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Aloe Ferox 117 Table 9: Phytochemical Constituents of Different Extracts of Aloe CIM- Sheetal Leaves 119 International Journal of Scientific & Engineering Research ISSN 2229-5518 1 Morphological, in vitro, Biochemical and Genetic Diversity Studies in Aloe species THESIS SUBMITTED TO OSMANIA UNIVERSITY FOR THE AWARD OF DOCTOR OF PHILOSOPHY IN GENETICS IJSER By B. CHANDRA SEKHAR SINGH DEPARTMENT OF GENETICS OSMANIA UNIVERSITY HYDERABAD - 500007, INDIA JULY, 2015 IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 2 DECLARATION The investigation incorporated in the thesis entitled “Morphological, in vitro, Biochemical and Genetic Diversity Studies in Aloe species’’ was carried out by me at the Department of Genetics, Osmania University, Hyderabad, India under the supervision of Prof. Anupalli Roja Rani, Osmania University, Hyderabad, India. I hereby declare that the work is original and no part of the thesis has been submitted for the award of any other degree or diploma prior to this date. IJSER Date: (Bhaludra Chandra Sekhar Singh) IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 3 DEDICATION I dedicateIJSER this work to my beloved and beautiful wife B. Ananda Sekhar IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 4 Acknowledgements This dissertation is an outcome of direct and indirect contribution of many people, which supplemented my own humble efforts. I like this opportunity to mention specifically some of them and extend my gratefulness to other well wisher, known and unknown. I feel extremely privileged to express my veneration for my superviosor Dr. Anupalli Roja Rani, Professor and Head, Department of Genetics, Osmania University, Hyderabad. Her whole- hearted co-operation, inspiration and encouragement rendered throughout made this in carrying out the research and writing of this thesis possible. I am extremely lucky to execute this under the exceptional guidance. Working under her gave me a true sence of freedom, has obtained my scientific outlook and inculcated a spirit of positive attitude in me. Without her unflinching support and active guidance, the research work presented in the thesis would not have been possible. I extend myIJSER deep sense of reverence and heartful gratitude to Dr. B. Rama Rao, Senior Principal Scientist, Natural Products Laboratory, Indian Institute of Chemical Technology (IICT), Hyderabad for his valuable inspiration. I express my deep regards and fathomless gratitude to him not only for his keen interest and meticulous analysis during the entire period of investigation but also for the great sense of appreciation and encouragement whenever I felt disappointed and distracted. I shall always remain deeply indebted to him for his warmth and constructive criticism. I would like to sincerely thank Dr. Pratibha Nallari, Professor, Department of Genetics, Osmania University for the active support and help provided during the entire study. I would also like to place on record my gratitude to Dr. Smita C. Pawar, Chairperson, Board of IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 5 Studies in Genetics and all the faculty members of Department of Genetics, Osmania University for their encouragement and support. I am highly thankful to all supporting staff, Department of Genetics, who helped me from time to time. Thanks to administrative staff of Osmania University for smooth completion of official formalities. I deem it privilege to extol my profound gratitude and sincere thanks to the management of Indian Immunologicals Ltd; Hyderabad for financial support and encouragement during the period of research work. Fervently and modestly I am pleased to place my profound sense of gratitude with respectful regard and immense pleasure to Prof. P. Prakash Babu, Department of Biotechnology, University of Hyderabad, Hyderabad, Prof. P. B. Kirti, Prof. G. Padmaja, Department of Plant Sciences, University of Hyderabad, Hyderabad, Dr. K. Rajender Rao, Scientist, National Institute of Nutrition (NIN), Hyderabad, Dr. Iliyas Ahemed, Senior Principal Scientist and Dr.B. Ramesha, Senior Scientist, Directorate of Rice Research (DRR), Hyderabad, IJSERDr. PH. Zaidi, Senior Scientist, ICRISAT, Hyderabad, Dr. K. Prabhakar, Senior scientist and Dr. Sherma, Scientist, Indian Institute of Chemical Technology (IICT), Hyderabad for their encouragement and support. I owe a very important debt to Dr. Y. Hari, Scientist, Regional Research Station, Warangal for his co-operation and encouragement. I am equally grateful to Dr. S. D. Basha, Junior Scientist, Kaveri Seeds Company (P) Ltd, Hyderabad and Mrs. T. Latha, Department of Plant Sciences, University of Hyderabad, Hyderabad for their encourage throughout the research. IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 6 I express my heartfelt gratitude to Mr. Chakrapani Pullagummi for his suggestions while planning and executing the experiments in bringing sense of direction to the whole investigation. Each research effort however individual it may be, can be successful, only if it is backed by a good team support Mr. K. Venkatesh, Mr. P. Prem, Mrs. P. Amareswari, Mrs. B. Arun Jyothi, Mrs. J. Saraswathi has actively supported me during a significant period of the research work. I would like to thank profusely Mrs. K. C. Sai Bindu, Mr. Ch. Sudhakar, Mrs. B. Shrawana Kumari and Mr. K. Suresh for their kind support during the course of the study. I am very much beholden and profoundly indebted to my affectionate and beloved wife B. Ananda Sekhar who endures me with patience and perseverance, and my unbounded gratitude forIJSER her moral support and co-operation during the course of the study. I don‟t know what my life would be without her. She believed in me even when I wanted to give up and has taken the load off my shoulder; there I found the first strong evidence of her love and support. I would like to thank my beloved daughter Chi. B. Naina Himavarsha (Cherry) who is always ready to cheer me up and deserve special mention for her unconditional love and support during the course of the study. The active support and encouragement received from my friend Mr. K. Rajeswar Rao has been immense and words fail me to express my gratefulness adequately for the patient discussions, valuable counsel, constant help and co-operation during the couse of the study. IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 7 The blessings received throughout my life from my mother B. Pushpalatha bai and my father Late Sri. B. Nagmal Singh who lighted my life with all sort of happiness and without whose support I would not have reached this stage. I acknowledge my special thanks to my in-laws Dr. S. Madhusudhan Singh and Smt. S. Naina Madhu for their kind assistance during my study period. Finally I take this opportunity to thank one and all who has directly or indirectly helped in completing this task. Last but not least, gratitude to God for blessing my life throughout…. (B. Chandra Sekhar Singh) Place: HyderabadIJSER Date: July, 2015 IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 8 TABLE OF CONTENTS CHAPTER Page No. Chapter 1. Introduction 1 - 16 Chapter 2. Review of Literature 17 - 65 Chapter 3. Material and Methods 66 - 100 Chapter 4. Results 101 - 160 Chapter 5. Discussion 161 - 195 Chapter 6. Summary 196 - 203 Chapter 7. Conclusions 204 - 206 Chapter 8. References 207 - 245 PublicationsIJSER IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 9 LIST OF TABLES Table No. Title of Table Page No. Table 1: Constituents of Aloe vera 28 Table 2: List of RAPD Primers 99 Table 3: List of analyzed ISSR Primers 100 Table 4: Details of Aloe vera accessions used in morphological, in vitro and diversity analysis 106 Table 5 (a): Morphological studies of Aloe accessions 107 Table 5 (b): Morphological studies of Aloe accessions 108 Table 6: Shoot proliferation response from shoot tips cultured on medium with different concentrations of growth regulators 110 Table 7: Effect of auxins on root induction from in vitro regenerated shoots of Aloe 111 Table 8: PhytochemicalIJSER constituents of ethyl acetate and methanol extract of Aloe ferox 117 Table 9: Phytochemical constituents of different extracts of Aloe CIM- Sheetal leaves 119 Table 10: Aloe ferox leaf ethyl acetate and methanol extracts fractions 120 Table 11: Paw volume in control group animals 122 Table 12: Paw volume in standard group animals 122 Table 13: Paw volume in Aloe ferox methanol extract 250mg/kg treated animals 123 Table 14: Paw volume in Aloe ferox methanol extract 500 mg/Kg treated animals 123 Table 15: Aloe ferox leaf extract with Carrageenan Induced Paw Edema in Rats anti- inflammatory activity of Aloe ferox, leave values expressed as Mean ± SD; Number of animals in each group: 6 125 IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 10 Table 16: Antidiabetic study for various groups of SD Rats using Aloe CIM-Sheetal 146 Table 17: Effect of Aloe CIM–Sheetal leaf extracts on body weights (in grams) in control and experimental group rats. 147 Table 18: Jaccard‟s similarity coefficient values of 12 Aloe vera accessions based on pooled data of RAPD and ISSR primers. 157 IJSER IJSER © 2018 http://www.ijser.org International Journal of Scientific & Engineering Research ISSN 2229-5518 11 LIST OF ILLUSTRATIONS Figure No. Title of Figure Page No. Fig 1: Aloe vera (Aloe barbadensis Mill) Plant 02 Fig 2: Aloe ferox plant with red flowers 14 Fig 3: Components of Aloe vera 27 Fig 4: Medicinal uses of Aloe vera 32 Fig 5-8: Aloin (Barbaloin), Isobarbaloin, Aloesin, aloeresin and Aloe emodin 62 Fig 9: Flow diagram of overall procedure of micro propagation 73 Fig 10: Phytochemical extractions of Aloe sps by different organic solvents 79 Fig 11: Chemical structure of Acemannan 82 Fig 12: Column chromatography extractions 84 Fig 13: Different accessions of Aloe vera 103-105 Fig.a.
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