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"13598","6","archive","1",,,"disk0/00/01/35/98","2015-02-26 22:58:35","2016-07-08 09:49:31","2015-02-26 22:58:35","thesis",,,"show",,,,"","","","","","","","","","",,,,"Hao","Yanwei","","","","","Génomique et Biotechnologie des Fruits - GBF (Castanet-Tolosan, France)","","","","","","",,"","Auxin-mediated fruit development and ripening: new insight on the role of ARFs and their action mechanism in tomato (S. lycopersicum)","unpub",,,"","INPT","","public",,,"Tomato - ARFs - Fruit - Ethylene - Ripening - Development - Topless - TPL - Auxin response factors - Protein–protein interaction - Two-hybrid screening - Phenotype - Repressor - Histone - Hormone",,"harvest from ethesis","The plant hormone auxin coordinates plant development through the regulation of a specific set of auxin-regulated genes and Auxin Response Factors (ARFs) are transcriptional regulators modulating the expression of auxin-response genes. Recent data demonstrated that members of this gene family are able to regulate fruit set and fruit ripening. ARFs are known to act in concert with Aux/IAA to control auxin-dependent transcriptional activity of target genes. However, little is known about other partners of ARFs. The main objective of the thesis research project was to gain more insight on the involvement of ARFs in fruit development and ripening and to uncover their interaction with other protein partners beside Aux/IAAs. Mining the tomato expression databases publicly available revealed that among all tomato ARFs, SlARF2 displays the highest expression levels in fruit with a marked ripening-associated pattern of expression. This prompted us to uncover the physiological significance of SlARF2 and in particular to investigate its role in fruit development and ripening. Two paralogs, SlARF2A and SlARF2B, were identified in the tomato genome and transactivation assay in a single cell system revealed that the two SlARF2 proteins are nuclear localized and act as repressors of auxin-responsive genes. In fruit tissues, SlARF2A is ethylene-regulated while SlARF2B is auxin-induced. Knock-down of SlARF2A or SlARF2B results in altered ripening with spiky fruit phenotype, whereas simultaneous down-regulation of SlARF2A and SlARF2B leads to more severe ripening inhibition suggesting a functional redundancy among the two SlARF2 paralogs during fruit ripening. Double knock-down fruits produce less climacteric ethylene and show delayed pigment accumulation and higher firmness. Exogenous ethylene treatment cannot reverse the ripening defect phenotypes suggesting that SlARF2 may act downstream of ethylene signaling. The expression of key ethylene biosynthesis and signaling genes is dramatically disturbed in SlARF2 down-regulated fruit and major regulators of the ripening process, like RIN, CNR, NOR, TAGL1, are under-expressed. The data support the notion that SlARF2 is instrumental to fruit ripening and may act at the crossroads of auxin and ethylene signaling. Altogether, while ethylene is known as a key hormone of climacteric fruit ripening, the ripening phenotypes associated with SlARF2 down-regulation bring unprecedented evidence supporting the role of auxin in the control of this developmental process. To further extend our knowledge of the molecular mechanism by which ARFs regulate the expression of auxin-responsive genes we sought to investigate interactions SlARF and putative partners, mainly Aux/IAAs and Topless co-reppressors (TPLs) reported to be key players in gene repression dependent on auxin signaling. To this end, genes encoding all members of the tomato TPL family were isolated and using a yeast-two-hybrid approach comprehensive protein-protein interaction maps were constructed. The study revealed that Aux/IAA interact preferentially with activator SlARFs while Sl-TPLs interact only with repressor SlARFs. The data support the hypothesis that activator ARFs recruit Sl-TPLs co-repressors via Aux/IAAs as intermediates, while repressor ARFs can physically interact with Sl-TPLs. Further investigation indicated that SlARFs and Sl-TPLs can interact with polycomb complex PRC1 PRC2 components, VRN5 and LHP1, known to be essential players of epigenetic repression of gene transcription through the modification of histones methylation status. These data establish a potential link between ARFs and epigenetic regulation and thereby open new and original perspectives in understanding the mode of action of ARFs. Altogether, the thesis work provides new insight on the role of ARFs and their underlying action mechanisms, and defines SlARF2 as a new component of the regulatory network controlling the ripening process in tomato.","2014-11-14",,,,,,,,,,,,,,,,"Institut National Polytechnique de Toulouse","phd",,,,,,,,,"","http://ethesis.inp-toulouse.fr/archive/00002863/","","",,"","",,,,,,"",,,,,,,"","Bouzayen","Mondher","","","",,,,,"","",,,,,"","",,"oai:ethesis.inp-toulouse.fr:2863","2015-02-26","",,,,,,,,,
"13598",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"Zouine","Mohamed","","",,,,,,,,,,,,,,,,,,,,,,,,,,
"10989","102","archive","1",,,"disk0/00/01/09/89","2014-02-18 22:58:23","2016-07-08 09:54:10","2014-02-18 22:58:23","thesis",,,"show",,,,"","","","","","","","","","","0",,,"Wang","Xinyu","","","","","Génomique et Biotechnologie des Fruits - GBF (Castanet-Tolosan, France)","","","","","","",,"","Characterization of protein-protein interactions involved in auxin signaling pathway in tomato","unpub",,,"","INPT","","public",,,"Tomato – Protein-protein interaction – IAA – ARF – TPL – Auxin",,"harvest from ethesis","The plant hormone auxin plays a central role in plant growth and development. The specific Aux/IAAs and Auxin Response Factors (ARFs) interactions are involved in auxin signaling pathway to regulate the auxin-responsive gene expression. Studies in Arabidopsis showed that TOPLESS family (TPLs) also was recruited by some Aux/IAAs to repress the function of ARFs. The whole machinery of the auxin signaling pathway is not clear yet, and most of this knowledge comes from the research on Arabidopsis. As a reference for Solanaceae and fleshy fruit plant, tomato (Solanum lycopersicon) is a good alternative model to better understand general traits of the auxin regulation process. In our work, we first established in our labs three experimental protocols – Yeast two-Hybrid, Pull-down and Bifluorescence complementation to unravel protein-protein interactions. These methods were first challenged on specific Aux/IAA and ARF proteins that were already characterized as major actors in fruit tomato development or ripening (SlIAA9, SlARF8, SlIAA3, SlARF4, SlIAA27). This also enabled us to build an ARF-Aux/IAA interaction map. In a second part, taking advantage of the tomato genome sequence, we carried a whole-genome study on tomato TOPLESS family. This investigation included gene cloning and characterization, protein sequence analysis, phylogenetic analyses, expression pattern and construction of protein-protein interaction maps. In a last part, we developed tools to start a non-targeted approach aiming at identifying new potential partners or protein complex involved in auxin signaling pathway using BY-2 tobacco cell protoplasts transiently expressing tagged-proteins. Although this study is still preliminary, it demonstrated the importance of nucleus integrity for Aux/IAA stability even in absence of auxin.","2013-12-03",,,,,,,,,,,,,,,,"Institut National Polytechnique de Toulouse","phd",,,,,,,,,"","http://ethesis.inp-toulouse.fr/archive/00002526/","","",,"","",,,,,,"",,,,,,,"","BOUZAYEN","Mondher","","","",,,,,"","",,,,,"","",,"oai:ethesis.inp-toulouse.fr:2526","2014-02-18","",,,,,,,,,
"10989",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"VAN DER REST","Benoît","","",,,,,,,,,,,,,,,,,,,,,,,,,,