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Molecular Phylogenetics and Medicinal Plants of Asclepiadoideae from India

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Molecular Phylogenetics and Medicinal Plants of Asclepiadoideae from India See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/29843773 Molecular phylogenetics and medicinal plants of Asclepiadoideae from India Article · January 2008 Source: OAI CITATION READS 1 315 1 author: Siddharthan Surveswaran The University of Hong Kong 17 PUBLICATIONS 568 CITATIONS SEE PROFILE All content following this page was uploaded by Siddharthan Surveswaran on 04 March 2015. The user has requested enhancement of the downloaded file. Molecular phylogenetics and medicinal plants of Title Asclepiadoideae from India Author(s) Surveswaran, Siddharthan Citation Issue Date 2008 URL http://hdl.handle.net/10722/53090 Rights unrestricted MOLECULAR PHYLOGENETICS AND MEDICINAL PLANTS OF ASCLEPIADOIDEAE FROM INDIA SIDDHARTHAN SURVESWARAN Ph.D. THESIS The University of Hong Kong 2007 Abstract of the thesis entitled MOLECULAR PHYLOGENETICS AND MEDICINAL PLANTS OF ASCLEPIADOIDEAE FROM INDIA submitted by Siddharthan Surveswaran for the degree of Doctor of Philosophy at The University of Hong Kong in November 2007 Medicinal plants are rich sources of antioxidants and their antioxidant poten- tial is an important factor for disease treatment. Traditional Indian medicine sys- tems employ thousands of herbs in their formulations. A large scale evaluation of antioxidants in Indian medicinal plant species has not been done. In the first part of this study, 133 medicinal plants from 64 families used in Indian tradi- tional medicine systems were systematically screened for antioxidant activities using ABTS (2,2-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid), DPPH (1,1-diphenyl- 2-picrylhydrazyl) and FRAP (Ferric reducing antioxidant power) assays. Signifi- cant and positive linear correlations were found between total antioxidant capacities and phenolic contents (R=0.89–0.97), indicating that phenolic compounds were the dominant antioxidant constituents in the tested medicinal plants. The samples with highest antioxidant activities had high levels of hydrolysable tannins and gallic acid. It was found from the preliminary screening that subfamilies Asclepiadoideae and Periplocoideae (Apocynaceae) had moderately high antioxidant activities (> 5 to 20 mmol TEAC/100 g DW by ABTS assay). Twelve species of medicinal plants belonging to Asclepiadoideae and Periplocoideae were further surveyed for antioxidants, xanthine oxidase inhibition activity and hydroxyl radical scavenging activity. The principal phenolic phytochemicals from these plants were identified by LC-MS, including flavonoids, phenolic acids and phenolic terpenoids. Chlorogenic acid and rutin were detected in almost all the plant samples. The LC-MS analysis provided full fingerprints of the principal phenolic compounds which will also be useful in the authentication and quality evaluation of these medicinal herbs. The family Apocynaceae was revised recently and the former Asclepiadaceae was subsumed into it based on molecular data. Earlier works concentrated on tribal and subtribal divisions of Apocynoideae, and the sampling of Asclepiadoideae was limited. In this study, the phylogeny of Asclepiadoideae was further investigated with improved sampling of the taxa. In the family level analysis of Apocynaceae s.l. using rbcL gene data, the subfami- lies, Asclepiadoideae, Secamonoideae and Periplocoideae, were well resolved. Within Asclepiadoideae, the three tribes were also well resolved. Tribe Asclepiadeae, which is the largest and comprising seven subtribes, was polyphyletic, whereas tribes Mars- denieae and Ceropegieae were monophyletic. Subtribal classification within tribe Asclepiadeae is discussed based on previous morphological work. Molecular systematics of three genera, Ceropegia, Brachystelma and Caralluma of the tribe Ceropegieae (Asclepiadoideae), were studied in detail based on samples collected from India using the nuclear ribosomal internal transcribed spacer (ITS) region, chloroplast trnL, trnT-L, and trnL-F intron and intergenic spacers. The Western Ghats Ceropegia were separated into two major clades that can be differ- entiated based on broad or narrow leaves. Ceropegia was polyphyletic and closely related to Brachystelma. The study of the genus Caralluma and allies showed several clades supporting the revised morphological classification of the genus. MOLECULAR PHYLOGENETICS AND MEDICINAL PLANTS OF ASCLEPIADOIDEAE FROM INDIA by Siddharthan Surveswaran B.Sc. Botany, Loyola College, Madras, India M.Sc. Plant Science, University of Madras, India A thesis submitted in partial fulfillment of the requirements for the Degree of Doctor of Philosophy at The University of Hong Kong November 2007 Declaration I declare that this thesis represents my own work, except where due acknowledge- ment is made, and that it has not been previously included in a thesis, dissertation or report submitted to this University or to any other institution for a degree, diploma or other qualifications. Signed....................................................................... (Surveswaran Siddharthan) i Acknowledgements First of all I would like to express gratitude to my supervisors Dr. Mei Sun and Dr. Harold Corke for their insightful advice, encouragement, guidance and support throughout my PhD studentship period. I am indebted to Dr. Yi-Zhong Cai for his expert guidance in my research and in preparation of manuscripts. I wish to thank the Faculty of Science and Graduate School of the University of Hong Kong for my postgraduate studentship. I would like to express my thanks to Prof. Shrirang Yadav and Dr. Mayur Kamble, Shivaji University, Kolhapur, for their invaluable help in the collection of specimens and discussion of the results. I am also thankful to Prof. T. Pullaiah and Dr. S. Karuppusamy of Sri Krishnadevaraya University, Anantapur, for helping me in collection of specimens. I wish to thank Juan Manuel Laulh´eof www.xerics.com for sending two specimens from the Canaries. Thanks are due to friends at Shivaji University, Nilesh, Mansingh, Shankar, Nilesh, Girish for their loving help during collection. Many thanks to present and past labmates and friends, Dr. Huang Junchao, Ms. Wuyang Huang, Ms. Shan Bin, Dr. Lu Bei, Ms. Vivian Luk, Ms. Zhongquan Sui, Mr. Xiang Li Kong and Dr. Anil Gunaratne, for their help and support during my PhD period. Thanks are due to the timely technical assistance of Ms. M.W. Fong. My thanks to Vijaykrishna Dhanashekar and Belle Damodara Shenoy for teach- ing me phylogenetic analysis. Special thanks go to my friend Senthil Kumar for teaching me LATEX and guiding me in typesetting this thesis with wonderful open source software. ii I express my gratefulness to my wife Vatsala Mrinalini for her love and patience during my study. Finally, I express my gratitude to my parents who have loved and guided me all through my life, without whom my PhD study would not have been possible. I wish to thank all my relatives and friends both in India and Hong Kong who have shared happy moments with me and greatly helped me to achieve my goal. iii Contents Declaration ................................. i Acknowledgements ............................. ii Abbreviations ................................xiv 1 Review of Literature 1 1.1 Free radicals, Antioxidants and Health . ... 1 1.1.1 Antioxidants and their role in cancer . 3 1.2 Phenolic compounds and antioxidation . .. 4 1.3 Alternativemedicine ........................... 5 1.4 Traditional Indian medicinal systems . ... 5 1.5 Antioxidants from Indian medicinal plants . .... 8 1.6 Medicinal plants from Asclepiadoideae and Periplocoideae ...... 9 1.6.1 Antioxidants from Asclepiadoideae and Periplocoideaeplants . 9 1.7 Classification of the family Apocynaceae s.l. ..... 11 1.8 Classification of the subfamily Asclepiadoideae . ....... 12 1.8.1 Salient features of Asclepiadoideae . 13 1.8.2 Molecular systematics of Apocynaceae s.l. .. 16 1.8.3 Recent updates on Apocynaceae s.l. 17 1.9 Classification and phylogeny of the tribe Ceropegieae (Asclepiadoideae) 17 1.9.1 The genus Ceropegia ....................... 17 1.9.2 The genus Caralluma ....................... 26 1.10 Objectives ofthisresearchproject . .... 28 iv 2 Antioxidant properties of Indian medicinal plants and their pheno- lic compounds 31 2.1 Introduction................................ 31 2.2 MaterialsandMethods . 32 2.2.1 Samplecollection . 32 2.2.2 Chemicalsandreagents. 32 2.2.3 Extractpreparation. 33 2.2.4 ABTSassay............................ 33 2.2.5 DPPHAssay ........................... 34 2.2.6 FRAPassay............................ 34 2.2.7 Determination of total phenolic content . .. 35 2.2.8 RP-HPLCanalysis . .. .. 35 2.2.9 Statisticalanalysis . 35 2.3 Results................................... 36 2.3.1 Total antioxidant capacity and phenolic content . .... 36 2.3.2 Relationships among total antioxidant capacities by ABTS, DPPH,andFRAPassays . 50 2.3.3 Relationship between total antioxidant capacity and phenolic content............................... 51 2.3.4 Preliminary identification and analysis of phenolic compounds 52 2.3.5 Discussion............................. 65 3 Antioxidant properties and principal phenolic phytochemicals of Indian medicinal plants from subfamilies Asclepiadoideae and Periplo- coideae 69 3.1 Introduction................................ 69 3.2 MaterialsandMethods . 70 3.2.1 Plantmaterial........................... 70 3.2.2 Chemicalsandreagents. 70 3.2.3 Extractpreparation. 70 v 3.2.4 ABTSassay............................ 70 3.2.5 FRAPassay............................ 71 3.2.6 Xanthine oxidase
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