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. graf
Article
in English
| LILACS
| ID: biblio-839240
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.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Polyesters
/
Azospirillum brasilense
/
Plant Roots
/
Herbaspirillum
/
Flow Cytometry
/
Hydroxybutyrates
Language:
English
Journal:
Braz. j. med. biol. res
Journal subject:
Biology
/
Medicine
Year:
2017
Type:
Article
Affiliation country:
Brazil
Institution/Affiliation country:
Universidade Federal do Paraná/BR
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