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Cobeta, P.

Publications and source records attributed to Cobeta, P..

4 recordsLinked to original sources

Drift-driven microbiome simplification generates reconstructable and ecologically cohesive microbial communities

Engineering simplified microbial communities that retain function and ecological cohesiveness remains a major challenge because the taxa and interactions required for community establishment are rarely known a priori. Here, we experimentally evaluated drift-driven microbiome simplification as an alternative strategy for generating reduced and reconstructable microbial consortia. Using the tomato rhizosphere as a model system, three source microbial communities were subjected to different dilution bottlenecks and serial propagation. Increasing dilution was the main determinant of community composition and simplification, while repeated passage produced additional reductions in ASV richness, phylogenetic diversity, and evenness. Importantly, diversity loss was not accompanied by a uniform deterioration in bacterial colonization or plant performance, indicating that substantial simplification can occur without a parallel collapse in these system-level properties. Candidate Minimal Microbiome prototypes were selected from endpoint communities, reconstructed as synthetic communities, and their ecological cohesiveness was evaluated through invasion experiments. The reconstructed communities strongly restricted the establishment of the original complex microbial fractions, although invasion success depended markedly on invader identity. The drift-derived communities were at least as resistant to invasion as an independently designed bottom-up synthetic community. Together, these results provide experimental support for drift-driven simplification as a strategy to generate reduced microbial communities whose dominant members can be isolated, reconstructed, and experimentally evaluated. By allowing ecological assembly to generate candidate community configurations before cultivation and reconstruction, this approach provides a complementary route to rational bottom-up design and function-directed top-down microbiome engineering.

microbiology↗

Combinatorial community coalescence in early tomato assembly reveals a rhizosphere attractor in composition and abundance architecture

The rhizosphere microbiome plays fundamental roles in plant health and productivity, yet the ecological rules governing microbiome assembly remain poorly understood. Here, we investigated early rhizosphere community assembly in tomato using a replicated combinatorial community coalescence framework, in which seven distinct natural bacterial communities were inoculated individually and in all possible pairwise and triplet combinations. Single-inoculum communities clustered according to inoculum identity, indicating a strong effect of source community composition on assembly trajectories. However, when all communities were analyzed jointly, samples formed a continuous compositional landscape with no clear evidence of discrete community states. Despite major differences in source community composition, rhizosphere communities consistently converged toward a highly similar uneven rank-abundance structure, with two ASVs accounting for 50% and a median of nineteen ASVs for 90% of total abundance. While assembly was dominated by a very small number of Pseudomonas ASVs, limited evidence of alternative dominant states was observed. Increasing inoculum complexity did not increase stochasticity but instead promoted stronger convergence toward a global rhizosphere compositional centroid. Moreover, dominance hierarchies emerging from community coalescence closely mirrored the distance of source communities to this centroid. The two most dominant communities originated from orchard soils, suggesting that historical contingency and prior adaptation to horticultural crop rhizospheres may influence competitive success. Together, these results are consistent with the existence of a canonical rhizosphere attractor in composition and abundance architecture, with patterns consistent with assembly occurring under a limited number of dominant ecological niches imposed by the tomato rhizosphere.

microbiology↗

Early rhizosphere assembly during the onset of photosynthesis reveals inoculum-constrained succession with increased phylogenetic clustering, filtering and diversity

The rhizosphere microbiome, one of the most diverse and metabolically active microbial ecosystems known, plays fundamental roles in plant health and productivity. However, the ecological dynamics occurring during the transition between germination and the establishment of the first true leaves, a developmental window associated with the onset of active photosynthesis and rapid root expansion, remain poorly understood. Here, we investigated rhizosphere microbiome assembly during the first four weeks of tomato development by sampling communities arising from seven distinct natural soil inocula twice weekly to obtain fine-scale temporal resolution. Bacterial load, richness, evenness and phylogenetic diversity all increased significantly during plant development, indicating progressive increases in rhizosphere ecosystem complexity. In addition, diverse initial microbial communities differentially influenced both host plant development and the bacterial carrying capacity of the resulting rhizosphere ecosystem. Although temporal effects on rhizosphere microbiome composition were significant, assembly trajectories remained strongly constrained by the initial inoculum. Temporal analysis nevertheless revealed significant taxonomic turnover despite limited global compositional restructuring. In particular, Proteobacteria and Pseudomonadaceae decreased over time, whereas Actinobacteria, Acidobacteria and Streptomycetaceae increased. However, communities did not become progressively more similar or divergent over time. Altogether, our results indicate that early rhizosphere microbiome assembly involves rapid ecological succession within inoculum-constrained compositional trajectories, with early copiotrophic Proteobacteria progressively giving rise to more diverse and phylogenetically structured communities. These findings suggest that the first weeks of plant development may represent a critical ecological window for microbiome-based manipulation strategies in agriculture.

microbiology↗

Modular automated high-throughput isolation and phylogenetic identification of bacteria from complex microbiomes

Metagenomic analysis can generate hypotheses about microbiome interactions and function, yet mechanistic understanding is only possible through precise experimentation manipulating its microbiota composition. The high-throughput isolation of microbiome members thus represents a core resource in this field of research. Here, we present and test a culturomics pipeline based on the use of the limiting dilution method with multi-well plates, an optical plate reader, and barcoded sequencing for phylogenetic identification, and offer modularity by proposing different protocols and possibilities along the pipeline. Most importantly, we provide all scripts required for process automation using an affordable pipetting robot, along with associated estimates of financial and labor costs.

microbiology↗