Carbon starvation of Pseudomonas aeruginosa biofilms selects for dispersal insensitive mutants
| Autor: | Sujatha Subramoni, Wee Han Poh, Tim Tolker-Nielsen, Harikrishnan A. S. Nair, Staffan Kjelleberg, Nabilah Taqiah Binte Hasnuddin, Scott A. Rice, Martin Tay, Michael Givskov, Diane McDougald |
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| Přispěvatelé: | School of Biological Sciences, Interdisciplinary Graduate School (IGS), Singapore Centre for Environmental Life Sciences and Engineering (SCELSE) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
Bioflm Development Mutant Population Biology medicine.disease_cause Microbiology Biofilm development Green fluorescent protein Bacterial Proteins medicine education 06 Biological Sciences 07 Agricultural and Veterinary Sciences 11 Medical and Health Sciences education.field_of_study Bioreporter Pseudomonas aeruginosa Research Wild type Biofilm Biological sciences [Science] Gene Expression Regulation Bacterial Nutrients Dispersal biochemical phenomena metabolism and nutrition Image-based quantification Carbon QR1-502 C-di-GMP Complementation Starvation Biofilms Mutation Biological dispersal Morphotypic variants |
| Zdroj: | BMC Microbiology, Vol 21, Iss 1, Pp 1-10 (2021) Nair, H A S, Subramoni, S, Poh, W H, Hasnuddin, N T B, Tay, M, Givskov, M, Tolker-Nielsen, T, Kjelleberg, S, McDougald, D & Rice, S A 2021, ' Carbon starvation of Pseudomonas aeruginosa biofilms selects for dispersal insensitive mutants ', BMC Microbiology, vol. 21, 255 . https://doi.org/10.1186/s12866-021-02318-8 BMC Microbiology |
| DOI: | 10.1186/s12866-021-02318-8 |
| Popis: | Background Biofilms disperse in response to specific environmental cues, such as reduced oxygen concentration, changes in nutrient concentration and exposure to nitric oxide. Interestingly, biofilms do not completely disperse under these conditions, which is generally attributed to physiological heterogeneity of the biofilm. However, our results suggest that genetic heterogeneity also plays an important role in the non-dispersing population of P. aeruginosa in biofilms after nutrient starvation. Results In this study, 12.2% of the biofilm failed to disperse after 4 d of continuous starvation-induced dispersal. Cells were recovered from the dispersal phase as well as the remaining biofilm. For 96 h starved biofilms, rugose small colony variants (RSCV) were found to be present in the biofilm, but were not observed in the dispersal effluent. In contrast, wild type and small colony variants (SCV) were found in high numbers in the dispersal phase. Genome sequencing of these variants showed that most had single nucleotide mutations in genes associated with biofilm formation, e.g. in wspF, pilT, fha1 and aguR. Complementation of those mutations restored starvation-induced dispersal from the biofilms. Because c-di-GMP is linked to biofilm formation and dispersal, we introduced a c-di-GMP reporter into the wild-type P. aeruginosa and monitored green fluorescent protein (GFP) expression before and after starvation-induced dispersal. Post dispersal, the microcolonies were smaller and significantly brighter in GFP intensity, suggesting the relative concentration of c-di-GMP per cell within the microcolonies was also increased. Furthermore, only the RSCV showed increased c-di-GMP, while wild type and SCV were no different from the parental strain. Conclusions This suggests that while starvation can induce dispersal from the biofilm, it also results in strong selection for mutants that overproduce c-di-GMP and that fail to disperse in response to the dispersal cue, starvation. |
| Databáze: | OpenAIRE |
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