u Ottawa l.'Univcrsilc! cnnnrlicwu- Cnnodn's univcrsiiy FACULTE DES ETUDES SUPERIEURES ls=l FACULTY OF GRADUATE AND ET POSTOCTORALES U Ottawa POSDOCTORAL STUDIES L*University canadiennc Canada's university Vicky J. Filion AUTEUR DETATHISET/ AUTHOR OF THESIS" M_.Sc. (Biology) GRADE/DEGREE Department of Biology TAMTE"TCOLirDpARTEMENT~^ACUITY, S*CHOdi~DEWRTMENT" A Novel Phytochemical and Ecological Study of the Nunavik Medicinal Plant Rhodiola rosea L. TITRE DE LA THESE / TITLE OF THESIS _____„„____Dr._ J. Arnason A. Cuerrier EXAMINATEURS (EXAMINATRICES) DE LA THESE / THESIS EXAMINERS Dr. C. Charest Dr. J. Kerr Dr. N. Cappuccino _Garjr\V^SJatCT_ Le Doyen de la Faculte des etudes superieures et postdoctorales I Dean of the Faculty of Graduate and Postdoctoral Studies A Novel Phytochemical and Ecological Study of the Nunavik Medicinal Plant Rhodiola rosea L. Vicky J. Filion Thesis submitted to the Faculty of Graduate and Postdoctoral Studies University of Ottawa in partial fulfillment of the requirements for the M.Sc. degree in the Ottawa-Carleton Institute of Biology These soumise a la Faculte des etudes superieures et postdoctorales Universite d'Ottawa en vue de l'obtention de la maitrise es sciences Institut de biologie d'Ottawa-Carleton © Vicky J. 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The author retains copyright L'auteur conserve la propriete du droit d'auteur ownership and moral rights in et des droits moraux qui protege cette these. this thesis. Neither the thesis Ni la these ni des extraits substantiels de nor substantial extracts from it celle-ci ne doivent etre imprimes ou autrement may be printed or otherwise reproduits sans son autorisation. reproduced without the author's permission. In compliance with the Canadian Conformement a la loi canadienne Privacy Act some supporting sur la protection de la vie privee, forms may have been removed quelques formulaires secondaires from this thesis. ont ete enleves de cette these. While these forms may be included Bien que ces formulaires in the document page count, aient inclus dans la pagination, their removal does not represent il n'y aura aucun contenu manquant. any loss of content from the thesis. Canada ABSTRACT Canadian populations of Rhodiola rosea L. growing inNunavik, Quebec, were mapped and examined for the presence of medicinal phytochemicals for the first time. Four relevant phytochemicals salidroside and rosavins, including rosavin, rosarin and rosin, were detected in various Canadian collections. Quantitative comparison of chemical profiles, from different circumpolar samples, revealed significant variations between European and Canadian populations and among Nunavik local populations. In Canada, plants from the Mingan Islands showed the highest quantity of phytochemicals. Salidroside concentration in Nunavik plants was significantly affected by herbivory and gender whereas amount of rosavins remained stable. The ecological survey of Nunavik populations indicated an irregular distribution of R. rosea along Ungava Bay. Distribution maps were created and a predictive distribution model was generated. Plant abundance was observed along the east coast as opposed to near absence on the west coast. Various environmental factors and historical contingency could have influenced this distribution. n RESUME Cette etude porte sur la phytochimie et Tecologie des populations canadiennes du Rhodiola rosea L. au Nunavik. Les quatre composes actifs cles de la plante, soit salidroside et les rosavines, incluant rosavine, rosarine et rosine, ont ete retrouves chez toutes les populations canadiennes etudiees. Le profil phytochimique des populations europeennes et canadiennes a ete compare. Les resultats ont montre que les composes medicinaux variaient significativement entre les populations. Au Canada, la quantite la plus elevee de principes actifs a ete retrouvee chez les populations des iles Mingan. De plus, seul le salidroside contenu dans les plantes du Nunavik a ete affecte par la presence d'acariens et le sexe de la plante. 1/ analyse ecologique a revele que le R. rosea du Nunavik etait distribue inegalement sur les cotes de la Baie d'Ungava (Quebec, Canada), celui-ci etant plus abondant sur la cote est. Plusieurs facteurs environnementaux et historiques peuvent expliquer cette difference. in TABLE OF CONTENT ABSTRACT ii RESUME iii TABLE OF CONTENTS iv LIST OF TABLES vii LIST OF FIGURES viii LIST OF MAPS x ACKNOWLEDGEMENTS xi CHAPTER 1: GENERAL INTRODUCTION: RHODIOLA ROSEA OVERVIEW 1 1.1 General Introduction 2 1.2 Literature Review 3 Plant description 3 Biological properties 6 Phytochemistry 9 Commercialized products 11 1.3 Research Rationale 13 1.4 Research Objectives 13 Phytochemical objectives 13 Ecological objectives 14 CHAPTER 2: PHYTOCHEMICAL ANALYSIS OF RHODIOLA ROSEA L 15 2.1 Introduction 16 2.2. Phytochemical Analysis 17 iv 2.2.1 Material and Methods 17 2.2.2 Results 21 Inter-regional variation ofR. rosea from circumpolar collections.... 21 Inter-population variation ofNunavik R. rosea 24 Variation according to latitude 27 Soil nutrient impact 27 Herbivory impact 27 Effect of plant gender 31 2.2.3 Discussion 31 CHAPTER 3: ECOLOGICAL ANALYSIS OF NUNAVIK RHODIOLA ROSEA L 42 3.1 Introduction 43 3.2 Material and Methods 45 Geographical distribution ofR. rosea 45 Maxent distribution model 46 Habitat characterization in Nunavik 47 Investigation for plant mite infestation 48 Experimental plantation in Nunavik 48 3.3 Results and Discussion 49 Geographical distribution ofR. rosea 49 1. Rhodiola rosea distribution in Nunavik 49 2. Deformed Rhodiola rosea distribution in Nunavik. 56 3. "Worldwide distribution of herbarium specimens 58 Maxent distribution model 62 v Habitat characterization in Nunavik 62 Investigation for plant mite infestation 67 Experimental plantation in Nunavik 72 CHAPTER 4: GENERAL DISCUSSION 75 4.1 Major findings and claims to originality. 76 4.2 Comparison with published literature on R. rosea 77 4.3 Future work 79 APPENDICES 82 Al: Analytical Method Development Manuscript 82 A2: Phytochemical Analysis of Commercialized Rhodiola rosea 89 REFERENCES 95 vi LIST OF TABLES Table 1.1. Six important phytochemical groups and their compounds found in R. rosea....10 Table 2.1. Newly developed HPLC analytical method. Timetable of solvent gradient according to time (minutes) and flow rate (mL/min) 19 Table 2.2. Mean phytochemical concentration of salidroside and rosavins (mg/ g) from four Nunavik soil analyses in relation to three essential soil nutrients: nitrate, phosphorus and potassium 30 Table 3.1. Ecological data compiled from Nunavik collection set 1, August expedition 2005, UngavaBay, QC 64 Table 3.2. Ecological data compiled from Nunavik collection set 2, August expedition 2006, Ungava Bay, QC 65 Table 3.3. Nutrient content (N-P-K) and soil pH level of four soil samples collected in Nunavik, QC, and analyzed according to the Mehlich 3 method, by AgriDirect 68 Table 3.4. Survival rate of the Nunavik experimental plantation established in Kangiqsualujjuaq, August 2006 74 Table A2-1. List of commercialized products ofRhodiola rosea analysed phytochemically for their concentration in the four markers. Products names are arranged in alphabetical order 92 Table A2-2. Product accession numbers were randomly assigned to products. One gram of product was extracted from a homogenized mixture of material. Product # 20 was under liquid form (volume of 50 mL) and subsequently evaporated to dryness and weighted before analysis 93 Table A2-3. Mean phytochemical concentration of salidroside and rosavins (mg/ g) found in twenty commercialized natural products of R. rosea. Accession numbers were attributed randomly 94 vii LIST OF FIGURES Figure 1.1. Morphology of roseroot (Saule 2002) 4 Figure 1.2. Molecular structures of four important phytochemicals present in Nunavik Rhodiola rosea , 12 Figure 2.1. Mean concentration (mg/ g) of salidroside (A) and rosavins (B) with SEM for eight regionally distinct circumpolar collections of Eurasia and Canada 22 Figure 2.2. Concentration (mg/ g) of salidroside and rosavins present in eight circumpolar collections of Eurasia and Canada 23 Figure 2.3. Mean concentration (mg/ g) of salidroside (A) and rosavins (B) with SEM for different populations of Rhodiola rosea collected in August 2005 during the Nunavik collection set 1 expedition 25 Figure 2.4. Mean concentration (mg/ g) of salidroside (A) and rosavins (B) with SEM for different populations of Rhodiola rosea collected in August 2006 during the Nunavik