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Use of biotechnological tools as support to domestications and breeding of Argylia radiata (L.) D. Don. Pablo Andrés Morales Tapia 2019 Pontificia Universidad Católica de Chile Facultad de Agronomía e Ingeniería Forestal Use of biotechnological tools as support to domestications and breeding of Argylia radiata (L.) D. Don. Pablo Andrés Morales Tapia Thesis to obtain the degree of Doctor in Ciencias de la Agricultura Santiago, Chile, March 2019 Thesis presented as part of the requirements for the degree of Doctor in Ciencias de la Agricultura, approved by the Thesis Committee Prof. Gloria Montenegro, Advisor Dr. Marina Gambardella, Co-Advisor Dr. Rodrigo Barba González Dr. Ursula Steinfort Santiago, March 2019 Dedication To my family, especially my parents Manuel and Cecilia, and my sisters, Gabriela and María José. Thank for to believe in this “project”, to be with me during the complete process, for your support, cares, and help in the hard times. For all the hours on the road and the travels to La Serena to collect plants and seeds. To my professors María Eliana Varas and Fernando Cosio, who inspired and taught me to love and to appreciate the Chilean Native flora. Finally, I want to dedicate this work, in a very especially way, to the memory of my grandparents, who always were a source of inspiration, courage, and support. Thanks to funding sources This work was supported by CONICYT through its Grant for Chilean Ph.D. programs, grant number 21150714, and CIDI Grupo Hijuelas sponsorship with facilities and reagents. We want also thank to the team of the Dirección de Investigación y Postgrado, Facultad de Agronomía e Ingeniería Forestal UC because of its monetary help and its continuous support during all this research. Acknowledgements I want to thank my family, parents, and sisters. Thank for all your help and patience. Thank for supporting me during all this process. Thank my professors Gloria and Marina, because of their help and teachings, which awakened in me the love for the research and the innovation. Thank Rodrigo for welcoming me during my internship at México, for all the input that he made to this work and also for accepting to be part of this commission. Thank Ursula because of her continuous support and encouragement. Thank you to my friends and colleagues from Hijuelas Group and from the lab of plant biotechnology of FAIF UC, who always helped me with the lab work and encouraged me during difficulties. Also, thank my classmates. Thank you for making it easier for me, to study and to live in Santiago. Finally, I want to thank the team of DIP FAIF UC, because of all their support and help during all this process. Thank a lot to all of you! Pablo Morales Tapia Contents Chapter 1: General introduction, theoretical framework and research project ........... 3 1. Abstract ....................................................................................................................................... 4 2. Introduction................................................................................................................................. 4 2.1. Genus Argylia, description and relevant characteristics ...................................................... 5 2.3. Argylia radiata (L.) D. Don. Xerophytic species with huge ornamental potential ............... 6 2.4. Polyploidization: definitions and importance during domestication and plant breeding ... 8 2.5. Effect of polyploidization over plant development and production .................................. 11 2.6. Polyploid induction ............................................................................................................. 13 2.6.1. Colchicine .................................................................................................................... 13 2.6.2. Oryzalin........................................................................................................................ 13 2.6.3. Nitrous oxide ............................................................................................................... 14 2.6.4. Protoplast fusion ........................................................................................................ 15 2.6.5. Other factors inducing polyploidy ............................................................................... 16 2.7. Polyploidization and plant breeding of ornamental crops ................................................. 17 3. Work hypothesis ....................................................................................................................... 20 4. Objectives .................................................................................................................................. 21 4.1. General objective ............................................................................................................... 21 4.2. Specific objectives .............................................................................................................. 21 5. References chapter 1 ................................................................................................................ 22 Chapter 2: Morpho-Anatomical adaptations of Argylia radiata (L.) D. Don to an arid environment ................................................................................................................... 29 Abstract ......................................................................................................................................... 30 Keywords ....................................................................................................................................... 31 1. Introduction............................................................................................................................... 31 2. Materials and methods ............................................................................................................. 32 2.2. Description of tuberous roots ............................................................................................ 33 2.3. Sample preparation and analysis ....................................................................................... 33 2.4. Starch accumulation ........................................................................................................... 34 3. Results ....................................................................................................................................... 34 3.1. Root .................................................................................................................................... 35 1 3.2. Stems .................................................................................................................................. 38 3.3. Leaves ................................................................................................................................. 39 3.4. Floral stalk .......................................................................................................................... 41 4. Discussion .................................................................................................................................. 42 5. Conclusions................................................................................................................................ 47 Acknowledgements ....................................................................................................................... 48 References ..................................................................................................................................... 48 Chapter 3: Control of shoot tips necrosis during Argylia radiata (L.) D. Don micropropagation .......................................................................................................... 54 Abstract ......................................................................................................................................... 55 Key words: ..................................................................................................................................... 55 Acknowledgements ....................................................................................................................... 56 Authors contribution statement ................................................................................................... 56 Introduction .................................................................................................................................. 56 Materials and Method .................................................................................................................. 57 Results ........................................................................................................................................... 59 Discussion ...................................................................................................................................... 69 Conclusion ..................................................................................................................................... 73 Conflict of Interest: ....................................................................................................................... 74 References ..................................................................................................................................... 74 Chapter 4: General Discussion ..................................................................................... 79 References
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