bioRxiv Science⌕ Search

Biology subjects

Cadot, S.

Publications and source records attributed to Cadot, S..

6 recordsLinked to original sources

Development of a Polyelectrolyte Complex Scaffold and its specific cell seeding method as a tool for liquid cancers drug screening

This study focuses on the development of 3D culture model dedicated to liquid cancers drug screening. The challenge addressed was to effectively retain non adherent small cells within a 3D-scaffold with tailorable mechanical properties, while proposing a fast and effective tool for drug screening. To that aim, we developed a macroporous alginate-chitosan polyelectrolyte complex (PEC) scaffold combined with a low-viscosity alginate (LVA) cell seeding solution. We hypothesized that LVA could undergo in situ pore gelation via calcium ions retained from the PEC fabrication process, enabling effective retention and homogeneous cell distribution, leading to an improved platform for drug screening and personalized medicine. First, we evaluated scaffold suitability for LVA infiltration and gelation. Microtomography revealed a highly porous architecture (98%) enabling LVA homogeneous penetration and complete gelation within 30 min, as confirmed by SEM, microscopy, rheology, and micro-rheology. Next, we assessed cell retention and biocompatibility using primary human chronic lymphocytic leukemia (CLL) cells. LVA-assisted seeding increased cell density 2.6-fold compared to medium alone, with homogeneous distribution, >80% viability over 7 days, and preserved differentiation into nurse-like cells. Finally, we demonstrated a proof of concept for drug screening. The Alginate-PEC scaffold (A-PEC scaffold) supported both qualitative live/dead imaging and rapid quantitative viability measurement with the Alamar Blue assay. Drug responses reproduced microenvironment-dependent protection effects observed in vivo. This integrated scaffold and seeding method provides a promising 3D platform for in vitro liquid cancer studies and drug screening on patient-derived hematological cancer cells. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=67 SRC="FIGDIR/small/722037v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@9b71d4org.highwire.dtl.DTLVardef@14e1dd0org.highwire.dtl.DTLVardef@1876a56org.highwire.dtl.DTLVardef@15656bc_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Soil iron drives beneficial maize microbiome feedbacks inrotations with wheat

BackgroundPlants change their surrounding soil microbiome by root exudates and such conditioned microbiomes impact the performance of the present as well as the next plant generation as for example in crop rotations. The big challenge is that such microbiome feedbacks are highly context-dependent, i.e. they vary in strength and direction dependent on the local soil environment - of which the driving factor(s) remain unknown. Including maize in crop rotations involves benzoxazinoids (BXs), which are exuded from roots and alter the soil microbiome, which in turn affects growth and defence of the following crop. ResultsHere, we grew wild-type and BX-depleted maize in the field to differentially condition their soil microbiome and we found varying feedbacks on wheat performance dependent on the local physicochemical soil parameters. Using multivariate, correlation and modelling approaches and including additional data from two previous field experiments, we identified plant-available (PA) iron to explain BX-dependent microbiome feedbacks on wheat. The BX-conditioned soil microbiome caused wheat to grow taller at low levels of soil PA-iron but smaller at high levels. This finding was generalized testing these maize microbiome feedbacks on the model plant Arabidopsis thaliana using soil batches containing different levels of iron. Consistent with wheat, a significant inverse relationship between soil PA-iron levels and plant growth was found. This relationship was experimentally validated with Arabidopsis grown at low levels of soil iron where iron supplementation abolished the beneficial feedback of the BX-conditioned soil microbiome. ConclusionTogether these findings revealed that beneficial microbiome feedbacks occur at low levels of plant-available iron, i.e. when plants grow in a suboptimal soil, but they are lost when plants are nutritionally well supported. These results underscore the importance of iron availability in soil for beneficial microbial feedbacks on plant growth and predict agronomic benefits of incorporating maize in crop rotations on low iron soils.

microbiology↗

A TNL receptor mediates microbiome feedbacks in Arabidopsis

Plant performance depends on the soil microbiome. While microbiome feedbacks are well documented, the mechanisms by which plants perceive and mediate these feedbacks remain unclear. We established a framework using two distinct microbiomes in the same soil, where one led to enhanced growth of the Arabidopsis thaliana accession Col-0. Screening 410 accessions revealed substantial variation in growth feedbacks, which we used for genome-wide association mapping. We identified the immune receptor Mediator of Microbiome Feedback 1 (MMF1) as a candidate gene involved in microbiota feedbacks. Characterisation in the reference accession Col-0 revealed that mmf1 mutants lost the beneficial growth feedback, had an altered root bacterial community, and failed to induce a defence-related transcriptional response observed in wild-type plants. The discovery of MMF1 implies that integration of microbial signals optimises host microbiome composition and immune status to enhance growth.

plant biology↗

Soil chemical and microbial gradients determine accumulation of root exuded secondary metabolites and plant-soil feedbacks in the field

IntroductionHarnessing positive plant-soil feedbacks via crop rotations is a promising strategy for sustainable agriculture. Plants can influence soil properties including microbes by exuding specialized metabolites. However, the effects are often context dependent and variable. If and how local soil heterogeneity may explain this variation is unknown. Benzoxazinoids are specialized metabolites that are released in high quantities by cereals such as wheat and maize. Benzoxazinoids can alter rhizosphere microbiota and the performance of plants subsequently growing in the exposed soils and are thus an excellent model to study agriculturally relevant plant-soil feedbacks in the field, and to assess how soil factors affect their outcome. Materials & methodsTo understand the importance of local variation in soil properties on benzoxazinoid-mediated plant-soil feedbacks, we conditioned plots with wild-type maize and benzoxazinoid-deficient bx1 mutant plants in a grid pattern across an arable field. We then grew winter wheat across the entire field in the following season. We determined accumulation of benzoxazinoids, root-associated microbial communities, abiotic soil properties and wheat performance in each plot. We also determined benzoxazinoid conversion dynamics in a labelling experiment under controlled conditions, and then assessed associations between soil chemical variation and benzoxazinoid-mediated plant-soil feedbacks. ResultsAcross the field, we detected a marked gradient in soil chemical and microbial community composition. This gradient resulted in significant differences in benzoxazinoid accumulation. These differences were explained by differential benzoxazinoid degradation rather than exudation. Benzoxazinoid exudation modulated alpha diversity of root and rhizosphere bacteria and fungi during maize growth, but not during subsequent wheat growth, while the chemical fingerprint of benzoxazinoid accumulation persisted. Averaged across the field, we detected no significant feedback effects of benzoxazinoid conditioning on wheat performance and defence, apart from a transient decrease in biomass during vegetative growth. Closer analysis however, revealed pronounced feedback effects along the chemical and microbial gradient of the field, with effects gradually changing from negative to positive along the gradient. ConclusionOverall, this study revealed that plant-soil feedbacks differ in strength and direction within a field, and that this variation can be explained by standing chemical and microbial gradients, which strongly affect benzoxazinoid accumulation in the soil. Understanding within-field soil heterogeneity is crucial for the future exploitation of plant-soil feedbacks in sustainable precision agriculture.

plant biology↗

Soil composition and plant genotype determine benzoxazinoid-mediated plant-soil feedbacks in cereals

Plant-soil feedbacks refer to effects on plants that are mediated by soil modifications caused by the previous plant generation. Maize conditions the surrounding soil by secretion of root exudates including benzoxazinoids (BXs), a class of bioactive secondary metabolites. Previous work found that a BX- conditioned soil microbiota enhances insect resistance while reducing biomass in the next generation of maize plants. Whether these BX-mediated and microbially driven feedbacks are conserved across different soils and response species is unknown. We found the BX-feedbacks on maize growth and insect resistance conserved between two arable soils, but absent in a more fertile grassland soil, suggesting a soil-type dependence of BX feedbacks. We demonstrated that wheat also responded to BX-feedbacks. While the negative growth response to BX-conditioning was conserved in both cereals, insect resistance showed opposite patterns, with an increase in maize and a decrease in wheat. Wheat pathogen resistance was not affected. Finally and consistent with maize, we found the BX-feedbacks to be cultivar specific. Taken together, BX- feedbacks affected cereal growth and resistance in a soil and genotype dependent manner. Cultivar-specificity of BX-feedbacks is a key finding, as it hides the potential to optimize crops that avoid negative plant-soil feedbacks in rotations.

plant biology↗

Specific and conserved patterns of microbiota-structuring by maize benzoxazinoids in the field

BackgroundPlants influence their root and rhizosphere microbial communities through the secretion of root exudates. However, how specific classes of root exudate compounds impact the assembly of these root-associated microbiotas is not well understood. Maize roots secrete benzoxazinoids (BXs), a class of indole-derived defense compounds, and thereby impact the assembly of their microbiota. Here, we investigated the broader impacts of BX exudation on root and rhizosphere microbiotas of adult maize plants grown under natural conditions at different field locations in Europe and the US. We examined the microbiotas of BX-producing and multiple BX-defective lines in two genetic backgrounds across three soil types. ResultsOur analysis showed that the secretion of BXs affected community composition of rhizosphere and root microbiota, with the most pronounced effects observed for root fungi. The impact of the two genetic backgrounds was weaker than that of the presence or absence of BXs, suggesting that BX exudation is a key trait by which maize structures its associated microbiota. BX-producing plants were not consistently enriching microbial lineages across the three soil types. Instead, BX exudation consistently depleted Flavobacteriaceae and Comamonadaceae, and enriched various plant pathogenic fungi in the roots. ConclusionsThese findings reveal that BXs have a selective impact on root and rhizosphere microbiota composition across different field locations. Taken together, this study identifies the BX pathway as an interesting breeding target to manipulate plant-microbiome interactions.

plant biology↗