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Role of Nattrassia Sp. in Fruit Orchard Decline And

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Role of Nattrassia Sp. in Fruit Orchard Decline And 1 ROLE OF NATTRASSIA SP. IN FRUIT ORCHARD DECLINE AND DETERMINATION OF KEY FACTORS FOR ITS MANAGEMENT FAISAL SOHAIL FATEH 94-arid-224 Department of Plant Pathology Faculty of Crop and Food Sciences Pir Mehr Ali Shah Arid Agriculture University Rawalpindi Pakistan 2018 2 ROLE OF NATTRASSIA SP. IN FRUIT ORCHARD DECLINE AND DETERMINATION OF KEY FACTORS FOR ITS MANAGEMENT by FAISAL SOHAIL FATEH (94-arid-224) A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Plant Pathology Department of Plant Pathology Faculty of Crop and Food Sciences PirMehr Ali Shah Arid Agriculture University Rawalpindi Pakistan 2018 3 4 5 6 7 "In the Name of Allah, the most Beneficent, the most Merciful" 8 DEDICATION To MyAffectionate, Loving Parents & My Sweet Family 9 CONTENTS Page List of Tables xii List of Figures xvii List of Acronyms xx Acknowledgements xxi ABSTRACT xxii 1 GENERAL INTRODUCTION 1 1.1 EXPORT OF TARGET FRUITS 1 1.2 AREA AND PRODUCTION OF TARGET 2 ORCHARDS IN PAKISTAN 1.3 FRUIT FACING DECLINE 2 1.3.1 Citrus Decline 2 1.3.2 Mango Decline 4 1.3.2.1 Progressive or slow decline or dieback 4 1.3.2.2 Quick dieback 4 1.3.2.3 Mango sudden death (MSD) 5 1.3.2 Guava Decline 5 1.4 PATHOGENS INVOLVED IN DECLINE 6 1.4.1 Nattrassia mangiferae 6 1.4.2 Synonyms 6 1.4.3 Taxonomy 6 1.4.4 Host Range 7 1.4.5 Symptoms 7 1.4.6 Biology and Management 7 2 OCCURRENCE AND SYMPTOMATOLOGY OF CITRUS, 9 MANGO AND GUAVA DECLINE IN PUNJAB, PAKISTAN 10 2.1 INTRODUCTION 9 2.2 REVIEW OF LITERATURE 12 2.2.1 Decline of Fruit Orchards 12 2.2.1.1 Citrus decline 12 2.2.1.2 Mango decline 14 2.2.1.3 Guava decline 15 2.3 MATERIALS AND METHODS 17 2.3.1 Survey of Target Fruit Orchards for Disease Assessment 17 2.3.1.1 Profile of areas surveyed 18 2.3.1.1.1 Sargodha 18 2.3.1.1.2 Multan 18 2.3.1.1.3 Northern Irrigated Plan (Zone Iva) 19 2.3.2 Disease Incidence 35 2.3.3 Disease Severity 35 2.3.4 Disease Index 35 2.3.5 Symptomatology 36 2.4 RESULTS 36 2.4.1 Disease Assessment 36 2.4.1.1 Citrus decline assessment 36 2.4.1.2 Mango decline assessment 38 2.4.1.3 Guava decline assessment 47 2.4.2 Symptomatology 51 2.5 DISCUSSION 62 3 PREVALENCE OFNATTRASSIASP. FROM CITRUS, 64 MANGOAND GUAVA ORCHARDS ALONG WITH OTHER DECLINE CAUSING FUNGI 3.1 INTRODUCTION 64 3.2 REVIEW OF LITERATURE 66 11 3.3 MATERIALS AND METHODS 68 3.3.1 Sampling Methodology 68 3.3.2 Isolation and Identification of Pathogens 69 3.3.2.1 Potato dextrose agar medium (PDA) 69 3.3.2.2 Malt extract agar (MEA) 69 3.3.2.3 Stem decoction glucose agar (SDGA) 69 3.3.2.4 Sterilization 70 3.3.2.5 Isolation from infected tissues 70 3.3.3 Microscopy 70 3.3.4 Determination of Fungal Frequency Percantage 71 3.3.5 Purification of Fungal Cultures 71 3.4 RESULTS 71 3.4.1 Plant Parts Wise Fungal Frequency Percentage 71 3.4.1.1 Mycoflora from citrus decline affected trees 71 3.4.1.2 Mycoflora from mango decline affected trees 72 3.4.1.3 Mycoflora from guava decline affected trees 72 3.5 DISCUSSION 93 4 ROLE OFNATTRASSIA MANGIFERAEAND 97 OTHERDECLINE CAUSING FUNGI IN SYMPTOMS DEVELOPMENT 4.1 INTRODUCTION 97 4.2 REVIEW OF LITERATURE 99 4.3 MATERIALS AND METHODS 101 4.3.1 Pathogenicity Tests 101 4.3.1.1 Flap method 101 4.3.1.2 Root injury method 101 12 4.4 RESULTS 103 4.5 DISCUSSION 111 5 STUDIES ON FACTORS AFFECTING DEVELOPMENT 114 OFFUNGI WITH SPECIAL REFERENCE TO NATTRASSIA SP. 5.1 INTRODUCTION 114 5.2 REVIEW OF LITERATURE 114 5.3 MATERIALS AND METHODS 117 5.3.1 Effect of Temperature, Light, pH and Culture Media 117 onFungal Growth 5.3.1.1 Effect of culture media 117 5.3.1.2 Effect of temperature 118 5.3.1.3 Effect of light 118 5.3.1.4 Effect of Ph 118 5.4 RESULTS 119 5.4.1 Effect of Culture Media on Radial Mycelial Growth 119 5.4.2 Effect of Temperature on Radial Mycelial Growth 120 5.4.3 Effect of Light on Radial Mycelial Growth 122 5.4.4 Effect of pH on Radial Mycelial Growth 122 5.5 DISCUSSION 129 6 MORPHOLOGICAL VARIABILITY AMONG ISOLATES 132 OFCOMMONLY FOUND FUNGUS (NATTRASSIA MANGIFERAE)FROM DECLINE AFFECTED CITRUS, MANGO AND GUAVA TREES 6.1 INTRODUCTION 132 6.2 REVIEW OF LITERATURE 134 6.3 MATERIALS AND METHODS 136 6.3.1 Fungal Isolates 136 6.3.2 Morphological Characterization 136 6.4 RESULTS 136 6.5 DISCUSSION 145 7 FACTORS FAVOURING THE DECLINE DISEASE 147 13 ANDDEVELOPMENT OF STRATEGIES FOR ITS MANAGEMENT 7.1 INTRODUCTION 147 7.2 REVIEW OF LITERATURE 148 7.3 MATERIALS AND METHODS 150 7.3.1 Surveys for Factor Favoring Decline in Fruit Orchards 150 7.3.2 Statistical Analysis 151 7.4 RESULTS 151 7.5 DISCUSSION 191 8 GENERAL DISCUSSION 198 8.1 ASSESSMENT OF DECLINE IN CITRUS, MANGO 198 AND GUAVA 8.2 COMMON SYMPTOMS OF DECLINE IN CITRUS, 201 MANGO AND GUAVA 8.3 MYCOFLORA ISOLATED FROM DECLINE 202 AFFECTED TREES PATHOGENICITY OF THE IMPORTANT DECLINE 203 8.4 CAUSING FUNGI 8.5 FACTORS AFFECTING THE GROWTH AND 203 DEVELOPMENT OF NATTRASSIA SP. 8.6 MORPHOLOGICAL CHARACTERISTICS OF 204 NATTRASSIA SP. FROM CITRUS, MANGO AND GUAVA SUMMARY 206 LITERATURE CITED 210 14 List of Tables Table No. Page 2.1 Areas surveyed for citrus decline assessment in district Sargodha 22 2.2 Areas surveyed for mango decline assessment in various districts 25 of Punjab 2.3 Areas surveyed for guava decline assessment in various districts of 31 Punjab 2.4 Mean citrus decline incidence, severity and disease index 39 indifferent tehsils of district Sargodha 2.5 Mean citrus decline incidence, severity and disease index 39 indifferent location of tehsil Bhalwal of district Sargodha 2.6 Mean citrus decline incidence, severity and disease index 40 indifferent location of tehsil KotMomin of district Sargodha 2.7 Mean citrus decline incidence, severity and disease index 41 indifferent location of tehsil Sargodha of district Sargodha 2.8 Mean citrus decline incidence, severity and disease index 42 indifferent location of tehsil Sahiwal of district Sargodha 2.9 Mean citrus decline incidence, severity and disease index 43 indifferent location of tehsil Shahpur of district Sargodha 2.10 Mean citrus decline incidence, severity and disease index 43 indifferent location of tehsil Sillanwali of district Sargodha 2.11 Mean mango decline incidence, severity and disease index 46 indifferent districts of Punjab province 2.12 Mean mango decline incidence, severity and disease index 46 indifferent location of district Khanewal 2.13 Mean mango decline incidence, severity and disease index 48 indifferent location of district Multan 2.14 Mean mango decline incidence, severity and disease index 49 indifferent location of district Muzaffar Garh 2.15 Mean mango decline incidence, severity and disease index 49 indifferent location of district Rahim Yar Khan 15 2.16 Mean mango decline incidence, severity and disease index 50 indifferent location of district Bahawalpur 2.17 Mean guava decline incidence, severity and disease index 53 indifferent districts of Punjab province 2.18 Mean guava decline incidence, severity and disease index 53 indifferent location of district Kasur 2.19 Mean guava decline incidence, severity and disease index 54 indifferent location of district Sheikhupura 2.20 Mean guava decline, severity and disease index in 55 differentlocation of district Nankana Sahib 2.21 Decline symptoms development percentages in citrus, mangoand 61 guava trees 4.1 Response of citrus plants against artificial inoculation of N. 105 mangiferaeandB. theobromaeusing flap method 4.2 Response of citrus plants against artificial inoculation of N. 105 mangiferaeandB. theobromaeusing root injury method 4.3 Response of mango plants against artificial inoculation 106 ofCeratotocystissp.,N. mangiferae,B. theobromaeandFusariumsp. using flap method 4.4 Response of mango plants against artificial inoculation 107 ofCeratotocystissp.,N. mangiferae,B. theobromaeandFusarium sp. using root injury method 4.5 Pathogenicity of fungi associated with guava decline by usingflap 108 method inoculation 4.6 Pathogenicity of fungi associated with guava decline by usingroot 109 injury method of inoculation 5.1 Effect of culture media on radial mycelial growth of the 121 fungiisolated from Citrus on PDA medium 5.2 Effect of culture media on radial mycelial growth of the 121 fungiisolated from mango on PDA medium 5.3 Effect of temperature on radial mycelial growth of the fungi 123 isolated from guava on PDA medium 16 5.4 Effect of temperature on radial mycelial growth of the fungi 123 isolated from citrus on PDA medium 5.5 Effect of temperature on radial mycelial growth of the 124 fungiisolated from mango on PDA medium 5.6 Effect of temperature on radial mycelial growth of the 124 fungiisolated from guava on PDA medium 5.7 Effect of light on radial mycelial growth of the fungi isolated from 125 citrus on PDA medium 5.8 Effect of light on radial mycelial growth of the fungi isolatedfrom 125 mango on PDA medium 5.9 Effect of light on radial mycelial growth of the fungi isolatedfrom 127 guava on PDA medium 5.10 Effect of pH on radial mycelial growth of the fungi isolated 127 fromcitrus on PDA medium 5.11 Effect of pH on radial mycelial growth of the fungi isolated 128 frommango on PDA medium 5.12 Effect of pH on radial mycelial growth of the fungi isolated 128 fromguava on PDA medium 6.1 Morphological characteristics of N.
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