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Genetic Diversity, Nutritional and Biological Activity of Momordica

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Genetic Diversity, Nutritional and Biological Activity of Momordica Genetic diversity, nutritional and biological activity of Momordica cochinchinensis (Cucurbitaceae) A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy (Applied Biology and Biotechnology) Dilani Chathurika Wimalasiri BSc (Hons) (Institute of Chemistry, Sri Lanka) BSc (Spectrum Institute of Science and Technology, Sri Lanka) School of Applied Sciences College of Science Engineering and Health RMIT University August, 2015 Declaration I certify that except where due acknowledgement has been made, the work is that of the author alone; the work has not been submitted previously, in whole or in part, to qualify for any other academic award; the content of the thesis is the result of work which has been carried out since the official commencement date of the approved research program; any editorial work, paid or unpaid, carried out by a third party is acknowledged; and, ethics procedures and guidelines have been followed. Dilani Chathurika Wimalasiri 31 August 2015 ii ACKNOWLEDGEMENTS I wish to express my greatest gratitude to my supervisor, Dr Tien Huynh, for her mentoring, guidance and constant encouragement throughout the years. I’m also thankful to her for collecting the plant samples for me to work on. I would like to thank my co supervisors Associate Professor Terrence Piva and Dr Sylvia Urban for their patience, guidance and helpful discussions throughout this project. I’m thankful for Professor Ann Lawrie for her comments on the project and my thesis chapters. I am grateful for the help from Dr Robert Brkljača and his assistance in the nutritional analysis section of my project. I am also thankful for Mr Paul Morrison’s (School of Applied Sciences - Applied Chemistry, RMIT Unniversity) guidance in UPLC analysis. I would like to thank Layla Mehdi Alhasan for kindly providing the breast cancer cells and assistance in cell culture techniques. I am also thankful for Mr Griffin D’Costa, Mr Alaa Banjar and Xian Yang Chang for providing the melanoma cells, normal human fibroblast cells (NHDF) and the support in the laboratory. iii I would like to thank Dr Chaitali Dekiwadiya from RMIT microscopy and microanalysis facility (RMMF) for helping me with the TEM experiment. I would like to thank Dr Julie Quach from the confocal laser scanning microscopy facility, for training me for on the instruments. I would like to thank Miss Lillian Chuang from Food Technology & Nutrition, School of Applied Sciences, RMIT University for allowing me to use the Konico Minolta, Chroma meter – CR 400. I am grateful for the support from Thu Nga Nguyen and Huong Nguyen in Hanoi University of Agriculture for collection assistance, Duc Nguyen (Hanoi University of Agriculture) for access and use of lab facilities, Cuong Nguyen (Hanoi University of Agriculture) for agronomy advice, Nonnipa Labbunruang and Wanisika Phadungsii (Tammasat University) for collection assistance, Rattanapong Charntawong (Thailand gac farm) and Thuy Nguyen for collection assistance. I’m thankful for Dr Sophie Parks of Department of Primary Industries, New South Wales, Australia for providing M. cochinchinensis aril and leaf samples. Huge thanks for Alison Fong for the friendship, constant encouragement and most of all, for being the motivation to continue my work. I’m very grateful that we shared this three year journey together. iv Special thanks, to my mama, dada brother Delan and sister Kanchana whose confidence in me kept me going until the very end of this project. Finally, I would like to thank my dear husband, Kalana, for his love, understanding and constant help. No words can describe how much I appreciated your support throughout this journey. v DEDICATION This thesis is dedicated to my mother J. P Wimalasiri, father I. D Wimalasiri, my brother Delan Wimalasiri and my husband Kalana Katuwawala vi TABLE OF CONTENTS Acknowledgements ........................................................................................................ iii Dedication ....................................................................................................................... vi Table of Contents .......................................................................................................... vii List of Tables ................................................................................................................ xiv List of Figures .............................................................................................................. xvi List of Abbreviations .................................................................................................... xx Thesis Publications .................................................................................................... xxiv Thesis Abstract ........................................................................................................... xxv CHAPTER 1 .................................................................................................................... 1 Background and review of literature ............................................................................ 1 1.1 Cucurbitaceae family ............................................................................................ 1 1.1.1 Economic importance ................................................................................. 2 1.1.2 Taxonomical relationship within cucurbit crops ........................................ 2 1.2 Momordica genus and representatives .................................................................. 4 1.2.1 Origin and distribution ............................................................................... 9 1.3 Momordica cochinchinensis ............................................................................... 10 1.3.1 Distribution and vernacular names ........................................................... 10 1.3.2 Cultivation and propagation ..................................................................... 11 1.3.3 Botany ....................................................................................................... 12 1.3.4 Diversity ................................................................................................... 15 1.3.4.1 Morphological diversity ..................................................................... 15 1.3.4.2 Molecular markers ............................................................................. 17 1.3.4.3 Genetic diversity ................................................................................ 18 1.4 Phytochemicals ................................................................................................... 20 vii 1.4.1 Carotenoids ............................................................................................... 20 1.4.1.1 Carotenoids in M. cochinchinensis .................................................... 25 1.4.1.1.1 Pro-vitamin A activity ................................................................. 27 1.4.1.1.2 Antioxidant activity ..................................................................... 28 1.4.1.2 Factors affecting carotenoid content .................................................. 29 1.4.1.2.1 Genetic influences ....................................................................... 29 1.4.1.2.2 Eco-geographical influences ....................................................... 30 1.4.1.2.3 Stage of maturity ......................................................................... 31 1.4.1.2.4 Post-harvest processing ............................................................... 32 1.4.1.2.5 Extraction .................................................................................... 33 1.4.1.2.6 Analysis ....................................................................................... 34 1.4.1.2.6.1 Chromatography .................................................................. 34 1.4.1.2.6.2 UV-Visible spectrophotometry ............................................ 36 1.4.1.2.6.3 Colorimetry .......................................................................... 38 1.4.2 Fatty acids ................................................................................................. 39 1.4.3 Phenolic compounds ................................................................................. 40 1.4.3.1 Flavonoids .......................................................................................... 40 1.4.3.2 Phenolic acids ..................................................................................... 41 1.5 Medicinal importance ......................................................................................... 41 1.5.1 Traditional medicinal uses of M. cochinchinensis.................................... 41 1.5.2 Cancer ....................................................................................................... 44 1.5.2.1 Breast cancer ...................................................................................... 45 1.5.2.2 Skin cancer ......................................................................................... 45 1.5.2.3 Hallmarks of cancer ........................................................................... 47 1.5.2.4 Cell death ........................................................................................... 48 1.5.2.4.1 Morphological aspects of cell death ............................................ 49 viii 1.5.2.4.2 Biochemical aspects of apoptosis ................................................ 50 1.5.2.4.3 Analysis methods for cell
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