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Scientific topics: Plant Genetics, Plant Molecular Biology, Biotechnology, Cell Biology, Developmental Biology, Functional Genomics, Plant Genetics, Plant Molecular Biology, Biotechnology, Cell Biology, Developmental Biology, Functional Genomics.

 

Keywords : Sexual reproduction, Mutant, Mitochondria, Plastids, PPR proteins, Nucleo-cytoplasmic male sterility, Arabidopsis thaliana, Rapeseed (Brassica napus), Raphanus , Cytonuclear coadaptation.

 
Doctoral school affiliationt : ED 145 Sciences du végétal, Université Paris-Sud 11, Orsay

Contacts :

Institut Jean-Pierre Bourgin, UMR1318 INRA-AgroParisTech
Bâtiment 7
INRA Centre de Versailles-Grignon
Route de St-Cyr (RD10)
78026 Versailles Cedex France

tél : +33 (0)1 30 83 31 80- fax : +33 (0)1 30 83 33 19

   

Group leader
Françoise Budar
Senior scientist
Cytonuclear co-adaptation in Arabidopsis thaliana
Evolution of restoration genes in the Ogura Cytoplasmic Male Sterility system

Martine Quadrado
Technician

 

Céline Dargel-Graffin
Technicien

Noelya Planchard
PhD student

  

Hakim Mireau
Senior scientist
Functional and biochemical study of PPR proteins.

Noémie Dehaene
PhD student

Chuande Wang
PhD student



Tan-Trung Nguyen
Post-Doc

Tan-Trung.Nguyen@inra.fr


Summary :

Fluorescence image of an anther from an Arabidopsis thaliana transgenic plant producing a mitochondria-targeted GFP specifically in the tapetum. The red fluorescence is the auto-fluorescence of chlorophyll, the green fluorescence corresponds to GFP.

Our research programs focus mainly on the analysis of the functional relationship existing between the plant cell nucleus and 2 types of cellular organelles, the mitochondria and the chloroplasts. Besides the comprehension of these phenomena at the molecular level, we are interested in understanding the effects of this crosstalk on plant sexual reproduction. Cytoplasmic male sterilities (CMS) have long illustrated the peculiar involvement of mitochondria in plant sexual reproduction. CMS, which condition the inability to produce functional pollen, are being used in various plant species to facilitate the production of hybrid seeds in self-pollinating crop species. They actually rely on a double genetic determinant: a mitochondrial gene that induces the sterility and a counter-acting nuclear gene restoring the male fertility.
The analysis of nuclear factors regulating the expression of chloroplast and mitochondrial genes is also based on the biochemical characterization of PPR proteins (Pentatricopeptide Repeat). A fair number of PPR proteins have been shown to take part in various aspect of post-transcriptional regulation of gene expression in these organelles. Our interest for this family of proteins is also motivated by the fact that many identified genes that act as restorers of fertility in CMS systems encode PPR proteins.
We recently developed a project aiming at exploring the natural diversity in cytoplasmic genomes of A. thaliana and at using this diversity in order to study cyto-nuclear coadaptation.

 



Main Results :

We contributed to the identification of the Rfo gene, the restorer of fertility in the Ogura CMS system (Desloire et al, 2003). Using biochemical approaches, we showed that the restoring function of the Rfo gene product, a PPR protein, is correlated with its capacity to interact with the sterility gene mRNA (Uyttewaal et al, 2008b). We analyzed the sequence of a non-restoring allele of the Rfo locus and compared the PPR genes of this allele with those of the restorer one. We found that intra- and inter-genic recombinations occurred during the evolution of the Rfo locus (Hernandez Mora et al, 2010)
We recently decrypted the function of a mitochondria-targeted Arabidopsis PPR protein and revealed its essential role in the stabilization of the nad4 messenger RNA, encoding a respiratory complex I subunit (Haili et al, 2013). Analysis of its mode of action showed that this protein binds to the last 20 nucleotides of the nad4 3’ untranslated region and prevents its degradation by exoribonucleases. These results prove that the mRNA stability mechanism in plant mitochondria involving barrier proteins attached to mRNA extremities is conserved with the chloroplast, at least for what concerns the protection of mRNA 3’ ends. 

We also described cytoplasmic diversity among 95 accessions of Arabidopsis thaliana, and built a phylogeny of A. thaliana cytotypes (Moison et al, 2010). Our results also suggest that there is co-adaptation between nuclear and organelle genomes in this species. In collaboration with the Arabidopsis thaliana Resource Centre, we built genetic resources, named cytolines, dedicated to the study of cyto-nuclear coadaptation in this species. Cytolines possess the nuclear genome from one natural accession and cytoplasmic genomes (chloroplasts and mitochondria) from another. Their phenotypes are currently being characterized at different integrated levels.

We recently identified a cryptic CMS in A. thaliana, which is unlikely to be presently active in natural populations but strongly suggests a gynodioecious period in the history of the species (Gobron et al, 2013).

 


Selected Publications :

Primary papers

Gobron N, Waszcak C, Simon M, Hiard S, Boivin S, Charif D, Ducamp A, Wenes E, Budar F (2013) A Cryptic Cytoplasmic Male Sterility Unveils a Possible Gynodioecious Past for Arabidopsis thaliana. PLoS ONE 8(4):e62450.

Haïli N, Arnal N, Quadrado M, Amiar S, Tcherkez G, Dahan J, Briozzo P, Colas des Francs Small C, Vrielynck N, Mireau H (2013) The pentatricopeptide repeat MTSF1 protein stabilizes the nad4 mRNA in Arabidopsis mitochondria. Nucl Acids Res 41(13).

Hernandez-Mora, J.R., E. Rivals, H. Mireau and F. Budar (2010) « Sequence analysis of two alleles reveals that intra-and intergenic recombination played a role in the evolution of the radish fertility restorer (Rfo). » BMC Plant Biol 10 :35

Moison M, Roux F, Quadrado M, Duval R, Ekovich M, Lê DH, Verzaux M, Budar F(2010) Cytoplasmic phylogeny and evidence of cyto-nuclear co-adaptation in Arabidopsis thaliana. Plant J 63:728-738.

Duroc, Y., S. Hiard, N. Vrielynck, S. Ragu and F. Budar (2009). The Ogura sterility-inducing protein forms a large complex without interfering with the oxidative phosphorylation components in rapeseed mitochondria. Plant Mol Biol 70(1-2) :123-37.

Uyttewaal, M., N. Arnal, M. Quadrado, A. Martin-Canadell, N. Vrielynck, S. Hiard, H. Gherbi, A. Bendahmane, F. Budar and H. Mireau (2008b). "Characterization of Raphanus sativus Pentatricopeptide Repeat Proteins Encoded by the Fertility Restorer Locus for Ogura Cytoplasmic Male Sterility." Plant Cell 20(12): 3331-45.

Berthome, R. Thomasset, M. Maene, M. Bourgeois, N. Froger, N. Budar, F. (2008) pur4 mutations are lethal to the male, but not the female gametophyte and affect sporophyte development in Arabidopsis Plant Physiol 147(2) 650-60

Gonzalez-Melendi, P. Uyttewaal, M. Morcillo, C. N. Hernandez Mora, J. R. Fajardo, S. Budar, F. Lucas, M. M. (2008) A light and electron microscopy analysis of the events leading to male sterility in Ogu-INRA CMS of rapeseed (Brassica napus). J Exp Bot 59(4) 827-38

Uyttewaal M, Mireau H, Rurek M, Hammani K, Arnal N, Quadrado M, Giege P. (2008a) PPR336 is associated with polysomes in plant mitochondria. J Mol Biol 375(3) 626-36.

Giancola, S., Y. Rao, S. Chaillou, S. Hiard, A. Martin-Canadell, G. Pelletier and F. Budar (2007) Cytoplasmic suppression of Ogura cytoplasmic male sterility in European natural populations of Raphanus raphanistrum. Theor Appl Genet, 114:1333-1343.

Arnal N, Alban C, Quadrado M, Grandjean O, Mireau H. (2006) The Arabidopsis Bio2 protein requires mitochondrial targeting for activity. Plant Mol Biol 62 : 471-479.

Duroc, Y, Gaillard, C., Hiard, S., Tinchant, C, Berthomé, R, Pelletier, G and Budar, F (2006) « Nuclear expression of a cytoplasmic male sterility gene modifies mitochondrial morphology in yeast and plant cells. » Plant Science 170 :755-767. (pdf -intranet uniquement)


Review articles

Dahan J, Mireau H (2013) Rf and Rf-like PPRs in higher plants, an intriguing subclass of PPR proteins. RNA Biology, 10(9)..

Budar, F. & Roux, F., 2011. The role of organelle genomes in plant adaptation: Time to get to work! Plant signaling & behavior, 6(5), Pp. 635–639.

Pelletier, G. and F. Budar (2007) The molecular biology of cytoplasmically inherited male sterility and prospects for its engineering. Curr Opin Biotechnol 18(2): 121-125.

Budar F, Berthomé R. (2007) Cytoplasmic male sterilities and mitochondrial gene mutations in land plants. Logan, D. C. Plant Mitochondria. Blackwell Publishing 31 278-307.

Budar F, Touzet P & Pelletier G "Cytoplasmic male sterility" In Flowering and its manipulation, Annual Plant Reviews, vol 20 (C. Ainsworth Ed) Blackwell Publishing 2006.


Book Chapters

Budar, F. & Fujii, S., 2012. Cytonuclear Adaptation in Plants. In L. Marechal-Drouard, ed. Mitochondrial Genome Evolution, Advances in Botanical Research Volume 63. Elsevier, Pp. 99–126.

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