SYSTEMATICS OF THE GENUS EULOPHIA R. Br. (ORCHIDACEAE) IN GHANA WILLIAMS BENJAMIN DARKO (S.ID NO. 10563658) A THESIS PRESENTED TO THE DEPARTMENT OF PLANT AND ENVIRONMENTAL BIOLOGY IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF PHILOSOPHY OF THE UNIVERSITY OF GHANA DEPARTMENT OF PLANT AND ENVIRONMENTAL BIOLOGY UNIVERSITY OF GHANA LEGON JANUARY, 2023 University of Ghana http://ugspace.ug.edu.gh i DECLARATION This is to certify that the work presented in this thesis: “SYSTEMATICS OF THE GENUS EULOPHIA R. Br. ex Lindl. (ORCHIDACEAE) IN GHANA” was carried out under the supervision of Professor Alex Asase and Dr. Matthew Kweku Essilfie, Department of Plant and Environmental Biology, University of Ghana, Legon. I hereby declare that no part of this thesis has been presented for another degree of this University or elsewhere. Williams Benjamin Darko (Student) Date: 31-01-2023 Prof. A. Asase (Supervisor) Date: 31-01-2023 Dr. M. K. Essilfie (Supervisor) Date: 31-01-2023 University of Ghana http://ugspace.ug.edu.gh ii DEDICATION I dedicate this work to God almighty for His grace, guidance and protection, and to my mother, Ms. Rebecca Asmah, my sister, Ms. Mariam A. Williams and my guardian, Mr. Ebenezer Sackey who have supported me immensely in my entire education. University of Ghana http://ugspace.ug.edu.gh iii ACKNOWLEDGEMENTS For the favour, health and patience during this project, I am most grateful to God. I wish to express my sincere appreciation to my supervisors, Professor Alex Asase and Dr. Matthew Kweku Essilfie for making it possible for me to complete this work. Their guidance was very instructive. My earnest appreciation goes to Mr. Francis Asamoah and Mr. Patrick Eyiah for their companionship during my field work. My sincere thanks to Ms. Diana Banyeh, Ms. Anna Teye, Ms. Gladys Aboagye Asamoah, Ms. Christiana Peprah Oppong and Ms. Jacqueline Amoakoah Coffie for their encouragement and contributions during my analysis of data. I am very grateful to Professor Gabriel Komla Ameka for all his encouragement throughout the dissertation writing and for constructive criticisms of this work. University of Ghana http://ugspace.ug.edu.gh iv TABLE OF CONTENTS 1.0 INTRODUCTION................................................................................................................... 1 1.1 Background ............................................................................................................................. 1 1.2 Problem statement .................................................................................................................. 2 1.3 Justification ............................................................................................................................. 3 1.4 Objectives of the study............................................................................................................ 4 2.0 LITERATURE REVIEW ...................................................................................................... 5 2.1 Systematics and Taxonomy .................................................................................................... 5 2.2 Orchidaceae Juss. .................................................................................................................... 6 2.2.1 Taxonomy of the Orchidaceae ....................................................................................... 11 2.2.2 Distribution and Ecology of Orchidaceae .................................................................... 16 2.3 Orchidaceae in West Africa including Ghana.................................................................... 17 2.4 Genera of Orchids in Ghana ................................................................................................ 18 2.4.1 Aerangis Rchb.f. .............................................................................................................. 19 2.4.2 Ancistrorhynchus Finet ................................................................................................... 19 2.4.3 Ancistrochilus Rolfe ........................................................................................................ 20 2.4.4 Angraecum Bory ............................................................................................................. 20 2.4.5 Ansellia Lindl. ................................................................................................................. 21 2.4.6 Auxopus Schltr. ............................................................................................................... 21 2.4.7 Bolusiella Schltr. ............................................................................................................. 22 2.4.8 Brownleea Harv. ex Lindl. ............................................................................................. 22 2.4.9 Bulbophyllum Thouars. .................................................................................................. 23 2.4.10 Calyptrochilum Kraenzl. ............................................................................................... 23 2.4.11 Corymborkis Thouars. .................................................................................................. 24 2.4.12 Cheirostylis Blume ......................................................................................................... 24 2.4.13 Cyrtorchis Schltr. .......................................................................................................... 24 2.4.14 Diaphananthe Schltr. .................................................................................................... 25 2.4.15 Didymoplexis Griff. ....................................................................................................... 26 2.4.16 Disa P.J. Bergius ........................................................................................................... 26 2.4.17 Disperis Sw..................................................................................................................... 26 2.4.18 Eggelingia Summerh. ................................................................................................... 27 2.4.19 Epipogium Borkh. ......................................................................................................... 28 2.4.20 Eulophia R. Br............................................................................................................... 28 2.4.21 Manniella Rchb.f. .......................................................................................................... 31 2.4.22 Microcoelia Lindl. ......................................................................................................... 31 University of Ghana http://ugspace.ug.edu.gh v 2.4.23 Polystachya Hook. ......................................................................................................... 32 3.0 MATERIALS AND METHODS ......................................................................................... 33 3.1 Morphological Studies .......................................................................................................... 33 3.2 Ecological and Distributional Studies ................................................................................. 37 3.3 Taxonomic Studies ................................................................................................................ 39 3.4 Data Analysis ......................................................................................................................... 39 4.0 RESULTS .............................................................................................................................. 41 4.1 Morphological Studies .......................................................................................................... 41 4.1.1 Leaf and Stem morphology ............................................................................................ 41 4.1.2 Inflorescence, Flower and Fruit Morphology .............................................................. 41 4.2 Ecology and Distribution ...................................................................................................... 46 4.3 Taxonomic Studies ............................................................................................................. 51 4.3.1 Identification Key to Eulophia speicies ......................................................................... 51 4.3.2 Checklist of Eulophia species in Ghana ........................................................................ 52 5.0 DISCUSSION ........................................................................................................................ 71 5.1 Eulophia species Delimitation .............................................................................................. 71 5.2 Morphology and Phylogeny of Eulophia species ................................................................ 72 5.3 Ecology of Eulophia species.................................................................................................. 73 6.0 SUMMARY, CONCLUSION, AND RECOMMENDATIONS ....................................... 76 6.1 Summary ................................................................................................................................ 76 6.2 Conclusion ............................................................................................................................. 76 6.3 Recommendations ................................................................................................................. 77 REFERENCES ............................................................................................................................ 78 APPENDICES ............................................................................................................................. 88 University of Ghana http://ugspace.ug.edu.gh vi LIST OF FIGURES Figure Page Fig 4.1: Distribution of Eulophia species in Ghana ...................................................................... 47 Fig 4.2: Distribution of Eulophia species in the vegetation zones of Ghana (following the species alphabetical order) ........................................................................................................................ 49 University of Ghana http://ugspace.ug.edu.gh file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page48 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page48 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page50 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page50 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page50 vii LIST OF PLATES Plate Page Plate 3.1 Morphological studies of herbarium specimens ............................................................ 33 Plate 4.1: image of a voucher specimen of Eulophia alta…………………………………………….51 Plate 4.2: image of a voucher specimen of Eulophia angolensis ................................................. 54 Plate 4.3: image of a voucher specimen of Eulophia buettneri .................................................... 56 Plate 4.4: image of a voucher specimen of Eulophia cristata ...................................................... 58 Plate 4.5: image of a voucher specimen of Eulophia cucullata .................................................... 60 Plate 4.6: image of a voucher specimen of Eulophia euglossa..................................................... 62 Plate 4.7: image of a voucher specimen of Eulophia flavopurpurea ............................................ 64 Plate 4.8: image of a voucher specimen of Eulophia milnei......................................................... 66 University of Ghana http://ugspace.ug.edu.gh file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page34 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page34 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page55 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page55 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page57 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page57 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page59 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page59 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page61 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page61 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page63 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page63 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page65 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page65 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page67 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page67 viii LIST OF TABLES Table Page Table 3.1: Morphological characters assessed, source and number of species ............................ 34 Table 3.2: Species of Eulophia, sources of information and collection sites ............................... 37 Table 4.1: Quantitative vegetive morphological characters of Eulophia species ......................... 42 Table 4.2: Qualitative vegetive morphological characters of Eulophia species ........................... 42 Table 4.3: Quantitative reproductive morphological characters of Eulophia species .................. 43 Table 4.4: Qualitative reproductive morphological characters of Eulophia species .................... 44 Table 4.5: Habitat types of Eulophia species growing in Ghana .................................................. 45 Table 4.6: Flowering and fruiting periods of Eulophia species .................................................... 46 University of Ghana http://ugspace.ug.edu.gh file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page35 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page35 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page38 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page38 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page43 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page43 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page43 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page43 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page44 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page44 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page45 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page45 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page46 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page46 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page47 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page47 ix APPENDICES Appendix Page APPENDIX A: Data collected from herbarium labels on Eulophia species ................................ 84 APPENDIX B: One way ANOVA of the leaf length of the species of Eulophia ........................ 87 APPENDIX C: One way ANOVA of the leaf width of the species of Eulophia ......................... 87 APPENDIX D: One way ANOVA of the tepal length of the species of Eulophia ...................... 88 APPENDIX E: One way ANOVA of the tepal width of the species of Eulophia........................ 88 APPENDIX F: One way ANOVA of the tepal length of the species of Eulophia ....................... 88 APPENDIX G: Scatterplot for cluster analysis ............................................................................ 89 APPENDIX H: Mean ±standard error of the morphological characters measured ...................... 90 APPENDIX I: Cluster data of morphological characters measured ............................................. 91 APPENDIX J: Distribution of Eulophia species in Ghana (GBIF, 2022) .................................... 96 APPENDIX K: Phenology of Eulophia species in Ghana............................................................ 97 APPENDIX L: Dendrograms of the species of Eulophia ............................................................. 98 APPENDIX M: t-tests of sample means between E. cucullata and E. dilecta ........................... 101 University of Ghana http://ugspace.ug.edu.gh file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page85 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page85 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page88 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page88 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page88 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page88 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page89 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page90 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page90 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page91 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page91 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page92 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page92 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page97 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page97 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page98 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page98 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page99 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page99 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page102 file:///C:/Users/Ghana%20Herbarium/Downloads/10563658_THESIS.doc%23page102 x LIST OF ABBREVIATIONS GBIF………………………………………... Global Biodiversity Facility Information POWO………………………………………. Plants of the World Online AOO…………………………………………. area of occupation EOO…………………………………………extent of occurrence IUCN………………………………………. International Union for Conservation of Nature SD…………………………………………. standard deviation GC………………………………………… Ghana Herbarium KUU……………… herbarium in the Kwame Nkrumah University of Science and Technology CCG…….................................................... herbarium in the University of Cape Coast PROTA……………………………………Plant Resources of Tropical Africa University of Ghana http://ugspace.ug.edu.gh xi ABSTRACT The family Orchidaceae includes 850 genera and 24, 000 species, and it is arranged into 70 subtribes, 22 tribes, and five subfamilies on the basis of anther number and position. Many orchids are utilized in the traditional system of medication as medicines for several illnesses. There is no complete taxonomic revision of the genus Eulophia in Africa. A previous study based on morphological, ecological, and biogeographic characters of Eulophia cristata in Ghana has been reported by Lock and Profita (1975). However, the extent of morphological variations in species of the genus in Ghana has not been analyzed. The overall goal of this study is to undertake a systematic study of the genus Eulophia in Ghana for their sustainable utilization and conservation. 50 herbarium specimens of the genus Eulophia (35 specimens at GC, 10 specimens at CCG, and 5 specimens at KUU) were examined for morphological characters. Information from herbarium labels of the specimens examined were retrieved to understand aspects of the ecology such as phenology, habitat types and pollinator-flower relationship for the species of the genus Eulophia. Taxonomic studies focused on development of an authoritative checklist and construction of dichotomous keys for identification of species of the genus. Morphological studies on the species of Eulophia from Ghana suggested eight species but records from the herbaria in Ghana provided nine species. The members of the Eulophia range from small to large herbs, lacking woody stems; plant height ranges from 560–2000mm, 2– 16mm in width; has leaves that are often alternately arranged. Leaves are plicate, erect, often linear to lanceolate. Flowers are mostly in the shades of yellow and red, except shades of green in E. euglossa and E. flavopurpurea. Generally, the species of Eulophia have a cosmopolitan distribution across the country from the herbarium records. University of Ghana http://ugspace.ug.edu.gh 1 CHAPTER ONE 1.0 INTRODUCTION 1.1 Background The family Orchidaceae includes 850 genera and 24, 000 species, and it is organized into 70 subtribes, 22 tribes, and five subfamilies based on anther number and position (Kowsalya et al., 2017). The Apostasioideae and Cypripedioideae are considered as group sister to the rest of the monadrous clade of Orchidoideae (including Spiranthoideae) and Epidendroideae (Cameron et al., 1999; Kowsalya et al., 2017). Unlike the Epidendroideae and Orchidoideae, the Vanilloideae lack pollinia (Shririam et al., 2014). Epidendroideae is the largest of the five subfamilies (Shririam et al., 2014). Plastid gene rbcL shows that Orchidaceae consists of five major monophyletic clades, and these clades, with a few exceptions, correspond to presently recognized subfamilies (Cameron et al., 1999; Kowsalya et al., 2017). For many orchids, photosynthesis is the primary source of carbon. However, myco- heterotrophic species, which make up a small percentage of the Orchidaceae, include saprophytic species (Zhang et al., 2015c). The CAM group, which is most common at low elevations and within the epiphytic clade, makes for around 10% of all orchid species in Panama and Costa Rica; C3 photosynthesis being the ancestral condition (Sailo et al., 2014; Zhang et al., 2018). Most orchids, especially species with thin leaves, assimilate CO2 through the C3 pathway. Those plants have fewer layers of smaller mesophyll cells and a larger number of stomata than the thick-leaved species. They also have high CO2 compensation points and active glycolic acid activity, all of which are characteristics of plants with high rates of photorespiration (Sailo et al., 2014; Zhang et al., 2018). University of Ghana http://ugspace.ug.edu.gh 2 The thick-leaved orchids usually have features typical of CAM plants, such as leaf and cell succulence, diurnal fluctuations in titratable acidity and nocturnal CO2 fixation, and inverted stomatal physiology. Those genera include Vanilla Plum. ex Mill., Cattleya Lindl., Thunia Rchb.f., Coelogyne Lindl., Laelia Adans., Dendrobium Sw., Calanthe R.Br., Bulbophyllum Thouars, Aerides Lour., Phalaenopsis Blume, Aranda Anon., and Aranthera Anon. (Sailo et al., 2014; Zhang et al., 2018). Members of the Orchidaceae have a cosmopolitan distribution. Major genera of the family include Ophrys L., Oncidium Sw., Peristylus Blume, Stanhopea J.Frost ex Hook., Disa P.J.Bergius, and Vanilla Plum. ex Mill. (Maridass et al., 2008). Many orchids are utilized in the traditional system of medication as medicines for several illnesses. For example, the tubers and pseudobulbs of Orchis latifolia L., Orchis mascula (L.) L., Cymbidium aloifolium (L.) Sw., and Zeuxine strateumatica (L.) Schltr. are used as a restorative and in the treatment of many diseases. The seeds of Cymbidium aloifolium are used for the healing of injuries (Maridass et al., 2008). Vanilla flavoring is extracted from the fruits of Vanilla planifolia Andrews. The leaves of one species of Anoectochilus Blume are sold as a vegetable, and the leaves of Dendrobium salaccense (Blume) Lindl. are cooked as a seasoning with rice. In certain parts of the Asian tropics, the tubers of some species of Gastrodia R.Br. are eaten like potatoes. The family is of great economic importance because of its numerous ornamentals Cattleya Lindl., Dendrobium Sw., Epidendrum L., Paphiopedilum Pfitzer, Phalaenopsis Blume, Vanda R.Br., Brassia R.Br., Cymbidium Sw., but to mention a few (Panwar et al., 2012). 1.2 Problem statement The genus Eulophia R.Br. ex Lindl. (Orchidaceae, Cymbidieae) is the most diverse in the subtribe Eulophiinae. The genus consists of about 164 terrestrial species and shows a University of Ghana http://ugspace.ug.edu.gh 3 pantropical distribution mainly in Africa (Central and Southern), Madagascar, and Asia, with seven species in Australasia and five in America (Ortúñez et al., 2020). There is no complete taxonomic revision of the genus Eulophia in Africa (Martos et al., 2014). However, a taxonomic study on Eulophia in Nigeria, Cameroon, Equatorial Guinea, Gabon, and the Republic of the Congo has been conducted (Ortúñez et al., 2020). The genus has also been examined in Madagascar and South Africa (Peter & Johnson, 2009). But due to the wide distribution of the Eulophia, there is a high degree of interspecific morphological variation in both vegetative and reproductive characters of the genus (Ortúñez et al., 2020). Furthermore, the lack of complete preserved material of many taxa of the genus in herbaria has led to misidentifications and an infrageneric classification that is not firmly settled at present (Panwar et al., 2012). Owing to these gaps of knowledge and the high degree of interspecific morphological variation in the genus, a revision is needed. 1.3 Justification Preliminary studies at the Ghana Herbarium in the Department of Plant and Environmental Biology at the University of Ghana show that there are ten species of Eulophia occurring in Ghana. Studies on members of the genus are scarce in Ghana. A previous study based on morphological, ecological, and biogeographic characters of Eulophia cristata in Ghana has been reported (Lock & Profita, 1975). However, the extent of morphological variations in species of the genus in Ghana has not been analyzed given that infra-generic classification of the genus Eulophia is complicated by morphological diversity (Cieslicka, 2006). Furthermore, distributional ranges of the species in Ghana have not been investigated but according to Mota-Vargas & Octavio (2012), understanding biodiversity and making conservation decisions are fundamentally impacted by the delineation of species distribution University of Ghana http://ugspace.ug.edu.gh 4 ranges. Also, understanding species' ability to colonize new habitats is a key knowledge that aid to predict species' survival in the changing landscapes (Vazačová & Münzbergová, 2014). 1.4 Objectives of the study The overall goal of this study is to undertake a systematic study of the genus Eulophia in Ghana for their sustainable utilization and conservation. Specifically, the objectives of the study were to: (1) determine the range of morphological variations within the genus in order to clarify the delimitation of the taxa in Ghana, (2) evaluate aspects of the ecology and distribution of the genus in Ghana (3) provide an updated identification key and checklist of species of the genus in Ghana. University of Ghana http://ugspace.ug.edu.gh 5 CHAPTER TWO 2.0 LITERATURE REVIEW 2.1 Systematics and Taxonomy Systematics is the study of the current diversity of biological life on earth as well as its evolutionary past. The framework, or categorization, that other biologists use to discuss species is also provided by systematics. This study is crucial to our comprehension of and communication about the natural world. The fundamental tasks of systematics include naming and classification, which are the oldest human approaches to handling data about the natural world. Systematics rely on a wide variety of species for organic materials like food, shelter, fiber, clothing, paper, medicines, tools, and dyes. The search for prospective commercially important plants is guided by systematics knowledge. The most fundamental and all- encompassing branch of biology is systematics since it serves as a reference framework for all of biology (Simpson, 2010; Pandey & Kasana, 2021). Systematics is fundamental because it takes some level of classification to identify and name organisms before they can be addressed in a scientific context. Because it compiles and integrates all information on the features of species, including geographic, morphological, physiological, genetic, ecological, and molecular information, systematics covers the most ground (Pandey & Kasana, 2021). Taxonomy is concerned with the concepts and methods of classification. Based on certain similarities and differences, taxa are identified, described, classified, and given new names in this biological discipline. De Candolle (1813) first used the Greek terms taxis (meaning arrangement) and nomos (meaning law, rule) to create the phrase "taxonomy." The word "taxonomy" literally means "order by rules" or "lawful arrangement of things." Description, University of Ghana http://ugspace.ug.edu.gh 6 Identification, Nomenclature, and Classification are the four main parts of taxonomy (Pandey & Kasana, 2021). Singh (2010) considers taxonomy and systematics to be synonymous while Pandey & Kasana (2021) considers systematics significantly more inclusive than taxonomy, which is frequently mistaken for it. In this study, the viewpoint of Singh (2010) will be consistently used. 2.2 Orchidaceae Juss. One of the two largest families of flowering plants, Orchidaceae is possibly second only to Asteraceae in size (Chase et al., 2015). In these families, new species are being discovered at a pace of about 500 each year. There are currently 736 genera of orchids known to science, and roughly 13 new genera are described each year (Chase et al., 2015). Most long-lived, evergreen or deciduous herbs are orchids. Some plants, like those of the Cypripedium calceolus, can live for 30 to 100 years. Most pre-flowering vegetative periods of the species typically last four to seven years, but they may endure considerably longer (Wang et al., 2010; Cribb & Angell, 2014). They may have a naturally slow growth rate and a low photosynthetic capability, which could explain their extended lifespan. One of two growth patterns, monopodial or sympodial, is typically followed by orchids. For monopodial orchids, a single bud develops into a stem that lengthens and produces leaves from the apex every year. The adjacent shoots of sympodial orchids grow in a sequence until they bloom and are then replaced (Sailo et al., 2014). The life form of the orchid can be terrestrial, saprophytic, epiphytic, or lithophytic. The epiphytic orchids, which grow on rocks or in tree canopies, differ greatly from terrestrial orchids, which grow in soil, in terms of their roots, stems, and leaves. Epiphytic orchids have modified aerial roots that can occasionally reach a length of more than 1 m. They also possess University of Ghana http://ugspace.ug.edu.gh 7 a velamen made of dead cells. With the exception of the tip, this velamen covers the entire root and serves to quickly absorb moisture and nutrients from the humid environment around it (Zhang et al., 2018). Although certain species in the subfamily Vanilloideae display a reticulate venation, orchids typically have simple leaves with parallel veins. However, epiphytic orchids differ from terrestrial species in that they have smaller stomata and leaves with dense cell walls, cuticles, and a tiny substomatal chamber; thick, succulent leaves (Sailo et al., 2014). Despite the fact that orchid blooms are bilaterally symmetrical, the inferior ovary, or pedicel, typically rotates 180 degrees so that the labellum is placed on the lower section of the flower, creating a platform for pollinators. While flower color is greatly divergent across species of Paphiopedilum, the shape of the lip staminode and petal, as well as the width of the petal, are phylogenetically conserved (Zhang et al., 2016). For many orchids, photosynthesis is the primary source of carbon. However, myco- heterotrophic species, which make up a small percentage of the Orchidaceae, include saprophytic species (Zhang et al., 2015c). The CAM group, which is most common at low elevations and within the epiphytic clade, makes for around 10% of all orchid species in Panama and Costa Rica; C3 photosynthesis being the ancestral condition (Sailo et al., 2014; Zhang et al., 2018). Most orchids, especially species with thin leaves, assimilate CO2 through the C3 pathway. Those plants have fewer layers of smaller mesophyll cells and a larger number of stomata than the thick-leaved species. They also have high CO2 compensation points and active University of Ghana http://ugspace.ug.edu.gh 8 glycolic acid activity, all of which are characteristics of plants with high rates of photorespiration (Sailo et al., 2014; Zhang et al., 2018). The thick-leaved orchids usually have features typical of CAM plants, such as leaf and cell succulence, diurnal fluctuations in titratable acidity and nocturnal CO2 fixation, and inverted stomatal physiology. Those genera include Vanilla, Cattleya, Thunia, Coelogyne, Laelia, Dendrobium, Calanthe, Bulbophyllum, Aerides, Phalaenopsis, Aranda, and Aranthera (Sailo et al., 2014; Zhang et al., 2018). Orchids are well known for their economic importance and widely cultivated for ornamental purposes. Orchids are cosmopolitan in distribution. Due to its importance as a source of vanillin, a flavoring used in food products, Vanilla planifolia is an economically significant orchid. Orchids are frequently used in traditional Chinese medicine. Several of them have been the subject of pharmacological and phytochemical investigations (Kasulo et al., 2009). Some work has been carried out in India on chemical analysis of some medicinally useful orchids. Eulophia campestris, Orchis latifolia, Vanda roxburgii are some important plants to mention. Dendrobium macraei is another important orchid from Ayurvedic point of view as it is reported to be source of Jivanti. Cypripedium parviflora is widely used as aphrodisiac and nervine tonic in Western Herbalism (Singh & Duggal, 2009). Many medicinal orchids are reported to contain alkaloids. Antimicrobial activities of some orchids have been suggested although detailed investigations are still warranted. Recent works have reported isolation of anthocyanins, stilebnoids and triterpenoids from orchids. University of Ghana http://ugspace.ug.edu.gh 9 Orchinol, hircinol, cypripedin, jibantine, nidemin and loroglossin are some important phytochemicals reported from orchids (Singh & Duggal, 2009). The Indian Vanda orchid does indeed express antiproliferative effects against various types of cancers, including those from choriocarcinoma (cancer of germ cells), lung cancers, and stomach cancers (Ho & Chen, 2003). Still other orchids have been used in the treatment of epilepsy, flatulence, rheumatism and spasms. They have also been used for sedatives and flavor enhancers and for everything from cramps to increased virility (Dash et al., 2008). The present investigation has also brought to light the therapeutic value of orchids in curing scorpion and snake bites, as well as in the treatment of stomach disorders, chest pains, arthritis, tuberculosis, pregnancy, syphilis, paralysis, cholera, acidity, eczema, tumor, menstrual disorder, spermatorrhea, leucoderma, wounds and sores, diahorrhea, muscular pain, blood dysentery, bone fractures, rheumatism, asthma, malaria, earache and sexually transmitted diseases (Dash et al., 2008). Orchid cultivation which started as a hobby in the early part of 20th century transformed into a commercial enterprise in late sixties. They rank among the top ten sought after plant for cut flower production (Vij, 2002). The flowers of some modern hybrids have a shelf life of 8- 12 weeks, surpassing all other flowers in this respect. The international trade in floriculture product is rapidly developing especially orchids which command a high price in world market. South East Asian countries are the major player in orchid trade. Thailand as the largest producer with an export destination to USA and Europe earning as high as 4 million US dollar by 1995 (Imchen & Imchen, 2011). University of Ghana http://ugspace.ug.edu.gh 10 There was a 1.3-billion-unit plant stock demand for orchids grown for the manufacture of cut flowers and potted plants in 1995. More than $500 million was spent on orchids (potted plants) worldwide in 2000. China, Germany, Japan, The Netherlands, Taiwan, Thailand, and the United States are among the nations that produce potted orchids on an extensive scale. Recently, India, Costa Rica, and the Dominican Republic started growing orchids for export (Laws, 2004). Many orchids primarily obtain carbon through photosynthesis. But saprophytic species, which make just a small percentage of the orchid family (Orchidaceae), are myco-heterotrophic (Zhang et al., 2015c). Most frequently found at low elevations and within the epiphytic clade, the CAM group accounts for around 10% of all orchid species in Panama and Costa Rica. C3 photosynthesis is the ancestral state (Sailo et al., 2014; Zhang et al., 2018). The most expensive crop sold in pots in The Netherlands is now Phalaenopsis; the number of potted orchids sold at Dutch flower auctions increased from 50,000 to 18 million pots between 1983 and 2003. (Laws, 2004; Wang, 2004). Orchids were regarded as a small crop by the U.S. Department of Agriculture (USDA), and data on output was not gathered until 1997. The value of U.S. potted orchid production climbed by 170% between 1996 and 2004, with an estimated $128 million in wholesale sales in that year (USDA, 2005). The second most valuable flowering potted crop in the United States right now is orchids, of which 70 to 90 percent are Phalaenopsis. Over 85% of all orchids sold in the United States in 2004 were grown by farmers in California, Florida, and Hawaii, with wholesale prices reported at $50, $47, and $17 million, respectively (USDA, 2005). University of Ghana http://ugspace.ug.edu.gh 11 The majority of orchids are cultivated for their aesthetically pleasing blossoms. The majority of orchid genera' flowering physiology in controlled conditions has only received a little amount of published research, though. Therefore, it is impossible to time the flowering process to coincide with specified sales dates, with the exception of a few species (such as Phalaenopsis and some Cattleya) (Bullough et al., 2021). 2.2.1 Taxonomy of the Orchidaceae Dressler (1993) recognized five subfamilies, but he split Spiranthoideae from Orchidoideae and included Vanilla and relatives in Epidendroideae. Vanilloideae into two tribes, Pogonieae and Vanilleae. No changes were made to the four tribes of the Orchidoideae: Cranichideae, Diurideae, Orchideae, and Codonorchideae. In Epidendroideae, there has been a great deal of change in the generic limits in many tribes/subtribes since Chase et al. (2003). Little has changed in the groups with mostly mealy (primitive) pollinia, the formerly recognized Neottioideae. In Triphoreae, we recognize two subtribes in parallel with the treatment in Pridgeon et al. (2005), with the addition of heteromycotrophic Pogoniopsis. Likewise, in Nervilieae, Nervilia was recognized to be in a separate subtribe from the rest. Major taxa within the family are as follows. The Cattleya genus includes 48 species that are native to tropical areas of Central and South America. From sea level to 1500 m in height, epiphytic plants can be found growing on top of trees in moist and wet woods. The growth of many genera of tropical orchids is accelerated by high temperatures. Different times of the year are when different Cattleya species and hybrids bloom. Some Cattleya species begin to form flower buds in the spring as a result of the low temperatures and short photoperiod, but the buds do not grow and attain anthesis until the fall or winter (Lopez & Runkle, 2005). University of Ghana http://ugspace.ug.edu.gh 12 There are roughly 44 species in the genus Cymbidium, which is endemic to Australia, the tropical parts of southeast Asia, and the Himalayas. For rapid growth and pseudobulb maturity, the terrestrial, epiphytic, lithophytic, and semi-epiphytic species need long days and day/night temperatures of 30/25 °C. Several orchid taxa, including Cymbidium, are frequently transferred from lowland production sites to higher altitudes in Japan during the warm summer. This method exposes plants to milder temperatures and lighter for blossom initiation, preventing high-temperature stress during vegetative growth (Lopez & Runkle, 2005). Cymbidium have a wide geographic distribution; hence they are frequently categorized into the three horticultural groups of chilly, moderate, and warm depending on how well they tolerate different temperatures. A noticeable chilly period during which the nighttime temperature is maintained at between 10 °C and 14 °C encourages the majority of the large-flowered Cymbidium species from the Himalayas and China to flower. Summer is the typical flowering season for Cymbidium orchids that are native to warm climates like Taiwan and Southern China. In Cymbidium ensifolium var. misericors, pseudobulb development and flower production were sped up at warm day/night temps of 30/25 °C and 25/20 °C, but were slowed down at 20/15 °C. Plants with 1- to 2-year-old mature pseudobulbs produced 2.3, 1.6, and 1.1 inflorescences at 30/25 °C, 25/20 °C, and 20/15 °C, respectively (Lopez & Runkle, 2005). There is no evidence from research that photoperiod affects floral induction. Cymbidium niveomarginatum Lindl. was able to blossom in vitro thanks to a combination of root excision, cytokinin (6-benzylaminopurine), restricted nitrogen, and enhanced phosphorus (Lopez & Runkle, 2005). University of Ghana http://ugspace.ug.edu.gh 13 The second-largest genus in the family of orchids, Dendrobium (also known as the spray orchid), has more than 1,200 epiphytic species that are indigenous to tropical and subtropical Asia, Australia, and a number of Pacific Islands. Rapid vegetative development occurs for the majority of Dendrobium orchids between 24 °C and 30 °C (Lopez & Runkle, 2005). Miltoniopsis is a genus of six species that is either epiphytic or lithophytic and is found from Peru to Costa Rica's moist cloud forests (610 to 2100 m). Miltoniopsis thrives in environments with light intensity between 150 and 300 mol m -2 s -1 . When Miltoniopsis is young, flowering doesn't start until the pseudobulbs are ready to detect inductively low temperatures (Lopez & Runkle, 2005). More than 50 species under the genus Phalaenopsis are native to tropical and subtropical regions of Asia and the South Pacific Islands. From southern India to Australia, their range includes China, Taiwan, and the Philippines. Phalaenopsis orchids may withstand temperatures as high as 32 °C to 35 °C for brief durations before showing indications of heat stress, but they remain vegetative above 27 °C to 29 °C (Lopez & Runkle, 2005). For all developmental phases, the predicted base temperature for Phalaenopsis is between 10.8 and 11.2 °C. Phalaenopsis displayed indications of chilling injury, such as sunken, yellow, water- soaked patches on upper leaf surfaces, after being subjected to 2, 4, or 7 °C for 1, 2, 4, or 8 hours. Phalaenopsis must be exposed to temperatures around 26 °C for a certain amount of time in order to start spiking, or the beginning of the inflorescence. The third and frequently the fourth node beneath the apical leaf is where the inflorescence typically appears. If the environment is favorable, the spike begins to form flower buds when it is about 5 cm long (Lopez & Runkle, 2005). University of Ghana http://ugspace.ug.edu.gh 14 About 20 species of the sympodial, terrestrial, and epiphytic South American genus Zygopetalum, also known as the ladybird orchid, can be found there. They are native to neotropical mid-elevation mountains (1300 to 1700 m) in Brazil, Guiana, Venezuela, and Peru, However, a temperature of 23 °C is recommended to balance growth pace with optimal development features because vegetative growth is most rapid at 26 °C (e.g., long leaves). When light intensity is more than 500 μ.mol.m -2 s -1 , black necrotic lesions can develop (Lopez & Runkle, 2005). The Old-World tropics of Africa, Asia, and Australasia are home to the subtribe Eulophiinae (Cymbidieae, Epidendroideae, Orchidaceae), which has about 270 species grouped into nine genera and few taxa in the Neotropics (Bone et al., 2015). This extraordinarily diverse population lacks recognizable diagnostic traits (Pridgeon et al., 2009). Phylogenetic investigations of molecular data, however, are starting to clarify relationships in Eulophiinae and between this and related subtribes of Cymbidieae (Bone et al., 2015). Of the nine currently accepted Eulophiinae genera, four (Cymbidiella, Eulophiella, Grammangis and Paralophia) are endemic to Madagascar. Each comprises fewer than seven species that are mostly large, lush epiphytes, including Eulophiella roempleriana (Rchb.f) Schltr., described by Hermans (Pridgeon et al., 2009) as ‘one of the giants of the orchid world’ with plants exceeding 2 m in height. Cymbidiella and Eulophiella each comprise a single terrestrial species. Hermans & Cribb (2005) recognized Eulophia epiphytica P. J. Cribb, Du Puy & Bosser and E. palmicola H. Perrier; as a new genus, Paralophia. The remainder of the subtribe comprises five terrestrial genera (Acrolophia Pfitzer, Eulophia R. Br. ex. Lindl., Geodorum Jacks., Oeceoclades Lindl. and Orthochilus Hochst. Ex A. Rich) that occupy a range of habitats, from the semi-arid conditions of sub-Saharan Africa and the Arabian Peninsula, deciduous woodland and University of Ghana http://ugspace.ug.edu.gh 15 seasonally inundated grasslands of south-central Africa, to a range of habitats on Madagascar, including humid pockets on karst limestone (‘tsingy’), and dry, littoral and mesic forests (Cribb & Hermans, 2009). Of these genera, Eulophia is the most diverse, representing c. 60% of species richness in the subtribe. Martos et al. (2014) focused on South African Eulophia and including comprehensive sampling of the South African endemic genus Acrolophia, their study found evidence for a clade of morphologically well characterized taxa of Eulophia that corresponded to the ‘nigricans alliance.’ Eulophia nevertheless remains the largest genus of the subtribe (164 species) and is a conspicuous element of the terrestrial African orchid flora. No complete taxonomic account for the genus exists and an infrageneric classification is required to provide a practical framework for the identification and further classification of species (Bone et al., 2015). The next largest genus after Eulophia is Oeceoclades (39 species), which has a distribution that is primarily Madagascan but also includes continental eastern Africa, the western Indian Ocean islands, and beyond away. Many have broad, plicate leaves that are more resemblant of Eulophia, which are xerophytes with thick, coriaceous leaves (Bone et al., 2015). This latter, widely distributed species has been referred to by various authors as Eulophia pulchra (Thouars) Lindl. and Oeceoclades pulchra, but Martos et al. (2014) resolved it as sister to a monophyletic Oeceoclades and suggested that it should be treated as a member of this genus until broader taxon sampling could be finished for more in-depth analysis. University of Ghana http://ugspace.ug.edu.gh 16 2.2.2 Distribution and Ecology of Orchidaceae Numerous ecological variables including the emergence of holobionts, which are crucial for colonization and the development of ecological communities, have an impact on the distribution and population dynamics of orchids. Mycorrhizal fungi are essential for orchid species' seed germination, seedling establishment, reproduction, and survival, and cheating by mycorrhizal fungus in photosynthetic orchids is becoming more common. As a result, changes in the diversity and composition of orchid mycorrhizal fungus can have a significant impact on the distribution and fitness of orchids (McComick et al., 2018; Li et al., 2021). Amazing morphological features in orchids, including the labella and modified petals, show how adaptable they are to their surroundings (Zhang et al., 2018). Orchid reproduction is aided by interactions with particular pollinators, but soil exploitation needs the presence of symbiotic fungus. The diversity of orchid species typically rises as a result of increased environmental heterogeneity and a larger range of resource availability. As a result, understanding how environmental factors affect the distribution and abundance of orchids is necessary for the effective conservation of these species (McComick et al., 2018). Djordjevi & Tsiftsis (2020) have conducted a systematic analysis of the relationships between environmental conditions and the distribution, abundance, and richness of orchid species. For instance, the rainfall and light regime of the habitat have a direct impact on the flowering cycles of orchids and population dynamics, and the physical and chemical characteristics of the soil (such as pH, soil moisture, nutrients, etc.) have a big impact on how well orchid populations do. Curiously, there is mounting evidence that suggests coexisting orchid species typically display strongly spatially segregated distribution patterns due to strong clustering within individual species and little overlap between species, and that they are frequently associated with various orchid mycorrhizal fungi communities, which is largely explained by variations in soil moisture and pH. University of Ghana http://ugspace.ug.edu.gh 17 2.3 Orchidaceae in West Africa including Ghana There are 401 orchid species in West Africa, which includes Cameroon, Nigeria, and the nations to the west. However, many of these have special scientific and horticultural value. The key determinants of their distribution are moisture availability, which depends on rainfall, air humidity, the length of the dry season, and the presence of undisturbed areas. These parameters are mainly influenced by man and his agricultural and logging operations (Sanford, 1969; Bothalia, 2013). West Africa is extremely variable in moisture availability. Furthermore, dense population, shifting agriculture, and timber exploitation, have made the area very variable in vegetation cover. For these reasons, the orchids of the region often show discontinuous distributions. Of the 401 species of orchids recorded in West Africa, 19 species, or 4.7 per cent of the total, are found only in Nigeria, and these should receive particular attention for conservation. Thirteen of these are epiphytes of extremely limited distribution, while the remaining 6 are terrestrial species. In general, the epiphytes are more demanding as to habitat, more easily destroyed by man's activities, and consequently in greater danger than the terrestrial species (Sanford, 1969; Bothalia, 2013). Habenaria keayi and H. nigerica are quite widespread and common. Another group of orchids has a slightly less restricted distribution, found in Nigeria and one other country as well. In all but five instances, the other country is in West Africa. The exceptions are particularly interesting: Nervilia reniformis is found in Tanzania, Angraecum egertonii and Rangaeris trilobata are in Gabon, Microcoelia microglossa is in Uganda, and Eulophia adenoglossa is in Kenya. There are 27 species, or 6.7 per cent of the West African total, belonging to this category. These include 19 epiphytic and 8 terrestrial species. Only one, the terrestrial Nervilia reniformis, is actually common enough to give no cause for conservation worry, although University of Ghana http://ugspace.ug.edu.gh 18 Habenaria lelyi and Eulophia sordida, also terrestrial, are quite widespread within Nigeria (Sanford, 1969; Bothalia, 2013). Eight of the species, which are unique to Nigeria, are found in the northern highlands of Zaria Province (Jos and Bauchi), four are found in the eastern highlands (Obudu and Oban), two are found in the Idanre Hills, two are found in Sapoba, three are found in the Cross River region (Calabar and Itu), and two are found in Lagos and Eket, the last of which is in the Calabar region. The Obudu-Mambila Plateau and the Oban Hills are part of the eastern highlands, which continue the Cameroon highlands (Sanford, 1969; Bothalia, 2013). The 31 species that are exclusive to a single adjacent nation are present in Cameroon-Fernando Poo. The highlands that extend from Zaria Province in the east through the Jos and Bauchi Plateau and into Cameroon, running south through Bamenda to Mt. Isabel (Clarence Peak) in Fernando Poo, with western branches into southeastern Nigeria from Obudu to the Mambila plateau area and, further south, into the Oban Hills, are without a doubt the area of highest priority for orchid conservation. It is environmentally intriguing that such a region would be abundant in orchids and house several extinct species due to the region's overall West African distribution (Sanford, 1969; Bothalia, 2013). In Ghana, orchids have a cosmopolitan distribution. Several genera like Disa P.J. Bergius, Disperis Sw., Eulophia R. Br., and Bulbophyllum Thouars. can be found in 12 of the 16 regions in the country. A few like Polystachya Hook. and Cheirostylis Blume can be found in only 3 regions. 2.4 Genera of Orchids in Ghana According to records from the Ghana Herbarium, there are 23 genera of orchids in Ghana. These are Aerangis Rchb.f., Ancistrorhynchus Finet, Ancistrochilus Rolfe, Angraecum Bory, Ansellia Lindl., Auxopus Schltr., Bolusiella Schltr., Brownleea Harv. ex Lindl., Bulbophyllum University of Ghana http://ugspace.ug.edu.gh 19 Thouars., Calyptrochilum Kraenzl., Corymborkis Thouars., Cheirostylis Blume, Cyrtorchis Schltr. Diaphananthe Schltr., Didymoplexis Griff., Disa P.J. Bergius, Disperis Sw., Eggelingia Summerh., Epipogium Borkh., Eulophia R. Br., Manniella Rchb.f., Microcoelia Lindl., and Polystachya Hook. Owing to little research carried out on the orchidaceae of Ghana, information of the genera of orchids in Ghana were obtained from the Ghana Herbarium and the Plants of the World Online. Each of the genera have been reviewed below. 2.4.1 Aerangis Rchb.f. There are 3 species in Ghana, these orchids are mostly distributed in the Eastern, Greater Accra, and Ashanti regions. They are mostly epiphytic plants with woody stems that are either short or tall and have several long aerial roots at the base. Epiphytic plants have woody stems that are either short or tall and have several long aerial roots at the base. Distinctive, fleshy or coriaceous, basally sheathing, mostly oblanceolate, occasionally linear or ligulate, and unequally 2-lobed at the apex leaves. Racemose, lateral, few to many-flowered inflorescence. White, occasionally tinged with green, brown, or pink resupinate flowers. Whether free, spreading, or reflexed, sepals and petals. Lip whole, frequently resembling tepals, spurred at base. Pollinia 2, sessile on a single stipe; viscidium variously shaped; stigma an oval or rhomboid sticky depression. Column short and stout or elongated and small, frequently narrow at the base and enlarged at the level of the stigma. Androclinium straight or sloping, the anther cap occasionally beaked (POWO, 2022). 2.4.2 Ancistrorhynchus Finet The genus Ancistrorhynchus has 5 species in Ghana mostly distributed in Eastern, Western, and Ashanti regions. They are mostly herbs with monopod epiphytes. Short stems that are sheathed in old leaf bases; roots grouped at stem base. Imbricate, ligulate, or tapering leaves with occasionally dentate lobes that are unequally 2-lobed at the apex. Bracts are papery and University of Ghana http://ugspace.ug.edu.gh 20 typically as long as the flowers; inflorescences are sessile, axillary, and grow from the lower leaves or the bases of the leaves. Flowers are white, occasionally with a green lip. Free, subsimilar sepals and petals; oblong orbicular, whole or 3-lobed, ecallose lip; straight, sigmoid or geniculate spur; occasionally enlarged at apex and/or mouth. Anthers are hemispherical with a short, truncate appendage in front; the column is short; the rostellum projects downward before suddenly rising in the apical half; the stipites are either two and subspathulate or one and bifid; and the viscidium is one (POWO, 2022). 2.4.3 Ancistrochilus Rolfe Ancistrochilus is a monospecific genus in Ghana which is mostly distributed in the Oti region. Members of this genus are small epiphytic herbs. Pseudobulbs clustered, orbicular to conical, ± dorsiventrally flattened, bearing 1–2 apical leaves. Leaves thin-textured, plicate. Inflorescences arising from base of pseudobulbs, arcuate-spreading, 1–5-flowered, pubescent. Flowers showy, white to rose-pink or purple, marked with brown on column. Sepals spreading, subsimilar, pubescent on outer surface. Petals smaller but similar in shape. Lip 3-lobed, recurved apically, with erect side lobes on each side of column. Column clavate, porrect, pubescent, with a short foot; pollinia 8, of two sizes, lacking a stipe and viscidium; rostellum pointed (POWO, 2022). 2.4.4 Angraecum Bory There are 8 species of this genus in Ghana, these are mostly distributed from the Eastern to Ashanti regions. Members of this genus are herbs that are small to large and are epiphytic, lithophytic, or very rarely terrestrial. Short to elongate, branching or unbranched, upright to pendent stems that are covered by leaf bases. Unevenly bilobed at the apex, thin-textured, fleshy or coriaceous, flattened or infrequently iridiform, frequently twisted at the base, articulated to persistent leaf-bases. Axillary, one to many-flowered, racemose, or infrequently University of Ghana http://ugspace.ug.edu.gh 21 paniculate inflorescences. Large to tiny, fleshy flowers that are frequently stellate; sessile or pedicellate ovary that is frequently rotated through 180 degrees. Petals and sepals are separate and comparable. Concave, whole to three-lobed, and spurred at the base, the lip encloses the column at the base. Pollinia 2, globose, sulcate, joined to a common viscidium or each attached to a distinct viscidium; column fleshy, very short, missing a foot; clinandrium somewhat shallow, split in front with a tiny tooth-like rostellum in the sinus (POWO, 2022). 2.4.5 Ansellia Lindl. Ansellia is monospecific genus in Ghana with a distribution from the Western to the Eastern regions. Plants range from medium to large-sized epiphytic, lithophytic, or very rarely terrestrial herbs. Pseudo-bulbs are grouped together, fusiform, several-noded, ridged as they age, leafy in the apical half, and are encircled by many tall bristly roots at the base. In the apical region of the pseudobulb are numerous, large, plicate leaves. Axillary, paniculate, paniculately arcuate, and laxly many-flowered inflorescence from apical nodes. Large, beautiful, pale greenish yellow or pale to rich yellow flowers with red or dark maroon spots that are frequently heavy. petals and sepals are similar but stretch out. Three-lobed, upright lip with numerous longitudinal ridges; upright side lobes on both sides of column. Column porrect; pollinia, four in two pairs, ovoid and of two sizes, attached to viscidium that is difficult to discern (POWO, 2022). 2.4.6 Auxopus Schltr. There are 2 species from this genus in Ghana, these are mostly distributed in the Western, Eastern, and Ashanti regions. Members of this genus are herbs without leaves; achlorophyllose saprophytes and have globular to narrowly cylindrical tubers. Erect, unbranched, thin, and barely flexible stems. Small, fugacious, unspurred, yellow or brown flowers are in bloom. Tepals produce a brief tube by partially adnating. Simple, without a lip, and with a thin claw. Anthers declinate, pollinia 4, caudicle, and viscid disc absent; column elongated; little University of Ghana http://ugspace.ug.edu.gh 22 expanded at apex; arms and foot absent; stigma close to column apex. The pedicel grows significantly longer as the erect capsule develops. Racemose, terminal, and single-flowered inflorescences (POWO, 2022). 2.4.7 Bolusiella Schltr. There are 4 species in Ghana. They are mostly distributed towards the border of Togo. Plants are monopodial epiphytic herbs with few, fine basal roots and short stems. Equidistant, imbricate, fan-shaped, meaty leaves. At the base of the plant, inflorescences that are densely or laxly several- to many-flowered emerge from the axils of old leaf sheaths. White, tiny resupinate flowers. Free, equal tepals. Lip spurred, generally shorter than the lip, and whole or subtly 3-lobed. Pollinia 2; stipites 2; viscidium 1; column oblong, fleshy, constricted above, without a foot; rostellum subulate, hooked; anther quadrate, cucullate (POWO, 2022). 2.4.8 Brownleea Harv. ex Lindl. Brownleea is a monospecific genus in Ghana with a narrow distribution in the Ashanti region. They are terrestrial herbs with sessile undivided tubers. Stems unbranched, leafy. Leaves few to several. Inflorescence terminal, 1- to many-flowered, lax or dense. Flowers resupinate, white, rose, lilac or purple. Dorsal sepal hooded, extending into a dorsal conical or narrowly conical spur, the laterals spreading, free or united in the lower part. Petals erect, adherent to the edges of the dorsal sepal, united at their base to the column. Lip very small, entire, often curved upwards and adpressed to the column, ecalcarate. Column short; anther reclinate, horizontal or reflexed, the loculi parallel, canals short; pollinaria 2, each with sectile pollinium, caudicle and viscidium; auricles on each side of rostellum, ± rugulose; stigmas united into a cushion in front on the anther, rostellum erect, ± 2-lobed. Capsules cylindrical or ellipsoid, ribbed (POWO, 2022). University of Ghana http://ugspace.ug.edu.gh 23 2.4.9 Bulbophyllum Thouars. This is the largest genus in terms of number of species (26) in Ghana. They are widely distributed in the Western, Eastern, Ashanti, Bono East, Ahafo, Greater Accra, and Volta regions. Members in this genus are epiphytic or lithophytic herbs with sympodial growth. Rhizomes creeping, often woody, sometimes branched. Pseudobulbs each comprising 1 internode (representing the main axis of a sympodial branch), clustered or spaced out along the rhizome, 1–2(3)-leaved at the apex. Leaves mostly coriaceous or fleshy, rarely thin-textured. Rhachis sometimes swollen or flattened; flowers white, cream, yellow, green, orange or purple, ± fleshy. Inflorescences arising from base of the pseudobulb, racemose or rarely umbellate, (1) few- to many-flowered. Sepals usually free and subequal, the lateral sepals united at the base to the column foot to form an obscure mentum. Petals usually smaller than the sepals. Lip often much smaller than the sepals, hinged to end of the column foot, usually motile, often fleshy and tongue-like, sometimes fringed with long or short hairs. Column short, usually with apical lateral extensions (stelidia), and often winged. Pollinia 2 or 4 in 2 pairs (POWO, 2022). 2.4.10 Calyptrochilum Kraenzl. There are 2 species of this genus in Ghana. They are mostly located in the Volta region. Members of this genus are monopodial, epiphytic herbs with long woody stems. Leaves alternate, fleshy, twisted at the base to lie in one plane. Inflorescences short; flowers resupinate, white. Sepals and petals free, similar, spreading, the petals slightly shorter than the sepals. Lip 3-lobed, spurred, the spur inflated at the apex. Column short and stout with a small foot. Rostellum prominent; anther apiculate; pollinia 2, globose, joined to a long linear caudicle; viscidium 1, large, triangular, grooved at base to clasp the rostellum (POWO, 2022). University of Ghana http://ugspace.ug.edu.gh 24 2.4.11 Corymborkis Thouars. Corymborkis is monospecific genus in Ghana, they are distributed mostly in the Atewa range (Eastern region). These are plants that grow on land with short rhizomes. Several, rigid, tall, and leafy stems. Distinctive but frequently spiral-appearing leaves that are sessile to quickly petiolate, plicate, lanceolate to elliptic. Inflorescences 1-4 few-many-flowered axillary panicles. White to greenish white flowers. Tepals are long, linear, subequal, and connivent at the base. Similar lips, but with a broad oval peak. Pollinia 2, thin, sectile, on a long, slender caudicle attached to a peltate viscidium falling behind the column; stigma large, deeply 2- lobed; rostellum upright, bifid; column long, slender, straight; dilated at apex with 2 lateral auricles. Perianth and column remains are preserved in the capsule (POWO, 2022). 2.4.12 Cheirostylis Blume This is a monospecific genus mostly distributed in the Eastern region. Plants are small terrestrial herbs with upright stems and leaves develop from robust rhizomes. petiolate leaves with a basal coating and thin texture. an upper stem bearing a terminal, erect inflorescence with few sheaths. little flowers that are mostly white. Petals cling to the dorsal sepal, which is joined for about half of its length. A lip that is erect, joined to the base of the column, saccate at the base with 2 calli, and lobed at the apex with a diverging lobe or a whole tooth. It should also be longer than the tepals. Short, upright column with apical appendages parallel to the long rostellum and two lateral stigmas. Viscidium is oblong, caudicle is short, and the dorsal anther is acuminate in pollinia two (POWO, 2022). 2.4.13 Cyrtorchis Schltr. There are 5 species of this genus, they are distributed mostly towards the border of Togo. Members of this genus are herbs; mostly epiphytic or, less frequently, lithophytic, having short to long, erect or pendent stems, distichous leaf bases, and roots that emerge all along the stem University of Ghana http://ugspace.ug.edu.gh 25 and branch out along it. Distinctive, coriaceous or fleshy, flat or conduplicate leaves that are longer than they are wide, unevenly bilobed at the apex, articulated at the base to a sheathing base, and deciduous, exposing the obvious sharp edges of the leaf bases. Inflorescences are often one to numerous, shorter than the leaves, few to many-flowered, and axillary on the upper part of the stem. The peduncle is small and frequently covered in massive, typically sterile sheathing bracts. White, stellate, sweet-smelling flowers with the occasional green, pink, or brown spur. In general, petals and sepals are recurved, free, linear-lanceolate, and acuminate. Lips are similar, full, or highly obscure It has three lobes, is ecallose, and has a sigmoid- or slightly incurved-shaped long tapering spur at the base. Pollinia 2; stipites 2; viscidium either hyaline or consisting of an indurated saddle-shaped top part and a hyaline lower part; anther- cap elongated at apex. Column short, fleshy; rostellum 3-lobed, pendent, outer lobes considerably longer than the mid-lobe, frequently papillate towards apex. Fruit a capsule with 3–6 wings (POWO, 2022). 2.4.14 Diaphananthe Schltr. There are 6 species of this genus in Ghana, with a distribution from the Eastern region towards the border of Togo. Members of this genus are monopodial herbs that are ephiphytic and rarely lithophytic. Long or short stem that is coated in the old leaf bases. Long, generally numerous, and frequently clearly visible white streaked roots that emerge along the stem. Distichous leaves with an uneven 2-lobed apex, articulated to a sheathing base, and frequently twisted to lie in a single plane. One to more inflorescences, typically with several to many flowers. Translucent, mostly small-sized, white, greenish, pale yellow, straw-orange, or, sporadically, pinkish and purplish flowers are also common. free of sepals and petals. Pollinia 2, stipites 2, viscidia 1 or 2, spurred lip that is whole or obscurely lobed, pollinia 2, and stipites 2. Rostellum was unique (POWO, 2022). University of Ghana http://ugspace.ug.edu.gh 26 2.4.15 Didymoplexis Griff. In Ghana, there is one species from this genus, they are mostly found in the Eastern region. Plants from this genus are herbs with no leaves that are saprophytic and lack chlorophyll; horizontal, meaty tubers. Unbranched, tall stems; racemose, terminal inflorescences with one to several flowers. Tepals create a short tube at the base of the little flowers' free-floating lip, which is attached to the foot of the column. Stigma close to apex; anther declinate; column long, broader at apex, with thin arms. Pollinia 4 is caudicle-less. As the capsule develops, the pedicel quickly lengthens (POWO, 2022). 2.4.16 Disa P.J. Bergius There 2 species of Disa in Ghana, mostly distributed in Ashanti and Eastern regions. Members of this genus are tuberous-rooted herbs that grow on the ground. Unbranched, leafy stems. leaves dispersed on many sterile shoots or along the flowering stalk. terminal inflorescence with one to several flowers. Flowers reseed and come in a variety of colors. The dorsal sepals are free, the laterals are spreading, and the dorsal is upright, hooded, or helmet-shaped. The base petals are irregularly shaped, adnate to the column, and frequently incorporated in the dorsal sepal. Lips are typically thin and without any spurs. Pollinaria 2; each with sectile pollinium, caudicle, and naked viscidium; column short; anther erect, horizontal, or reflexed; loculi parallel; anther short; stigmas united into a cushion below the rostellum; rostellum small; 3-lobed; middle lobe small; folded; side lobes short; fleshy; often adnate to petals. Twisted and nearly terete ovary. cylindrical, club-shaped, or narrowly elliptical in shape (POWO, 2022). 2.4.17 Disperis Sw. There are two species of the genus Disperis in Ghana, they are mostly distributed in the Eastern region. Members of this genus are erect largely tiny terrestrial herbs that grow typically in extremely shallow soil or leaf-mold in forests. They develop from little tubers. stems at the University of Ghana http://ugspace.ug.edu.gh 27 base of which have one or more sheathing scale-leaves (cataphylls). Leaves are sparse, alternate or opposite, and usually never almost obsolete. Flowers are tiny, typically less than 2.5 cm long, white, yellow, green, pink, or magenta, solitary, or in many-flowered racemes; bracts resemble leaves. Lateral sepals have prominent spurs or pouches on the inner margins, while the dorsal (middle) sepal is fused with the petals to form a structure that can range in shape from a nearly flat limb to an extended spur. Differently formed petals that are occasionally auriculate at the base and frequently falcate, obliquely acute, or lobed at the apex. Lip strikingly modified, its claw attached to the column's face and rising above it, varyingly curved into the spur if present, frequently dilated into a smooth or papillate, straight or reflexed limb, and typically bearing a simple or 2-lobed appendage, the shape of which varies greatly between species. Column erect, mostly stout; rostellum large, membranous, 2-lobed, produced in front into 2 rigid cartilaginous arms (fitting into the lateral sepal-pouches when in bud) holding the glands of the pollinia at their apices; anther-bearing part of column horizontal or ascending; anther-loculi distinct, parallel, approximate; pollinia-granule stigma two lobes, with lobes situated on either side of the adnate claw of the lip (POWO, 2022). 2.4.18 Eggelingia Summerh. This is a monospecific genus which is mostly distributed in the Eastern region. Plants in this genus are mostly herbs that are epiphytic or very rarely lithophytic have long monopodial stems, few branches, and are leafy along their length. The roots in the base are aerial and elongate. Distichous, spreading, ligulate, unevenly bilobed at apex, articulated to persistent leaf-sheaths, opposite leaf's leaf-sheath sporting a hair-like ligule. Very brief and sparsely flowered axillary inflorescences. Small, pedicellate flowers. Petals and sepals are separate and comparable. At the base, the lip may be whole or obscurely 3-lobed, ovate, sharp, ecallose, or spurred; the spur may be cylindric or subclavate and be as long as the lip. The rostellum is deflexed, triangular, and deeply bifid; the column is short and fleshy; the anther-cap is University of Ghana http://ugspace.ug.edu.gh 28 hemispherical and is produced in front; the pollinia are two and oval or subpyriform-ovoid; the stipes are ligulate and dilated at the apex; and the viscidium is rectangular-elliptic (POWO, 2022). 2.4.19 Epipogium Borkh. This is a monospecific genus which is mostly distributed in the Eastern and Ashanti regions. Members of the genus are saprophytic, leafless, achlorophyllose herbs with scapes that emerge from tuberous rhizomes. Simple, upright scape with a few basal sheaths. Lax, terminal inflorescence. Flowers form pedicels. Subequal and free tepals. Concave, spurred, and with beautifully warted crests, the lip is wider than the tepals. Short column; short, broad anther; two sectile pollinia, each with a thin, curved caudicle linked to a single, broad viscidium; prominent, broad stigma; no rostellum. Ovoid and pendulous capsule (POWO, 2022). 2.4.20 Eulophia R. Br. Little research has been conducted on the genus Eulophia in Ghana. There are eight species currently according to POWO (2022) but nine species were recorded in the Ghana Herbarium. In 1975, Lock and Profita conducted a survey to investigate the pollination patterns of E. cristata in southern Ghana. They confirmed that the inflorescence is approximately 1 m. tall, and produces up to 30 flowers. The flower is zygomorphic, held horizontally at maturity. It is resupinate, reversing quickly after pollination. The reflexed sepals, the upper petals and the spur are pinkish-violet, variable in depth of colour. The centre of the labellum and the ovary are deep violet, and the inverted boundaries of the labellum are greenish (Lock & Profita, 1975). The genus Eulophia R. Br. Lindl is one of the most notable representatives of the Orchidaceae family. More than 230 species make up this group, which is found in southern and tropical University of Ghana http://ugspace.ug.edu.gh 29 Africa, Madagascar, Australia, subtropical and tropical Asia, and 1 species in tropical America. 500 of the 723 entries for Eulophia in the International Plant Names Index (IPNI) are synonyms, according to Cieslicka, (2006). Members of the genus can be found in a several habitat ranges. Most of them grow on land in the savanna and scrub, but epiphytic species can also occur in grasslands and in the lowland and equatorial forest. A large number of species exhibit sympodial growth, except for Eulophia epiphytica from Madagascar, which is monopodial and should maybe be placed in a separate genus (Cieslicka, 2006). The morphological diversity of the genus further complicates its infrageneric classification. Their perennating organs mostly are pseudobulbous or tube-like (Cieslicka, 2006). The leaves are oblong and plicate, narrow and grass-like, or thin yet robust. Some species are saprophytic and lack green leaves. Eulophia is home to two different kinds of flowers. The sepals and petals of the first kind are remarkably alike in size, shape, and color. In the second, the sepals frequently recurve and are much reduced than the petals. The lip continues into a spur, which can take on a wide variety of shapes, in both types. The adaptation to various pollinators leads to modified floral components, particularly spurs (Cieslicka, 2006). The genus Pteroglossaspis Rchb.f. was distinguished from Eulophia based on the lack of a spur. Lips typically have three lobes, crests, papillae, and basal callus appendages on the top surface. With or without a column foot, an arcuate lip can be connected to the column. An important character in an infrageneric treatment of Eulophia appears to be the presence or lack of a column foot. Pollinia are numbered four or two, similarly in the genus Cymbidium Sw. (Szlachetko, 2003). Genus Eulophia was formally described by John Lindley in 1823 within Botanical Register. Few years later, in 1833, he suggested three sections. Section Genuinae Lindl. consisted of University of Ghana http://ugspace.ug.edu.gh 30 19 species, from which three were moved to the genus Graphorkis Thouars, and one species, Eulophia artrovirens Lindl. was added to the genus Oeceoclades Lindl. Four species from section Desciscentes Lindl. are now a part of the genus Acrolophia Pfitzer. Three species from section Aphylae Lindl. are now synonyms of Eulophia dabi (D. Don) Hochr (Cieslicka, 2006). In 1833, Lindley described genus Cyrtopera, which was differentiated from Eulophia based on the presence of a column foot. Cyrtopera consisted of 9 species: C. plantaginea Lindl., C. woodfordii Sims, C. scabrilinguis Lindl., C. flava Lindl., C. obtusa Lindl., C. bicarinata Lindl., C. plicata Lindl., C. pedicellata (L.f.) Lindl. and C. gigantea (L.f.) Lindl. Currently, Cyrtopera has been added to Eulophia, even though some authors recommended that it should keep, the status of a section. Kraenzlin in 1897 described section Pulchra that contained Eulophia pulchra (Thouars) Lindl. and similar species. List of names for that section was published by Garay and Taylor in 1976, although most of them are currently considered as synonyms of E. pulchra (Cieslicka, 2006). The most current infrageneric classification was proposed by Perrier de la Bâthie in 1935, who reviewed the Madagascan Eulophia species. He differentiated three sections. First section, Saprolophia, contained just 1 species, Eulophia hologlossa Schltr.; second section Lissolophia, was composed of 7 species. Third section, Eulophia, contained 25 species, with most of them presently included in the genera Graphorkis and Oeceoclades (Cieslicka, 2006). In 1941, Perrier de la Bâthie moved all Eulophia species to the genus Lissochilus, and raised sections Eulophidium Kraenzl. and Lissolophia H. Perrier to the subgeneric rank. He also differentiated subgenus Stiriolophia. At present, taxa from section Lissolophia are found in section Eulophia; Eulophidium is a synonym of Oeceoclades, and species from the subgenus Stiriolophia are now placed into Eulophia and Graphorkis. Since the review by Perier de la University of Ghana http://ugspace.ug.edu.gh 31 Bâthie, there were no attempts to propose a different infrageneric treatment of the genus (Cieslicka, 2006). 2.4.21 Manniella Rchb.f. There are 2 species of this genus in Ghana, with a narrow distribution mostly in the Ashanti region. Members of this genus are mostly herbs that grow on the ground with thick, meaty roots. With a rosette of radical leaves and very small stems. Membranous, long-petioled leaves. A few membrane sheaths are present on the erect, long, and many-flowered scape. fewer than 1 cm long in the flowers. For at least half of their length, the lateral sepals are connected to one another, the petals, and the lip. The base of the free section has two reflexed marginal appendages, and the lip is attached to the sepals and has a thin claw. Anthers are subsessile between the column wings, broad, and concave, and the pollinia are loosely granular. The column is long, attached to the perianth tube, and free at the apex (POWO, 2022). 2.4.22 Microcoelia Lindl. This is a monospecific genus in Ghana, they are mostly distributed in the Volta region. Plants are mostly short-stemmed, leafless epiphytic or, occasionally, lithophytic herbs. Unbranched or with a few branches, typically elongate roots that are firmly or loosely linked to the substrate, generally thick, terete or less frequently dorso-ventrally flattened, smooth or rarely verrucose. Acute to rostrate scales on the stem protect the stem apex. Few- to many-flowered inflorescences that are axillary, racemose, and concentrated at the tip of the stem; long or short peduncles; rhachis that are terete or angular, smooth or with processes; and bracts that are sheathed or not. Flowers on spur and other segments are small to minute, sessile or pedicellate, and often white with varying green, brown, or pink tints. Petals and sepals are separate and comparable. Lips closed or masked; spurs can be globose, cylindrical, or have differently shaped swellings. They are 3-lobed, free, and typically include fleshy calli at the base on either University of Ghana http://ugspace.ug.edu.gh 32 side of the mouth of the spur. Column fleshy, androclinium short to long, anther-cap hemispherical, frequently elongated at the apex, pollinia 2 to pyriform, stipes linear to oblanceolate, whole or bifid at the apex, viscidium linear or oblong, short to long, and rostellum bifid, short to as long as the column (POWO, 2022). 2.4.23 Polystachya Hook. There are 25 species of this genus in Ghana, they are mostly distributed in the Eastern, Bono East, Volta, and Western regions. Members of this genus are perennial herbs that are epiphytic, occasionally lithophytic, or terrestrial. On a creeping rhizome, stems that typically form pseudobulbs are clustered or less frequently spaced apart, occasionally branched or superposed (emerging from nodes above the base of the preceding pseudobulb), and have one to several nodes and leaves. Linear, lanceolate, oblanceolate, or elliptic leaves with a variety of textures. An inflorescence can range from one to many blooms, and it can be simple or branched. Setiform, lanceolate, ovate or obovate, acute or acuminate to mucronate, floral bracts that are upright to reflexed. The majority of flowers are non-resupinate, small to medium in size, very rarely large, fragrant, frequently pubescent, and colored green, yellow, white, pink, mauve, or orange, very never red. The lateral sepals are oblique and connected to the column-foot to form a mentum; the dorsal sepal is oblong to ovate. Petals are obovate to linear. Lips are often 3-lobed but occasionally entire, glabrous, pubescent, or farinose, sometimes meaty, and challenging to flatten without breaking. They may also have a basal callus. Pollinia 2, ovoid; stipe 1, square or subtriangular to oblong or linear; viscidium small to large, spherical or ellipsoid; rostellum often obscure, bifid in front, rarely somewhat elongated or beak-like (POWO, 2022). University of Ghana http://ugspace.ug.edu.gh 33 CHAPTER THREE 3.0 MATERIALS AND METHODS 3.1 Morphological Studies Morphological studies were based mainly on herbarium specimens of the genus Eulophia collected in Ghana. Three herbaria, namely, Ghana Herbarium (GC) at the University of Ghana, herbarium in the University of Cape Coast (CCG), and herbarium in the Kwame Nkrumah University of Science and Technology (KUU) were visited to undertake the morphological studies. Herbarium code follows Index Herbariorum (http://www.sweetgum.nybg.org/science/ih/herbarium-list/). 50 herbarium specimens of the genus Eulophia were examined for morphological characters. 35 specimens at GC, 10 specimens at CCG, and 5 specimens at KUU. The number of specimens examined per species ranged from 2 specimens for E. angolensis to 15 specimens for E. cristata (Appendix A). From studies under the dissecting microscope (Leica zoom 2000 model) and using 30cm transparent ruler, eleven quantitative characters were measured (Table 3.1) and seven qualitative characters assessed (Table 3.1). Selection of the characters for assessment was based largely on the of work Ortúñez et al. (2020). Additional morphological data were gathered from online databases such as the Plants of the World Online (https://powo.science.kew.org/), Global Biodiversity Information Facility (https://www.gbif.org/) as well as taxonomic literature (Ortúñez et al., 2020; Hutchinson & Dalziel, 1963). University of Ghana http://ugspace.ug.edu.gh http://www.sweetgum.nybg.org/science/ih/herbarium-list/ https://powo.science.kew.org/ https://www.gbif.org/ 34 Plate 3.1 Morphological studies of herbarium specimens University of Ghana http://ugspace.ug.edu.gh 35 Table 3.1: Morphological characters assessed, source and number of species Character Source of information Number of species Leaf length Herbarium voucher specimens 8 Leaf width Herbarium voucher specimens 8 Tepal length Herbarium voucher specimens 8 Tepal width Herbarium voucher specimens 8 Interfloral spaces Herbarium voucher specimens 8 Fruit length POWO, 2022 8 Fruit width POWO, 2022 8 Bract length Herbarium voucher specimens 8 Interbractal spaces Herbarium voucher specimens 8 Stem width Herbarium voucher specimens 8 Lip shape Herbarium voucher specimens 8 Midlobe shape Ortúñez et al. (2020) 7 Interveinal length Herbarium voucher specimens 8 Perennating organ Ortúñez et al. (2020) & POWO, 2022 8 Leaf shape Herbarium voucher specimens 8 Tepal colour Ortúñez et al. (2020) & POWO, 2022 8 Leaf base Herbarium voucher specimens 8 lip ridge number Ortúñez et al. (2020) 7 University of Ghana http://ugspace.ug.edu.gh 36 Spur colour Ortúñez et al. (2020) & POWO, 2022 8 Leaf apex Herbarium voucher specimens 8 Spur shape Ortúñez et al. (2020) & POWO, 2022 8 Leaf margin Herbarium voucher specimens 8 Spur apex Ortúñez et al. (2020) 7 Leaf surface Herbarium voucher specimens 8 Anther cap shape Ortúñez et al. (2020) 7 Leaf arrangement Herbarium voucher specimens 8 Lip colour Ortúñez et al. (2020) & POWO, 2022 8 Midlobe margin Ortúñez et al. (2020) 7 lip pappillae colour Ortúñez et al. (2020) 7 callus shape Ortúñez et al. (2020) 7 callus colour Ortúñez et al. (2020) 7 lip ridge colour Ortúñez et al. (2020) 7 Table 3.1 continued University of Ghana http://ugspace.ug.edu.gh 37 3.2 Ecological and Distributional Studies Field work was conducted at Aburi Botanical Gardens, Shai Hills, Achimota Forest and Legon Hills based on information obtained from voucher specimens about occurrence of species of the genus in Ghana. However, none of the Eulophia species were found after careful consideration of the phenology and the help of local guides. As such, information from herbarium labels of the specimens examined were retrieved to understand aspects of the ecology such as phenology, habitat types and pollinator-flower relationship for the species of the genus Eulophia. Data were also downloaded from Global Biodiversity Facility Information (GBIF) and Plants of the World Online (POWO). Species distribution maps were developed using ArcGIS Pro 2.8 software for the genus and individual species of Eulophia in Ghana. Data on species collection localities on herbarium specimens and those from GBIF as well locality data from local field guides in Ghana (Hall, 1980; Hawthorne, 1990) were used. In total, 55 locality points (20 from herbarium data, 25 from GBIF and 10 from field guides) were used after removing duplicate points for species and georeferencing to ensure accuracy of the points. The number of georeferenced points for each species is shown in Table 3.2. University of Ghana http://ugspace.ug.edu.gh 38 Table 3.2: Species of Eulophia, sources of information and collection sites Species Source of locality information Collection sites E. alta Herbarium voucher specimens, GBIF 2 E. angolensis GBIF 1 E. buettneri Herbarium voucher specimens 2 E. cristata Herbarium voucher specimens, GBIF 19 E. cucullata Herbarium voucher specimens, GBIF 10 E. euglossa Herbarium voucher specimens, GBIF 5 E. flavopurpurea Herbarium voucher specimens, GBIF 8 E. milnei Herbarium voucher specimens, GBIF 7 In order to estimate the area of occupancy (AOO) and extent of occurrence (EOO) for each species, a rapid geospatial conservation assessments was conducted using GeoCat (http://geocat.kew.org/). Rapid geospatial analysis for Red List assessment was also performed through the open source, browser-based GeoCat. The methodology, which was created to employ spatially referenced primary occurrence data, concentrates on two characteristics of a taxon's geographic range: the extend of occurrence (EOO) and the region of occupation (AOO). The IUCN Red List includes these measures as part of its categories and criteria, although obtaining them in a precise, dependable, and reproducible manner has frequently proven difficult. University of Ghana http://ugspace.ug.edu.gh http://geocat.kew.org/ 39 3.3 Taxonomic Studies Taxonomic studies focused on development of an authoritative checklist and construction of dichotomous keys for identification of species of the genus. In order to develop a checklist of the species of Eulophia in Ghana, type specimens, nomenclature, ethnobotanical uses and vernacular names besides that of morphology, ecology and distributional are essential (Utteridge, 2016). Type specimens of the various species of Eulophia especially holotypes were studied from available online databases, mainly Biodiversity Heritage Library (https://www.biodiversitylibrary.org/) and Global Plants on JSTOR (https://plants.jstor.org/). Information on nomenclature changes were obtained Plants of the World online (https://powo.science.kew.org/), International Plant Names Index (https://www.ipni.org/), and Catalogue of Life (https://www.catalogueoflife.org/). Ethnobotanical information and vernacular names were retrieved from PROTA (2022) and iNaturalist (2022). To construct dichotomous key for identification of species, a data matrix of the various taxonomic characters examined was used. The matrix consisted of species plotted against 35 characters. Good characters whose attributes were contrasting and could separate the taxa into two groups were selected. Couplets with contrasting statements ended in an identified taxon or the next set of couplets to visit. The bracketed key was used in order to exhaust every alternative statement for each couplet. A total of 7 couplets were used to delimit the species of Eulophia occurring in Ghana. 3.4 Data Analysis Cluster analysis was employed to analyse the various samples of each species to verify significant patterns of the various morphological quantitative characters using STATA University of Ghana http://ugspace.ug.edu.gh https://www.biodiversitylibrary.org/ https://plants.jstor.org/ https://powo.science.kew.org/ https://www.ipni.org/ https://www.catalogueoflife.org/ 40 version 15 and R/R-studios (version 4.2.2). The significance of variations in morphological characters among the Eulophia species was determined by subjecting the data to one-way and two-way analysis of variance (ANOVA) using R/R-studios (version 4.2.2). All data were recorded in Microsoft Excel 2019 for preliminary data analysis before imported into R/R- studios (version 4.2.2) for final analyses. University of Ghana http://ugspace.ug.edu.gh 41 CHAPTER FOUR 4.0 RESULTS 4.1 Morphological Studies 4.1.1 Leaf and Stem morphology The members of the Eulophia range from slender to large herbs, lacking woody stems; plant height ranges from 560–2000mm, 2–16mm in width; has leaves that are often alternately arranged. Leaves are plicate, erect, often linear to lanceolate except in in E. alta, E. cristata and E. euglossa where the leaves are elliptical. Leaves are either acute or acuminate at the apex, ca. 90–730 × 2–60 mm in size. Leaves have 4–9 veins with margin entire. Leaf surfaces are aciculate to striate except in E. alta where leaves are glaucous. Leaf sheaths ca. 23–70 mm; most members have closed leaf sheaths expect in E. angolensis, E. cucullata, and E. euglossa, where leaf sheaths are opened (Tables 4.1 & 4.2). 4.1.2 Inflorescence, Flower and Fruit Morphology Inflorescence are often densely flowered ranging from 3–21 flowers except in E. buettneri, E. cucullata, E. dilecta, and E. euglossa where flowers are mostly above 21. Peduncle diameter ranges from 4–12mm except for smaller diameters (2–3mm) observed for E. milnei. Bracts are reflexed, often linear except in E. buettneri and E. cristata where bracts can be lanceolate to elliptical. Bract is acute to acuminate at the apex, 5–25 mm long except for longer bracts observed for E. euglossa (27–29mm). Distances between successive bracts range from 4–80 mm (Tables 4.3 & 4.4). Flowers are mostly in the shades of yellow and red, except shades of green in E. euglossa and E. flavopurpurea. The distance between successive flowers ranges from 6–45 mm. Sepal colour is mainly yellow but green in E. cristata. The shape of dorsal sepal is obovate- University of Ghana http://ugspace.ug.edu.gh 42 lanceolate to obovate with a narrow base, ca. 10–18 ×4–7 mm, apex acuminate while lateral sepals are obovate-lanceolate, asymmetrical, ca. 16–20 × 3–7 mm. The spur is pale green to rose, with red spots in E. milnei, saccate to conical and subnull to conical at the apex. Petals range from deep pink to bright yellow, occasionally with brown spots, oblong-ligulate except linear-lanceolate in E. cristata and E. euglossa, acute to acuminate at the apex, ca. 5–29 × 1.5– 20 mm. Lip is trilobed near the apex, pale yellow to dark rose often white rose in E. alta and E. buettneri, ca. 5.5–7.5 × 4–5 mm, ridges yellow to purple, 3–11; midlobe oblong to elliptical but semi-circular in E. alta, margins undulate to crenulate but entire in E. buettneri. Fruit pendent, ca. 10–50 × 4–22 mm (Tables 4.3 & 4.4). University of Ghana http://ugspace.ug.edu.gh 43 Table 4.1: Quantitative vegetative morphological characters of Eulophia species SPECIES Leaf length (cm) Intervenal length (cm) Leaf width (cm) Stem width (cm) Leaf size (cm 2 ) Range mean±SD Range mean±SD Range mean±SD Range mean±SD range mean±SD E. alta 50.1-61.8 56.14±6.47 0.25-0.75 0.50±0.25 5.59-5.89 5 .64±0.25 0.69-1.17 0.93±0.24 313.10-320.78 316.63±3.56 E. angolensis 55.3-63.2 59.95±4.42 0.31-0.41 0.36±0.05 0.95-1.68 1.22±0.39 0.41-0.49 0.45±0.04 68.67-75.12 71.78±3.74 E. buettneri 24.3-34.7 29.45±5.45 0.15-0.19 0.17±0.02 0.42-0.64 0.53±0.11 0.34-0.56 0.45±0.11 11.23-20.15 15.35±4.54 E. cristata 29.34-33.65 31.56±2.25 0.48-0.54 0.51±0.03 2.54-2.76 2.65±0.11 0.78-0.90 0.84±0.06 61.34-65.45 63.78±1.56 E. cucullata 21.76-27.89 24.67±3.38 0.51-0.61 0.56±0.05 2.33-3.08 2.72±0.39 0.69-1.01 0.85±0.16 61.87-67.53 64.65±2.76 E. dilecta 23.12-27.67 25.17±2.25 0.28-0.32 0.30±0.02 1-1.2 1.10±0.10 0.70-0.84 0.77±0.07 25.37-28.35 27.12±1.71 E. euglossa 21.34-25.51 23.36±2.20 0.12-0.16 0.14±0.02 0.31-0.37 0.34±0.03 0.46-0.52 0.49±0.03 7.02-9.34 7.82±1.79 E. flavopurpurea 17.87-19.06 18.44±0.74 0.08-0.12 0.10±0.02 0.33-0.43 0.38±0.05 0.79-0.89 0.84±0.05 6.77-6.90 6.84±0.06 E. warneckeana 46.50-52.54 49.52±3.02 0.39-0.47 0.43±0.04 1.52-1.84 1.68±0.16 1.0-1.26 1.13±0.13 77.86-80.21 78.40±2.65 Table 4.2: Qualitative vegetat