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Accueil > Plateformes > Plateformes IMAGERIE-GIF : Cytométrie, Microscopie Electronique & Microscopie Photonique > Publications & Communications



  • L. Becker, S. Bellow, V. Carré, G. Latouche, A. Poutaraud, D. Merdinoglu, S. C. Brown, Z. G. Cerovic, et P. Chaimbault, « Correlative Analysis of Fluorescent Phytoalexins by Mass Spectrometry Imaging and Fluorescence Microscopy in Grapevine Leaves », Analytical Chemistry, juin 2017.
    Résumé : Plant response to their environment stresses is a complex mechanism involving secondary metabolites. Stilbene phytoalexins, namely resveratrol, pterostilbene, piceids and viniferins play a key role in grapevine (Vitis vinifera) leaf defense. Despite their well-established qualities, conventional analyses such as HPLC-DAD or LC-MS lose valuable information on metabolite localization during the extraction process. To overcome this issue, a correlative analysis combining mass spectroscopy imaging (MSI) and fluorescence imaging was developed to localize in situ stilbenes on the same stressed grapevine leaves. High-resolution images of the stilbene fluorescence provided by macroscopy were supplemented by specific distributions and structural information concerning resveratrol, pterostilbene, and piceids obtained by MSI. The two imaging techniques led to consistent and complementary data on the stilbene spatial distribution for the two stresses addressed: UV-C irradiation and infection by Plasmopara viticola. Results emphasize that grapevine leaves react differently depending on the stress. A rather uniform synthesis of stilbenes is induced after UV-C irradiation, whereas a more localized synthesis of stilbenes in stomata guard cells and cell walls is induced by P. viticola infection. Finally, this combined imaging approach could be extended to map phytoalexins of various plant tissues with resolution approaching the cellular level.
    Mots-clés : IMAGIF, PF, PHOT.

  • S. C. Brown, M. Bourge, N. Maunoury, M. Wong, M. W. Bianchi, S. Lepers-Andrzejewski, P. Besse, S. Siljak-Yakovlev, M. Dron, et B. Satiat-Jeunemaître, « DNA remodelling by Strict Partial Endoreplication in orchids, an original process in the plant kingdom », Genome Biology and Evolution, avr. 2017.
    Résumé : DNA remodelling during endoreplication appears to be a strong developmental characteristic in orchids. In this study, we analysed DNA content and nuclei in 41 species of orchids to further map the genome evolution in this plant family. We demonstrate that the DNA remodelling observed in 36 out of 41 orchids studied corresponds to strict partial endoreplication. Such process is developmentally regulated in each wild species studied. Cytometry data analyses allowed us to propose a model where nuclear states 2C, 4E, 8E, etc. form a series comprising a fixed proportion, the euploid genome 2C, plus 2 to 32 additional copies of a complementary part of the genome. The fixed proportion ranged from 89% of the genome in Vanilla mexicana down to 19% in V. pompona, the lowest value for all 148 orchids reported. Insterspecific hybridisation did not suppress this phenomenon. Interestingly, this process was not observed in mass-produced epiphytes. Nucleolar volumes grow with the number of endocopies present, coherent with high transcription activity in endoreplicated nuclei. Our analyses suggest species-specific chromatin rearrangement. Towards understanding endoreplication, V. planifolia constitutes a tractable system for isolating the genomic sequences that confer an advantage via endoreplication from those that apparently suffice at diploid level.
    Mots-clés : BIOCELL, CYTO, cytogenetics, cytometry, DYNBSJ, endoreplication, genome imbalance, Genome Size, IMAGIF, MINION, PF, PHOT, Vanilla.

  • E. Dambroise, M. Simion, T. Bourquard, S. Bouffard, B. Rizzi, Y. Jaszczyszyn, M. Bourge, P. Affaticati, A. Heuzé, J. Jouralet, J. Edouard, S. Brown, C. Thermes, A. Poupon, E. Reiter, F. Sohm, F. Bourrat, et J. - S. Joly, « Postembryonic Fish Brain Proliferation Zones Exhibit Neuroepithelial-Type Gene Expression Profile: Features of Neuroepithelial Cells in Fish », STEM CELLS, 2017.
    Mots-clés : BMgif, CYTO, IMAGIF, NGS, PF, PHOT.

  • J. Marion, R. Le Bars, B. Satiat-Jeunemaitre, et C. Boulogne, « Optimizing CLEM protocols for plants cells: GMA embedding and cryosections as alternatives for preservation of GFP fluorescence in Arabidopsis roots », Journal of Structural Biology, 2017.
    Mots-clés : Arabidopsis, BIOCELL, Correlative microscopy, DYNBSJ, GFP, GMA resin, IMAGIF, MET, PF, PHOT, Tokuyasu, Transmission electron microscopy.

  • J. Nikolic, R. Le Bars, Z. Lama, N. Scrima, C. Lagaudrière-Gesbert, Y. Gaudin, et D. Blondel, « Negri bodies are viral factories with properties of liquid organelles », Nature Communications, vol. 8, nᵒ 1, p. 58, juill. 2017.
    Résumé : Replication of Mononegavirales occurs in viral factories which form inclusions in the host-cell cytoplasm. For rabies virus, those inclusions are called Negri bodies (NBs). We report that NBs have characteristics similar to those of liquid organelles: they are spherical, they fuse to form larger structures, and they disappear upon hypotonic shock. Their liquid phase is confirmed by FRAP experiments. Live-cell imaging indicates that viral nucleocapsids are ejected from NBs and transported along microtubules to form either new virions or secondary viral factories. Coexpression of rabies virus N and P proteins results in cytoplasmic inclusions recapitulating NBs properties. This minimal system reveals that an intrinsically disordered domain and the dimerization domain of P are essential for Negri bodies-like structures formation. We suggest that formation of liquid viral factories by phase separation is common among Mononegavirales and allows specific recruitment and concentration of viral proteins but also the escape to cellular antiviral response.Negative strand RNA viruses, such as rabies virus, induce formation of cytoplasmic inclusions for genome replication. Here, Nikolic et al. show that these so-called Negri bodies (NBs) have characteristics of liquid organelles and they identify the minimal protein domains required for NB formation.
    Mots-clés : IMAGIF, PF, PHOT, RHABDO, VIRO.

  • J. - A. Pedroza-García, C. Mazubert, I. del Olmo, M. Bourge, S. Domenichini, R. Bounon, Z. Tariq, E. Delannoy, M. Piñeiro, J. A. Jarillo, C. Bergounioux, M. Benhamed, et C. Raynaud, « Function of the Plant DNA Polymerase Epsilon in Replicative Stress Sensing, a Genetic Analysis », Plant Physiology, vol. 173, nᵒ 3, p. 1735-1749, 2017.

  • P. Pétriacq, L. de Bont, L. Genestout, J. Hao, C. Laureau, I. Florez-Sarasa, T. Rzigui, G. Queval, F. Gilard, C. Mauve, F. Guérard, M. Lamothe-Sibold, J. Marion, C. Fresneau, S. Brown, A. Danon, A. Krieger-Liszkay, R. Berthomé, M. Ribas-Carbo, G. Tcherkez, G. Cornic, B. Pineau, B. Gakière, et R. De Paepe, « Photoperiod Affects the Phenotype of Mitochondrial Complex I Mutants », Plant Physiology, vol. 173, nᵒ 1, p. 434-455, 2017.

  • Z. M. Song, L. Bouchab, E. Hudik, R. Le Bars, O. Nüsse, et S. Dupré-Crochet, « Phosphoinositol 3-phosphate acts as a timer for reactive oxygen species production in the phagosome », Journal of Leukocyte Biology, p. jlb.1A0716-305R, janv. 2017.


  • M. Benincasa, Q. Barrière, G. Runti, O. Pierre, M. Bourge, M. Scocchi, et P. Mergaert, « Single Cell Flow Cytometry Assay for Peptide Uptake by Bacteria », BIO-PROTOCOL, vol. 6, nᵒ 23, 2016.
    Mots-clés : CYTO, IMAGIF, MICROBIO, PBI, PF.

  • C. Chaintreuil, D. Gully, C. Hervouet, P. Tittabutr, H. Randriambanona, S. C. Brown, G. P. Lewis, M. Bourge, F. Cartieaux, M. Boursot, H. Ramanankierana, A. D'Hont, N. Teaumroong, E. Giraud, et J. - F. Arrighi, « The evolutionary dynamics of ancient and recent polyploidy in the African semiaquatic species of the legume genus Aeschynomene », The New Phytologist, vol. 211, nᵒ 3, p. 1077-1091, août 2016.
    Résumé : The legume genus Aeschynomene is notable in the ability of certain semiaquatic species to develop nitrogen-fixing stem nodules. These species are distributed in two clades. In the first clade, all the species are characterized by the use of a unique Nod-independent symbiotic process. In the second clade, the species use a Nod-dependent symbiotic process and some of them display a profuse stem nodulation as exemplified in the African Aeschynomene afraspera. To facilitate the molecular analysis of the symbiotic characteristics of such legumes, we took an integrated molecular and cytogenetic approach to track occurrences of polyploidy events and to analyze their impact on the evolution of the African species of Aeschynomene. Our results revealed two rounds of polyploidy: a paleopolyploid event predating the African group and two neopolyploid speciations, along with significant chromosomal variations. Hence, we found that A. afraspera (8x) has inherited the contrasted genomic properties and the stem-nodulation habit of its parental lineages (4x). This study reveals a comprehensive picture of African Aeschynomene diversification. It notably evidences a history that is distinct from the diploid Nod-independent clade, providing clues for the identification of the specific determinants of the Nod-dependent and Nod-independent symbiotic processes, and for comparative analysis of stem nodulation.
    Mots-clés : Aeschynomene, CYTO, dysploidy, genome downsizing, IMAGIF, PF, PHOT, Polyploidy, stem nodulation, Symbiosis.

  • E. Galli, M. Poidevin, R. Le Bars, J. - M. Desfontaines, L. Muresan, E. Paly, Y. Yamaichi, et F. - X. Barre, « Cell division licensing in the multi-chromosomal Vibrio cholerae bacterium », Nature Microbiology, vol. 1, nᵒ 9, p. 16094, juin 2016.

  • P. Maisonnasse, E. Bouguyon, G. Piton, A. Ezquerra, C. Urien, C. Deloizy, M. Bourge, J. - J. Leplat, G. Simon, C. Chevalier, S. Vincent-Naulleau, E. Crisci, M. Montoya, I. Schwartz-Cornil, et N. Bertho, « The respiratory DC/macrophage network at steady-state and upon influenza infection in the swine biomedical model », Mucosal Immunology, vol. 9, nᵒ 4, p. 835-849, 2016.

  • J. A. Pedroza-Garcia, S. Domenichini, C. Mazubert, M. Bourge, C. White, E. Hudik, R. Bounon, Z. Tariq, E. Delannoy, I. del Olmo, M. Piñeiro, J. A. Jarillo, C. Bergounioux, M. Benhamed, et C. Raynaud, « Role of the Polymerase ϵ sub-unit DPB2 in DNA replication, cell cycle regulation and DNA damage response in Arabidopsis », Nucleic Acids Research, p. gkw449, mai 2016.


  • C. Bourbousse, I. Mestiri, G. Zabulon, M. Bourge, F. Formiggini, M. A. Koini, S. C. Brown, P. Fransz, C. Bowler, et F. Barneche, « Light signaling controls nuclear architecture reorganization during seedling establishment », Proceedings of the National Academy of Sciences, vol. 112, nᵒ 21, p. E2836-E2844, mai 2015.
    Mots-clés : CYTO, IMAGIF, PF, PHOT.

  • M. Bourge, C. Fort, M. - N. Soler, B. Satiat-Jeunemaître, et S. C. Brown, « A pulse-chase strategy combining click-EdU and photoconvertible fluorescent reporter: tracking Golgi protein dynamics during the cell cycle », New Phytologist, vol. 205, nᵒ 2, p. 938-950, 2015.
    Mots-clés : 5-ethynyl-2′-deoxyuridine (EdU), Arabidopsis, cell cycle, Cell Proliferation, Click Chemistry, Copper, CYTO, Deoxyuridine, Fluorescence, Fluorescent Dyes, fluorescent proteins, G1 subcompartments, Golgi Apparatus, Golgi synthesis, Green Fluorescent Proteins, IMAGIF, Kaede pulse-chase, Luminescent Proteins, Molecular Imaging, PF, PHOT, Plant Proteins, Plants, Genetically Modified, Protoplasts, Tobacco, tobacco (Nicotiana tabacum) BY2 cells.

  • I. Guefrachi, O. Pierre, T. Timchenko, B. Alunni, Q. Barrière, P. Czernic, J. - A. Villaécija-Aguilar, C. Verly, M. Bourge, J. Fardoux, M. Mars, E. Kondorosi, E. Giraud, et P. Mergaert, « Bradyrhizobium BclA Is a Peptide Transporter Required for Bacterial Differentiation in Symbiosis with Aeschynomene Legumes », Molecular plant-microbe interactions: MPMI, vol. 28, nᵒ 11, p. 1155-1166, nov. 2015.
    Résumé : Nodules of legume plants are highly integrated symbiotic systems shaped by millions of years of evolution. They harbor nitrogen-fixing rhizobium bacteria called bacteroids. Several legume species produce peptides called nodule-specific cysteine-rich (NCR) peptides in the symbiotic nodule cells which house the bacteroids. NCR peptides are related to antimicrobial peptides of innate immunity. They induce the endosymbionts into a differentiated, enlarged, and polyploid state. The bacterial symbionts, on their side, evolved functions for the response to the NCR peptides. Here, we identified the bclA gene of Bradyrhizobium sp. strains ORS278 and ORS285, which is required for the formation of differentiated and functional bacteroids in the nodules of the NCR peptide-producing Aeschynomene legumes. The BclA ABC transporter promotes the import of NCR peptides and provides protection against the antimicrobial activity of these peptides. Moreover, BclA can complement the role of the related BacA transporter of Sinorhizobium meliloti, which has a similar symbiotic function in the interaction with Medicago legumes.
    Mots-clés : Bacterial Proteins, Bradyrhizobium, CYTO, Fabaceae, Flow Cytometry, Genetic Complementation Test, Host-Pathogen Interactions, IMAGIF, Medicago, Membrane Transport Proteins, MICROBIO, Microscopy, Confocal, Molecular Sequence Data, Mutation, PBI, Peptides, PF, Phylogeny, Polyploidy, Root Nodules, Plant, Sinorhizobium meliloti, Symbiosis.

  • A. Hajrudinović, S. Siljak-Yakovlev, S. C. Brown, F. Pustahija, M. Bourge, D. Ballian, et F. Bogunić, « When sexual meets apomict: genome size, ploidy level and reproductive mode variation of Sorbus aria s.l. and S. austriaca (Rosaceae) in Bosnia and Herzegovina », Annals of Botany, vol. 116, nᵒ 2, p. 301-312, août 2015.
    Résumé : BACKGROUND AND AIMS: Allopolyploidy and intraspecific heteroploid crosses are associated, in certain groups, with changes in the mating system. The genus Sorbus represents an appropriate model to study the relationships between ploidy and reproductive mode variations. Diploid S. aria and tetraploid apomictic S. austriaca were screened for ploidy and mating system variations within pure and sympatric populations in order to gain insights into their putative causalities. METHODS: Flow cytometry was used to assess genome size and ploidy level among 380 S. aria s.l. and S. austriaca individuals from Bosnia and Herzegovina, with 303 single-seed flow cytometric seed screenings being performed to identify their mating system. Pollen viability and seed set were also determined. KEY RESULTS: Flow cytometry confirmed the presence of di-, tri- and tetraploid cytotype mixtures in mixed-ploidy populations of S. aria and S. austriaca. No ploidy variation was detected in single-species populations. Diploid S. aria mother plants always produced sexually originated seeds, whereas tetraploid S. austriaca as well as triploid S. aria were obligate apomicts. Tetraploid S. aria preserved sexuality in a low portion of plants. A tendency towards a balanced 2m : 1p parental genome contribution to the endosperm was shared by diploids and tetraploids, regardless of their sexual or asexual origin. In contrast, most triploids apparently tolerated endosperm imbalance. CONCLUSIONS: Coexistence of apomictic tetraploids and sexual diploids drives the production of novel polyploid cytotypes with predominantly apomictic reproductive modes. The data suggest that processes governing cytotype diversity and mating system variation in Sorbus from Bosnia and Herzegovina are probably parallel to those in other diversity hotspots of this genus. The results represent a solid contribution to knowledge of the reproduction of Sorbus and will inform future investigations of the molecular and genetic mechanisms involved in triggering and regulating cytotype diversity and alteration of reproductive modes.
    Mots-clés : Apomixis, Bosnia and Herzegovina, Cell Nucleus, CYTO, cytotypes, DNA, Plant, Endosperm, Flow Cytometry, Genome Size, Geography, IMAGIF, PF, PHOT, Ploidies, Pollen, Polyploidy, reproduction, reproduction mode, Rosaceae., Seeds, sexuality, Sorbus, Sorbus aria, Sorbus austriaca.

  • T. Jégu, S. Domenichini, T. Blein, F. Ariel, A. Christ, S. - K. Kim, M. Crespi, S. Boutet-Mercey, G. Mouille, M. Bourge, H. Hirt, C. Bergounioux, C. Raynaud, et M. Benhamed, « A SWI/SNF Chromatin Remodelling Protein Controls Cytokinin Production through the Regulation of Chromatin Architecture », PLOS ONE, vol. 10, nᵒ 10, p. e0138276, oct. 2015.
    Mots-clés : Alkyl and Aryl Transferases, Arabidopsis, Arabidopsis Proteins, Carrier Proteins, cell cycle, Chromatin, Chromatin Assembly and Disassembly, Chromosomal Proteins, Non-Histone, CYTO, Cytokinins, DNA, Plant, Epigenesis, Genetic, Genetic Loci, Histones, IMAGIF, Meristem, PF.

  • T. - P. Vu Manh, J. Elhmouzi-Younes, C. Urien, S. Ruscanu, L. Jouneau, M. Bourge, M. Moroldo, G. Foucras, H. Salmon, H. Marty, P. Quéré, N. Bertho, P. Boudinot, M. Dalod, et I. Schwartz-Cornil, « Defining Mononuclear Phagocyte Subset Homology Across Several Distant Warm-Blooded Vertebrates Through Comparative Transcriptomics », Frontiers in Immunology, vol. 6, juin 2015.
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par Laetitia Besse - publié le , mis à jour le