Diversity of endophytic fungi possessing bioactive compounds isolated from selected medicinal plants Madira Coutlyne Manganyi C9orcid.org/0000-0002-0209-5547 Dissertation submitted in fulfilment of the requirements for the degree Doctor of Philosophy in Biology (Molecular Microbiology) at the Mafikeng Campus of the North-West University Promoter: Professor CN A TEBA Co-promoters: Professor T REGNIER (TUT) Professor CC BEZUIDENHOUT (NWU) Graduation October 2018 Student number: 26853795 http://dspace.nwu.ac.za/ DECLARATION I, Madira Coutlyne Manganyi, declare that the thesis entitled "Diversity of endophytic fungi possessing bioactive compounds isolated from selected medicinal plants", hereby submitted for the degree of Doctor of Science in Biology (Molecular Microbiology) , has not previously been submitted by me for a degree at this or any other university. I further declare that this is my work in design and execution and that all materials contained herein have been duly acknowledged . Signed ... ......... ... ...... ...... ......... this the ... ........ ... ....... .... day of ...... ... ..... .... 2017 Signature:...... .... ..... ............................... Date : .. ... ......... ... ...... ......... ......... ...... MC Manganyi (Student) Signature :... .. ............... ......... ....... .. ..... .... Date : ...... ... ....... ...... ... ........... ........ .. .. Prof CN Ateba (Supervisor) Signature :............... .............................. .. Date : .... .. ............ .. .. ..... ... ............... .. Prof T Regnier (Co-supervisor) Signature :.. ..... ........ .... .. ......... ....... ......... Date : ... ........ ...... ......... ..... .... .. .. .. ...... Prof CC Bezuidenhout (Co-superviso r) DEDICATION I would like to dedicate this PhD thesis to my parents Fridah and John Manganyi for their endless support, care and love. Honour Your Father and Mother, so that it may be well with you , and that you may live long on the earth . This is the first commandment with a promise Ephesians 6:2-3 ACKNOWLEDGEMENTS From the bottom of my heart, I wish to acknowledge the following people. Since , they have contributed towards the success of this research project: • Prof. CN Ateba, my supervisor, for his consistent motivation , amazing ded ication and commitment. Opening my eyes to priceless opportunities. I deeply appreciation and admire his devotion to his students, his guidance and world-class expertise. He has been a Father Figure and Mentor to all his students. May the Almighty continue to bless you ; • Prof. T Regnier, my co-supervisor, for seeing the potential in me. For your valuable support and guidance. His door is always open for me and ready to help. His marvellous passion and love for his research field made the study with wide experience; • Prof. CC Bezuidenhout, my co-supervisor, for his willingness to supervise me. The support and guidance resulted in the success of the project; • I want also to express my deepest appreciation and my special thanks to my ALL colleagues in the Department of Microbiology, Mafikeng, North West University, for the support and encouragements; • My friends and sisters (Dikeledi, Lydia, Maphapa Manganyi, Sefora Moloi) your support means the world to me. This achievement is for you as well; • My brothers (Lesetja and Lucky Moneyla) we have set the bar and the next generation will follow; • My family, this is our achievement, thanks for all your endless love and support; • Dr Adriaana Riana Jacobs and Mrs Grace Kwinda at Agricultural Research Council , PPRI , Mycology for their support and guidance with fungal identification and preservation. • Christ Donald Kaptchouang, Peter Montso and Daniel Matlou (North-West University, Department of Microbiology, Mafikeng) belong ing to the same research group (Molecular Microbiology) . They assistance and motivation throughout this study made it a success ; iii • Dr Guy Kamatou and Prof. Alvaro Viljoen {Tshwane University of Technology, Pharm aceutical Sciences) for their expert assistance in analytical instruments; • Prof Sandra Combrinck, (Tshwane University of Technology, Pharmaceutical Sciences) her knowledge on aromatic plants lead to the selection of the investigated medicinal plants ; • Shimadzu analytic group (Jim Malaza, Thabiso Khaeeane and others) guidance in the GC­ MS experiments ; • Mr Sizwe Loyilani (North-West Un iversity, Chemistry Department) for introducing me to the Shimadza family and their equipment; • Prof Oziniel Ruzvidzo, Prof Thami Sithebe (NWU , Mafikeng, School of Biolog ical Sciences) , thank you for your support, advice and motivation ; • Dr. Dave Kawadza (North-West University, Department of Microbiology), your continuous smile, motivation and Language editing skills assisted in the success of th is project; • Prof Olubukola Babalola (North-West University, Department of Microbiology) for allowing me to utilize her greenhouse and for her consistence motivation; • Prof. Eugene Olivier (Tshwane University of Technology, Pharmaceutical Sciences) for introducing me to my Father (Prof. Regnier) and guiding me throughout my Master project. • Dr. Tshegofatso Dikobe, Dr. Ramokoni Gopane, Prof. Lebo Katata-Seru, Dr. Zimbili Mkhize (North-West University, Mafikeng), Miss Ethel Mogale (Tshwane University of Technology, Biotechnology), Mrs Adoration Shubane (Agricu ltural Research Council, PPR! , Nematology) . You are all beautiful, strong , motivating , supportive, encouraging , black women in Science. You have all contributed in the success for this study. Ndza nkhensa Ndzi khense ngopfu lnkomu swinene 'IT TAKES A VILLAGE TO RAISE A CHILD' May the Almighty GOD continue to BLESS YOU ALL. j\l "The fear of the LORD is the beginning of knowledge, but fools despise wisdom and instruction." The Bible, Proverbs 1:7 My Lord, You are my Pinar of strength TABLE OF CONTENTS Contents DECLARATION ........................................................... .. ...... ........ ..... ...... ..... ... ... .. ..... .... ....... .... .... .... ...... i DEDICATION .. ... .. .......... .. ... ..................................... ..... ... .... .. .... ..... .. ....... ... ... ... ............. .... .. ... ...... .. ... ... .ii ACKNOWLEDGEMENTS ......... .... ... .. .. .... .. ...... .. .. ...... ... ..... .. ......... ............. ...... ..... ....... ...... .................... iii TABLE OF CONTENTS .. .. ......... .. ..... ... ..... ... ... ... ..... ........ ............... .. ... ....... ........................... .... ..... ........ vi LIST OF FIGURES ............. ......... ..... ... ... .. .......................... .......... .. .... ..... ..... .. ...... ................................ xii LIST OF TABLES .... ... .. ....... .. .. ... ........ ....... ...... .... .. .. .......... ... .... ...... .... .... ...... ...... ..... .. ... ... ... .. ........ ..... ... xv LIST OF APPENDICES ............· ............... .............................................. ... ..... ..... ................................ xvii LIST OF ABBREVIATIONS, ACRONYMS AND SYMBOLS ............................................. .... .................. xviii DEFINITION OF CONCEPTS ............................... ........ .. ........ ........... .. ...... .......... .......... ... .................... xix CONFERENCES AND CONFERENCE PROCEEDINGS .............. .. .. ..................... ...... .... ....................... xxi ARTICLES PUBLISHED AND MANUSCRIPTS SUBMITTED .... ...... .... ............... .... ...... .. ........ ..... ........... xxii SUMMARY ....... ... .......... .. ........... ...... ...... ... .. .. ... ......... ......... ..... ... .... .......... .. .. ..... .... ........ ................... xx iii CHAPTER 1 ......... .. ... ...... .. ..... ........ ........... ........................................ ................... ................................ 2 INTRODUCTION AND PROBLEM STATEMENT .. .. ...... .... ....................... .. ........ .... .... .. ................... .. ....... 2 1. GENERAL INTRODUCTION ................................................ ............................. ............. .. ..... .... ..... 2 1.1 Introduction and problem statement ........................ ...... .. .... ...... ... .. ..... ... ........ .. .... .. ..................... 2 1.2 Research Problem .. .. .. ................ .. ......... .. .. .. .... .... ........ ........ ............ .. ........................................ 6 1.3 Hypotheses .............. .. ......... ... ...................... .. ........... .... ....... .. .......... ....... ........ ............. .......... .. 6 1.4 Research aim and objectives .. ................................ .. .... .................. .... ..... .. ................................. 6 1.4.1 Aim .... ... .. .................... ............ ......................... ... ............................................ ... .. ................... 6 1.4.2 Objectives .. ........ .. .... ......... .... .. .. .... ............................. .. ............................ ................................ 6 CHAPTER 2 ... ..... .... .... ... ............. .... .... ... .. ........................ ....... ... ... ....... ... ....... ......... ..... ........................ 8 LITERATURE REVIEW .................................................................. .... .... .................. .. ............................ 8 2.1 INTRODUCTION .................................................................... ......................................... .. ... ..... 8 2.2 MEDICINAL PLANTS ..........................................
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