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Baseline Study of Coconut Scale, Aspidiotus Destructor Signoret (Hemiptera: Diaspididae) and Its Management on Mango (Mangifera Indica L.)

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Baseline Study of Coconut Scale, Aspidiotus Destructor Signoret (Hemiptera: Diaspididae) and Its Management on Mango (Mangifera Indica L.) BASELINE STUDY OF COCONUT SCALE, ASPIDIOTUS DESTRUCTOR SIGNORET (HEMIPTERA: DIASPIDIDAE) AND ITS MANAGEMENT ON MANGO (MANGIFERA INDICA L.) By SALAHUDDIN A dissertation submitted for partial fulfillment of the requirement for the degree of DOCTOR OF PHILOSOPHY IN ENTOMOLOGY DEPARTMENT OF ENTOMOLOGY FACULTY OF AGRICULTURE GOMAL UNIVERSITY DERA ISMAIL KHAN 2016 © 2016 SALAHUDDIN DEDICATED TO MY PARENTS ACKNOWLEDGEMENT All glory be to Allah, the beneficial, the merciful. I would like to thank Almighty Allah who enabled me to complete this painstaking study of PhD well in time. I would like to express the deepest appreciation to my supervisor Dr. Habib ur Rahman, co- supervisors Dr. Inamullah Khan and Dr. Muhammad Daud Khan for their substantial guidance, continuous inspiration, invaluable assistance and suggestions from initial planning of experiments to the preparation of manuscripts in bringing this dissertation to its present form. I would like to thank my IRSIP program supervisor Dr. Steven P. Arthurs at Mid- Florida Research and Education Center, IFAS, University of Florida, USA for his cooperation to conduct research in his laboratory and proof reading and correction of my PhD dissertation. His deep knowledge and experience helped me to learn advance skills in Entomology and improve my scientific writing. I am grateful to my other supervisory committee members Professor Dr. Muhammad Ayaz Khan, Agha Shah Hussain and Dr. Muhammad Mamoon ur Rashid for their excellent guidance and suggestions on how to improve my work. Dr. Gillian W. Watson, Senior Insect Biosystematist, California Department of Food and Agriculture, Plant Pest Diagnostic Center, Sacramento California, U.S.A., is gratefully acknowledged for identification of Aspidiotus destructor. I would like to acknowledge the Higher Education commission (HEC), Islamabad, Pakistan for providing financial assistance for the duration of my PhD research through Indigenous 5000 PhD fellowship program and Department of Agriculture Extension, Government of Khyber Pakhtunkhwa for granting three years study leave. I am thankful to The Nuclear Institute for Food and Agriculture (NIFA), Peshawar, Kohat University of Science and Technology, Kohat and Mid-Florida Research and Education center (MREC), Apopka, University of Florida, USA for allowing me to work in their laboratories. I am thankfull to Arid Zone Research Institute (AZRI), Ratta Kulachi, D. I. Khan for providing metereological data during 2011-12 and 2012-13. I thank Imran Khan (AHO) and his staff to allow me to work in Fruit Nursery Farm and provided technical and labor help in growing and handling mango nursery plants. I am grateful for the technical help of Dr. Hafiz Mumtaz Khan, Dr. Farmanullah, Dr. Kalimullah, Tariq Nawaz, Waqas Yousafzai and Muhammad Suleman Shah. I would like to deeply appreciate Mishkatullah, Muhammad Nisar, Ejaz Malook, Saeed Hassan, Syed Fayaz Ali Shah, Luis Aristizábal, Robert Leckel, Nastaran Tofangsazi, Runquian Mao and Yingfang Xiao for supporting me as a lab assistant and as a friend. I iv would like to thank all of them for making Peshawar, Kohat and Florida a home away from home. I appreciate the love and support that I received from friends Shah Namir Khan, Muhammad Gul, Imran Khan, H. Attaullah, Muhammad Riaz, H. Nabi Gul, Adnan khan, M. Tariq Khan, Sher Zaman, Waliullah, and all others whose names are much important but not written here. I have no words at command of acknowledging my late parents who taught me values, hard work and truthfulness in life, without whom I would not be the person as today. My love and pray for them is eternal. Their sincere prayers empowered me to achieve success in every sphere of life. May their soul rest in peace. Ameen. I appreciate the determination of my wife and kids, Rimsha Gul, Hammad Ahmad Khan, Iffat Gul and Fanila Gul who endured my long absence from home. I will never forget the moment when I would go to Peshawar for research work, my son would sit angry in the corner of home that “why Dad is going to Peshawar”. Their love provided me inspiration and was my driving force. I owe them everytime and wish I could show them how much I love them. I would like to thank my brothers Dr. Razaman Bittan, Dr. Ibrahim Gul Bittani, Advocate Noor zaman Bittani, my sister, my nephews Humaira, Sawaira, Afaq, Jawad and Hurrain for their long distance support. Salahuddin Bittani v List of Contents Chapter Contents Page No. ACKNOWLEDGEMENT iv LIST OF CONTENTS vi LIST OF TABLES xiii LIST OF FIGURES xv LIST OF ABBREVIATIONS xvii ABSTRACT xix 1. GENERAL INTRODUCTION AND REVIEW OF LITERATURE 1 1.1. Economic impact of mango 1 1.2. Pests of mango 1 1.3. Overview of scale insects 2 1.4. Armored scales 3 1.5. Aspidiotus destructor 3 1.5.1. Distribution 4 1.5.2. Economic importance 5 1.5.3. Use of common names of A. destructor 5 1.5.4. Seasonal population dynamics of A. destructor 7 1.5.5. Natural enemies of A. destructor 7 1.5.6. Reproduction in A. destructor 8 1.5.6.1. Common feature of A. destructor 8 1.5.6.2. Morphological description 9 1.5.7. Physiological, biochemical and molecular changes in mango 10 leaves under scale insects infestation 1.5.7.1. Ions analysis 11 1.5.7.2. Physiological changes in mango plant under insect 12 infestation 1.5.7.3. Biochemical changes in mango plant under insect 13 infestation 1.5.7.4. Molecular changes in mango plant under insect infestation 15 1.5.8. Development of generic doses of irradiation for the growth inhibition of A. destructor 16 vi 1.5.9. Physiochemical analysis and sensory evaluation of mango fruits 18 1.5.10. Efficacy of different Insecticides against A. destructor 21 1.6. Aims 23 2. GENERAL MATERIALS AND METHODS 24 2.1. Study area at a glance 24 2.2. Raising mango nursery plants and release of A. destructor crawlers on test plants 24 2.3. Collection and Identification of scale insects on mango plants 24 2.4. Natural efficiency of parasitoids on A. destructor: 25 2.5. Biology of A. destructor on mango plants under laboratory and greenhouse conditions 25 2.6. Scale-induced, physiological, biochemical and molecular changes in mango leaves 25 2.6.1. Physiological changes in mango leaves under A. destructor infestation 26 2.6.2. Biochemical changes in mango leaves under A. destructor infestation 26 2.6.3. Molecular changes in mango leaves under A. destructor infestation 26 2.7. Effect of irradiation on pest mortality and development inhibition 26 2.8. Effect of irradiation on physiochemical analysis and sensory quality of mango fruit 27 2.9. Evaluation of different insecticides against A. destructor on mango nursery plants 27 2.10. Evaluation of different insecticides against A. destructor on mature mango trees 28 2.11. Experimental design / statistical analysis 28 3. INCIDENCE OF ASPIDIOTUS DESTRUCTOR IN RELATION TO ABIOTIC FACTORS AND NATURAL EFFICIENCY OF ITS 29 PARASITOIDS 3.1. Introduction 29 3.2. Materials and Methods 30 vii 3.2.1. Population dynamics of A. destructor on four mango c cultivars at five locations 30 3.2.2. Natural efficiency of parasitoids on A. destructor 30 3.3. Results 31 3.3.1. Population dynamics of A. destructor on four mango 31 cultivars 3.3.2. Population dynamics of A. destructor at five locations on 33 mango plants 3.3.3. Relation of A. destructor population fluctuation with abiotic 37 factors 3.3.4. Natural efficiency of A. melinus and Anagyrus sp. on A. 38 destructor 40 3.4. Discussion and conclusion 4. BIOLOGY OF A. DESTRUCTOR ON MANGO PLANTS (MANGIFERA SP.) UNDER LABORATORY AND GREENHOUSE 43 CONDITIONS 4.1. Introduction 43 4.2. Materials and Methods 44 4.2.1. Laboratory study 44 4.2.2. Greenhouse study 45 4.3. Results 45 4.3.1. Laboratory study 45 4.3.1.1. Aspidiotus destructor developmental stages and morphometry 45 4.3.1.2. Development of male Aspidiotus destructor 48 4.3.1.3. Development of female Aspidiotus destructor 48 4.3.2. Greenhouse study 49 4.4. Discussion and conclusion 51 viii 5. PHYSIOLOGICAL AND BIOCHEMICAL CHANGES IN MANGO LEAVE UNDER A. DESTRUCTOR STRESS 54 5.1. Introduction 54 5.2. Materials and Methods 56 5.2.1. Raising nursery of test plants 56 5.2.2. Determination of leaf biomass, leaf area and photosynthetic pigments 56 5.2.3. Cell viability and cell injury 57 5.2.3.1. Cell viability 57 5.2.3.2. Membrane leakage 57 5.2.4. Determination of various ions (Na+, K+, Ca++) 58 5.2.5. Determination of stress markers 58 5.2.6. Determination of total soluble proteins (TSP) and antioxidants 59 5.2.7. Proline accumulation in mango leaves under scale insects infestation 59 5.3. RESULTS 60 5.3.1.Physiological changes in mango leaves under scale 60 infestation 5.3.2. Changes in photosynthetic pigments 61 5.3.3. Changes in cell viability under scale insect infestation 62 5.3.3.1. TTC method 62 5.3.3.2. Electrolyte leakage under scale insect infestation 63 5.3.4. Scale-induced ionic changes in mango leaves 63 5.3.5. Effect of scale insect infestation on total soluble protein (TSP) contents and proline accumulation 64 5.3.6. Changes in stress markers under scale insects infestation 65 5.3.7. Changes in the antioxidant enzymes of mango leaves under 66 scale insects infestation 5.4. Discussion and conclusion 68 ix 6. MOLECULAR CHANGES IN MANGO LEAVES UNDER SCALE INSECT INFESTATION 74 6.1. Introduction 74 6.2. Materials and Methods 74 6.2.1. DNA damage analysis 74 6.2.1.1.
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