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Tolerance Strategies of Atriplex Atacamensis and A. Halimus in Response to Multiple Abiotic Stresses 2019

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Tolerance Strategies of Atriplex Atacamensis and A. Halimus in Response to Multiple Abiotic Stresses 2019 Tolerance strategies of Atriplex atacamensis and A. halimus in response to multiple abiotic stresses Fabiola Alejandra Orrego Márquez 2019 1 Pontificia Universidad Católica de Chile Facultad de Agronomía e Ingeniería Forestal Tolerance strategies of Atriplex atacamensis and A. halimus in response to multiple abiotic stresses Fabiola Alejandra Orrego Márquez Thesis to obtain the degree of Doctor en Ciencias de la Agricultura Santiago, Chile, April 2019 2 Thesis presented as part of the requirements for the degree of Doctor in Ciencias de la Agricultura, approved by the Thesis Committee _____________________ Dr. Rosanna Ginocchio, Advisor _____________________ Dr. Claudia Ortíz-Calderón _____________________ Dr. José Miguel Fariña Santiago, April 2019 3 Financial funding support for Fabiola Orrego and all thesis activities was obtained from CONICYT Doctoral Grant (21141059/2014), Center of Applied Ecology & Sustainability (CAPES) by CONICYT PIA/BASAL FB0002 (2014) and Facultad de Agronomía e Ingeniería Forestal UC. 4 Acknowledgements I would like to thank mi thesis advisor, Rosanna Ginocchio for her support and dedication during this process, and for teaching me to take care of myself as much as I care for what I do. To mi co-advisor Claudia, for her precise comments on plant physiology and biochemistry that shed light on my understanding of plants. I thank my family, for accompanying me with curiosity and enthusiasm on each step of my academic path. To my beloved Diego, for his permanent support during this process and for being a man I can look up to. To all my professional and scientific colleagues in the RESUME lab, especially to Luz María de La Fuente, that taught me how to work properly with plants. To every one of my friends in FAIF and FCB, thanks for help me broaden my perspectives, and being allies in this beautiful path of knowledge. 5 Contents General introduction .............................................................................................. 11 Hypothesis and objectives ..................................................................................... 26 Chapter 1 Diversidad de halófitas chilenas: distribución, origen y hábito .............................. 29 Introducción ........................................................................................................... 31 Materiales y métodos ............................................................................................ 33 Resultados ............................................................................................................ 34 Discusión y conclusiones ...................................................................................... 38 Agradecimientos .................................................................................................... 44 Referencias ........................................................................................................... 44 Chapter 2 Effect of single and combined Cu, NaCl and water stress on three Atriplex species with phytostabilization potential ............................................................................. 55 Abstract ................................................................................................................. 56 Introduction............................................................................................................ 57 Materials and methods .......................................................................................... 60 Results .................................................................................................................. 63 Discussion ............................................................................................................. 67 Conclusion............................................................................................................. 75 6 Acknowledgments ................................................................................................. 76 References ............................................................................................................ 76 Chapter 3 Growth and physiological effects of combined Copper, NaCl and water stresses on Atriplex atacamensis and A. halimus ..................................................................... 87 Abstract ................................................................................................................. 88 Introduction............................................................................................................ 89 Materials and methods .......................................................................................... 91 Results .................................................................................................................. 96 Discussion ........................................................................................................... 104 Conclusion........................................................................................................... 112 Acknowledgments ............................................................................................... 113 References .......................................................................................................... 113 General Conclusions ........................................................................................... 130 7 Table Index Chapter 1 TABLA 1. Distribución de la riqueza de especies halófitas de Chile según su familia. .............................................................................................................................. 36 TABLA 2. Riqueza, hábito y origen de las especies halófitas presentes en Chile según su distribución geográfica ........................................................................... 40 ANEXO 1. Listado de especies halófitas presentes en Chile según la base de datos eHALOPH.. ............................................................................................................ 48 Chapter 2 Table 1. Shoot length (SL), dry root weight (DRW) and dry shoot weight (DSW) of Atriplex atacamensis (AA), A. halimus (AH) and A. nummularia (AN) seedlings subjected to available Cu, NaCl and a PEG-induced water stress for seven days.86 Chapter 3 Table 1. Root length (cm.) and root and shoot fresh weight (g) of Atriplex atacamensis and A. halimus seedlings exposed for 10 days to single and combined Cu, NaCl and PEG stresses. .............................................................. 125 Table 2. Total chlorophyll and carotenoid content of Atriplex atacamensis and A. halimus subjected to single and combined Cu, NaCl and PEG stresses. ............ 126 Table 3. Root and shoot water content (WC in %) of A. atacamensis and A. halimus seedlings exposed for 10 days to single and combined Cu, NaCl and PEG stresses ............................................................................................................... 127 Table 4. Sodium and K concentration (in mg∙g-1) in roots and leaves of A. atacamensis and A. halimus seedlings exposed for 10 days to single and combined Cu, NaCl and PEG stresses. .............................................................. 128 8 Table 5. Reduced and oxidized glutathione (nmol g FW -1) in leaves and roots of Atriplex atacamensis and A. halimus exposed for 10 days to single and combined Cu, NaCl and PEG stresses.. .............................................................................. 129 9 Figure Index Chapter 2 Figure 1. Root length increase of seedlings of A. atacamensis, A. halimus and A. nummularia subjected to increasing available Cu, NaCl and a decrease of solute potential. ................................................................................................................ 82 -1 Figure 2. Plant water content (g H2O g ) of Atriplex atacamensis, A. halimus and A. nummularia seedlings subjected to increasing available Cu, NaCl and a decrease of solute potential................................................................................... 83 Figure 3. Increase in root length (cm) of Atriplex atacamensis and A. halimus subjected to increasing single Cu concentrations and its combination with NaCl and PEG. ............................................................................................................... 84 Figure 4. Picture of A. halimus and A. atacamensis seedlings under control, Cu and Cu+PEG treatments of combined stress assay.. ............................................ 85 Chapter 3 Figure 1. Visual symptoms of single and combined stresses on Atriplex seedlings. (a) Loss of turgor and leaf reddening on Atriplex halimus leaves subjected to 7.85 mM PEG. ............................................................................................................. 120 Figure 2. Leaf osmotic potential (MPa) of A. atacamensis (left) and A. halimus (right) seedlings exposed for 10 days to single and combined Cu, NaCl and PEG stresses. .............................................................................................................. 121 Figure 3. Leaf proline (mg g-1 FW) of A. atacamensis (left) and A. halimus (right) seedlings exposed for 10 days to single and combined Cu, NaCl and PEG stresses (n=4; mean± EE). ................................................................................................ 122 Figure 4. Copper concentration (mg∙g-1) in roots and leaves of A. atacamensis (upper) (n=5; means± EE) and A. halimus (lower). ............................................
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