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Biology subjects

Fournier, S.

Publications and source records attributed to Fournier, S..

9 recordsLinked to original sources

A root pathogen drives rhizosphere enrichment of antagonistic Pseudomonas and induces production of an antimicrobial metabolite

In plants, the development of soil-borne diseases has been shown to trigger the recruitment of beneficial microbes, which contribute to defense against pathogens. However, the underlying mechanisms driving this recruitment remain elusive. Here, we used a gnotobiotic system combined with a synthetic bacterial community (SynCom) derived from the Medicago truncatula rhizosphere to dissect microbiota pathogen interaction and its role in the development of root rot caused by Aphanomyces euteiches, a devastating soilborne oomycete pathogen of legumes. Through integrated metabarcoding, metabolomics and transcriptomics, we reveal that pathogen infection restructures the bacterial SynCom, selectively enriching the microbial community with specific Pseudomonas spp. strains displaying anti A. euteiches activity. This shift alleviates root rot symptoms, triggers the biosynthesis of the antibiotic 2,4-diacetylphloroglucinol (DAPG), a Pseudomonas specialized metabolite inhibiting A. euteiches growth, and amplifies the plant endogenous isoflavonoid defense responses. Unexpectedly, we found that A. euteiches directly activates DAPG production in beneficial bacteria independently of the plant, through the production of a heat-stable, high-molecular-weight (30 - 100 kDa) extracellular components. These findings uncover a novel mechanism whereby a pathogen inadvertently activates antibiotic production in beneficial bacteria, extending the plant immune system. Our research underscores the critical role of microbial interactions in the rhizosphere in determining root disease outcomes, paving the way for microbiome-based strategies to combat root diseases.

plant biology↗

A cold-water coral garden with co-occurring Antipathella subpinnata and Dendrophyllia cornigera in the mesophotic zone of the northern Bay of Biscay

Large aggregations of cold-water corals (CWC), often termed CWC gardens, are considered as biodiversity hotspots and recognized as vulnerable marine ecosystems. Their three-dimensional structure creates important habitat complexity, providing refuge, nursery and feeding areas for many deep-water species. Their vulnerability and limited current knowledge highlight the importance of describing the distribution and habitats of these gardens. Using remotely operated vehicles (ROV), we describe an unprecedented large aggregation of the co-occurring black (Antipathella subpinnata) and yellow (Dendrophyllia cornigera) coral gardens in the mesophotic in the northern Bay of Biscay. These distributions acknowledged their latitudinal and bathymetric distributions, in particular for A. subpinnata. Black and yellow corals presented mean densities of 2.0 {+/-} 1.8 colonies.10 m-2 and 1.5 {+/-} 1.3 colonies.10 m-2, respectively. Following the complex topography of this area, they were mostly fixed on hard substrates, particularly in narrow crests close to or surrounded by sand or mud planes. These findings are essential to develop appropriate local spatial management plans, ultimately contributing to the conservation of these vulnerable ecosystems.

ecology↗

Comparing fluorometric methods (in vivo vs extracted) for cyanoHAB monitoring in six Rhode Island ponds

Harmful algal blooms caused by cyanobacteria (cyanoHABs) are detrimental to human and environmental health and can be difficult to monitor without specialized training and equipment. A variety of instruments have been developed to measure cyanoHAB indicators (i.e., chlorophyll a or phycocyanin) that do not require advanced laboratory processes (e.g., pigment extraction). We compared measurements from five in vivo fluorometers (Turner Trilogy in-vivo module, Turner Fluorosense, Turner Cyanofluor, bbe AlgaeTorch, and bbe Phycoprobe) to results from solvent-based extractions for chlorophyll a and phycocyanin at six different waterbodies in Rhode Island. We found a strong relationship between extracted phycocyanin and in vivo fluorometers (R2 ranging from 0.78-0.96). We found less consistency between in vivo measurements of chlorophyll a and the extracted results (R2 between 0.34 and 0.82). Some variability in the chlorophyll a results can be explained by differences in the phytoplankton community across the different sampling sites. Phycocyanin results from in vivo fluorometry were also strongly related with cell counts, which implies that phycocyanin measurements from these instruments can be a good proxy for cell counts. Many federal, state, and local entities use cell counts of cyanobacteria to determine when to issue health or contact advisories for waterbodies. Producing accurate cell counts requires highly specialized training/equipment, processing time, and counts can vary greatly between technicians. The results from this study encourage further adoption of in vivo fluorometry and phycocyanin for cyanoHAB monitoring efforts.

ecology↗

Guiding ATR and PARP inhibitor combinations with chemogenomic screens

Combinations of inhibitors of Ataxia Telangiectasia- and Rad3-related kinase (ATRi) and poly(ADP-ribose) polymerases (PARPi) synergistically kill tumor cells through modulation of complementary DNA repair pathways, but their tolerability is limited by hematological toxicities. To address this we performed a genome-wide CRISPR/Cas9 screen to identify genetic alterations that hypersensitize cells to a combination of the ATRi RP-3500 with PARPi, including deficiency in RNase H2, RAD51 paralog mutations or the Alternative Lengthening of Telomeres telomere maintenance mechanism. We show that RP-3500 and PARPi combinations kill cells carrying these genetic alterations at doses sub-therapeutic as single agents. We also demonstrate the mechanism of combination hypersensitivity in RNase H2-deficient cells, where we observe an irreversible replication catastrophe, allowing us to design a highly efficacious and tolerable in vivo dosing schedule. Altogether, we present a comprehensive dataset to inform development of ATRi and PARPi combinations and an experimental framework applicable to other drug combination strategies.

cancer biology↗

Cannabinoids vs. whole metabolome: relevance of cannabinomics in analyzing Cannabis varieties

Cannabis sativa has a long history of domestication both for its bioactive compounds and its fibers. This has produced hundreds of varieties, usually characterized in the literature by chemotypes, with {Delta}9-THC and CBD content as the main markers. However, chemotyping could also be done based on minor compounds (phytocannabinoids and others). In this work, a workflow, which we propose to name cannabinomics, combines mass spectrometry of the whole metabolome and statistical analysis to help differentiate C. sativa varieties and deciphering their characteristic markers. By applying this cannabinomics approach to the data obtained from 20 varieties of C. sativa (classically classified as chemotype I, II, or III), we compared the results with those obtained by a targeted quantification of 11 phytocannabinoids. Cannabinomics can be considered as a complementary tool for phenotyping and genotyping, allowing the identification of minor compounds playing a key role as markers of differentiation.

biochemistry↗

CCNE1 amplification is synthetic-lethal with PKMYT1 kinase inhibition

Amplification of the gene encoding cyclin E (CCNE1) is an oncogenic driver in several malignancies and is associated with chemoresistance and poor prognosis. To uncover therapeutic targets for CCNE1-amplified tumors, we undertook genome-scale CRISPR/Cas9-based synthetic lethality screens in cellular models of CCNE1 amplification. Here, we report that increasing CCNE1 dosage engenders a vulnerability to the inhibition of the PKMYT1 kinase, a negative regulator of CDK1. To inhibit PKMYT1, we developed RP-6306, an orally bioavailable and selective inhibitor that shows single-agent activity and durable tumor regressions when combined with gemcitabine in models of CCNE1-amplification. RP-6306 treatment causes unscheduled activation of CDK1 selectively in CCNE1 overexpressing-cells, promoting early mitosis in cells undergoing DNA synthesis. CCNE1 overexpression disrupts CDK1 homeostasis at least in part through an early activation of the FOXM1/MYBL2/MuvB-dependent mitotic transcriptional program. We conclude that PKMYT1 inhibition is a promising therapeutic strategy for CCNE1-amplified cancers.

cancer biology↗

Phytohormone production by the arbuscular mycorrhizal fungus Rhizophagus irregularis

Arbuscular mycorrhizal symbiosis is a mutualistic interaction between most land plants and fungi of the glomeromycotina subphylum. The initiation, development and regulation of this symbiosis involve numerous signalling events between and within the symbiotic partners. Among other signals, phytohormones are known to play important roles at various stages of the interaction. During presymbiotic steps, plant roots exude strigolactones which stimulate the fungus, and favour the initiation of symbiosis. At later stages, different plant hormone classes can act as positive or negative regulators of the interaction. Although the fungus is known to reciprocally emit regulatory signals, its potential contribution to the phytohormonal pool has received little attention, and has so far only been addressed by indirect assays. In this study, using mass spectrometry, we analyzed phytohormones released into the medium by germinated spores of the arbuscular mycorrhizal fungus Rhizophagus irregularis. We detected the presence of a cytokinin (isopentenyl-adenosine) and an auxin (indole-acetic acid). In addition, we identified a gibberellin (gibberellic acid 4) in spore extracts. We also used gas chromatography to show that R. irregularis produces ethylene from methionine and the -keto {gamma}-methylthiobutyric acid pathway. These results highlight the possibility for AM fungi to use phytohormones to interact with their host plants, or to regulate their own development.

plant biology↗

DIVA: natural navigation inside 3D images usingvirtual reality

As three-dimensional microscopy becomes commonplace in biological re-search, there is an increasing need for researchers to be able to view experimental image stacks in a natural three-dimensional viewing context. Through stereoscopy and motion tracking, commercial virtual reality headsets provide a solution to this important visualization challenge by allowing researchers to view volumetric objects in an entirely intuitive fashion. With this motivation, we present DIVA, a user-friendly software tool that automatically creates detailed three-dimensional reconstructions of raw experimental image stacks that are integrated in virtual reality. In DIVAs immersive virtual environment, users can view, manipulate and perform volumetric measurements on their microscopy images as they would to real physical objects. In contrast to similar solutions, our software provides high-quality volume rendering with native TIFF file compatibility. We benchmark the software with diverse image types including those generated by confocal, light-sheet and electron microscopy. DIVA is available at https://diva.pasteur.fr and will be regularly updated.

bioinformatics↗

MS-CleanR: A feature-filtering approach to improve annotation rate in untargeted LC-MS based metabolomics

Untargeted metabolomics using liquid chromatography-mass spectrometry (LC-MS) is currently the gold-standard technique to determine the full chemical diversity in biological samples. This approach still has many limitations, however; notably, the difficulty of estimating accurately the number of unique metabolites being profiled among the thousands of MS ion signals arising from chromatograms. Here, we describe a new workflow, MS-CleanR, based on the MS-DIAL/MS-FINDER suite, which tackles feature degeneracy and improves annotation rates. We show that implementation of MS-CleanR reduces the number of signals by nearly 80% while retaining 95% of unique metabolite features. Moreover, the annotation results from MS-FINDER can be ranked with respect to database chosen by the user, which improves identification accuracy. Application of MS-CleanR to the analysis of Arabidopsis thaliana grown in three different conditions improved class separation resulting from multivariate data analysis and lead to annotation of 75% of the final features. The full workflow was applied to metabolomic profiles from three strains of the leguminous plant Medicago truncatula that have different susceptibilities to the oomycete pathogen Aphanomyces euteiches; a group of glycosylated triterpenoids overrepresented in resistant lines were identified as candidate compounds conferring pathogen resistance. MS-CleanR is implemented through a Shiny interface for intuitive use by end-users (available at: https://github.com/eMetaboHUB/MS-CleanR).

bioinformatics↗