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Prospects of Arbuscular Mycorrhizal Technology on Growth, Mineral Nutrient Acquisition and Fixed Oil Profile of Selected Sunflow

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Prospects of Arbuscular Mycorrhizal Technology on Growth, Mineral Nutrient Acquisition and Fixed Oil Profile of Selected Sunflow PROSPECTS OF ARBUSCULAR MYCORRHIZAL TECHNOLOGY ON GROWTH, MINERAL NUTRIENT ACQUISITION AND FIXED OIL PROFILE OF SELECTED SUNFLOWER (Helianthus annuus L.) HYBRIDS AT VARIOUS LEVELS OF ROCK PHOSPHATE IN MARGINAL SOIL BY SAYEDA SARAH MUBARAK DEPARTMENT OF BOTANY UNIVERSITY OF PESHAWAR SESSION: 2012-2013 PROSPECTS OF ARBUSCULAR MYCORRHIZAL TECHNOLOGY ON GROWTH, MINERAL NUTRIENT ACQUISITION AND FIXED OIL PROFILE OF SELECTED SUNFLOWER (Helianthus annuus L.) HYBRIDS AT VARIOUS LEVELS OF ROCK PHOSPHATE IN MARGINAL SOIL The thesis submitted to Department of Botany University of Peshawar in partial fulfillment of the requirements for the Degree of Doctor of Philosophy in Botany BY SAYEDA SARAH MUBARAK DEPARTMENT OF BOTANY UNIVERSITY OF PESHAWAR SESSION: 2012-2013 DEDICATION Dedicated to my loving parents, who have been an intimate source of inspiration throughout my life, in whatever I am and whatever I do. UNIVERSITY OF PESHAWAR PESHAWAR PROSPECTS OF ARBUSCULAR MYCORRHIZAL TECHNOLOGY ON GROWTH, MINERAL NUTRIENT ACQUISITION AND FIXED OIL PROFILE OF SELECTED SUNFLOWER (Helianthus annuus L.) HYBRIDS AT VARIOUS LEVELS OF ROCK PHOSPHATE IN MARGINAL SOIL A dissertation submitted in partial satisfaction of the requirement for the degree of Doctor of Philosophy In BOTANY By Sayeda Sarah Mubarak Supervisor: Prof. Dr. Mohammad Ibrar Graduate Studies Committee 1. Prof. Dr. S. Bashir Ahmad, Member 2. Prof. Dr. Muhammad Ibrar, Member 3. Prof. Dr. Muhammad Nafees, Member 4. Dr. Zahir Muhammad, Member 5. Prof. Dr. Siraj ud Din, Convener APPROVAL This thesis, entitled “Prospects of arbuscular mycorrhizal technology on growth, mineral nutrient acquisition and fixed oil profile of selected sunflower (Helianthus annuus L.) hybrids at various levels of rock phosphate in marginal soil” submitted by Ms. Sayeda Sarah Mubarak is hereby approved and recommended as partial fulfillment for the award of Degree of Doctor of Philosophy in BOTANY, _________________________ (External Examiner) Prof. __________________ (Supervispr) Prof. Dr. Mohammad Ibrar Research supervisor Department of Botany, University of Peshawar. Dated:___________ March 21st , 2016 PUBLICATION OPTION I hereby reserved the rights of publication, including right to reproduce this thesis in any form for a period of 5 years from the date of submission. Sayeda Sarah Mubarak VITAE November 14, 1986, Born, Mardan M.phil: Department of Botany University of Peshawar, 2012 M.Sc: Department of Botany University of Peshawar, 2008 B.Sc: F.G. Degree College for women Peshawar, 2006 F.Sc: F.G. Degree College for women Peshawar, 2004 SSc: F.G. Girls Public High School Peshawar Cantt, 2002 Lecturer in Botany, Govt. Frontier College For women Peshawar, 2014---Present. Major field Botany (Mycorrhizal Technology). Botany Courses studied Teacher 1. Agrostology Prof. Dr. Siraj-ud-Din 2. Plant molecular systematic Prof. Dr. Siraj-ud-Din 3. Fresh water algae Prof. Dr. Nadeem 4. Environmental Health and Problems Dr Ghulam dastagir 5. Vegetation ecology Dr. Lal badshah 6. Physiology of plant under Stress Dr. Barkat-ullah TABLE OF CONTENTS S. No. Contents Page No. Acknowledgment i-ii Abstract iii-iv CHAPTER 1 INTRODUCTION 1-19 1.1 Mycorrhiza 1 1.2 Classification of mycorrhiza 2 1.3 Arbuscular mycorrhiza 2 1.4 Distribution of AM fungi 3 1.5 Taxonomy of AM fungi 3 1.6 Morphology of AM fungi 4 1.7 Significance of AM fungi 6 1.7.i VAM as bio-Fertilizer 6 1.7.ii Phosphorus (P) uptake 6 1.7.iii Rock phosphate (RP) uptake 7 1.7.iv Nitrogen (N) uptake 8 1.7.v Potassium (K) uptake 9 1.7.vi Carbon (C) 10 1.7. vii Other macronutrients uptake 10 1.7. viii Zinc (Zn) uptake 10 1.7.ix Other micronutrients uptake 10 1.7.x Disease and pathogen 11 1.7.xi Soil aggregation 11 1.7.xii Hormone production 11 1.7.xiii Salinity 12 1.7.xiv Drought resistance 12 1.7.xv Succession 13 1.7.xvi Phytoremediation 13 1.7.xvii Land rehabilitation 13 1.7.xviii Weed control 14 1.8 Mycorrhizal dependency 14 1.9 Test plant sunflower (Helianthus annuus L.) 14 1.9.i Local names 14 1.9.ii Systematic position 14 1.9.iii Cultivation 16 1.9.iv Botanical description 16 1.9.v Chemical constituents 16 1.9.vi Uses 17 1.9.vii Nutritional values of sunflower oil 17 1.10 Objectives of present study 18 1.11 Expected outcomes 19 CHAPTER 2 LITERATURE REVIEW 20-46 2.1 Effect of AM fungal inoculation on plant growth, yield and nutrient 20 acquisition 2.2 Effect of AMF on spores density and root colonization 31 2.3 Effect of AM fungal inoculation on plant growth, yield and nutrient 36 acquisition in relation to fertilizer application 2.4 Effect of AM fungal inoculation on percent root colonization and 41 spores density in relation to fertilizer application 2.5 Mycorrhizal dependency (MD) 44 CHAPTER 3 MATERIALS AND METHODS 47-59 3.1 Field work 47 3.1.i Equipment and reagents 47 3.1.ii Plant material 48 3.1.iii Experimental site 48 3.1.iv Soil 48 3.1.v Application of AMF inoculum 48 3.1.vi Fertilizer application 49 3.1.vii Experimental design, treatments and replications 49 3.1.vii Evaluation 50 3.1.ix Mycorrhizal dependency 51 3.1.x Statistical analysis 51 3.2 Analysis of plant 51 3.2.i Mineral analysis 51 Wet digestion method for nitrogen 51 Wet digestion for other elements 52 Preparation of stock/standard solutions (100 ppm) 52 Preparation of stock/standard solutions (2.5,5,10 ppm) 52 Instrumental condition for different elements in the samples 52 Procedure 52 3.2.ii Proximate analysis 53 Determination of moisture 53 Determination of ash 53 Determination of crude protein 53 Determination of crude fiber 54 Determination of crude fats 55 Determination of carbohydrates 55 3.2.iii Oil fatty acid profile 55 3.3 Laboratory work 56 Equipment and reagents 56 3.3.i Assessment of roots 56 Collection and preservation of roots 56 Preparation of preservative 56 Preservation 56 Preparation of KOH solution (10%) 56 Preparation of acid fuchsin stain 56 Staining procedure 57 Assessment of root colonization 57 3.3.ii Extraction of spores 57 Collection of soil samples 57 Mounting of spores 58 Calculation of density of spores 58 Identification of spores 58 CHAPTER 4 RESULTS AND DISCUSSION 60- 4.1 Growth parameters 61 4.1.i Plant height 61 4.1.ii Root length 65 4.1.iii Number of fresh leaves 68 4.1.iv Number of wilted leaves 72 4.1.v Leaf length 75 4.1.vi Leaf width 78 4.1.vii Dry weight of plant 81 4.1.viii Head (Capitulum) diameter 85 4.1.ix Number of seeds per head 88 4.1.x Weight of seeds per head 92 4. 2 Oil fatty acid profile 95 4. 2.i Oil content (%) 95 4.2. ii Fatty acid profile 99 Unsaturated fatty acids 99 Linoleic acid (C18:2) 99 Oleic acid (C18:1 cis-9) 103 Saturated fatty acids 106 Palmitic acid (C16:0) 106 Stearic acid (C18:0) 109 4.3 Proximate analysis 118 4.3.i Crude protein (%) 118 4.3.ii Ash content (%) 122 4.3.iii Moisture content (%) 125 4.3.iv Crude fats (%) 128 4.3.v Crude fibers 131 4.3.vi Carbohydrate contents 134 4.4 Mineral composition 137 4.4.i Macronutrients 137 Phosphorus (P) 137 Nitrogen (N) 141 Potassium (K) 144 4.4.ii Micronutrient 147 Zinc (Zn) 147 4.5 Mycorrhizal dependency 151 4.6 Effect of indigenous AMF and rockphosphate levels on spore 153 density and root colonization 4.6.i AMF spore density 153 4.6.ii AMF species 154 4.6.iii AMF colonization in roots 160 CHAPTER 5 CONCLUSIONS AND RECOMMENDATIONS 165-167 5.1 Conclusion 165 5.2 Recommendations 167 CHAPTER 6 REFERENCES 168-225 LIST OF TABLES Table Title Page No. No. 1. Biological materials, time schedule and cultural practice 59 2. Plant Height (cm) in different hybrids of Helianthus annuus L. at various levels of 63 rock phosphate (RP). 3. Root Length (cm) in different hybrids of Helianthus annuus L. at various levels of 66 rock phosphate (RP). 4. Number of Fresh Leaves/plant in different hybrids of Helianthus annuus L. at 70 various levels of rock phosphate (RP). 5. Number of Wilted Leaves/plant in different hybrids of Helianthus annuus L. at 73 various levels of rock phosphate (RP). 6. Leaf Length (cm) in different hybrids of Helianthus annuus L. at various levels of 76 rock phosphate (RP). 7. Leaves Width (cm) in different hybrids of Helianthus annuus L. at various levels of 79 rock phosphate (RP). 8. Dry Weight (gm) in different hybrids of Helianthus annuus L. at various levels of 83 rock phosphate (RP). 9. Head (capitulum) diameter (cm) in different hybrids of Helianthus annuus L. at 86 various levels of rock phosphate (RP). 10. Number of seeds/ Head in different hybrids of Helianthus annuus L. at various levels 90 of rock phosphate (RP). 11. Seeds weight/ Head in different hybrids of Helianthus annuus L. at various levels of 93 rock phosphate (RP). 12. Oil content (%) in different hybrids of Helianthus annuus L. at various levels of rock 97 phosphate (RP). 13. Linoleic acid (%) in different hybrids of Helianthus annuus L. at various levels of 101 rock phosphate (RP). 14. Oleic acid (%) in different hybrids of Helianthus annuus L. at various levels of rock 104 phosphate (RP) 15. Palmatic acid (%) in different hybrids of Helianthus annuus L. at various levels of 107 rock phosphate (RP). 16. Stearic acid (%) in different hybrids of Helianthus annuus L. at various levels of rock 110 phosphate (RP).
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