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Llrg Onv'gvajooof 'o'r{d'u"urys'u selll tr 'c'qd'uosuqol'J llrg 'cl'qd'uosuqol Ielu€c'Y rosr^pY ''G'qd 'uoJ; 'v ueoFll€) :ooilrurruoJ Eurururexg leqc''q'qd'Eu€lX (,{uua1) un1-nlb :,,(g pe,ro;ddy g1g7 erdselllD "I {l1ug lq3t.tddo3 eurloJeJ rIUoN'ua1eg-uo1sut16 0I0z dentr fEolorg AHdOSO'IIHd dO UOJf,OC ;o esr8eq aqtr roJ sluerue;mbed orllJo lueruilUlnC plu€d ul SSJNSIJS ONV SJdY dO TOOHJS EJVNC\rUC AJISUSAINN JSIUOC E)VA\ 3o fi1nceg epnperD aql ol pogprqns uop€Uessrq V !IIdS!I'ITIC'T A'IIWII Ag (sgovrc r d oISSyJ oNV'gvaJooof"IlHd'lrvfl cloJrua :iIVAf,Y3ruO SUSHJVIIH ONV STIIUNV'I IIHT CO SEIOOJS CIJIINSOOTAHd ii ACKNOWLEDGEMENTS I would like to thank my committee members, Drs. Kathleen A. Kron (WFU), A. Daniel Johnson (WFU), Erik C. Johnson (WFU), Miles R. Silman (WFU), and Qiu-Yun (Jenny) Xiang (NC State U.), for their ongoing support of my academic training. I called on them many times during the past five years, and I was always met with enthusiasm and genuine interest. Particular thanks to Kathy, for being a kind and patient advisor, for tolerating my mysterious work hours and my desire to study things like Russian, and for exchanging funny cat stories. I am most grateful for the opportunity to have learned from an expert in my field. Funding for this study was provided by NSF grant DEB-023043 and Wake Forest University as part of a broader study of the Rhododendron clade. The Department of Biology provided a Vecellio Grant in order to facilitate my trips to Duke University to learn plant anatomy techniques. I received Cocke Travel Grants from the WFU Biology Department, Alumni Travel Grants from the WFU Graduate School of Arts and Sciences and Student Travel Grants from The Association of Southeastern Biologists to attend numerous meetings. The following herbaria loaned material for this project: Harvard U. Herbaria (HUH), Swedish Museum of Natural History Herbarium (S), Museum of Nature and Human Activities, Japan (HYO), Royal Botanic Garden Edinburgh (RBGE), U. Alaska Museum of the North (ALA), Royal Botanic Gardens, Kew (RBGK), U. Washington (WTU), the Heather Society, Rhododendron Species Foundation (RSF), Florida Museum of Natural History (FLAS), California Academy of Sciences (CAS) and the Smithsonian iii Institution (US). The University of British Columbia Botanical Garden allowed me to collect plants on site. The following individuals collected or facilitated collections of plant material for these studies: Drs. Jiang Hongyuan and Du Fengguo (Forestry College of Beihua U.), Salih Terzioğlu (Karadeniz Technical U.), Robert Meinke and Aaron Liston (Oregon State U.) and Richard Abbott (Florida Museum of Natural History). Drs. Rick Ree (Field Museum), Jun Wen (Smithsonian Institution), David Boufford (Harvard U.), Peter Fritsch and Bruce Bartholomew (California Academy of Sciences) generously provided plant material from the Gaoligongshan Biodiversity Survey collections for the Cassiope project. Drs. Dick White and Mel Turner (Duke U.) graciously agreed to have me visit their lab to learn various plant anatomy techniques, and Dr. Anita McCaulay (Wake Forest U.) helped me with microscopy techniques. Support staff in the WFU Department of Biology helped keep things running, including our IT guru, Mr. Jeff Muday, and our wonderful secretaries, Ms. Cindy Davis and Ms. Zella Johnson. Numerous individuals in addition to those listed above assisted me with technical aspects of the project, made valuable suggestions and were happy to talk formal and informal ‘shop’ with some frequency. These include Drs. Torsten Eriksson (Swedish Museum of Natural History), Randy Small (UT-Knoxville), Zack Murrell (Appalachian State U.), Ann Powell (U. Evansville), Henry Wilbur (U. Virginia), Peter Stevens (Missouri Botanical Garden), Gary Wallace (Rancho Santa Ana Botanic Garden), Helen Murphy and Michelle DaCosta (Wake Forest U.), Jim Petranka (UNC-Asheville), Yu Yan (Sichuan U.), and Ms. A.J. Harris (Oklahoma State U.). These folks and many other iv regional biologists and herbarium curators provided feedback, guidance, encouragement and friendship along the way. Many thanks to all. v TABLE OF CONTENTS LIST OF FIGURES………..…………………………………………..…………..... vi LIST OF TABLES AND APPENDICES…………………………………………… viii ABSTRACT………………………………………………………….……………… ix Chapter I. INTRODUCTION……………………………………………..........……….….. 1 II. MOLECULAR PHYLOGENETIC RELATIONSHIPS AND A REVISED CLASSIFICATION OF THE SUBFAMILY ERICOIDEAE (ERICACEAE)...…………………………………………………………………. 9 Published in Molecular Phylogenetics and Evolution, 2010 III. PHYLOGENETIC RELATIONSHIPS AND HISTORICAL BIOGEOGRAPHY OF THE PHYLLODOCEAE (ERICACEAE) ………………………………….. 48 IV. PRELIMINARY PHYLOGENETIC ANALYSIS OF CASSIOPE AND EVALUATION OF THE EVOLUTION OF SOME MORPHOLOGICAL AND WOOD ANATOMY CHARACTERS….………… 99 V. CONCLUSIONS…………………………………………………….………….. 136 VITA…………………………………………………………………………............ 141 vi LIST OF FIGURES FIGURE 2.1 Total combined chloroplast data (matK, ndhF, rbcL and 38 trnS-G spacer) analysis of the Ericoideae FIGURE 2.2 Total combined nuclear data (nrITS and waxy) analysis of 39 the Ericoideae FIGURE 2.3 Total combined molecular data (nrITS, waxy, matK, ndhF, 40 rbcL and trnS-G spacer) analysis of the Ericoideae FIGURE 2.4 Phylogram of Ericoideae showing branch lengths resulting 41 from Bayesian analysis of total combined molecular data FIGURE 3.1 Total combined chloroplast data (matK, ndhF, rbcL and 88 trnS-G spacer) analysis of the Phyllodoceae FIGURE 3.2 Total combined nuclear data (nrITS and waxy) analysis of 89 the Phyllodoceae FIGURE 3.3 Total combined molecular data (nrITS, waxy, matK, ndhF, 90 rbcL and trnS-G spacer) analysis of the Phyllodoceae FIGURE 3.4 Phylogram of Phyllodoceae showing branch lengths 91 resulting from Bayesian analysis of total combined molecular data FIGURE 3.5 Reconstruction of ancestral areas of the Phyllodoceae in S- 92 DIVA FIGURE 3.6 Estimation of relative node ages with node bars in the 93 Phyllodoceae FIGURE 3.7A Estimation of absolute node ages with node error bars in the 94 Phyllodoceae FIGURE 3.7B Estimation of absolute mean node ages in the Phyllodoceae 95 with node bars removed FIGURE 4.1 Chloroplast data (trnS-G spacer) analysis of Cassiope. 124 FIGURE 4.2 Total combined nuclear data (nrITS and waxy) analysis of 125 Cassiope. FIGURE 4.3 Total combined molecular data (trnS-G spacer, nrITS and 126 waxy) analysis of Cassiope. vii FIGURE 4.4 Phylogram of Cassiope showing branch lengths resulting 127 from Bayesian analysis of total combined molecular data FIGURE 4.5 Parsimony reconstruction of leaf form in Cassiope traced 128 onto the combined molecular phylogeny. FIGURE 4.6 Parsimony reconstruction of abaxial leaf stomata in Cassiope 129 traced onto the combined molecular phylogeny. FIGURE 4.7 Parsimony reconstruction of number of growth rings in 130 Cassiope traced onto the combined molecular phylogeny. FIGURE 4.8 Parsimony reconstruction of mean elevation in Cassiope 131 traced onto the combined molecular phylogeny. FIGURE 4.9 Parsimony reconstruction of stem diameter in Cassiope 132 traced onto the combined molecular phylogeny. FIGURE 4.10 Parsimony reconstruction of vessel density (vessels per mm2) 133 in Cassiope traced onto the combined molecular phylogeny. viii LIST OF TABLES AND APPENDICES TABLE 2.1 Abbreviated taxonomic history of the Ericoideae according 42 to Copeland (1943), Cox (1948), Stevens (1971), Kron et al. (2002) and the current study TABLE 2.2 Gene region, aligned length, number of potentially 44 parsimony informative characters (and %), and number of missing taxa (and %) in a phylogenetic study of the Ericoideae APPENDIX 2.1 Taxa, voucher information and Genbank accessions for a 45 phylogenetic study of the Ericoideae APPENDIX 2.2 DNA regions, evolutionary models, primer sequences and 47 PCR protocols for studies in the Ericoideae, Phyllodoceae and Cassiope TABLE 3.1 Gene region, evolutionary model, aligned length, number of 96 potentially parsimony informative characters (and %), and number of missing taxa (and %) in a phylogenetic study of the Phyllodoceae APPENDIX 3.1 Taxa, voucher information and Genbank accesions for a 97 phylogenetic study of the Phyllodoceae TABLE 4.1 Gene region, evolutionary model, aligned length, number of 134 potentially parsimony informative characters (and %), and number of missing taxa (and %) in a phylogenetic study of Cassiope APPENDIX 4.1 Taxa, voucher information and Genbank accession numbers 135 for a phylogenetic study of Cassiope ix ABSTRACT The current study explores phylogenetic studies within the laurel and heather clades within the Ericaceae. The two groups are members of two closely related clades, the Ericoideae and Cassiope. Phylogenetic relationships within the subfamily Ericoideae were investigated first in order to establish its main clades. Five main clades were recovered. The first four are the tribes Ericeae (African and European heathers), Empetreae (crowberries and Diplarche, a Rhododendron-like species), Rhodoreae (Rhododendron and relatives) and Bryantheae, comprised of two poorly understood genera, Bryanthus and Ledothamnus. The fifth (Phyllodoceae) includes plants such as the mountain laurels (Kalmia), the mountain heathers (Phyllodoce) and their close relatives. The Phyllodoceae was the subject of the second part of the research. The goal here was to establish a species-level phylogeny and to examine historical biogeography of the tribe. DIVA analysis was used to infer ancestral areas of nodes, and putative vicariance events. Identification of these events was followed by relative
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