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Biology of the Cell and Regeneration
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Keywords : Organogenesis, somatic embryogenesis, in vitro culture, gene transfer

Doctoral school affiliation : ED 145 (Sciences du végétal) Université Paris-Saclay

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
Pierre Hilson
Senior Scientist

 

Liudmila Chelysheva
Engineer

Philippe Rech
Lecturer Paris VI

Camille Soulhat

Camille Soulhat
Technician

 

Manon Louis
Engineer (project LabEx SPS MISEDIT)
from 1/9/16 to 31/8/18

 

Catherine Pannetier
Senior Scientist CIRAD

Oumaya Bouchabké-Coussa
Engineer
Genetic transformation of Brachipodium distachon

Marco Da Costa
Lecturer Paris VI

 

Fatima Aissa Abdi
PhD student
from 1/9/2014 to 31/8/2017

 

 

Summary :


Our team studies the developmental programs that control the plant regeneration process, either via organogenesis or somatic embryogenesis. Furthermore, we seek to improve in vitro culture methods with the aim to alleviate bottlenecks in crop regeneration, transformation and genome engineering.

We work on two model plant species, Arabidopsis thaliana and Brachypodium distachyon, as well as on breadwheat (Triticum aestivum) and, in collaboration with CIRAD, on cotton (Gossypium hirsutum).


 


Main Results :

 

1. Brachypodium transgenesis and gene function analysis (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

 

 

2. Somatic embryogenesis and genetic engineering of cotton (C. Pannetier)

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

 

 

3. Organogenesis and meristem plasticity (M. Da Costa, P. Rech)

Lateral root meristems (LRM) can be converted into shoot meristems (SM) following treatment with phytohormones. We study this process in the model plant Arabidopsis thaliana to better understand the plasticity of organogenesis, with a focus on the molecular factors that determine whether plant cells are totipotent or differentiate.

The transformation of a LRM into a SM is rapid and involves a major reorganization of the stem cell niches. We are developing tools to track the cellular events occurring during this conversion. We search for the genetic and epigenetic factors involved and we examine their spatio-temporal activity. Our long-term goal is to model organogenetic reprogramming to better control the regeneration process.

 


 


4. Arabidopsis protoplast regeneration (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)


 


Selected Publications :


Articles:
Fernandez A, Hilson P and Beeckman T (2013) GOLVEN peptides as important regulatory signaling molecules of plant development. J. Exp. Bot. 64, in press.

Bhosale R, Jewell JB, Hollunder J, Koo AJ, Vuylsteke M, Michoel T, Hilson P, Goossens A, Howe GA, Browse J and Maere S (2013) Predicting gene function from uncontrolled expression variation among individual wild-type Arabidopsis plants. Plant Cell 25, in press.

Chupeau MC, Granier F, Pichon O, Renou JP, Gaudin V and Chupeau Y (2013) Characterization of the early events leading to totipotency in an Arabidopsis protoplast liquid culture by temporal transcript profiling. Plant Cell 25, Epub ahead of print.

Bouchabké-Coussa O, Obellianne M, Linderme D, Montes E, Maia-Grondard A, Vilaine F and Pannetier C (2013) Wuschel overexpression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro. Plant Cell Rep. 32: 675-86..

Fernandez A, Drozdzecki A, Hoogewijs K, Nguyen A, Madder A, Beeckman T and Hilson P (2013) Transcriptional and functional classification of the GLV/RGF/CLEL signaling peptides reveals their role in lateral root and hair formation. Plant Physiol. 161: 954-970.

Karimi M, Inzé D, Van Lijsebettens M and Hilson P (2013) Gateway vectors for transformation of cereals. Trends Plant Sci. 18: 1-4.

Baerenfaller K, Massonnet C, Walsh S, Baginsky S, Bühlmann P, Hennig L, Hirsch-Hoffmann M, Howell K, Kahlau S, Radziejwoski A, Russenberger D, Rutishauser D, Small I, Sulpice R, Stekhoven D, Wuyts N, Stitt M, Hilson P, Granier C and Gruissem W (2012) Systems-based analysis of Arabidopsis leaf growth reveals adaptation to water deficit. Mol. Syst. Biol. 8: 606.

Bouvier d'Yvoire M, Bouchabké-Coussa O, Voorend W, Antelme S, Cézard L, Legée F, Lebris P, Legay S, Whitehead C, McQueen-Mason SJ, Gomez LD, Jouanin L, Lapierre C and Sibout R (2012) Disrupting the cinnamyl alcohol dehydrogenase 1 gene (BdCAD1) leads to altered lignification and improved saccharification in Brachypodium distachyon. Plant J. 73: 496-508.

Vaucheret H, Chupeau Y (2012) Ingested plant miRNAs regulate gene expression in animals. Cell Res. 22: 3-5.

Whitford R, Fernandez A, Tejos R, Cuéllar Pérez A, Kleine-Vehn J, Vanneste S, Leitner J, Abas L, Aerts M, Drozdzecki A, Hoogewijs K, Baster P, De Groodt R, Lin Y-C, Storme V, Van de Peer Y, Beeckman T, Madder A, Devreese B, Luschnig C, Friml J and Hilson P (2012) GOLVEN secretory peptides regulate auxin carrier turnover during plant gravitropic responses. Dev. Cell 22: 678-685.

Anami SE, Mgutu AJ, Taracha C, Coussens G, Karimi M, Hilson P, Van Lijsebettens M and Machuka J (2010) Somatic embryogenesis and plant regeneration of tropical maize genotypes. Plant Cell Tiss. Organ Cult. 102: 285-295.

Konan EK, Durand-Gasselin T, Kouadio JY, 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 Rep. 29: 1-13.

Rahantamalala A, Rech P, Martinez Y, Chaubet-Gigot N, Grima-Pettenati J and Pacquit V (2010) Coordinated transcriptional regulation of two key genes in the lignin branch pathway - CAD and CCR - is mediated through MYB- binding sites. BMC Plant Biol. 10: 130.

Atta R, Laurens L, Boucheron-Dubuisson E, Guivarc'h A, Carnero E, Giraudat-Pautot V, Rech P, Chriqui D (2009) Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grown in vitro. Plant J. 57: 626-644.

Fernandez A, Viron N, Alhagdow M, Karimi M, Jones M, Amsellem Z, Sicard A, Czerednik A, Angenent G, Grierson D, May S, Seymour G, Eshed Y, Lemaire-Chamley M, Rothan C and Hilson P (2009) Flexible tools for gene expression and silencing in tomato. Plant Physiol. 151: 1729-1740.

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.

Bouchabké-Coussa O, Quashie ML, Seoane-Redondo J, Fortabat MN, Gery C, Yu A, Linderme D, Trouverie J, Granier F, Téoulé E and Durand-Tardif M (2008) ESKIMO1 is a key gene involved in water economy as well as cold acclimation and salt tolerance. BMC Plant Biology 8: 125.

Bouchabké O, Chang F, Simon M, Voisin R, Pelletier G and Durand-Tardif M (2008) Natural variation in Arabidopsis thaliana as a tool for highlighting differential drought responses. PLoS One 3: e1705.

Whitford R, Fernandez A, De Groodt R, Ortega E and Hilson P (2008) Plant CLE peptides from two distinct functional classes synergistically induce division of vascular cells. Proc. Natl. Acad. Sci. USA 105: 18625-18630.

Reviews :
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.

 

 

 
 


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