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Species Diversity and Biological Activities of Some Lichens of District Mansehra Department of Botany Hazara University Mansehra

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Species Diversity and Biological Activities of Some Lichens of District Mansehra Department of Botany Hazara University Mansehra SPECIES DIVERSITY AND BIOLOGICAL ACTIVITIES OF SOME LICHENS OF DISTRICT MANSEHRA ASMAT ULLAH DEPARTMENT OF BOTANY HAZARA UNIVERSITY MANSEHRA 2019 HAZARA UNIVERSITY MANSEHRA Department of Botany SPECIES DIVERSITY AND BIOLOGICAL ACTIVITIES OF SOME SELECTED LICHENS OF DISTRICT MANSEHRA By Asmat Ullah This research study has been conducted and reported as partial fulfulment of the requirements of Ph.D degree in Botany awarded by Hazara University Mansehra, Pakistan Date: 30 August, 2019 ii SPECIES DIVERSITY AND BIOLOGICAL ACTIVITIES OF SOME LICHENS OF DISTRICT MANSEHRA Submitted by ASMAT ULLAH PhD Scholar Supervisor DR. ZAFAR IQBAL Assistant Professor Department of Botany Hazara University Mansehra Co-Supervisor DR. MUHAMMAD FIAZ Assistant Professor Department of Botany Hazara University Mansehra DEPARTMENT OF BOTANY HAZARA UNIVERSITY MANSEHRA 2019 iii iv v Plagiarism Undertaking I solemnly declare that research work presented in the thesis titled “Species Diversity and Biological Activities of Some Lichens of District Mansehra” is solely my research work with no significant contribution from any other person. Small contribution/help where taken has been duly acknowledged and that complete thesis has been written by me. I understand the zero tolerance policy of the HEC and Hazara University towards plagiarism. Therefore, I as an Author of the above titled thesis declare that no portion of my thesis has been plagiarized and any material used as reference is properly referred/cited. I undertake that if I am found guilty of any formal plagiarism in the above titled thesis even after award of Ph.D. degree, the University reserves the rights to withdraw/revoke my Ph.D. degree and that HEC and the University has the right to publish my name on the HEC/University Website on which names of students are placed who submitted plagiarized thesis. Student Signature: __________________ Student Name: Asmat Ullah vi AUTHOR’S DECLARATION I Asmat Ullah hereby state that my PhD thesis titled “Species Diversity and Biological Activities of Some Lichens of District Mansehra” is my own work and has not been submitted previously by me for taking any degree from this University (Hazara University Mansehra Pakistan) Or anywhere else in the country/world. At any time if my statement is found to be incorrect even after my Graduate the university has the right to withdraw my PhD degree. Asmat Ullah Date: 30-08-2019 vii DEDICATION To my parents viii ACKNOWLEDGEMENTS All praises be to Almighty Allah Who is The Creater of the universe and knows whatever is in it, hidden or evident, and Who bestowed upon me the intellectual ability and wisdom to look for its secrets. First, I would like to thank my Supervisor Dr. Zafar Iqbal, Department of Botany and Co-supervisor Dr. Muhammad Faiz, Department of Botany for their guidance, encouragement, and patience throughout this tenure. I also express my heartiest and sincerest gratitude to Dr. Fakhr-e-Abbas BRC Islamabad for his support and for giving me the opportunity to carry out my research work and thesis at Bioresource Research Center (BRC) Islamabad. I would like to thank Safia Janjua, Madeeha Manzoor, Kainat Willaim, Abdur Raziq, Fida Muhammad Khan and other BRC staff for their assistance in my research work. I would also like to thank my fellow graduate students Mazhar-ul-Islam, Rashid Ahmad, Ashfaq Ahmad, Amir Khan Afridi and Saeed Muhammad for all their time and help as their assistance, cooperation, and experience proved essential for the completion of the field work. My deep appreciation goes out to Syed Junaid Shah and Syed Mushrraf Shah, the locals of District Mansehra. Their excellent help during field surveys and collection of data made an invaluable contribution to my Ph.D work. I would like to thank various staff members and Ph.D students of the Senckenberg World of Biodiversity Frankfurt, Germany, with whom I had the opportunity to work; Dr. Christian Printzen, Viola Zilller, Heike Kappes, Tetiana Lutsak, Elisa Lagostina. They provided cooperative and a friendly atmosphere at work and also insightful comments on my work. ix I express my special thanks to Andre Aptroot Curator, ABL Herbarium, Soest, Netherlands, for his support in identification of lichen specimens. I would like to say special thanks to my special friends Muhammad Umar, Muhammad Khalil, Imran Khan and Salman Shahzad in Germany for their assistance and cooperation during my stay in Germany on IRSIP fellowship of Higher Education Commission, Pakistan. Finally, I also thank my mother and other family members who encouraged me and prayed for me throughout the time of my research. May the Almighty Allah richly bless all of you? ASMAT ULLAH x TABLE OF CONTENTS Title Page No. ACKNOWLEDGEMENTS ................................................................................. ix LIST OF TABLES .............................................................................................. xiv LIST OF FIGURES ............................................................................................ xvi LIST OF PLATES ................................................................................................ xx LIST OF SYMBOLS/ABBREVIATIONS .................................................... xxiii ABSTRACT ....................................................................................................... xxv Chapter 1 INTRODUCTION......................................................................... 1 1.1 Study Area ............................................................................................... 1 1.1.1 Geography ................................................................................. 1 1.1.2 Climate ....................................................................................... 1 1.1.3 Rivers ......................................................................................... 3 1.1.4 Lakes .......................................................................................... 3 1.1.5 Forests Types ............................................................................ 3 1.2 Lichens ...................................................................................................... 7 1.2.1 General Structure ..................................................................... 7 1.2.2 Occurrence and Distribution .................................................. 8 1.2.3 Climate ....................................................................................... 8 1.2.4 Morphology and Growth Form ............................................. 9 1.2.5 Chemistry ................................................................................ 10 1.2.6 DNA Barcoding ...................................................................... 10 1.2.7 Economic Importance ............................................................ 11 1.2.8 Biological Activities ............................................................... 12 xi 1.2.8.1 Antibacterial Activity ............................................................ 13 1.2.8.2 Antifungal Activity ................................................................ 13 1.2.8.3 Plants Cell Carcinogensis ...................................................... 13 1.3 Study Objectives .................................................................................... 14 Chapter 2 REVIEW OF LITERATURE ...................................................... 15 2.1 Species Diversity and Distribution ..................................................... 15 2.2 Biological Activities of Lichens ........................................................... 21 Chapter 3 MATERIALS AND METHODS............................................... 32 3.1 Field Survey ........................................................................................... 32 3.2 Characterization .................................................................................... 32 3.2.1 Morphological Characters .................................................... 32 3.2.2 Anatomical Characters .......................................................... 33 3.2.3 Chemical Characters .............................................................. 33 3.2.3.1 Spot Tests ................................................................................ 33 3.2.3.2 Thin Layer Chromatography ............................................... 34 3.2.4 DNA Barcoding ...................................................................... 35 3.2.4.1 Genomic DNA Extraction ..................................................... 35 3.2.4.2 Gel Electrophoresis ................................................................ 36 3.2.4.3 Polymerase Chain Reaction (PCR) ...................................... 37 3.3 Identification and Documentation ..................................................... 39 3.4 Distribution Pattern Studies ................................................................ 40 3.5 Biological Activities .............................................................................. 40 3.5.1 Preparation of Lichen Extracts ............................................. 40 3.5.1.1 Antimicrobial Assays ............................................................ 41 xii 3.5.1.2 Cytotoxicity Assay ................................................................. 41 3.5.1.3 Anticarcinogenic Assay ......................................................... 42 3.5.1.4 Antioxidant Assay ................................................................. 42 Chapter 4 RESULTS .....................................................................................
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