Grasses of the Neelum Valley Azad Jammu and Kashmir: Systematics, Anatomy and Physiology
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GRASSES OF THE NEELUM VALLEY AZAD JAMMU AND KASHMIR: SYSTEMATICS, ANATOMY AND PHYSIOLOGY BY KHAWAJA SHAFIQUE AHMAD Regd. No. 2011-ag-17 M. Phil. (UAAR) A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN BOTANY DEPARTMENT OF BOTANY FACULTY OF SCIENCES UNIVERSITY OF AGRICULTURE FAISALABAD PAKISTAN 2015 DECLARATION I hereby declare that the contents of the thesis “Grasses of the Neelum Valley Azad Jammu and Kashmir: Systematics, anatomy and physiology” are the product of my own research and no part has been copied from any published source (except the references, standard mathematical or genetic models/ equations/ formulae/ protocols, etc). I further declare that this work has not been submitted for award of any other degree/ diploma. The University may take action if the information provided is found inaccurate at any stage. (In case of any fault the scholar will be proceeded against as per HEC plagiarism policy). Khawaja Shafique Ahmad 2011-ag-17 i To The Controller of Examinations, University of Agriculture, Faisalabad. “We, the Supervisory Committee, certify that the contents and form of thesis submitted by Mr. Khawaja Shafique Ahmad, Regd. #. 2011-ag-17, have been found satisfactory and recommend that it be processed for evaluation by the external Examiner (s) for the award of degree” Supervisory Committee 1. Chairman ------------------------------- Dr. Mansoor Hameed 2. Member ------------------------------- Dr. Farooq Ahmad 3. Member ------------------------------- Dr. Bushra Sadia ii ACKNOWLEDGEMENT Firstly, I offer my adoration to ALLAH Almighty who gave me the strength and courage to complete my thesis work as well as the opportunity to thank all those people through whom his grace was delivered to me. Indeed the words at my command are not adequate to convey the depth of my feeling and gratitude to my supervisor Dr. Mansoor Hameed, Associate Professor, Department of Botany, University of Agriculture, Faisalabad, the most valuable and inspiring guidance, concrete suggestions, constant encouragement, expect evaluation, enormous help and constructive criticism throughout the course of this investigation and preparation of this thesis. With reverence and gratitude, I would like to thank Dr. Farooq Ahmad Department of Botany, University of Agriculture, Faisalabad, whose help, stimulating suggestions, and encouragement helped me throughout the course. I am very much thankful to Dr. Bushra Sadia, Centre of Agriculture Biochemistry and Biotechnology, University of Agriculture, Faisalabad for her help, guidance and motivation during all the research work. Sincere thanks to Dr. Abdul Wahid (Chairman), Dr. Muhammad Sajid Aqeel Ahmad, Dr. Mumtaz Hussain, Dr. Shehbaz, my lab members, Dr. Riffat Batool, Dr. Noreen Akhtar, Sana Fatima, Mehvish Naseer and my friends especially Muhammad Salman, Aqeel Butt, Muhammad Asif Shehzad, Waqas Ahmad, Aqil Sarwar, Sharif Abdullah, Abdul Hameed and Khawaja Baber for their kindness and moral support during my study. Thanks for the friendship and memories. I thank my parents, for striving hard to provide a good education for my siblings and I always fall short of words and felt impossible to describe their support. You have contributed irreversibly to the person I have become. I can’t thank you enough. My dear brothers Saeed Shad and Rafique Parwana, my dear sisters, Zubaida and Ayesha and my nephews, Saad and Abdullah who always are excited to hear of my success and that inspires me to perform better and be successful. I acknowledge my entire family for providing me a very educated atmosphere. For any errors or inadequacies that may remain in this work, of course, the responsibility is entirely my own. Khawaja Shafique Ahmad iii DEDICATED TO MY SUPERVISOR DR. MANSOOR HAMEED iv CONTENTS CHAPTER TITLE PAGE 1. INTRODUCTION 1 2. REVIEW OF LITERATURE 8 3. MATERIALS AND METHODS 21 4. RESULTS 31 5. DISCUSSION 175 6. SUMMARY 185 LITERATURE CITED 197 v Chapter List of contents Page 1. Introduction 1 2. Review of literature 13 2.1. Distributional range of grasses 13 2.1.1. Distribution in Pakistan 13 2.1.2. Worldwide distribution 14 2.2. Phytosociology 16 2.3. Systematics 17 2.4. Anatomy 19 2.5. Physiology 23 2.6. Modifications for environmental stresses 24 3. Materials and methods 27 3.1. Site selection 27 3.2. Collection and preservation of grasses 27 3.3. Edaphology 27 3.3.1. Soil collection 27 3.3.2. Soil texture 29 3.3.3. Saturation percentage 29 3.3.4. Soil ph and electrical conductivity (ece) 29 3.3.5. Organic matter (Walbley) 29 3.3.6. Total Nitrogen (Kjeldahl method) 31 3.3.7. Phosphorus (Olsen method) 31 3.3.8. Soil ionic content (Flame photometric method) 32 3.4. Phytosociology 32 3.4.1. Vegetation sampling 32 3.4.2. Species association 33 3.5. Morphological parameters 33 3.6. Anatomical parameters 34 3.6.1. Root anatomy 34 3.6.2. Stem anatomy 34 3.6.3. Leaf-sheath anatomy 35 3.6.4. Leaf blade anatomy 35 3.7. Physiological Parameters 35 3.7.1. Analysis of plant nutrients 35 3.7.2. Determination of shoot and root ionic content 36 3.7.3. Chlorophyll contents 36 3.8. Cladistic analysis of Andropogonae tribe 36 3.8.1. Morphological data and selection of outgroups 36 3.8.2. Gene bank data 37 3.8.3. Phylogenetic analysis 37 3.9. Statistical analysis 38 4. Results 39 4.1. Soil analysis 39 4.1.1. Soil physiochemical characteristics 39 4.1.2. Soil RDA analysis 41 4.2. Phytosociology 41 4.2.1. Taxonomic diversity 41 4.2.2. Relative density 45 4.2.3. CCA analysis 48 4.2.4. Relative frequency 48 4.2.5. CCA analysis 51 vi 4.2.6. Relative cover 53 4.2.7. CCA analysis 54 4.2.8. Importance value 56 4.2.9. CCA analysis 57 4.2.10. Species association analysis 59 4.3. Morphology 69 4.3.1. Plant height (cm) 69 4.3.2. Root length (cm) 69 4.3.3. Root dry weight (g) 72 4.3.4. Shoot dry weight (g) 72 4.3.5. Ligule length (cm) 72 4.3.6. Tiller Plant-1 72 4.3.7. Leaves Plant-1 74 4.3.8. Inflorescence length (cm) 74 4.3.9. Spike length (cm) 74 4.3.10. Spikelet numbers 74 4.3.11. Multivariate cluster analysis 74 4.4. Anatomy 75 4.4.1. Root anatomy 75 4.4.1.1. Root cross sectional area (µm2) 75 4.4.1.2. Root hair length (µm) 75 4.4.1.3 Epidermal thickness (µm) 77 4.4.1.4. Cortical thickness (µm) 77 4.4.1.5. Cortical cell area (µm2) 77 4.4.1.6. Endodermal cell area (µm2) 80 4.4.1.7. Pericyle thickness (µm) 80 4.4.1.8. Metaxylem vessel number 80 4.4.1.9. Metaxylem cell area (µm2) 80 4.4.1.10. Pith area (µm) 80 4.4.1.11 Pith cell area (µm)2 82 4.4.1.12. Multivariate cluster analysis 82 4.4.2. Stem anatomy 84 4.4.2.1. Stem cross sectional area (mm2) 84 4.4.2.2. Epidermal thickness (µm) 84 4.4.2.3. Sclerenchyma thickness (µm) 84 4.4.2.4. Cortical cell area (µm2) 86 4.4.2.5. Vascular bundle area (µm2) 86 4.4.2.6. Metaxylem area (µm2) 86 4.4.2.7. Phloem area 86 4.4.2.8. Multivariate cluster analysis 86 4.4.3. Leaf sheath anatomy 88 4.4.3.1. Sheath thickness (µm) 88 4.4.3.2. Sclerenchyma thickness (µm) 88 4.4.3.3. Epidermal cell area (µm2) 88 4.4.3.4. Vascular bundle area 91 4.4.3.5. Multivariate cluster analysis 91 4.4.4. Leaf blade anatomy 93 4.4.4.1. Leaf blade thickness (µm) 93 4.4.4.2. Lower epidermal thickness (µm) 93 4.4.4.3. Upper epidermal thickness (µm) 93 vii 4.4.4.4. Sclerenchyma thickness (µm) 93 4.4.4.5. Mesophyll thickness (µm) 95 4.4.4.6. Bundle sheath thickness (µm) 95 4.4.4.7. Vascular bundle area (µm2) 95 4.4.4.8. Metaxylem vessel area (µm2) 95 4.4.4.9. Phloem area (µm2) 97 4.4.4.10. Bulliform cell area (µm2) 97 4.4.4.11. Adaxial stomatal area (µm2) 97 4.4.4.12. Abaxial stomatal area (µm2) 97 4.4.4.13. Adaxial stomatal number 97 4.4.4.14. Abaxial stomatal number 99 4.4.4.15. Multivariate cluster analysis 99 4.4.4.16. Leaf structural modifications in grasses 101 4.5. Physiology 121 4.5.1. Root Sodium (mg g-1) 121 4.5.2. Shoot Sodium (mg g-1) 121 4.5.3. Root Potassium (mg g-1) 121 4.5.4. Shoot Potassium (mg g-1) 124 4.5.5. Root Calcium (mg g-1) 124 4.5.6. Shoot Calcium (mg g-1) 124 4.5.7. Root Magnesium (mg g-1) 124 4.5.8. Shoot Magnesium (mg g-1) 125 4.5.9. Root Phosphorous (mg g-1) 125 4.5.10. Shoot Phosphorous (mg g-1) 125 4.5.11. Total chlorophyll Contents 125 4.5.12. Multivariate Cluster analysis 128 4.5.13. CCA analysis of root nutrients 128 4.5.14. CCA analysis of shoot nutrients 131 4.6. Cladistic analysis of tribe Andropogoneae 131 5. Discussion 138 6. Summary 145 7. Literature cited 149 viii List of Figures Figure Title Page 1. Panoromic views of various study sites: a. Janawai, b. Taobut, c. Jura, 10 d, Kundal Shahi, e Sardari, and f. Halmat 2. Panoromic views of various study sites: a.