IJPB-Cell biology, plant regeneration

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JP Bourgin

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Cell Structures, Signals and Morphogenesis

Dynamics and Expression of plant Genomes

Adaptation of Plants to the Environment

Reproduction and Seeds

cytology and cell imaging

biochemistry

plant chemistry

phenotyping Arabidopsis

resources Arabidopsis

resources Brachypodium

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informatics

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Cell biology, plant regeneration

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Keywords :In vitro plant tissue culture - protoplasts - transfer of genes - transient expression- Agrobacterium

Doctoral school affiliation :

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 Catherine Pannetier Senior scientist CIRAD
	Yves Chupeau Senior scientist
	Camille Soulhat Technician

	Marie-Christine Chupeau Engineer Use and culture of protoplasts
	Oumaya Bouchabké-Coussa Engineer Genetic transformation of Brachipodium distachon
	Katleen Patron professional Licence 3 from 16/01/12 to 16/07/12

Summary :

The team is interested in the control of the regeneration process through organogenesis or somatic embryogenesis as well as in methods for the transfer of genes into plant cells cultivated in vitro. Our research relates to both fundamental processes (transitory activity, knock-out ...) and more narrowly focussed studies (resistance to abiotic stress, the use of bio mass...)

Our research centres on two model plants: Arabidopsis thaliana and Brachypodium distachyon in addition to cotton (Gossypium hirsutum) in collaboratiom with CIRAD and, more recently, oil palm (Elaeis guineensis, Elaeis oleifera).

Main Results :

Arabidopsis thaliana protoplasts (M.C Chupeau, Y. Chupeau) :

The work carried out on protoplasts allows pursuing and validating on the model plant Arabidopsis thaliana, our approaches developed on various species. With this species routinely in planta transformed via Agrobacterium, the main interest of in vitro and protoplasts culture is the possibility to manipulate large populations of cells in homogeneous conditions in order to select very rare events. We have improved the conditions for liquid and "en masse" culture of protoplasts of Arabidopsis (ecotype Columbia and WS) as well as for plant regeneration.

Shoot regeneration from Arabidopsis thaliana protoplast derived calli (ecotype WS)

Brachypodium genetic engineering (O. Bouchabke-Coussa, C. Soulhat)

We use Brachypodium distachyon as a tool for gene functional analysis in monocots. Routinely used approaches are geneover-expression, gene extinction (miR or RNAi) and localisation of gene expression. For this purpose we developed an efficient method of gene transfer via Agrobacterium tumefaciens and regeneration of transgenic plants via somatic embryogenesis, as well as series of binary vectors based on Gateway technology versatile and easy to handle to clone rapidly a large number of candidate genes (to know more). Our work deals mainly with functional analysis of genes involved in cell wall synthesis linked with “Primary wall” and “Secondary wall” teams at IJPB. We have also other projects in diverse subjects like nitrogen use or resistance to pathogens based on internal and external collaborations.

Plant regenerations on transformed embryogenic calli from immatures embryos

Cotton genetic engineering

Transgenic cotton plants are obtained through somatic embryogenesis on hypocotyl explants and Agrobacterium tumefaciens mediated transformation. Various methodological improvements have been developed through selection of plant material and culture condition optimization. (to know more)

Genes of interest are transferred into cotton genome and tools have been developed in order to carry out functional genomic studies on cotton fiber development.

In order to better understand and master the regeneration process, we are analyzing the effect of over expression of Arabidopsis genes involved in the apical meristem development

Using the LAM (Laser Assisted Micro-dissection) methodology we are performing the transcriptome of early steps of cotton somatic embryogenesis. This study is also carried out on organogenesis on the model plant Arabidopsis thaliana.

Participating to studies on the impact of transgenic cotton cultures for small-scale farmers, we have analyzed the possibility of cross-pollination between cultivated cotton (Gossypium hirsutum, tetraploid) and wild cotton (G. herbaceum, diploid. The evidence of non reduced gametes has been demonstrated. (to know more)

Agrobacterium tumefaciens mediated transformation and regeneration of cotton plants through somatic embryogenesis

Selected Publications :

Articles :
Vaucheret H, et Chupeau Y. (2011) Ingested plant miRNAs regulate gene expression in animal. Cell Research advance online publication 1 November 2011; doi:10.1038/cr.2011.Cell Research, (2011) :1-3.

Konan E.K, Durand-Gasselin T, Kouadio J.Y, Flori A, Rival A, Duval Y, Pannetier C. (2010). In vitro conservation of oil palm somatic embryos for 20 years on a hormone-free culture medium: characteristics of the embryogenic cultures, derived plantlets and adult palms. Plant cell reports, 29 (1) : 1-13. http://dx.doi.org/10.1007/s00299-009-0787-y

Tessadori F, Chupeau MC , Chupeau Y, Knip M, Germann S, van DrielL, R, Franz P, Gaudin V. (2007). Large-scale dissociation and sequential reassembly of pericentric heterochromatin in dedifferentiated Arabidopsis cells. J. Cell Sciences, 120, 1200-1208.

Mazier, M., Botton, E., Flamain, F., Bouchet, JP., CourtialL, B., Chupeau, MC. , Chupeau, Y., Maisonneuve, B., Lucas, H. (2007). Successful gene tagging in lettuce using the Tnt1 retrotransposon from tobacco. Plant Physiol, 144, 18-31.

Nicolas Le Moigne, Emilie Montes, Catherine Pannetier, Herman Höfte, Patrick Navard (2008) Gradient in Dissolution Capacity of Successively Deposited Cell Wall Layers in Cotton Fibres Macromolecular symposia, 262,1, 65-71.

Commun K, Mauro MC, Chupeau Y, Boulay M, Burrus M, Jeandet P (2003) Phytoalexin production in grapevine protoplasts during isolation and culture. Plant Physiol Bioch, 41: 317-323

Djenanne S, Chauvin JE, Quilleré I, Meyer C, Chupeau Y (2002). Introduction and expression of a deregulated tobacco nitrate reductase gene in potato lead to highly reduced nitrate levels in transgenic tubers. 11, 175-184. (PubMed)

Revues et vulgarisation :
Chupeau, Y., et Davey, M. (2007). Gene transfer to plants. pp 123–142. in « Functionnal Plant Genomics ». Morot-Gaudry, JF., Lea, P. Briat, JF., Eds Science Publishers Enfield, NH, USA.

Testard, J., et Chupeau, Y. (2007) OGM : quels risques. Editions Promethée- Bordeaux

Pannetier C (2006) Les cotonniers transgéniques résistant aux insectes ravageurs. Le Sélectionneur Français, 57,29-38.

Chupeau Y (2003) Transgenic plants towards healthier food. OGM et Sante, 7-18

Communications ou posters :
Bouchabké-Coussa O, Soulhat C, Bouvier d’Yvoire M, Antelme S, Le Bris P, Sibout R. (2011) Tools and methods for functional gene analysis in Brachypodium distachyon. Examples of genes involved in cell wall synthesis, 1st European Brachypodium Workshop proceedings, p.116.

Bouchabké-Coussa O, Bouvier d’Yvoire M, Sibout R, Darracq O, Antelme S, Jouanin L, Pollet B, Dalmais M, Chupeau M-C, Pannetier C, Chupeau Y. (2010) Brachypodium distachyon as a model system for studying genes involved in cell wall synthesis, 12th World Congress of the International Association for Plant Biotechnology and 2010 Meeting of the Society of In Vitro Biology, p.164.

Pannetier C, Montes E, Bouchabke O, Hamadeh B, Obellianne M, Maia-Grondard A, Kerhoas L. (2008) Embryogenèse somatique et transfert de gènes chez le cotonnier, Gossypium hirsutum L. poster. Abstract in: Actes du colloque BIOTECH 2008 : Biotechnologies végétales et gestion durables desrésistances aux stress biotiques et abiotiques, p 202. Rennes juillet 2008.

Pannetier C, Hofs JL, Montes E, Eber F, Coriton O, Huteau V, Hau B, Chevre AM (2009) Evidence of unreduced gamete production from interspecific crosses between Gossypium hirsutum and G. herbaceum. In International Conference on Polyploidy, Hybridization and Biodiversity May 17-20 St Malo France p 91.

Publications pour la jeunesse :
Le Quérré V, Chupeau Y, Legrand P, LeGuyader H (1999). La reine rouge (BD garantie avec OGM). INRA Editions.