Biology subjects

Remus, D. M.

Publications and source records attributed to Remus, D. M..

4 recordsLinked to original sources

bioRxiv2022

The influence of resource overlap on bacterial reproductive success in the phyllosphere

The phyllosphere is densely colonised by rich microbial communities, despite sparse and heterogeneously distributed resources. The limitation of resources is expected to drive bacterial competition resulting in exclusion or coexistence based on fitness differences and resource overlap between individual colonisers. We studied the impact of resource competition by determining the effects of different bacterial colonisers on the growth of the model epiphyte Pantoea eucalypti 299R (Pe299R). Resource overlap was predicted based on genome-scale metabolic modelling. By combining results of metabolic modelling and pairwise competitions in the Arabidopsis thaliana phyllosphere and in vitro, we found that ten resources sufficed to explain fitness of Pe299R. An effect of both resource overlap and phylogenetic relationships was found on competition outcomes in vitro as well as in the phyllosphere. However, effects of resource competition were much weaker in the phyllosphere when compared to in vitro experiments. When investigating growth dynamics and reproductive success at the single-cell resolution, resource overlap and phylogenetic relationships are only weakly correlated with epiphytic Pe299R reproductive success, indicating that the leafs spatial heterogeneity mitigates resource competition. Although the correlation is weak, the presence of competitors led to the development of Pe299R subpopulations that experienced different life histories and cell divisions. Surprisingly, in some in planta competitions, Pe299R benefitted from the presence of epiphytes despite high resource overlap to the competitor strain suggesting other factors having stronger effects than resource competition. This study provides fundamental insights into how bacterial communities are shaped in heterogeneous environments and provides a framework to predict competition outcomes.

microbiology↗

bioRxiv2020

Constitutively expressed fluorescent proteins allow to track bacterial growth and to determine relative fitness of bacteria in mixed cultures

Bacterial growth is classically assessed by measuring the increase in optical density of pure cultures in shaken liquid media. Measuring growth using optical density has severe limitations when studying multistrain interactions as it is not possible to measure the growth of individual strains within mixed cultures. Here we demonstrated that constitutively expressed fluorescent proteins can be used to track the growth of individual strains in different liquid media. Fluorescence measurements were highly correlated with optical density measurements and cell counts. This allowed us to assess bacterial growth not only in pure cultures, but also in mixed bacterial cultures and determine the impact of competitors on a focal strain, thereby assessing relative fitness. Furthermore, we were able to track the growth of two different strains simultaneously by using fluorescent proteins with differential excitation and emission wavelengths. Bacterial densities measured by fluorescence yielded more consistent data between technical replicates than optical density measurements. Our setup employs fluorescent microplate readers that allow for high throughput and replication. ImportanceWe expand on an important limitation of the concept of measuring bacterial growth which is classically limited to one strain at a time. By adopting this approach, it is possible to measure growth of several bacterial strains simultaneously in high temporal resolution and in a high throughput manner. This is important to investigate bacterial interactions such as competition and facilitation.

microbiology↗

bioRxiv2020

Characterisation of the biosurfactants from phyllosphere colonising Pseudomonads and their effect on plant colonisation and diesel degradation

Biosurfactant production is a common trait in leaf surface colonising bacteria that has been associated with increased survival and movement on leaves. At the same time the ability to degrade aliphatics is common in biosurfactant-producing leaf colonisers. Pseudomonads are common leaf colonisers and have been recognised for their ability to produce biosurfactants and degrade aliphatic compounds. In this study, we have investigated the role of biosurfactants in four non-plant plant pathogenic Pseudomonas strains by performing a series of experiments to characterise the surfactant properties, and their role during leaf colonisation and diesel degradation. The produced biosurfactants were identified using mass-spectrometry. Two strains produced viscosin-like biosurfactants and the other two produced Massetolide A-like biosurfactants which aligned with the phylogenetic relatedness between the strains. To further investigate the role of surfactant production, random Tn5 transposon mutagenesis was performed to generate knockout mutants. The knockout mutants were compared to their respective wildtypes in their ability to colonise gnotobiotic Arabidopsis thaliana and to degrade diesel. It was not possible to detect negative effects during plant colonisation in direct competition or individual colonisation experiments. When grown on diesel, knockout mutants grew significantly slower compared to their respective wildtypes. By adding isolated wildtype biosurfactants it was possible to complement the growth of the knockout mutants. ImportanceMany leaf colonising bacteria produce surfactants and are able to degrade aliphatic compounds, however, if surfactant production provides a competitive advantage during leaf colonisation is unclear. Furthermore, it is unclear if leaf colonisers take advantage of the aliphatic compounds that constitute the leaf cuticle and cuticular waxes. Here we test the effect of surfactant production on leaf colonisation and demonstrate that the lack of surfactant production decreases the ability to degrade aliphatic compounds. This indicates that leaf surface dwelling, surfactant producing bacteria contribute to degradation of environmental hydrocarbons and may be able to utilise leaf surface waxes. This has implications for plant-microbe interactions and future studies.

microbiology↗

bioRxiv2018

Chromatic Bacteria - A broad host-range plasmid and chromosomal insertion toolbox for fluorescent protein expression in bacteria

Differential fluorescent labelling of bacteria has become instrumental for many aspects of microbiological research, such as the study of biofilm formation, bacterial individuality, evolution, and bacterial behaviour in complex environments. We designed a variety of plasmids, each bearing one of eight unique, constitutively expressed fluorescent protein genes in conjunction with one of four different antibiotic resistance combinations. The fluorophores mTagBFP2, mTurquoise2, sGFP2, mClover3, sYFP2, mOrange2, mScarlet-I, and mCardinal, encoding for blue, cyan, green, green-yellow, yellow, orange, red, and far-red fluorescent proteins, respectively, were combined with selectable markers conferring tetracycline, gentamicin, kanamycin, and/or chloramphenicol resistance. These constructs were cloned into three different plasmid backbones: a broad host-range plasmid, a Tn5 transposon delivery plasmid, and a Tn7 transposon delivery plasmid. The utility of the plasmids and transposons was tested in bacteria from the phyla Actinobacteria, Proteobacteria, and Bacteroidetes. We were able to tag representatives from the phylum Proteobacteria at least via our Tn5 transposon delivery system. The here constructed plasmids are available to the community and provide a valuable tool to investigate bacteria-bacteria, bacteria-host, and bacteria-environmental interactions.

microbiology↗