Herbaspirillum seropedicae and Azospirillum brasilense. The method consists of three steps:
i) cell permeabilization, ii) Nile red staining, and iii) analysis by flow cytometry. The method was optimized step-by-step and can be carried out in less than 5 min. The final results indicated a high correlation coefficient (R2=0.99) compared to a standard method based on methanolysis and GC. This method was successfully applied to the quantification of PHB in epiphytic bacteria isolated from rice roots.
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Base de dados:
MEDLINE
Assunto principal:
Poliésteres
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Azospirillum brasilense
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Raízes de Plantas
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Herbaspirillum
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Citometria de Fluxo
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Hidroxibutiratos
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article