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The Role of Auxin in Cell Differentiation in Meristems Jekaterina Truskina

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The Role of Auxin in Cell Differentiation in Meristems Jekaterina Truskina The role of auxin in cell differentiation in meristems Jekaterina Truskina To cite this version: Jekaterina Truskina. The role of auxin in cell differentiation in meristems. Plants genetics. Université de Lyon, 2018. English. NNT : 2018LYSEN033. tel-01968072 HAL Id: tel-01968072 https://tel.archives-ouvertes.fr/tel-01968072 Submitted on 2 Jan 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Numéro National de Thèse : 2018LYSEN033 THESE de DOCTORAT DE L’UNIVERSITE DE LYON opérée par l’Ecole Normale Supérieure de Lyon en cotutelle avec University of Nottingham Ecole Doctorale N°340 Biologie Moléculaire, Intégrative et Cellulaire Spécialité de doctorat : Biologie des plantes Discipline : Sciences de la Vie Soutenue publiquement le 28.09.2018, par : Jekaterina TRUSKINA The role of auxin in cell differentiation in meristems Rôle de l'auxine dans la différenciation des cellules au sein des méristèmes Devant le jury composé de : Mme Catherine BELLINI, Professeure, Umeå Plant Science Center Rapporteure Mme Zoe WILSON, Professeure, University of Nottingham Rapporteure M. Joop EM VERMEER, Professeur, University of Zurich Examinateur M. Renaud DUMAS, Directeur de recherche, Université Grenoble Alpes Examinateur M. Teva VERNOUX, Directeur de recherche, ENS de Lyon Directeur de thèse M. Anthony BISHOPP, Chercheur, University of Nottingham Co-tuteur de thèse Thesis submitted to the University of Nottingham for joint examination with the ENS de Lyon for the degree of Doctor of Philosophy by Jekaterina Truskina The role of auxin in cell differentiation in meristems Rôle de l'auxine dans la différenciation des cellules au sein des méristèmes Date of submission : 11.06.2018 Date of public defence : 28.09.2018 The thesis jury is composed of : Mme Catherine Bellini, Professor, Umeå Plant Science Center Reporter Mme Zoe Wilson, Professor, University of Nottingham Reporter M. Joop EM Vermeer, Professor, University of Zurich Examiner M. Renaud Dumas, Research Director, Université Grenoble Alpes Examiner M. Teva Vernoux, Research Director, ENS de Lyon Thesis director M. Anthony Bishopp, University Research Fellow, University of Nottingham Thesis co-director ~ 1 ~ Abstract Auxin regulates plant growth and development through the transcription factors of the AUXIN RESPONSE FACTOR (ARF) gene family. Most notably in Arabidopsis thaliana ARF5, 6, 7, 8 and 19 activate expression of target genes in response to auxin. These five ARF activators control both variable and overlapping processes during plant development including regulation of growth at the root and the shoot apical meristems, lateral root and axillary shoot formation. Each of the five ARF activators shows unique tissue-specific expression patterns in the root and the shoot associated with their distinct functions. This tissue-specific expression is likely derived from the differences in the control of ARF activator transcription. In this study the upstream regulators of ARF5, 6, 7, 8 and 19 transcription were identified. This was achieved by utilizing a high-throughput yeast one-hybrid (Y1H) method. The transient protoplast assay revealed that each ARF activator is controlled by specific transcriptional regulators and that the majority of these regulators are repressors of ARF transcription in planta. Mutants of the regulatory transcription factors were utilized to additionally investigate the interactions in planta. These mutants display auxin-related developmental phenotypes in the root and the shoot including alternations in growth kinetics, emergence of lateral organs, responses to auxin and altered expression of ARF activators. Furthermore, this study additionally focuses on cross-talk between the auxin and cytokinin signaling pathways and its role in root and shoot development. One of the interactions identified in the Y1H screen is a repression of ARF7 by CRF10, a member of the Cytokinin Response Factors gene family. The importance of this interaction in maintaining architecture of the root apical meristem, in leaf senescence and in the phototropic response to blue light in hypocotyls is studied. ~ 2 ~ Résumé L'auxine régule la croissance et le développement des plantes grâce aux facteurs de transcription de la famille des "AUXIN RESPONSE FACTOR" (ARF). Chez Arabidopsis thaliana en particulier, ARF5, 6, 7, 8 et 19 activent l'expression de gènes cibles en réponse à l'auxine. Ces cinq ARF activateurs contrôlent de façon plus ou moins redondante des processus divers au cours du développement de la plante, notamment la régulation des croissances au niveau des méristèmes racinaires et aériens ainsi que la formation des racines latérales ou des méristèmes axillaires. Chacun de ces cinq ARF activateurs présente des patrons d'expression uniques dans chacun des tissus racinaires et aériens, en association avec leurs fonctions particulières. Il est probable que cette expression tissu-spécifique trouve son origine dans un contrôle différencié de leur transcription. Dans cette étude, des régulateurs amonts de la transcription de ARF5, 6, 7, 8 et 19 ont été identifiés par une méthode haut-débit de crible simple hybride en levure (Y1H). Une procédure d'expression transitoire en protoplastes a permis de confirmer que l'expression de chaque ARF activateur est contrôlée par des régulateurs spécifiques, dont la majorité se comportent comme des répresseurs de la transcription des ARF in planta. Parmi les régulateurs identifiés, les facteurs de transcription ont été étudiés grâce à des mutants pour préciser les interactions in planta. Ces mutants montrent des phénotypes développementaux typiques de perturbations de l'auxine dans les racines et les tiges : altérations des cinétiques de croissance, de l'émergence des organes latéraux ou de réponses à l'auxine et modification de l'expression des ARF activateurs. Par ailleurs, ce travail aborde également les dialogues entre les voies de signalisation de l'auxine et des cytokinines, et en particulier le rôle de ces interactions dans le développement des racines et des tiges. Une des interactions identifiées dans le crible Y1H est la répression de ARF7 par CRF10, un gène membre de la famille des "Cytokinin Response Factors". Nous avons mis en évidence l'importance de cette interaction pour le maintien de l'architecture du méristème apical racinaire, pour la sénescence des feuille et pour la réponse phototropique à la lumière bleue dans les hypocotyles. ~ 3 ~ Acknowledgments Teva and Tony, my most incredible supervisors, I own great gratitude to you both. You changed my life when you offered me this PhD position and since then, I spend awesome four years learning from you how to do science. When it comes to science your advices were always full of wisdom and experience guiding me through my thesis. I had amazing support from you both in my work but also in my life. I felt it from the start when you took this incredible effort to go through a huge pile of paperwork to organize this joint PhD. I felt it even more during my thesis when I was able to come up to any of you with my problems or difficulties at any time of the day, and you would always help me. Your kindness and friendliness made me feel great being a part of this project and I hope, both of you have become truly my friends. Tony, with your incredible patience, enthusiasm and kindness you are a perfect mentor that anyone can wish for. I felt your support throughout the whole thesis starting from the first day when you helped me to get to Sutton Bonington campus and all the way through. You had a young new team consisting of PhD students just starting their thesis, all fresh and inexperience like me, and it was great to see how everyone in the team had a very happy successful PhD thanks to your great supervision. Thank you for everything, Tony! Teva, it’s your endless energy, dedication and kindness that made you the amazing mentor. You are a very busy person managing a whole laboratory, and yet you always found time for me as much as I needed. I learned so much from you and your experienced team of researchers. You have such a great team and it was amazing to feel a part of it. Thanks for everything, Teva! Throughout my thesis I met many great people from all over the world and some of them became my friends. In Nottingham, I was part of a great PhD student community with John, George, Ben and Nicky. Thanks a lot guys for your support! Especially, a great thank you to Nicky with whom we shared great times as friends. I also would like to thank Kamal for your friendliness and help during my stay in the lab. Jingyi, thank you for helping me with experiments, I wish you a good start on your PhD thesis with Tony, I am sure it ~ 4 ~ will be great! I would like to thank Malcolm for your kindness and gentle personality; you treat every person, even simple student like me, with such respect and patience, making them feel welcomed and a part of the great scientific community. Finally, I would like to thank all people from the lab for the friendly atmosphere. In Lyon, I felt a true part of an incredible team. Carlos, thank you for all the great advices you gave me; you inspire everyone in the lab with your enthusiasm for science and your ingenious ideas. Stephanie, it was wonderfull to work with you together as a team. Antoine, Fabfab, Geraldine, Bihai, Jo and Guillaume, I thank you for all your support, great advices and friendliness during my thesis.
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