Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy

The potential of Pseudomonas putida KT2440 to act as a plant-growth promoter or as a bior-emediator of toxic compounds can be affected by desiccation. In the present work, the bacterial survival ratio (BSR) in response to air desiccation was evaluated for P. putida KT2440 in the presence of differen...

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Autores: Pazos-Rojas, L. A., Muñoz-Arenas, L. C., Rodríguez-Andrade, O., López-Cruz, L. E., López-Ortega, O., Lopes-Olivares, F., Luna-Suarez, S., Baez, Antonio, Morales-García, Y. E., Quintero-Hernández, V., Villalobos-López, M. A., Torre Zúñiga, Jesús de la, Muñoz-Rojas, Jesús
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/193117
Acceso en línea:http://hdl.handle.net/10261/193117
Access Level:acceso abierto
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spelling Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategyPazos-Rojas, L. A.Muñoz-Arenas, L. C.Rodríguez-Andrade, O.López-Cruz, L. E.López-Ortega, O.Lopes-Olivares, F.Luna-Suarez, S.Baez, AntonioMorales-García, Y. E.Quintero-Hernández, V.Villalobos-López, M. A.Torre Zúñiga, Jesús de laMuñoz-Rojas, JesúsThe potential of Pseudomonas putida KT2440 to act as a plant-growth promoter or as a bior-emediator of toxic compounds can be affected by desiccation. In the present work, the bacterial survival ratio (BSR) in response to air desiccation was evaluated for P. putida KT2440 in the presence of different protectors. The BSR in the presence of nonreducing disaccharides, such as trehalose, was high after 15 days of desiccation stress (occurring at 30C and 50% relative humidity), whereas in the absence of a protector the bacterial counts diminished to nondetectable numbers (ca 2.8 log CFU/mL). The LIVE/DEAD staining method showed that bacteria protected with trehalose maintained increased numbers of green cells after desiccation while cells without protection were all observed to be red. This indicated that nonprotected bacteria had compromised membrane integrity. However, when nonprotected bacteria subjected to 18 days of desiccation stress were rehydrated for a short time with maize root exudates or for 48 h with water (prolonged rehydration), the bacterial counts were as high as that observed for those not subjected to desiccation stress, suggesting that the cells entered the viable but nonculturable (VBNC) state under desiccation and that they returned to a culturable state after those means of rehydration. Interestingly an increase in the green color intensity of cells that returned to a culturable state was observed using LIVE/ DEAD staining method, indicating an improvement in their membrane integrity. Cellular activity in the VBNC state was determined. A GFP-tagged P. putida strain expressing GFP constitutively was subjected to desiccation. After 12 days of desiccation, the GFP-tagged strain lost culturability, but it exhibited active GFP expression, which in turn made the cells green. Furthermore, the expression of 16S rRNA, rpoN (housekeeping), mutL, mutS (encoding proteins from the mismatch repair complex), and oprH (encoding an outer membrane protein) were examined by RT-PCR. All evaluated genes were expressed by both types of cells, culturable and nonculturable, indicating active molecular processes during the VBNC state.This work is supported by: JMR: Apoyo Redes PRODEP 2015-2016 (CA-262 and CA-244); JMR: DITCo2016-3, JMR: DITCo2016-4; VIEP-BUAP-2018 (VQH: COL525704-VIEP2018; ABR: 100524554-VIEP2018, JMR: 100425788-VIEP2018). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer ReviewedPublic Library of ScienceConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2019201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/193117reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0219554Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1931172026-05-22T06:33:51Z
dc.title.none.fl_str_mv Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
title Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
spellingShingle Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
Pazos-Rojas, L. A.
title_short Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
title_full Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
title_fullStr Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
title_full_unstemmed Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
title_sort Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy
dc.creator.none.fl_str_mv Pazos-Rojas, L. A.
Muñoz-Arenas, L. C.
Rodríguez-Andrade, O.
López-Cruz, L. E.
López-Ortega, O.
Lopes-Olivares, F.
Luna-Suarez, S.
Baez, Antonio
Morales-García, Y. E.
Quintero-Hernández, V.
Villalobos-López, M. A.
Torre Zúñiga, Jesús de la
Muñoz-Rojas, Jesús
author Pazos-Rojas, L. A.
author_facet Pazos-Rojas, L. A.
Muñoz-Arenas, L. C.
Rodríguez-Andrade, O.
López-Cruz, L. E.
López-Ortega, O.
Lopes-Olivares, F.
Luna-Suarez, S.
Baez, Antonio
Morales-García, Y. E.
Quintero-Hernández, V.
Villalobos-López, M. A.
Torre Zúñiga, Jesús de la
Muñoz-Rojas, Jesús
author_role author
author2 Muñoz-Arenas, L. C.
Rodríguez-Andrade, O.
López-Cruz, L. E.
López-Ortega, O.
Lopes-Olivares, F.
Luna-Suarez, S.
Baez, Antonio
Morales-García, Y. E.
Quintero-Hernández, V.
Villalobos-López, M. A.
Torre Zúñiga, Jesús de la
Muñoz-Rojas, Jesús
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
description The potential of Pseudomonas putida KT2440 to act as a plant-growth promoter or as a bior-emediator of toxic compounds can be affected by desiccation. In the present work, the bacterial survival ratio (BSR) in response to air desiccation was evaluated for P. putida KT2440 in the presence of different protectors. The BSR in the presence of nonreducing disaccharides, such as trehalose, was high after 15 days of desiccation stress (occurring at 30C and 50% relative humidity), whereas in the absence of a protector the bacterial counts diminished to nondetectable numbers (ca 2.8 log CFU/mL). The LIVE/DEAD staining method showed that bacteria protected with trehalose maintained increased numbers of green cells after desiccation while cells without protection were all observed to be red. This indicated that nonprotected bacteria had compromised membrane integrity. However, when nonprotected bacteria subjected to 18 days of desiccation stress were rehydrated for a short time with maize root exudates or for 48 h with water (prolonged rehydration), the bacterial counts were as high as that observed for those not subjected to desiccation stress, suggesting that the cells entered the viable but nonculturable (VBNC) state under desiccation and that they returned to a culturable state after those means of rehydration. Interestingly an increase in the green color intensity of cells that returned to a culturable state was observed using LIVE/ DEAD staining method, indicating an improvement in their membrane integrity. Cellular activity in the VBNC state was determined. A GFP-tagged P. putida strain expressing GFP constitutively was subjected to desiccation. After 12 days of desiccation, the GFP-tagged strain lost culturability, but it exhibited active GFP expression, which in turn made the cells green. Furthermore, the expression of 16S rRNA, rpoN (housekeeping), mutL, mutS (encoding proteins from the mismatch repair complex), and oprH (encoding an outer membrane protein) were examined by RT-PCR. All evaluated genes were expressed by both types of cells, culturable and nonculturable, indicating active molecular processes during the VBNC state.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/193117
url http://hdl.handle.net/10261/193117
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0219554
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
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