Azospirillum brasilense viable cells enumeration using propidium monoazide-quantitative PCR.
Arch Microbiol
; 202(7): 1653-1662, 2020 Sep.
Article
en En
| MEDLINE
| ID: mdl-32274559
ABSTRACT
Azospirillum brasilense is a plant growth promoting bacteria used as an inoculant in diverse crops. Accurate analytical methods are required to enumerate viable cells in inoculant formulations or in planta. We developed a quantitative polymerase chain reaction (qPCR) assay associated to propidium monoazide (PMA) to evaluate the cell viability of A. brasilense in inoculant and in maize roots. A. brasilense was grown in culture medium and was exposed to 50 â. Maize roots were grown in vitro and harvested 7 days after inoculation. Quantification was performed by qPCR, PMA-qPCR, and plate counting. Standard curves efficiency values ranged from 85 to 99%. The limit of detection was 104 CFU per gram of fresh root. Enumeration obtained in maize roots by qPCR where higher than enumeration by PMA-qPCR and by plate counting. PMA-qPCR assay was efficient in quantifying inoculant viable cells and provides reliable results in a quickly and accurately way compared to culture-dependent methods.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Propidio
/
Azidas
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Microbiología Industrial
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Azospirillum brasilense
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Raíces de Plantas
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Viabilidad Microbiana
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Reacción en Cadena en Tiempo Real de la Polimerasa
País/Región como asunto:
America do sul
/
Brasil
Idioma:
En
Año:
2020
Tipo del documento:
Article