Biology subjects

Sehrt, M.

Publications and source records attributed to Sehrt, M..

5 recordsLinked to original sources

bioRxiv2026

Dual control of plasmid copy number in E. coli by combining temperature shifts and RNAI-based genetic circuits for tunable replication dynamics

Plasmid copy number (PCN) is a central determinant of recombinant gene expression levels and metabolic burden in bacterial cell factories. However, dynamic real-time control of PCN during microbial cultivation remains largely unexplored. In the ColE1/pUC19 replication system, PCN is governed by an antisense RNAI/RNAII circuit, but this native mechanism provides no external handle for process-level intervention. Here, we engineer and characterize a dual-layer control architecture that combines two orthogonal perturbations to direct plasmid replication in Escherichia coli populations i.e., temperature shifts, which modulate the secondary structure of RNAII and, consequently, its binding to RNAI and its inhibitory potency (ex vivo control), and arabinose-inducible overexpression of a supplementary RNAI cassette, integrated either on a secondary low-copy plasmid or directly on the high-copy target plasmid (in vivo control). Using a superfolder GFP validated fluorescent reporter and automated flow cytometry in continuous cultures, we track PCN dynamics at the population level in real time under chemostat conditions. Preliminary screening in microplates high-throughput cultivation device across three strains, two carbon sources, and three temperatures (30, 37, and 42C) reveals that the two control layers act synergistically. Neither temperature alone nor inducible RNAI alone achieves the dynamic range obtained by their combination. This synergistic effect is sustained during continuous cultivation at a fixed dilution rate of 0.1/h, with carbon source and temperature transitions producing predictable and reproducible shifts in population-level PCN. These results establish a scalable, dual-input framework for on-demand PCN tuning in continuous bioprocesses, with direct relevance to biopharmaceutical plasmid production and synthetic biology applications.

bioengineering↗

bioRxiv2026

Cellular responsiveness as a predictive indicator for population collapse and autonomous control in continuous cultures of Pseudomonas putida

How microbial populations respond to repeated environmental transitions determines both their ecological fitness and their utility in biotechnological applications. Using Pseudomonas putida KT2440 equipped with fluorescent biosensors and monitored by automated flow cytometry in the Segregostat platform, we show that exposure to benzoate, a plastic-derived aromatic feedstock, progressively reduces cellular responsiveness, defined as the fraction of cells that successfully activate a gene circuit following an environmental transition. Unlike classical switching costs, which promote phenotypic diversification, benzoate suppresses responsiveness without increasing population entropy, in a concentration-dependent and circuit-independent manner tightly correlated with fitness loss. A resource allocation model incorporating the competing demands of benzoate assimilation, toxicity, and tolerance reveals that this impairment emerges from a three-way competition for limited cellular resources. Above a critical benzoate load, insufficient resources remain available to sustain the adaptive reallocation required for circuit activation. In continuous culture, a non-responsive subpopulation accumulates as a leading indicator of population collapse. Exploiting this signal, we implement a two-stage connected bioreactor system in which benzoate feeding is autonomously regulated based on real-time population structure, enabling complete substrate consumption and stable operation at otherwise destabilizing concentrations. These results establish cellular responsiveness as a quantitative population variable and demonstrate that structure-aware feedback control, acting on population composition rather than bulk physiology, provides a principled route toward autonomous bioprocesses on challenging substrates.

Systems Biology↗

bioRxiv2025

A Fitness Entropy Compensation effect set the trade-off between growth and gene expression in cell populations

We present findings on a Fitness-Entropy Compensation (FEC) mechanism which offsets the activation of gene circuits that compromise survival. It counteracts the resulting fitness reduction by increasing the diversity in gene expression among individual cells within the population. This diversity, quantified by the Shannon entropy, enables cells with lower expression levels to support the survival of the entire population. We investigated the presence of FEC in a range of synthetic and stress-related genetic circuits in continuous culture. Our results reveal that it effectively stabilizes cell populations by mitigating the detrimental trade-offs between growth and gene expression. This stabilization is due to the reduced growth rate of the induced phenotype that leads to environmental changes, decreases induction strength, and promotes escape from unfit states. These findings suggest that the FEC mechanism may be a universal strategy for stabilization in various cellular systems and set the basis for a quantitative description of the trade-off between growth and gene expression and its consequences at the population level.

systems biology↗

bioRxiv2019

Less is more! Rapid increase in plant species richness after reduced mowing of urban grasslands

Urban lawns provide space for recreation in cities, and they are an important part of urban green infrastructures. However, most lawns are intensively managed. As only few plant species can survive the frequent mowing, urban lawns typically harbor only a limited number of plant species. To improve the biodiversity of urban lawns, it is often suggested to reduce the mowing frequency. Here, we studied the plant diversity of urban grasslands that have recently undergone management changes from mowing every few weeks to mowing only once or twice per season and compared them to intensively managed lawns. Within six years after the management changes, the grasslands with reduced mowing frequency indeed hosted 30% more plant species than intensively managed lawns, and they were more heterogeneous both within and between grasslands. Additionally, the species composition of less frequently mown grasslands shifted from common mowing-tolerant lawn species to typical meadow species. Our study thus shows that the reduction of mowing is a simple and effective tool for increasing the biodiversity in urban grasslands.

ecology↗

bioRxiv2019

Effects of forest management on the phenology of early-flowering understory herbs

Many organisms respond to anthropogenic environmental change through shifts in their phenology. In plants, flowering is largely driven by temperature, and therefore affected by climate change. However, on smaller scales climatic conditions are also influenced by other factors, including habitat structure. A group of plants with a particularly distinct phenology are the understorey herbs in temperate forests. In these forests, management alters tree species composition and stand structure and, as a consequence, light conditions and microclimate. Forest management should thus also affect the phenology of understorey herbs. To test this, we recorded the flowering phenology of 20 early-flowering herbs on 100 forest plots varying in management intensity, from near-natural to intensely managed forests, in Central and Southern Germany. We found that in forest stands with a high management intensity the plants flowered on average about two weeks later than in unmanaged forests. This was largely because management also affected microclimate (e.g. spring temperatures of 5.9 {degrees}C in managed coniferous, 6.7 in managed deciduous and 7.0 {degrees}C in unmanaged deciduous plots), which in turn affected phenology, with plants flowering later on colder and moister forest stands (+4.5 days per -1{degrees}C and 2.7 days per 10 % humidity increase). Among forest characteristics, the main tree species as well as the age, overall crown projection area, structural complexity and spatial distribution of trees had the greatest influence on microclimate. Our study demonstrates that forest management alters plant phenology, with potential far-reaching consequences for the ecology and evolution of understorey communities. More generally, our study suggests that besides climate change other drivers of environmental change, too, can influence the phenology of organisms.

ecology↗