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Thesis Title Thesis Title Physiological and Metabolic Responses of Two Rose Varieties to Plant Growth Regulators This thesis is presented for the degree of Doctor of Philosophy of Murdoch University By Mohammed Muhi Ibrahim School of Veterinary and Life Sciences Murdoch University Perth, Western Australia 2018 i DECLARATION I declare that this thesis is my own account of my research and contains as its main content work, which has not previously been submitted for a degree at any tertiary education institution. Signature: Mohammed Ibrahim Date: December 19, 2018 ii Acknowledgements “In the name of Allah, the most Beneficent and the Merciful” بسم هللا الرحمن الرحيم Allah Almighty, the creator of everything, the most beneficent, merciful, gracious and compassionate whose bounteous blessing gave me the potential thoughts, and guidance to complete my PhD research, write my thesis and to provide with the opportunity to make this humble effort and enable me to pursue and perceive higher ideas of life. First of all, I would like to thank my supervisor Professor Yonglin Ren Head of Postharvest Biosecurity and Food Safety Laboratory Murdoch, Western Australia. He was very kind and generous in his auspicious help, scholarly guidance, great encouragement and affectionate support. He has been a colossal source of inspiration and solace for me during my PhD studies. Secondly, it gives me great pleasure in acknowledging my associate supervisor Dr Manjree Agarwal Postharvest, Biosecurity and Food Safety Laboratory Murdoch, Western Australia. For her help during my research and her patience for many questions, valuable training, providing corrections and critical suggestions on experimental work and during my thesis writing stage. Thirdly, I take great pleasure to express my sincere and intense sense of gratitude to my associate supervisor Professor Giles Hardy Associate Dean Research/Professor in Forest Pathology in Murdoch University. For imparting all his experience, knowledge and skills in me, for his caring behaviour, moral support and his continuous efforts to accommodate a supportive environment throughout my research, and during writing stage despite his multifarious responsibilities. A sense of obligation compels me to express my cordial thanks to Professor Muslim Abdulhussein head of Horticulture Department & Landscape, Faculty of Agriculture, Kufa University, Najaf, Iraq, associate supervisor for helping me to prepare the required compounds and for critical suggestions and valuable feedback during my research. Many thanks are passed to Mr. Bo Du Postharvest, Biosecurity and Food Safety Laboratory Murdoch University for his help with technical support with laboratory instruments and his patience and time. I also gratefully pass my thanks to Mr. James Newman Postharvest, Biosecurity and Food Safety Laboratory Murdoch University for his help in providing plant materials, time, patience, and moral assistance during my research. I sincerely acknowledge iii and thank financial support from the Higher Committee for Education Development in Iraq (HCED) for providing me with the scholarship to undertake my doctoral study. Special thanks to my Uncle Younes Ibrahim for his encouragement, moral support, and many thanks to my cousin Marai Younes for his love, affection, support, help, and encouragement. It would have been a very tough job for me to complete this thesis without continuous motivation and inspiration. I am thankful to all my friends that I have known in Australia for giving the support and help in different ways, especially Charles Obiero for assistance with statistical analysis. I am also thankful to my relatives in Australia Mr. Faisal Al-Ramaid and his family especially his son Husam for their support. I take this opportunity to express my thanks to all my family members especially my affection mother, my wife Bushra, my daughter Elaf and my son Ameen “Their love, incredible patience and endless support during the entire period of my PhD study and throughout my life”, friends and my relatives in Iraq for their encouragement, moral support and good wishes. iv Abstract Rose (Rosa hybrida) is one of the most popular cut flowers with a worldwide production of more than 300 million stems per year. The perfume industry, which relies on a range of scented rose varieties, is also an important industrial application for roses. Among the numerous types of roses are some recent varieties including Hybrid Tea and Floribunda roses. The major problem in Australia is accelerated pre- and post-harvest flower drop and senescence, caused by deficiencies in endogenous plant growth regulators (PGRs). PGRs play important roles in the growth and development of flowers, especially in aromatic plants, stimulating the emission of volatile organic compounds (VOCs). The PGRs benzyladenine (BA) and naphthalene acetic acid (NAA) which belong to cytokinin and auxin group of PGRs respectively, are used by the floriculture industry as important growth regulators for promoting rose growth and development. However, it is still unknown that how these regulators and their application dosages influence rose plants. Therefore, this thesis aimed to evaluate and determine the efficacy of various concentrations of BA and NAA on arrange of morphological and physiological characteristics of roses to increase flowers longevity in two rose varieties (Floribunda and Hybrid Tea). In addition, this study was conducted to understand how different concentrations of BA and NAA effect the metabolic changes in different rose tissues together with a comparison of VOCs changes. This work in this thesis developed and optimized the headspace solid-phase microextraction (HS-SPME) with three-phase fibre 50/30µm divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) coupled with gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS) method for exploring and analysing VOCs emitted from the intact and excised flowers, leaves, and stems as well as from the rhizosphere and whole plants of the two rose varieties. This study has optimized different environmental factors involved in the performance of the two rose varieties was conducted, and three physiological characteristics photosynthesis rate, respiration rate, and chlorophyll content were assessed. Three different concentrations (0, 100 and 200 mg/L) of BA and NAA were applied to the two rose varieties, and different morphological and physiological characteristics were evaluated. For physiological effects (photosynthesis rate and chlorophyll index) studies, application of 200 mg/L of both BA and NAA were shown to increase plant height, numbers of branches and flowers, stem and flower diameters, length of flower stems and flower longevity, compared to the control. However, for v the respiration rate, the control plants had significantly superior performance to the plants treated with 100 and 200 mg/L BA and NAA, for both rose varieties. The VOC profiles of the two rose varieties were characterized by the optimized HS-SPME-GC method. The effects of different concentrations (0, 100 and 200 mg/L) of BA and NAA on the VOCs emitted from different rose tissues for the two rose varieties were determined. The highest amounts extracted, and evaluated from the sum of peak areas were achieved after the application of 200 mg/L BA and NAA in both varieties. Of the emitted VOCs, 20 were significantly different in treated compare to non-treated Floribunda and Hybrid Tea roses from different rose tissues. Moreover, five compounds 4-heptyn-2-ol, cis-muurola-4(14)5-diene, γ - candinene, y-muurolene and prenyl acetate increased significantly after applications of 200 mg/L of BA and NAA. These five compounds have great potential to develop commercially important new rose growth regulators. The actual dosages of BA applied to the leaves was determined using filter paper as 11.16 mg/cm2 and 7.17 mg/cm2 for Hybrid Tea and Floribunda respectively. In conclusion, the application to rose plants of different concentrations of BA and NAA can promote a number of changes to both morphological and physiological parameters, and in turn have a significant effect on metabolite changes in different rose tissues. Application BA and NAA method could be applied to other floriculture plants to increase the flowers production in rose or other ornamental plants. vi Table of Contents Contents Thesis Title ................................................................................................................................. i Acknowledgements ................................................................................................................ iii Abstract ..................................................................................................................................... v Table of Contents ................................................................................................................... vii Publications and Presentations .............................................................................................. xi Publications .......................................................................................................................... xi Poster Presentations ............................................................................................................ xi List of Abbreviations ............................................................................................................. xii Chapter 1 .................................................................................................................................
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