presentation
  access
  management
  job opportunities
  teaching resources
  publications
  contacts
  Morphogenesis, Signaling, Modeling
  Dynamics and Expression of plant Genomes
  Adaptation of Plants to the Environment
  Reproduction and Seeds
b
  Plant cell wall, function and utilization
  administrative team
  communication
  informatics
  workshop
  kitchen
  store
IJPB
blanc msm blanc deg blanc ape blancrg blancrg
blanc iNRA
présentation pôles laboratoires communs services communs intranet liens actualité
 
Cell differentiation and polarity
 presentation
 acces
 contact
 research groups
 publications

Keywords : Arabidopsis thaliana, Camelina sativa, Embryo development, Vascular tissues, Cell polarity, membrane dynamic, micro-domains, fatty acids, Sphingolipids.

Doctoral school affiliation : Sciences du végétal (ParisXI Orsay)
Areas of research and expertise : Plant Genetics, Plant Molecular Biology, Cell Biology and imaging, Developmental Biology, Lipid biochemistry   

Contacts :

Institut Jean-Pierre Bourgin, UMR1318 INRA-AgroParisTech
Bâtiment 2
INRA Centre de Versailles-Grignon
Route de St-Cyr (RD10)
78026 Versailles Cedex France

tél : +33 (0)1 30 83 30 00 - fax : +33 (0)1 30 83 33 19
   

Group leader
Jean-Denis Faure

Research director INRA
Professor AgroParisTech



Jean-Christophe Palauqui
Research Scientist INRA

 

Marine Froissard
Research Scientist INRA

Yannick Bellec
Technician INRA

Marianne Azopardi
Technician INRA


Kian Hématy
Research Scientist
Jean-Luc Cacas
Associate Professor AgroParisTech

Lionel Gissot
Engineer INRA

Abdelhak Fahiti
Post-Doc

Abdelhak.Fahiti@inra.fr

Kaori Sakai
Post-Doc

Kaori.Sakai@inra.fr

 

Summary

:

Plant cells are not mobile and their identity is inferred by their positions within the tissue. The different positional cues like hormonal or metabolic gradients, intercellular transport of transcription factors or mechanical stress to cite a few, are directly or indirectly mediated by a plasma membrane activity. Cell differentiation and cellular polarity could even be directly coupled in the case of auxin transport. Cell polarity is usually defined by the asymmetric distribution of morphological or molecular markers involving membrane constituents (plasma or endomembranes). Recent breakthrough in the understanding of plasma membrane lipid composition and their role in protein traffic and assembly provide now the basis for more in depth analysis of lipid function in plant plasma membrane structure and dynamics. The main questions addressed by our group are : (i) what cellular and subcellular mechanisms in particular those involving membrane lipids are governing cell polarity and (ii) how cell differentiation is inferred by cell polarity?


Main Results :

jpg
Lipids involvement in cell proliferation and differentiation
(Y Bellec, L Gissot, K Hematy, JD Faure). We have identified several mutants (pasticcino, pas) linking ectopic cell proliferation and tumor-like structures with fatty acid elongation (Faure et al. 1998; Vittorioso et al. 1998; Bellec et al. 2002; Baud et al. 2004; Bach et al. 2008) (Fig.1). The reduction of very-long-chain fatty acids (VLCFAs) occurred mainly in seed storage triacylglycerols, epicuticular waxes, and sphingolipids. VLCFAs depletion caused cell proliferation and several defects in tissue patterning  and cell polarity. In particular, The reduction of VLCFA in pas1 caused mistargetting of PIN1 and altered polar auxin transport during embryo organogenesis and lateral root development (Roudier et al., 2010). Among the different VLCFA-containing lipids, sphingolipids are known to be lipids involved in membrane trafficking and cell polarity.
Functional analysis of the ceramide synthase family in Arabidopsis thaliana demonstrated that very-long-acyl-chain (C > 18 carbons) but not long-chain sphingolipids are essential for plant development. Reduction of very-long-chain fatty acid sphingolipid levels leads in particular to auxin-dependent inhibition of lateral root emergence that is associated with defective targeting of polar auxin carriers (Markham et al., 2011).

 

jpgAcyl chain length and membrane dynamics (L Gissot, K Hematy, JD Faure).  Acyl chain length is thought to be crucial for biophysical properties of the membrane, in particular during cell division, when active vesicular fusion is necessary. Cell plate expansion was delayed and the formation of the endomembrane tubular network altered when VLCFA elongation was blocked (Bach et al., 2011). Inhibition of VLCFA-ceramide synthase induced also cytokinesis defects with specific marker retention at the cell plate associated with extended endosome interaction time. By using reconstructed liposomes, we demonstrated that glucosylceramide acyl chain length is directly involved in vesicle aggregation and fusion (Molino et al. in prep.).

 

jpgOPS regulates vascular differentiation (JC Palauqui). Vascular development is embedded into the developmental context of plant organ differentiation and can be divided into the consecutive phases of vascular patterning and differentiation of specific vascular cell types (phloem and xylem). To date, only very few genetic determinants of phloem development are known. We identified OCTOPUS (OPS) as a potentiator of phloem differentiation (Truernit et al., 2012). OPS is a polarly localized membrane-associated protein that is initially expressed in provascular cells and upon vascular cell type specification, becomes restricted to the phloem cell lineage. The ops mutants display discontinuous phloem differentiation suggesting that OPS integrates longitudinal signals that lead to the transformation of vascular initials into differentiating protophloem cells.

 

 


Development of new tools and methods
Sphingolipidomic. First, we developped an efficient sensitive HPLC assay to monitor long chain bases of sphingolipids (Bach et al., 2008; Lachaud et al., 2009, 2010; Melser et al., 2010; Roudier et al., 2010; Aubert et al., 2011; Bach et al., 2011; Markham et al., 2011).
We also set up a single injection mass spectrometry assay to analyze more than 350 type of sphingolipids (Tellier et al. Submitted).
jpg
High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression. Anoriginal staining method was developed for precisely imaging any kind of plant tissues that is now the basis for cellular phenotyping and modeling (Truernit et al., 2008; Truernit and Palauqui, 2009; Wuyts et al., 2010). This method allows also to visualize gene expression with GUS reporter genes.

Laser Assisted Microdissection of RNAs and lipids. In the frame of the ANR Génoplante project "REGENEOME" we have developped Laser Assisted Microdissection (LAM) for transcriptome and chromatin profiling of specific cells isolated at different stages of the regeneration process. We are now using this tool to provide a lipid maps of plant tissues.

 

jpgProtein-protein interactions in living cells. In the frame of the EU project AGRON-OMICS (http://www.agron-omics.eu/), we aim in deciphering protein interaction networks involved in leaf growth. We are thus developping medium to high throuput analysis of protein interaction by split-reporter assays like Bimolecular Fluorescence Complementation (BiFC) or split luciferase assay. We have also developped an optimized protocol for transient expression in Arabidopsis seedlings allowing efficient co-expression of fluorescent protein fusions in cotyledon cells (Marion et al. 2008). Subcellular distribution and co-localisation of protein can thus be directly adressed but also systematic BiFC studies (Fig.Marion.cover). These approaches were used in several studies vivo assays based on split reporter gene techniques (Bach et al., 2008; Marion et al., 2008; Roudier et al., 2010; Van Leene et al., 2010; Markham et al., 2011; Bernard et al., 2012).
To provide an easy and non invasive way to follow protein interaction in time and space, we are also developping the use of fluorescence anisotropy by confocal microscopy.


jpg
Translational research. Arabidopsis research on VLCFAs and sphingolipids provided several lines of potential interest for improving oil yield and quality.  To validate some of these findings, we used Camelina sativa, a Brassicae specie that is easy to transform and to extract oil from. We developped new vectors and optimized genetic transformation of Camelina. The fact that Camelina showed a very low ability of crosses to wild relatives provide an opportunity to also use this specie for translational research and field trials (Julie-Galau et al. 2013).



Our current projects - (Master and PhD projects available - Join us!)
Our ongoing projects
1. Functional analysis of OPS in plant development (JC Palauqui). The project focus on OPS family and OPS protein interactors.

b) Genetic dissection of lipid involvement in cell differentiation (Y Bellec, L Gissot, K Hematy, C Morineau). New genetic screen in plants and in yeast are currently ongoing to decipher the regulatory network associated with fatty acid elongation.

c) Structural role of lipids in membrane domains (K Hematy).

jpg In the last couple of decades, the plasma membrane (pm) has been shown to be more heterogenous and organized than initially thought with macrodomains (micrometer scale) and microdomains (nanometer scale). The localizations of relevant markers of macro- and micro-domains were characterized in different mutant backgrounds revealing the importance of sterols and VLCFA. Combination of molecular genetics, lipid chemistry, imaging and biophysics will be used to address the role of the different class of lipids in the organization of the pm.

 

 

jpgd) From cellular assymetry to tissue organization (JC Palauqui). In order to understand how cell orientation and subcellular organization is integrated into tissue organogenesis, we developped a multiscale approach describing early steps of embryo development. The spatio-temporal dynamics of cell divisions in early embryo based  on cellular size and shapes identifyed cellular assymetry governing cell fate (Trubuil et al., in preparation). The objective will be to integrate the different subcellular components (lipids, cytoskeleton…) into a 4D digital model to evaluate the role of polarity into this developmental process.

e) Lipid biosynthesis, enzymatic complexes and oil quality (Y Bellec, L Gissot, JD Faure). The existence of large protein-protein interactions among lipid biosynthetic enzymes as well as the discovery of the role of PAS1 as a putative scaffold protein suggest supramolecular protein organization in the ER (metabolon). Deciphering the different molecular interactions within fatty acid biosynthesis as well as their the molecular determinants will be provide a conceptual framework to improve oil quality. Modifying Camelina oil quality for nutritional and industrial use will be the aim of the project.


Selected Publications :

Morineau, C., Bellec, Y., Gissot, L., Kelemen, Z., Nogué, F. and Faure J.-D., (2017). Selective gene dosage by CRISPR-Cas9 genome editing in hexaploid Camelina sativa in press. doi:10.1111/pbi.12671
De Bigault Du Granrut, A. and Cacas, J.-L. (2016). How Very-Long-Chain Fatty Acids Could Signal Stressful Conditions in Plants? Front. Plant Sci. 7: 1–13.

Molino, D., Markham, J.E., Bellec, Y., Gissot, L., Palauqui, J.-C., Moreau, P., Napier, J.A., Faure, J.D., (2015). Very long chain fatty acids are required for cell polarity and organogenesis during plant development. Chem. Phys. Lipids 163, S17. doi:10.1016/j.chemphyslip.2010.05.054

Molino D, Van der Giessen E, Gissot L, Hématy K, Marion J, Barthelemy J, Bellec Y, Vernhettes S, Satiat-Jeunemaître B, Galli T and JD Faure (2014) Inhibition of very long acyl chain sphingolipid synthesis modifies membrane dynamics during plant cytokinesis. Biochim Biophys Acta 1842: 1422–30

Tellier F, Maia-Grondard A, Schmitz-Afonso I, Faure J-D (2014) Comparative plant sphingolipidomic reveals specific lipids in seeds and oil. Phytochemistry 103: 50–8
Julié-Galau S, Bellec Y, Faure J-D, Tepfer M (2013) Evaluation of the potential for interspecific hybridization between Camelina sativa and related wild Brassicaceae in anticipation of field trials of GM camelina. Transgenic Research, in press

Bach, L., Gissot, L., Marion, J., Tellier, F., Moreau, P., Satiat-Jeunemaitre, B., Palauqui, J.C., Napier, J.A., and Faure, J.D. (2011). Very long chain fatty acids are required for cell plate formation during cytokinesis in Arabidopsis thaliana. J. Cell Sci. 124:3223-3234.

Melser, S., Molino, D., Batailler, B., Peypelut, M., Laloi, M., Wattelet-Boyer, V., Bellec, Y., Faure, J.D., and Moreau, P. (2011). Links between lipid homeostasis, organelle morphodynamics and protein trafficking in eukaryotic and plant secretory pathways. Plant Cell Rep 30, 177-193.

Markham, J.E.*, Molino, D.*, Gissot, L., Bellec, Y., Hématy, K., Marion, J., Belcram, K., Palauqui, J.C., Satiat-Jeunemaitre, B., and Faure, J.D. (2011). Sphingolipids containing very-long chain fatty acids define a secretory pathway for specific polar plasma membrane protein targeting. Plant Cell 23:2362-78. * Co-premiers auteurs.

Aubert A, Marion J, Boulogne C, Bourge M, Abreu S, Bellec Y, Faure JD, Satiat-Jeunemaitre B (2011) Sphingolipids involvement in plant endomembrane differentiation: the BY2 case. Plant J 65: 958-971 (Pubmed)

Bach L, Gissot L, Marion J, Tellier F, Moreau P, Satiat-Jeunemaitre B, Palauqui JC, Napier JA, Faure JD (2011) Very-long-chain fatty acids are required for cell plate formation during cytokinesis in Arabidopsis thaliana. Journal of cell science 124: 3223-3234 (Pubmed)

Bach L, Michaelson L, Haslam R, Bellec Y, Gissot L, Marion J, Da Costa M, Boutin J-P, Miquel M, Tellier F, Domergue F, Markham J, Beaudoin F, Napier J, Faure J-D (2008) The plant very long chain hydroxy fatty Acyl-CoA dehydratase PASTICCINO2 is essential and limiting for plant development. Proc. Natl. Acad. USA 105: 14727-14731 .(Pubmed)

Baud S, Bellec Y, Miquel M, Bellini C, Caboche M, Lepiniec L, Faure JD, Rochat C (2004) gurke and pasticcino3 mutants affected in embryo development are impaired in acetyl-CoA carboxylase. EMBO Rep 5: 1-6 (PubMed)

Bernard A, Domergue F, Pascal S, Jetter R, Renne C, Faure JD, Haslam RP, Napier JA, Lessire R, Joubes J (2012) Reconstitution of Plant Alkane Biosynthesis in Yeast Demonstrates That Arabidopsis ECERIFERUM1 and ECERIFERUM3 Are Core Components of a Very-Long-Chain Alkane Synthesis Complex. The Plant cell 24: 3106-3118

Jolivet P, Acevedo F, Boulard C, d’Andrea S, Faure JD, Kohli A, Nesi N, Valot B, Chardot T (2013) Crop seed oil bodies: From challenges in protein identification to an emerging picture of the oil body proteome. Proteomics. doi: 10.1002/pmic.201200431

Julié-Galau S, Bellec Y, Faure J-D, Tepfer M (2013) Evaluation of the potential for interspecific hybridization between Camelina sativa and related wild Brassicaceae in anticipation of field trials of GM camelina. Transgenic research. doi: 10.1007/s11248-013-9722-7

Lachaud C, Da Silva D, Cotelle V, Thuleau P, Xiong TC, Jauneau A, Briere C, Graziana A, Bellec Y, Faure JD, Ranjeva R, Mazars C (2009) Nuclear calcium controls the apoptotic-like cell death induced by d-erythro-sphinganine in tobacco cells. Cell Calcium, 47, 1, 92-100 (Pubmed)

Marion J, Bach L, Bellec Y, Meyer C, Gissot L, Faure JD (2008) Systematic analysis of protein subcellular localization and interaction using high-throughput transient transformation of Arabidopsis seedlings. Plant J 56: 169-179 (Pubmed)

Markham JE, Molino D, Gissot L, Bellec Y, Hematy K, Marion J, Belcram K, Palauqui JC, Satiat-Jeunemaitre B, Faure JD (2011) Sphingolipids Containing Very-Long-Chain Fatty Acids Define a Secretory Pathway for Specific Polar Plasma Membrane Protein Targeting in Arabidopsis. Plant Cell 23: 2362-2378 (Pubmed)

Melser S, Batailler B, Peypelut M, Poujol C, Bellec Y, Wattelet-Boyer V, Maneta-Peyret L, Faure JD, Moreau P (2010) Glucosylceramide Biosynthesis is Involved in Golgi Morphology and Protein Secretion in Plant Cells. Traffic 11: 479-490 (Pubmed)

Roudier F, Gissot L, Beaudoin F, Haslam R, Michaelson L, Marion J, Molino D, Lima A, Bach L, Morin H, Tellier F, Palauqui JC, Bellec Y, Renne C, Miquel M, Dacosta M, Vignard J, Rochat C, Markham JE, Moreau P, Napier J, Faure JD (2010) Very-long-chain fatty acids are involved in polar auxin transport and developmental patterning in Arabidopsis. Plant Cell 22: 364-375 (pubmed)

Truernit E, Bauby H, Belcram K, Barthelemy J, Palauqui JC (2012) OCTOPUS, a polarly localised membrane-associated protein, regulates phloem differentiation entry in Arabidopsis thaliana. Development 139: 1306-1315

Truernit E, Bauby H, Dubreucq B, Grandjean O, Runions J, Barthelemy J, Palauqui JC (2008) High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression enables the study of Phloem development and structure in Arabidopsis. Plant Cell 20: 1494-1503 (Pubmed)

Truernit E, Palauqui JC (2009) Looking deeper: whole-mount confocal imaging of plant tissue for the accurate study of inner tissue layers. Plant Signal Behav 4: 151-152

Van Leene J, Hollunder J, Eeckhout D, Persiau G, Van De Slijke E, Stals H, Van Isterdael G, Verkest A, Neirynck S, Buffel Y, De Bodt S, Maere S, Laukens K, Pharazyn A, Ferreira PC, Eloy N, Renne C, Meyer C, Faure JD, Steinbrenner J, Beynon J, Larkin JC, Van de Peer Y, Hilson P, Kuiper M, De Veylder L, Van Onckelen H, Inze D, Witters E, De Jaeger G (2010) Targeted interactomics reveals a complex core cell cycle machinery in Arabidopsis thaliana. Mol Syst Biol 6: 397

Wuyts N, Palauqui JC, Conejero G, Verdeil JL, Granier C, Massonnet C (2010) High-contrast three-dimensional imaging of the Arabidopsis leaf enables the analysis of cell dimensions in the epidermis and mesophyll. Plant Methods 6: 17

 

Other publications

 


© INRA 2010
home IJPB