VEGETABLE SHEEP: A CHEMOSYSTEMATIC STUDY OF THE CASSINIINAE. By ALAN REID BSc, Victoria, University of Wellington, Wellington, New Zealand, 1986 BSc.(Hons.), Victoria, University of Wellington, Wellington, New Zealand, 1987 MSc, Victoria, University of Wellington, Wellington, New Zealand, 1989 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Department of Botany) We accept this thesis as conforming to the required standard THE UNIVERSITY OF BRITISH COLUMBIA DECEMBER 1995 ©Alan Reid, 1995 In presenting this thesis in partial fulfilment of the requirements for an advanced degree at the University of British Columbia, I agree that the Library shall make it freely available for reference and study. I further agree that permission for extensive copying of this thesis for scholarly purposes may be granted by the head of my department or by his or her representatives. It is understood that copying or publication of this thesis for financial gain shall not be allowed without my written permission. Department The University of British Columbia Vancouver, Canada Date DE-6 (2/88) VEGETABLE SHEEP: A CHEMOSYSTEMATIC STUDY OF THE CASSINIINAE It is well known that there are few places in the Asteraceae where generic limits are more difficult to apply in practice than in the tribe Gnaphalieae subtribe Cassiniinae. This is the subtribe of the paper daisies, the Edelweiss, the cudweeds. These species are characterised by discoid floral heads with papery involucral bracts. This thesis presents the results of an investigation of the flavonoid chemistry of the major species present in Australasia from which an attempt to infer taxonomic relationships among the species based on the combination of chemical, morphological and ecological characters is made. The exudate and vacuolar flavonoid profiles of six genera Cassinia R. Br., Ozothamnus R. Br., Raoulia Hook, f and Haeckeria F. Muell., Leucoqenes R. Br. and Lawrencella Anderberg were established. The exudate chemistry consists of chalcones, dihydrochalcones and flavanones, several of which lack B-ring oxygenation, and quercetin 3-0- and 7-0-methyl ethers. Vacuolar flavonoids include galangin, eriodictyol-7-O-methyl ether, and a series of kaempferol and quercetin glycosides. Several combinations of these compounds occur in the taxa studied. Species in this study form a number of relationships that show a strong correlation to geography, altitude and ecological conditions. The production of exudate flavonoids show a direct correlation to the amount of UV-B radiation. These relationships mimic the classical taxonomy proposed by Allan in 1961 for the New Zealand species and by Burbidge in 1958 for the Australian species. Differences in distribution of flavonoids are taken as evidence for recognition that the relationships between the genera are as complex as classical taxononomic studies suggest. TABLE OF CONTENTS Abstract ii Table of Contents iii List of Figures iv List of Tables vi Acknowledgments vii Dedication viii INTRODUCTION 1 Chapter 1: The Biogeography of the Southern End of the world. 15 Chapter 2: Description of the species in this study. 30 Chapter 3: Previous flavonoid studies of the Gnaphalieae, Plucheeae and Inuleae 62 Chapter 4: Materials and Methods. 80 Chapter 5: Results ecological, morphological and chemical characters forming the cladogram. 100 Chapter 6: Discussion Ultraviolet light and flavonoid profiles. 124 Chapter 7: Discussion Ultraviolet light and morphology. 133 Chapter 8: Summary and Conclusions 140 Literature Cited. 146 Appendix 1. Collection sites. 170 Appendix 2. Flavonoid spectra. 191 Appendix 3. Trivial and common names for flavonoids. 200 Appendix 4. Authority names. 203 iii LIST OF FIGURES Figure 1 Style and anther types found in the Gnaphalieae. 2 Figure 2 The strict consensus tree of the Cassiniinae sensu Anderberg. 4 Figure 3 The strict consensus tree of the Cassiniinae sensu Puttock. 7 Figure 4 Species relationships within the Australasian Gnaphalieae sensu Breitwieser and Ward. 9 Figure 5 The New Zealand Geological and Botanical region. 16 Figure 6 Vegetation map of New Zealand. 18 Figure 7 Vegetation map of Australia. 21 Figure 8 Coastlines of New Zealand. 25 Figure 9 Geological movements of the Australian Continent 27 Figure 10 New Zealand Cassinia species. 33 Figure 11 Distribution of Cassinia species in New Zealand 35 Figure 12 Altitudinal distribution of New Zealand Cassinia species across 41°S latitude. 36 Figure 13 Cassinia vauvilliersii 40 Figure 14 Cassinia longifolia 41 Figure 15 Cassinia denticulata 41 Figure 16 Cassinia rugata 41 Figure 17 Distribution of the Australian Cassinia species 42 Figure 18 Leaf detail of whipcord Qzothamnus species 47 Figure 19 Qzothamnus depressus 49 Figure 20 Qzothamnus diosmifolius 49 Figure 21 Distribution of the Australian Qzothamnus species 54 Figure 22 Distribution of the New Zealand Edelweiss 61 Figure 23 Floral morphology of the Compositae. 90 Figure 24 Ecological differences 101 Figure 25 Strict consensus cladogram of the New Zealand species 102 Figure 26 Strict consensus cladogram of the Australian species 103 Figure 27 Tomentum position in mat forming species of Raoulia. 138 Figure 28 Transverse section of R. glabra leaf lamina. 139 Figure 29 Collection sites: New Zealand 189 Figure 30 Collection sites: Australia 190 LIST OF TABLES Table 1 Species in this thesis. 13 Table 2 B-ring deoxyflavonoids of Gnaphalium species. 66 Table 3 Unsubstituted, methylated and glycosylated chalcones in Helichrysum species. 68 Table 4 C-prenyl and cyclised C-prenyl chalcones in Helichrysum species. 70 Table 5 O-prenylated and substituted chalcones in Helichrysum species. 70 Table 6 Methylated, glycosylated and unsubstituted flavanones in Helichrysum species. 71 Table 7 Substituted flavanones from Helichrysum species. 72 Table 8 B-ring deoxyflavones and flavonols from Helichrysum species. 73 Table 9 Data matrix of flavonoid, morphological and ecological characters. 91 Table 10 Flavonoids from Cassinia, Ozothamnus, Haeckeria, Lawrencella, Leucogenes and Raoulia. 94 Table 11 Morphological characters of Cassinia rugata and related species. 123 Acknowledgements I wish to acknowledge the financial contribution of N. S. E. R. C. (an operating grant to Dr. B. A. Bohm) and the contribution (financial, emotional and spiritual) of my parents. Drs. Bruce A Bohm, Jack Maze, Wilf Schofield, Gerald Straley and Gil Hughes for their time and patience and for including me as a colleague. The contribution of people you interact with on a daily basis must be acknowledged as they provided an encouraging place to work Andree Desrochers, Ann Eastman, Ken Marr, Ji Yong Yang, Santokh Singh, Mohammed Koupai, Sandra Millard, Anthony Fielding and Sonya Budge, Kelly Bannister, Dean Mulyk. Also to my most favourite room mate Anita Andree!!!!! Jean and Jeff White, Roni Bartram, The Keyes: Bob Deborah Lauren and Douglas listened to my toils and tribulations as if they were my parents and gave me a home to relax in. To all the people at the Burnaby Lake Rugby Football Club, especially to Jamie, Garry, John, Walt, Tanya Donaldson, Britta Hobkirk and the B. C. I. T. Cougars rugby team. Its been great coaching and playing with yah!! I wish to acknowledge the contributions of all my friends at St Christopher's Anglican Church West Vancouver, but especially those who put up with me the most and also trained my warblings into something a little more pleasant than before: Jack Butterfield, Ian and Ruth Whitelaw, Roy Holland, Charles McNeight, Kathryn McNeight, Richard McDiarmid, Simon Farrow, John (my tenor buddy) Wright, The Keyes: Bob Deborah Lauren and Douglas, Dr. George Robb, The Rev. Ray and Sadie Murrin, The Ven. Lou and Georgie Rivers, Margaret Manning, Sandra Murrata, Mariel Terry, Betty Phillips, Anne Wood, Barbara Cave, Joan Uren, Dr John and Kit Stevenson, Nathalie Djan-Chekar, Pat Bice, Shelly McCann, Susan Dumoulin, Judy Lowe, Judy Tang, Paul and Elizabeth Cox, Gillian Sutherland, Neil and Sheila Sutherland, Lesley Sutherland, Don and Carole Neilson. The people at Our Lady of Perpetual Help: Elizabeth Towers, Susan Crober, Patricia Macdonald, Kim Smyth, Francisco Salinas and especially for the support of Angela Uruski And lastly to Lesley (a real angel) and Neil, you taught me the life in music and the music of life. This thesis is dedicated to the memory of Neil Sutherland. He showed me the music of life and the life in music. Thank you for your gift. viii Introduction In terms of species numbers the Asteraceae (Compositae) is the largest family of plants. According to Bremer (1994) this family comprises 1535 genera and over 23,000 species, arranged in 3 subfamilies and 17 tribes. Of these 17 tribes it is particularly difficult to define the generic limits in the tribe Gnaphalieae (Jeffrey, 1969). The Gnaphalieae sensu Anderberg consists of five subtribes (Gnaphaliinae, Cassininae, Loricariinae, Relhaniinae and Angianthinae), 167 genera and more than 2000 species making it one of the largest tribes in the family. The tribe has a worldwide distribution and is well represented in Australia, New Zealand and southern Africa. Most species are cosmopolitan and weedy colonizers of disturbed habitats, e.g., Gnaphalium uliginosum L. Some are commonly used in dried flower arrangements including the everlasting daisies(e.g., Helichrysum bracteatum Vent) the Edelweiss (Leontipodium R. Br. ex Cass.), the cudweeds (Gnaphalium L.) and the pussytoes (Antennaria Gaert). Bentham