Anthurium Fragrance: Genetic and Biochemical Studies
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ANTHURIUM FRAGRANCE: GENETIC AND BIOCHEMICAL STUDIES A DISSERTATION SUBMITTED TO THE GRADUATE DIVISION OF THE UNIVERSITY OF HAWAII IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN HORTICULTURE DECEMBER 1997 By Nuttha Kuanprasert Dissertation Committee: Adelheid R. Kuehnle, Chairperson Chung-Shih Tang Catherine Cavaletto Richard Criley Richard M. Manshardt David T. Webb We certify that we have read this dissertation and that, in our opinion, it is satisfactory in scope and quality as a dissertation for the degree of Doctor of Philosophy in Horticulture. DISSERTATION COMMITTEE 1^-hXLu.AA fl 16^1—^ Chairperson M HL > L x6 © Copyright 1997 by Nuttha Kuanprasert All Rights Reserved iii ACKNOWLEDGEMENTS I am very grateful to my advisor, Dr. Adelheid R. Kuehnle for her valuable advice and support throughout the dissertation process and also for her assistance in editing my manuscript. I would like to thank my committee members: Dr. C. S. Tang for his guidance in chemical analysis, Ms. Catherine Cavaletto for her instruction in fragrance evaluation. Dr. Richard Criley for providing useful and relevant articles and abstracts. Dr. Richard M. Manshardt for his support and comments, and Dr. David T. Webb for his long-term assistance in histological study. I extend my sincere thanks to Emeritus Professor Haruyuki Kamemoto. I truly enjoyed the numerous hours we spent working in the nursery. Through this experience, I gained invaluable knowledge about general horticultural techniques and philosophy of life. 1 would like to extend my gratitude to Dr. Teresita Amore for her kind assistance and suggestions, Ms. Nellie Sugii, and lab members for their friendship and encouragement. A special thanks to Ms. Susan Takahashi for special arrangements in ordering chemicals and supplies. I also extend sincere thanks to my expert panelists, Ms. Natalie Nagai, Mr. Dale Evans, Mr. Paul Murakami, Dr. Tang, Dr. Kuehnle and Dr. Amore for their assistance in developing Anthurium fragrance descriptors. None of this would have been possible without the support of many friends. I would like to thank Cindy and Manoj for accompanying me during conducting surveys at night, Wei-fan, Jocelyne and Karl for their help in GC and GC-MS analysis. IV Theeranuch for assisting in graphic technique, and Tracie Matsumoto and Manoj Potapohn for their continuous support through the good times and bad times. My deepest appreciation is extended to my parents and my brother who provided me with the moral support and encouragement to complete my dissertation. ABSTRACT The total of 147 Anthurium species and hybrids at the University of Hawaii and the Missouri Botanical Garden germplasm collections was evaluated for the presence or absence of scent, type of fragrance, time of emission, daily occurrence, developmental stage of scent emission, color of spathe and spadix. A majority of inflorescences (76%) emitted scent. Scent was categorized as citrus, fishy, floral, foul, fruity, menthol, minty, pine, spicy and sweet. There was no relationship between scent production or scent quality with flower color or botanical section. A plurality of inflorescences emitted scent during the morning (45%) and at the pistillate stage (77%). Fragrance life of unharvested inflorescences varied from 2 to 3 days up to 4 weeks, whereas that of harvested inflorescences was only 1 or 2 days. Fi progenies of crosses between fragrant x fragrant and non-fragrant x fragrant parents were studied to determine whether a single gene or more than one gene governed presence of scent. Progenies from 24 crosses were tested by Chi-square analysis for a single dominant or recessive trait. None produced expected segregation ratios for a single gene trait, indicating that multiple genes likely govern presence of fragrance in Anthurium. Progeny also segregated for fragrance quality and included non-parental scents. Fragrance of seven Anthurium species and ten hybrids was analyzed by gas chromatography and mass spectrometry. Nineteen monoterpenes (lipids) and some alcohols, aldehydes and esters were identified. Limonene and 1,8-cineole were common vi to most samples along with a-pinene, p-pinene, myrcene and linalool. Hybrid UH1299, emitting a sweet and floral scent all day, showed fluctuation in amounts and types of compound during the daily cycle. Tepals were associated with fragrance production in Anthurium. Histological comparison between tepals of fragrant and non-fragrant spadices showed lipids and starches present in both fragrant and non-fragrant samples. However, in fragrant samples, the amount of lipids was significantly greater than that of non-fragrant ones, whereas the amount of starch was significantly greater in non-fragrant samples compared with fragrant ones. These data support the hypothesis that high levels of lipids were associated with fragrance production. Vll TABLE OF CONTENTS Acknowledgements ................................................................................................... iv Abstract....................................................................................................................... vi List of Tables............................................................................................................ xiii List of Figures ...........................................................................................................xv Chapter 1 Literature Review ..................................................................................... 1 1.1 Sense of smell ............................................................................................1 1.1.1 How do human beings perceive odor?............................................. 1 1.1.2 Terminology........................................................................................2 1.1.3 Factors affecting sense of smell.......................................................2 1.1.4 Flower scent........................................................................................3 1.1.5 Importance of flower scent in plant biology ................................. 4 1.2 Fragrance classification ..............................................................................6 1.3 Fragrance analysis........................................................................................8 1.3.1 The chemistry of volatile compounds.............................................. 8 1.3.2 Extraction............................................................................................8 1.3.3 Distillation...........................................................................................9 1.3.4 Headspace.........................................................................................10 1.3.5 Chemical analysis............................................................................. 11 1.4 Scent production in flowering plants...................................................... 12 1.4.1 Apiaceae............................................................................................14 1.4.2 Araceae............................................................................................. 14 viii 1.4.3 Asteraceae........................................................................................15 1.4.4 Labiatae............................................................................................16 1.4.5 Orchidaceae.....................................................................................17 1.4.6 Rosaceae............................................................................................17 1.5 Inheritance of fragrance............................................................................17 1.5.1 Chamomilla recutita......................................................................... 18 1.5.2 Mentha spp........................................................................................ 19 1.5.3 Ocimum basilicum........................................................................... 22 1.5.4 Perilla frutescens.............................................................................23 1.5.5 Oryza sativa......................................................................................24 1.5.6 Thymus vulgaris................................................................................25 1.6 Anthurium genetics and breeding ............................................................26 1.6.1 Spathe color......................................................................................26 1.6.2 Flower fragrance.............................................................................. 27 1.6.3 Value of crop and breeding............................................................28 Chapter 2 Survey of fragrant Anthurium germplasm at the University of Hawaii and the Missouri Botanical Garden ..........36 2.1 Abstract ......................................................................................................36 2.2 Introduction .............................................................................................. 37 2.3 Material and methods................................................................................38 2.3.1 Species, cultivars and hybrids survey............................................ 38 2.3.2 Fragrance life ................................................................................. 39 IX 2.4 Results and discussion .............................................................................39 2.4.1 Species survey................................................................................