Desiccation-induced viable but nonculturable state in Pseudomonas putida KT2440, a survival strategy.
Colony Count, Microbial
Desiccation
Green Fluorescent Proteins
/ metabolism
Humidity
Microscopy, Fluorescence
Oligonucleotides
Plant Roots
/ microbiology
Pseudomonas putida
/ physiology
RNA, Ribosomal
/ metabolism
RNA, Ribosomal, 16S
/ metabolism
Rhizosphere
Temperature
Trehalose
Zea mays
/ microbiology
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
04
01
2019
accepted:
26
06
2019
entrez:
20
7
2019
pubmed:
20
7
2019
medline:
10
3
2020
Statut:
epublish
Résumé
The potential of Pseudomonas putida KT2440 to act as a plant-growth promoter or as a bioremediator 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 30°C 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.
Identifiants
pubmed: 31323038
doi: 10.1371/journal.pone.0219554
pii: PONE-D-18-34522
pmc: PMC6641147
doi:
Substances chimiques
Oligonucleotides
0
RNA, Ribosomal
0
RNA, Ribosomal, 16S
0
Green Fluorescent Proteins
147336-22-9
Trehalose
B8WCK70T7I
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0219554Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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