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Multiple Approaches for Taxonomic Study of Selected Artemisia L

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MULTIPLE APPROACHES FOR TAXONOMIC STUDY OF SELECTED ARTEMISIA L. SPECIES FROM PAKISTAN By Muhammad Qasim Hayat Department of Plant Sciences Quaid-i-Azam University Islamabad, Pakistan 2011 MULTIPLE APPROACHES FOR TAXONOMIC STUDY OF SELECTED ARTEMISIA L. SPECIES FROM PAKISTAN A MANUSCRIPT PRESENTED TO THE QUAID-I-AZAM UNIVERSITY, ISLAMABAD IN THE PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN PLANT SCIENCES By Muhammad Qasim Hayat Department of Plant Sciences Quaid-i-Azam University Islamabad, Pakistan 2011 FOREIGN EVALUATORS OF THE THESIS 1. DR. SHAHINA A. GHAZANFAR Head of Temperate Regional Team Royal Botanical Garden, Kew Richmond, Surrey London TW9 3AB United Kingdom Email: [email protected] 2. DR. LORNA DAWSON Macaulay Institute Craigiebuckler Aberdeen AB15 United Kingdom Email: [email protected] 3. DR. PEI SHENGJI Professor Kunming Institute of Botany Chinese Academy of Sciences, Heilongtan Kunming 650204 China Email: [email protected] CERTIFICATION This thesis by Mr. Muhammad Qasim Hayat is accepted in its present form by the Department of Plant Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan as satisfying the thesis requirement for the degree of Doctor of Philosophy in Plant Sciences. Supervisor Dr. MUHAMMAD ASHRAF Professor of Cell Biology NUST Centre of Virology & Immunology National University of Sciences & Technology H-12 Islamabad, Pakistan External Examiner I Dr. M. K. LEGHARI Director Botanical Science Division Pakistan Museum of Natural History Garden Avenue, Shakarparian Islamabad, Pakistan External Examiner II Dr. MUQARRAB SHAH Professor & Dean Department of Microbiology Faculty of Health Sciences Hazara University Mansehra, Pakistan Chairperson Dr. ASGHARY BANO Professor & Chairman Department of Plant Sciences Quaid-i-Azam University Islamabad, Pakistan Date May, 24, 2011 i DEDICATION I DEDICATED MY THESIS TO MY SPIRITUAL MASTER “HAZOOR SUFI IQBAL MUHAMMAD” ii TABLE OF CONTENTS Chapter Title Page CERTIFICATION i DEDICATION ii TABLE OF CONTENTS iii LIST OF TABLES vi LIST OF PLATES vii LIST OF FIGURES ix LIST OF ABBREVIATIONS xi ACKNOWLEDGEMENT xii LIST OF PUBLICATIONS xiii ABSTRACT xiv 1 INTRODUCTION 1-25 1.1. Origin, development and diversification of Artemisia 1 in the geological past 1.2. Evolutionary relationships with other groups in the 1 family Asteraceae 1.3. Pollen Evolution 2 1.4. Kariology 3 1.5. Ecology, center of diversity and geographical 3 distribution 1.6. Life form and morphology 5 1.7. Phenology and reproductive biology 5 1.8. Historical developments in classification 5 1.9. Molecular phylogeny 8 1.10. Generic delimitation 9 1.10.1. Infrageneric classification 10 1.11. Economic importance 12 1.11.1. Medicinal importance 12 1.11.2. Insecticidal, repellent and insect antifeedent 14 properties 1.11.3. Ecological importance 14 1.11.4. Other uses 15 1.12. Perspectives in the study of Artemisia L. 15 1.13. Introduction to the study area 16 1.13.1. Location 16 1.13.2. Geology 16 1.13.3. Climate 17 1.13.4. Phytogeographic regions 17 1.14. Artemisia in Flora of Pakistan 19 1.15. Artemisia in Pakistani - Economic potentials 19 1.15. Justification of Present work 22 1.16. Objectives of Present Study 24 2 REVIEW OF LITERATURE 26-38 2.1. Ethnotaxonomy 26 2.2. Morphology 26 2.3. Leaf epidermal anatomy 31 2.4. Palynology 33 iii 2.5. Phytogeography 35 2.6. Molecular phylogeny 35 2.7. Chemotaxonomy based on Elemental Contents 38 3 MATERIALS AND METHODS 39-66 3.1. Ethnotaxonomy 39 3.2. Morphology 39 3.3. Leaf epidermal anatomy 44 3.4. Palynology 54 3.5. Phytogeography 56 3.6. Molecular phylogeny 58 3.6.1. DNA Extraction 58 3.6.1.1. CTAB method 58 3.6.1.2. DNeasy kit (QIAGEN) method 59 3.6.2. PCR 59 3.6.3. PCR products Purification 59 3.6.4. Sequencing PCR 59 3.6.5. Sequencing 59 3.6.6. Sequence processing 64 3.6.7. Phylogenetic Analysis 64 3.6.7. Bootstrapping 64 3.6.7. Maximum parsimonious analysis 64 3.6.7. Bayesian inference 64 3.6.7. Neighbor joining trees 65 3.6.8. Maximum Like-Hood Phylogenetic trees 65 3.7. Diversity in elemental contents 65 3.7.1. Sample preparation 65 3.7.2. Digestion 65 3.7.3. Atomic absorption Spectrophotometery 66 3.7.3. Statistical analysis 66 3.7.3.1 Principal Component Analysis (PCA 66 3.7.3.2 Cluster Analysis 66 4 RESULTS 67-164 4.1. Ethnotaxonomy 67 4.2. Morphology 88 4.3. Leaf epidermal anatomy 100 4.3.1. Leaf epidermal cells and stomata 100 4.3.2. Foliar trichomes 100 4.3.3. Phylogenetic relationships based on distribution of foliar trichomes in the genus Artemisia 102 4.4. Palynology 128 4.4.1 Phylogenetic analysis of Artemisia l. (Asteraceae) based on micromorphological traits of pollen grains 128 4.5. Phytogeography 139 4.6. Molecular phylogeny 144 4.6.1. Neighbor joining consensus trees 144 4.6.2. Phylogenetic tree with posterior probabilities 144 4.6.3. Maximum likehood phylogenetic tree 144 4.6.4 Maximum parsimonious tree 144 4.7. Chemotaxonomy based on elemental diversity 158 4.7.1 Cluster analysis 158 iv 4.7.2 Principal component analysis 158 5 DISCUSSION 165-180 5.1. Ethnotaxonomy 165 5.2. Morphology 166 5.3. Leaf epidermal anatomy 168 5.4. Palynology 171 5.5. Phytogeography 173 5.6. Molecular phylogeny 173 5.7. Diversity in elemental contents 175 5.8. Conclusions 179 5.9. Recommendations 180 REFERENCES 182-203 v LIST OF TABLES Sr. Title Page 1.1. Comparison among different infrageneric classifications of Artemisia 7 1.2. Phytogeographical Regions in Pakistan 20 2.1. Ethnobotanically identified Artemisia species in the world 27 2.2. Ethnobotanically identified Artemisia species in Pakistan 28 3.1. List of taxa studied and their herbarium vouchers 41-43 3.2. Morphological characters and character states of Artemisia for the phylogenetic analysis. 45-46 3.3. Data matrix used in phylogenetic analysis of Artemisia based on morphological features. 47-50 3.4. Character and character states of foliar trichomes for the Phylogenetic analysis of Artemisia 52 3.5. Data matrix used in phylogenetic analysis of Artemisia based on foliar trichomes 53 3.6. Characters and character states of pollen grains for the phylogenetic analysis of Artemisia. 55 3.7. Character state matrix used in phylogenetic analysis of Artemisia based on pollen features. 57 3.8. Composition of PCR reaction mixture per sample 61 3.9. PCR conditions for genomic amplification 61 3.10. Details of primers used in this study 62 3.11. Reaction mixture composition for sequencing PCR per Sample 63 3.12. Conditions for sequencing PCR 63 4.1. Stomatal characteristics in Artemisia species 103-104 4.2. Guard cells characteristics in Artemisia species 105-106 4.3. Stomatal comples characteristics in Artemisia species 107-108 4.4. Foliar epidermal cells characteristics in Artemisia species 109-110 4.5. Quantitative characteristics of glandular foliar trichomes of Artemisia 123 4.6. Quantitative characteristics of nonglandular foliar trichomes of Artemisia 124-125 4.7. Pollen quantitative characteristics of the studied taxa 129-130 4.8. Chromosome numbers (2n), basic numbers (X) and ploidy levels (PL) of the Pakistani Artemisia 140-141 4.9. Average concentration (ppm) (n=5) along with standard deviation of the elements in studied Artemisia species. 159-160 vi LIST OF PLATES Sr. Title Page 4.1. Artemisia dubia 76 4.2. A. moorcraftiana 76 4.3. A. vulgaris (A: young plants, B: Inflorescence) 77 4.4. A. maritima 77 4.5. A. scoparia 78 4.6. A. santolinifolia 79 4.7. A. herba-alba 80 4.8. A. dubia (A: Young plants, B: Flowers) 80 4.9. A. dracunculus 81 4.10. A. annua (A: Young plants, B: Flowers) 81 4.11. A. absinthium (A: Young plants, B: Flowers) 82 4.12. A. absinthium dry plant 82 4.13. S. brevifolium Population 83 4.14. S. brevifolium 83 4.15. A. roxburghiana 84 4.16. A. roxburghiana 84 4.17. A. tournifortiana 85 4.18. A. biannus inflorescence 86 4.19. A. japonica 87 4.20. A. japonica Leaves 87 4.21. Stomatal variations in Artemisia by means of SEM. 111 4.22. Stomatal variations in Artemisia by means of LM. 112 4.23. Adaxial leaf epidermal cells in Artemisia tangutica by means of SEM 113 4.24. Abaxial foliar surface variations in Artemisia by means of LM 114 4.25. Adaxial foliar surface variations in Artemisia by means of LM 115 4.26. Types of glandular foliar trichomes in Artemisia by means of LM 116 4.26. Types of glandular and nonglandular foliar trichomes in Artemisia by means of LM 117 4. 28. Types of glandular foliar trichomes in Artemisia by means of SEM 118 4.29. Types of nonglandular foliar trichomes in Artemisia by means of SEM 119 4.30. Types of glandular and nonglandular foliar trichomes in Artemisia by means of LM 120 4.31. Types of glandular and nonglandular foliar trichomes in Artemisia by means of SEM 121 vii 4.32. Types of glandular foliar trichomes in Artemisia by means of LM 121 4.33. Scanning electron micrographs showing the equatorial view of pollens of studied taxa 131 4.34. Scanning electron micrographs showing the polar view of pollens of studied taxa 132 4.35. Scanning electron micrographs showing the exine sculpture of studied Taxa 133 4.36. Light micrographs of pollen grains 135 4.37. Scanning electron micrographs of the taxa studied 137 4.38. Scanning electron micrographs of the taxa studied 138 4.39. Gel image of ETS PCR products of studied Artemisia Species 145 viii LIST OF FIGURES Sr.
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  • Phylogenetic Taxonomy of Artemisia L. Species from Kazakhstan Based On

    Phylogenetic Taxonomy of Artemisia L. Species from Kazakhstan Based On

    PROCEEDINGS OF THE LATVIAN ACADEMY OF SCIENCES. Section B, Vol. 72 (2018), No. 1 (712), pp. 29–37. DOI: 10.1515/prolas-2017-0068 PHYLOGENETIC TAXONOMY OF ARTEMISIA L. SPECIES FROM KAZAKHSTAN BASED ON MATK ANALYSES Yerlan Turuspekov1,5, Yuliya Genievskaya1, Aida Baibulatova1, Alibek Zatybekov1, Yuri Kotuhov2, Margarita Ishmuratova3, Akzhunis Imanbayeva4, and Saule Abugalieva1,5,# 1 Institute of Plant Biology and Biotechnology, 45 Timiryazev Street, Almaty, KAZAKHSTAN 2 Altai Botanical Garden, Ridder, KAZAKHSTAN 3 Karaganda State University, Karaganda, KAZAKHSTAN 4 Mangyshlak Experimental Botanical Garden, Aktau, KAZAKHSTAN 5 Al-Farabi Kazakh National University, Biodiversity and Bioresources Department, Almaty, KAZAKHSTAN # Corresponding author, [email protected] Communicated by Isaak Rashal The genus Artemisia is one of the largest of the Asteraceae family. It is abundant and diverse, with complex taxonomic relations. In order to expand the knowledge about the classification of Kazakhstan species and compare it with classical studies, matK genes of nine local species in- cluding endemic were sequenced. The infrageneric rank of one of them (A. kotuchovii) had re- mained unknown. In this study, we analysed results of sequences using two methods — NJ and MP and compared them with a median-joining haplotype network. As a result, monophyletic origin of the genus and subgenus Dracunculus was confirmed. Closeness of A. kotuchovii to other spe- cies of Dracunculus suggests its belonging to this subgenus. Generally, matK was shown as a useful barcode marker for the identification and investigation of Artemisia genus. Key words: Artemisia, Artemisia kotuchovii, DNA barcoding, haplotype network. INTRODUCTION (Bremer, 1994; Torrel et al., 1999). Due to the large amount of species in the genus, their classification is still complex Artemisia of the family Asteraceae is a genus with great and not fully completed.
  • Agriculture) Programme

    Agriculture) Programme

    St. PETER’S UNIVERSITY St. Peter’s Institute of Higher Education and Research (Declared under section 3 of UGC Act 1956) Avadi, Chennai – 600 054. B.Sc. (AGRICULTURE) PROGRAMME (3 Years) (I to VI SEMESTERS) REGULATIONS AND SYLLABI (REGULATIONS – 2017) (Effective from the Academic Year 2017-’18) B.Sc. (AGRICULTURE) PROGRAMME Regulations and Syllabi (Effective from the Academic Year 2017-’18) 1. Eligibility: Candidates who passed Higher Secondary Examination with Biology, Physics and Chemistry conducted by the Government of Tamil Nadu or an examination accepted by the University as equivalent thereto are eligible for admission to Three Year B.Sc. Programme in AGRICULTURE. 2. Duration: Three Years comprising 6 Semesters. Each semester has a minimum 90 working days with a minimum of 5 hours a day. 3. Medium: English is the medium of instruction and examinations except for the language subjects. 4. Eligibility for the Award of Degree: A candidate shall be eligible for the award of degree only if he/she has undergone the prescribed course of study in the University for a period of not less than three academic years (6 semesters), passed the examinations of all the six semesters prescribed carrying 150 credits and also fulfilled such conditions as have been prescribed thereof. 5. Choice Based Credit System: Choice Based Credit System is followed with one credit equivalent to 18 hours of study with a total of 25 credits in the Time Table in a semester. The total credit for the programme (6 semesters) is 150. [[ 6. Weightage for a Continuous and End Assessment: The weightage for Continuous Assessment (CA) and End Assessment (EA) is 25:75 unless the ratio is specifically mentioned in the Scheme of Examinations.
  • Study of Evaluation of Antimicrobial Property of Different Concentrations

    Study of Evaluation of Antimicrobial Property of Different Concentrations

    Indian Journal of Basic and Applied Medical Research; March 2019: Vol.-8, Issue- 2, P. 154 - 157 Original article: Study of evaluation of antimicrobial property of different concentrations of Artemisia pallens (Davana) extract against Streptococcus mutans serotype c (ATCC 25175) Dr Pankti Gajjar , Dr Rahul Deshpande , Dr Tarun Kukreja Department of Pedodontics , Dr D Y Patil Dental College and Hospital , Pimpri , Pune [ Dr DY Patil Vidyapeeth ] Corresponding author* Abstract Introduction: Caries is recognized as the greatest prevalent disorder existing around the world since times .Caries is nearly the common of all diseases and partly because of its relatively rapid progress, and correlation of various factors i.e bacteria, dental plaque ,saliva, fermentable carbohydrates and concentration of different ions in the environment surrounding the tooth determines the extent and the rate at which metabolic events may result in net loss of mineral and development and progression of the lesion. In adolsecents , caries is the particular cause of loss of teeth..” Methodology: The microbial inhibition assay was performed by standard norms and protocol by using the agar well diffusion method. Five different concentrations of Artemisia pallens (Davana) extract and 0.2% chlorhexidine as a gold standard was evaluated for their antimicrobial properties(Zone of Inhibition) in triplicate against streptococcus mutans serotype c(ATCC 25175). Adequate amount of Mueller Hinton Agar was evenly distributed over the surface of 15 cm diameter petridish to a thickness of 5 mm and was allowed to solidify under aseptic conditions. Results and Conclusion: This study evaluated the bacterial growth inhibition of the derived acetone extracts of Artemisia Pallens plant.