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DOCTORAL THESIS 2017 Hanan El Aou-Ouad

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DOCTORAL THESIS 2017 Hanan El Aou-Ouad DOCTORAL THESIS 2017 INTERACTIVE EFFECTS OF GRAPEVINE LEAFROLL ASSOCIATED VIRUS-3 (GLRaV-3) AND WATER STRESS ON THE GAS EXCHANGE, WATER USE EFFICIENCY, PLANT HYDRAULICS AND METABOLISM IN LOCAL GRAPEVINE CULTIVARS Hanan El Aou-ouad DOCTORAL THESIS 2017 Doctoral Programme in Plant Biology INTERACTIVE EFFECTS OF GRAPEVINE LEAFROLL ASSOCIATED VIRUS-3 (GLRaV-3) AND WATER STRESS ON THE GAS EXCHANGE, WATER USE EFFICIENCY, PLANT HYDRAULICS AND METABOLISM IN LOCAL GRAPEVINE CULTIVARS Hanan El Aou-ouad Thesis Supervisor: Josefina Bota Salort Thesis Supervisor: Alicia Pou Ph. Doctor by the Universitat de les Illes Balears الى امي٬ من اصدق قلبي أهديك الشكر و اﻻمتنان تمجيدا لعطائك الكريم وحبك الﻻمتناهي الى ابي٬شكراﻻنك وضعتني على الطريق الصحيح قبل ان ترحل٬ تاركا ايانا امرأة عظيمة تسير بنا الى بر اﻻمان Agradecimientos-Acknowledgements AGRADECIMIENTOS-ACKNOWLEDGEMENTS Cada historia tiene un final, y hoy ha llegado el tiempo de volver la mirada atrás y expresar mi agradecimiento a todas las personas que me han acompañado en este proyecto personal tan importante para mi. Espero que con las siguientes palabras pueda expresar mi más profundo agradecimiento a cada una de las personas con las que he podido compartir alguna experiencia durante todos estos años. Primero, gracias a Dios por permitirme vivir esta experiencia única en mi trayectoria académica y profesional. En segundo lugar, quiero expresar mi total agradecimiento a mis directoras, la Dra. Josefina Bota y la Dra. Alicia Pou (futura mami) por darme la oportunidad de realizar esta Tesis bajo su dirección y por toda la ayuda prestada. En especial, quiero darle las gracias a Josefina por acompañarme todos estos años, por su paciencia conmigo, por transmitirme su conocimiento, por el tiempo y el esfuerzo dedicados, por su apoyo y sus consejos, tanto en lo laboral como en lo personal. Por todos estos años mil gracias. Agradezco al Dr. Hipólito Medrano que se involucrara en la dirección de mi tesis desde el primer día, y por toda su enorme ayuda y por transmitirme sus brillantes ideas y consejos, además de su motivación y predisposición. También debo expresar mi agradecimiento a los Doctores/as, Javier Gulias (por su ayuda y paciencia desde que llegué a esta Isla), Jaume Flexas, Igor, Alex, Enrico (por tu amistad y tus ánimos), Cyril, Miquel Àngel, Xurxo, Pepe Escalona, Jeroni, Pep Cifre, Maria Jose, Elena, Perla, Alejandro, Sebastià, Carmen, Pedro, Xiong y Rafa por el gran apoyo que me prestaron durante esta etapa, disponibilidad para resolver mis dudas y por sus sabios consejos. Al Dr. Miquel Ribas por su apoyo incondicional y por haber confiado en mí y brindarme la oportunidad de participar en sus actividades academicas. También agradezco a las Doctoras. Arantxa Molins y Magdalena Tómas por su apoyo ilimitado, sus consejos, su valiosa amistad y por instarme a ser perseverante. Gracias a Belén (reina) por su amabilidad, por toda su ayuda y disponibilidad, Pep Sastre, Miquel Truyols e Issac por su apoyo en toda la parte experimental. i Agradecimientos-Acknowledgements Igualmente quiero expresar mi gratitud a toda la familia del ―Can Boom‖, que me han acompañado a lo largo de este proyecto, gracias a Cristina, Ocho, Veriozca (mi querida tan especial), Antonia (tus gestos de cariño), Amani, Esther (source de gentillesse et sympathie y por aguantarme en los buenos y malos momentos), Néstor (por contagiarme con tus ganas de trabajar), Paty, Marcel (notre artiste), Nacho (por poner siempre una sonrisa a la vida Can bomera), Miquel C, Miquel N, Alicia, Marc, Mateu y Toni, muchas gracias amigos por haber hecho de estos años momentos especiales e inolvidables. Me siento privilegiada de tener una familia al otro lado del Mediteraneo. Dr. Cyril Douthe, merci beaucoup pour ton aide, tes critiques constructives, ta disponibilité et ton écoute. Je serai eternellement reconaissante cher ami. I would like to state my gratitude to Dr. Alisdair Fernie for giving me the opportunity to work in his laboratory. Thank you very much to all colleagues of the Max Planck Institute in Postdam for helping me in the lab, specially Dr. Igor (por toda la ayuda desde mi primer dia en Mallorca) and Paula. Also I would like to thanks Dr. Toshihiro. Je remercie très intensément toutes les personnes que j‘ai côtoyé et devenus mes amies et au même temps ma famille que ce soit dans mon Pays ou bien à Majorque. Merci pour les talents de chacun, les discussions et les gestes d‘attention. En espérant n‘oublier personne, j‘adresse mes remerciements à mes amies: Hania, Hajar, Manal (et mon cher bébé Haroun), Hanane, Hanae, Bassima, Inssaf, Lamiae, Asmaa, Nawar, Kaoutar, Houda, Fadoua, Mehdi, Samar, Olfa et Fatema. Dans un registre plus personnel, je souhaite remercier tout particulièrement mes parents, ma sœur, Amal et son mari et finalement mon frère Jaber. Ils ont su m‘apporter l‘énergie et la confiance pour mener à terme ce travail. Je remercie aussi chaleureusement mes grands parents et toute ma famille qui s‘est toujours montré intéressé par l‘avancement de mes expériences, specialment la famille Mesali (Abdel ouahab, Souad, Lamiae, Bouchra). ii Agradecimientos-Acknowledgements Enfin, je tiens à remercie énormément mon mari, Simohamed. J‘ai eu la chance de te connaitre dans ce merveilleux parcours. Je te remercie pour ta gentillesse, ton encouragement, ton aide et ton support dans les pires conditions. Et merci tout particulièrement d'avoir remarquablement assumé ces dernières semaines……. Gracias a todos iii Agradecimientos-Acknowledgements iv Symbols and Abbreviations list SYMBOLS AND ABBREVIATIONS LIST Symbols Meaning α leaf absorptance ABA abscissic acid Ala alanine AN net CO2 assimilation AOX alternative oxidase Arg arginine Asp aspartic acid ATP adenosine triphosphate BL biochemistry limitation β partitioning of absorbed quanta between photosystems II and I C calose Ca atmospheric CO2 concentration Caf caffeic acid Cc chloroplast CO2 concentration CC companion cells Chl chlorophyll Chl a chlorophyll a Chl b chlorophyll b Ci sub-stomatal CO2 concentration CO2 carbon dioxide COX cytochrome oxidase CP coat protein CPC crystalline protein cluster CPm minor coat protein Ct threshold cycle CTV citrus tristeza virus CW cell wall Δa oxygen isotope fractionation of the alternative oxidase pathway c oxygen isotope fractionation of the cytochrome oxidase pathway DIECA na-diethyl-dithio-carbonate DL density of the leaves DTT dithiothreitol E leaf transpiration rate ELISA enzime-linked inmunosorbent assay ER endoplasmic reticulum ФCO2 apparent quantum efficiency of CO2 fixation ΦPSII photochemical efficiency of photosystem II Fo fluorescence signal when all reaction centers were open ƒias volume fraction of intercellular air spaces FM fluorescence signal when all reactions centers were closed FM’ maximum fluorescence during a light-saturating pulse Fs steady-state fluorescence v Symbols and Abbreviations list Symbols Meaning FV/FM maximum quantum efficiency of photosystem II Γ* CO2 compensation point in the absence of mitochondrial respiration GC-TOF-MS gas chromatography-time of flight-mass spectrometry GFkV grapevine fleck virus GFLV grapevine fanleaf virus GLD grapevine leafroll disease GLRaVs grapevine leafroll associated viruses Glu glutamic acid Gly glycine GRSPaV Grapevine rupestris stem pitting associated virus gs stomatal conductance gm mesophyll conductance GVA grapevine virus A GYSVd-1 grapevine yellow speckle viroid 1 His histidine hsp 70 heat shock protein 70 HRM high-resolution melting IC-PCR immuno-capture PCR Ile isoleucine JA jasmonic acid Jmax maximum capacity for electron transport rate Kc rubisco michaelis–menten constants for carboxylation Ko rubisco michaelis–menten constants for oxygenation LAMP loop-mediated amplification of nucleic acid LEDs light-emitting diodes Leu leucine LMA leaf mass area L-Pro leader protease Lys lysine M mitochondria Mal malate MAP mitogen-activated protein MCFI multicolour fluorescence imaging MCL mesophyll limitation MeOH methanol MEP methylerythritol phosphate Meth methionine mETC mitochondrial electron transport chain MRM multiple reaction monitoring MSTFA n-methyl-n-(trimethylsilyl) trifluoroacetamide MuLV moloney murine leukemia virus N nucleus NADH nicotinamide adenine dinucleotide O2 oxygen ORFs open reading frames PD plasmodesmata PEG polyethylene glycol Phen phenylalanine PPFD photosynthetically active photon flux density vi Symbols and Abbreviations list PPUs pore plasmodesmal units PR pathogenesis-related PSII photosystem II qPCR quantitative PCR Rd leaf dark respiration RdRp RNA dependent RNA polymerase ROS reactive oxygen species RT-PCR reverse transcription-PCR RuBP ribulose biphosphate RW rugose wood SA salicylic acid SDS sequence detection systems SE sieve elements SEL size exclusion limit Ser serine sgRNAs small guides RNAs SL stomatal limitation +ss positive sense single stranded SSCP single-strand conformation polymorphism τa electron partitioning through the alternative pathway TA titratable acidity TBE tris-borate-EDTA Thre threonine TL total limitation Tmes mesophyll thickness TSO2 total sulphur dioxide TSS transcriptional start sites Val valine Valt activity of the alternative oxidase pathway Vc,max maximum carboxylation capacity Vcyt activity of the cytochrome oxidase pathway VP vascular parenchyma cells Vt total oxygen uptake rate VTPU rate of triose-phosphate utilization ѡ width of leaf anatomical section PD predawn leaf water potential vii Symbols and Abbreviations list viii List of publications derived from the present thesis LIST OF PUBLICATIONS DERIVED
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