Taxonomic Revision of Selected Problem Groups of the Genus Clutia in Kwazulu-Natal, South Africa by Andani Robert Madzinge (218024650)
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COPYRIGHT AND CITATION CONSIDERATIONS FOR THIS THESIS/ DISSERTATION o Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. o NonCommercial — You may not use the material for commercial purposes. o ShareAlike — If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original. How to cite this thesis Surname, Initial(s). (2012). Title of the thesis or dissertation (Doctoral Thesis / Master’s Dissertation). Johannesburg: University of Johannesburg. Available from: http://hdl.handle.net/102000/0002 (Accessed: 22 August 2017). Taxonomic revision of selected problem groups of the genus Clutia in KwaZulu-Natal, South Africa By Andani Robert Madzinge (218024650) Dissertation Submitted in fulfilment of the requirements for the degree of Magister Scientiae (MSc) In Botany In the Faculty of Science at the University of Johannesburg South Africa Supervisor: Prof. A.N. Moteetee (UJ) Co-supervisor: Prof M. Van der Bank (UJ) Co-supervisor: Dr R.H. Archer (SANBI) January 2021 DECLARATION I, Andani Robert Madzinge, declare that this dissertation submitted by me for the degree of Masters of Science in Botany at the department of Botany and Plant Biotechnology in the faculty of Science at the University of Johannesburg is my own work in design and in execution. It has not been submitted before for any degree or examination at this or any other academic institution and that all the sources that I have used or quoted have been indicated and acknowledged by means of complete references. ………………………………. ………………………………… SIGNATURE DATE (MR A R MADZINGE) i Dedication I would like to dedicate this work to my parents, Nyambeni Lydia Madzinge and Azwihangwisi Edson Madzinge for all the support they have given me throughout my learning years from my first day of school, to the completion of this higher degree. I am also grateful to my daughter Uzwothe Madzinge for understanding when I didn’t have time for her while studying for this degree. I am thankful for all the support and encouragement throughout this project from all persons around me. ii ACKNOWLEDGEMENTS I would like to acknowledge institutions and persons who contributed to the success of this study by allowing me an opportunity, providing guidance and support in preparation of this dissertation, namely: University of Johannesburg (UJ), for accepting my application to study at the institution. Prof. A.N. Moteetee, my supervisor, for her guidance, advice and approval of the study project. Prof. Michelle van der Bank, my co-supervisor, for her assistance and guidance with molecular studies. Mr. Michael Pilusa, for his assistance with PCR and gene sequencing. Mr. Ross Stewart, for his training in gene sequence editing. Ms Asanda Mkiva, for her assistance with DNA extractions. South African National Biodiversity Institute (SANBI) for allowing me access to their herbarium to conduct my study. Dr. R.H. Archer, my co-supervisor, for his guidance, advice and assistance throughout this study. The Department of Agriculture, Land Reform and Rural Development, my employer, for allowing me to take time off to conduct my study. KwaZulu-Natal Ezimvelo, for granting me a collection permit to collect specimens in the province. iii Symposium presentations Madzinge, A.R, Archer, R.H., Van de Bank, M., and Moteetee A.N., 2018. Taxonomic revisions on selected problematic species of the genus Clutia in KwaZulu Natal. Oral presentation for the Post Graduate Symposium. Department of Botany and Plant Biotechnology, Faculty of Science, University of Johannesburg, iv Abstract The genus Clutia L. was previously placed in Euphorbiaceae but has recently been transferred to Peraceae. Clutia species are perennial herbs or shrubs with simple, alternate, thin and oval to round leaves that are covered with simple hairs or are sometimes glabrous. Clutia is closely related to Pera and the two are the largest genera in the family. Clutia occurs mainly in southern Africa but also extends to the east into the Democratic Republic of Congo and as far north as Arabia. In southern Africa, the genus was last revised a 100 years ago, making the identification of certain groups within the genus problematic. Although the genus is relatively large, a comprehensive revision for southern Africa is beyond the scope of this study. The aim of the study was firstly, to conduct detailed taxonomic revisions of selected problematic groups of species in the genus Clutia in the KwaZulu-Natal Province. The selected groups are restricted to the Province, except for Clutia pulchella which is known to occur outside KZN. The correct names, diagnostic characters, correct typification, known distribution ranges are provided. The second aim of the study was to infer phylogenetic relationships among the southern African species using molecular data. The hairs on the leaves and branches were studied under binocular microscope (Nikon model). A scanning electron microscope (Phenon Pure) was also used to study the type and length of hairs, while scanning electron micrographs of seed epidermal cell structure were studied for seeds of Clutia abyssinica subsp. abyssinica var. abyssinica and Clutia abyssinica subsp. “pondoensis”. Scanning electron micrographs were used to study the size and structure of seed epidermal cells. The leaves were softened with 70% ethanol and a piece was cut for scanning. Flowers collected from herbarium specimens were softened using 70% alcohol solution, after which they were dissected under a binocular microscope to observe the number of glands on sepals and petals. Seeds were scanned in their natural form and cut into vertical cross sections and scanned. The selected groups of species identified as having taxonomic problems are a) Clutia abyssinica and its varieties (C. abyssinica var. abyssinica, C. abyssinica var v pedicellaris and C. abyssinica var. usamabarica), which have been confused with the ‘Pondoland’ specimens, b) C. galpinii, which has been alleged to be the same as C. pulchella L. and c) C. platyphylla - a poorly known taxon that was originally described from one specimen which is currently missing. Vegetative and reproductive morphological characters were examined for all the selected specie. The length, width, shapes of the leaves and the number of glands found in the flowers were all studied to clearly describe the species and to formulate clear diagnostic characters. Clutia abyssinica in southern Africa was re-classified into two subspecies namely Clutia abyssinica subsp. abyssinica and the newly described Clutia abyssinica subsp. pondoensis. The latter is described based on ‘Pondoland’ specimens housed in the National Herbarium, Pretoria. The new classification of Clutia abyssinica was mainly based on flower and leaf morphology. The specimens belonging to Clutia abyssinica subsp. pondoensis were mistakenly identified as C. abyssinica subsp. abyssinica var. usambarica. However, closer examination has shown that the two taxa differ in several ways. Furthermore, Clutia abyssinica subsp. abyssinica var. usambarica is only known to occur in Kenya and Tanzania whereas C. abyssinica subsp. pondoensis only occurs in KwaZulu-Natal, South Africa. Clutia galpinii is formally reduced to synonymy under Clutia pulchella var. pulchella as there are no morphological characters, which can be used to satisfactorily separate them. The narrow ovate leaf and short petiole which were the characters used to describe Clutia galpinii are not consistent and reliable. Clutia platyphylla is reduced to synonymy under Clutia hirsuta. Abnormal specimens of Clutia hirsuta, which match the description of Clutia platyphylla were examined. It was concluded that the original description of Clutia platyphylla was based on an abnormal specimen of Clutia hirsuta. A phylogenetic analysis indicated that relationship could not be established between Clutia abyssinica subsp. abyssinica var. abyssinica and C. abyssinica subsp. pondoensis. Instead, a close relationship between Clutia abyssinica subsp. pondoensis and Clutia pulchella was highly supported by Bayesian inference (PP = 0.99) and moderately supported by bootstrap analysis (BP = 81). Even though the relationship between Clutia abyssinica subsp. pondoensis and Clutia pulchella received good support, morphological characteristics separate them convincingly. The results also indicated that Clutia pulchella is not monophyletic but rather vi paraphyletic. These results could be due to the limited sampling, this suggests that in future, a wider sampling of the genus beyond the KwaZulu-Natal Province should be undertaken. vii Table of contents DECLARATION……………………………………………………………… (i) DEDICATION…………………..……………………………………………. (ii) ACKNOWLEDGEMENT……………………………………………………. (iii) SYMPOSIUM PRESENTATIONS……………………………………….. (iv) ABSTRACT…………………………………………………………………… (v) LIST OF FIGURES…………………………………………………………….. xi LIST OF TABLES……………………………………………………………. (xii) LIST OF ABBREVIATIONS, ACRONYMS AND SYMBOLS…………….. (xiii) Chapter 1 GENERAL INTRODUCTION…………………………………… 1 1.1. The family Peraceae……………………………………………………. 1 1.2. The genus Clutia ………………………………………………………… 2 1.3. Phylogenetic relationships………………………………………………. 4 1.4. Aims and Objectives …………………………………………………….. 4 1.4.1. Research aims …………………………………………………………. 4 1.4.2. Research questions ……………………………………………………. 5 Chapter 2 TAXONOMICALLY PROBLEMATIC SPECIES FROM