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Induction and Termination of Diapause in Khapra Beetle, Trogoderma Granarium

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Induction and Termination of Diapause in Khapra Beetle, Trogoderma Granarium INDUCTION AND TERMINATION OF DIAPAUSE IN KHAPRA BEETLE, TROGODERMA GRANARIUM SUNIL SHIVANANJAPPA Bachelor of Science in Agriculture, University of Agricultural Sciences, 2016 A thesis submitted in partial fulfilment of the requirements for the degree of MASTER OF SCIENCE in BIOLOGICAL SCIENCES Department of Biological Sciences University of Lethbridge LETHBRIDGE, ALBERTA, CANADA © Sunil Shivananjappa, 2019 INDUCTION AND TERMINATION OF DIAPAUSE IN KHAPRA BEETLE, TROGODERMA GRANARIUM SUNIL SHIVANANJAPPA Date of Defence: August 12, 2019 Dr. R. Laird Associate Professor Ph.D. Dr. K. Floate Research Scientist Ph.D. Thesis Co-Supervisors Dr. P. Fields Research Scientist Ph.D. Thesis Examination Committee Member Dr. T. Burg Professor Ph.D. Thesis Examination Committee Member Dr. T. Russell Associate Professor Ph.D. Chair, Thesis Examination Committee DEDICATION To my parents, Shivananjappa and Manjula, for being incredibly supportive. iii ABSTRACT Larvae of the stored product pest, khapra beetle, Trogoderma granarium, are unusual in that they can enter a diapause state that confers protection against pest control methods. In Chapter 1, I reviewed the literature on general aspects of insect diapause induction and termination, particularly in Dermestidae. In Chapter 2, I assessed the effects of age of culture and diet quality on diapause termination. The results indicated that diapausing larvae can accumulate the nutrients required to terminate diapause and complete development when provided with an opportunity to do so. In Chapter 3, I assessed whether desiccation enhances cold tolerance in diapausing and non-diapausing larvae. Results showed that cold tolerance of diapausing larvae increased with greater desiccation stress. This finding suggests that the physiological mechanisms that protect diapausing larvae from desiccation may also increase cold-tolerance. Overall results of this research provide new information with potential application for control of this global pest. iv ACKNOWLEDGEMENTS I am beyond grateful to have had the experience of completing my Master's degree in Biological Sciences. First and foremost, I would like to express my sincere gratitude to a team of amazing people, Dr. Paul Fields, Dr. Kevin Floate, and Dr. Robert Laird, who provided me with the opportunity to undertake this project. If it were not for their constant support, guidance, patience and encouragement, I wouldn’t have made this far in completing this thesis. I would like to thank Dr. Sarma Mohan, without whom, I wouldn’t have come to Canada for pursuing my graduate program. His support and encouragement throughout this study has helped me enormously. I would also like to thank my lab mentor Paul Coghlin, for supplying all the necessary materials for my experiments and for looking into my non-academic needs. I greatly acknowledge Diana Wilches and Dr. Timothy Schwinghamer for their statistical guidance and inputs. I also thank my friends from India and fellow graduate students for their pep talks along the way. Finally, I would like to thank my parents and my sister Sowmyashree, for being my pillars of strength during this journey, and for always supporting my passion for entomology. Thank you all for this wonderful learning experience of a lifetime. v TABLE OF CONTENTS DEDICATION.............................................................................................................................. iii ABSTRACT .................................................................................................................................. iv ACKNOWLEDGEMENTS ......................................................................................................... v LIST OF TABLES ..................................................................................................................... viii LIST OF FIGURES ..................................................................................................................... ix LIST OF ABBREVIATIONS ...................................................................................................... x CHAPTER 1: GENERAL INTRODUCTION AND LITERATURE REVIEW .................... 1 1.1 Objectives ............................................................................................................................. 1 1.2 Introduction ......................................................................................................................... 2 1.3 The phenomenon of diapause ............................................................................................ 6 1.3.1 Diapause vs quiescence ................................................................................................. 6 1.3.2 Phases of diapause ......................................................................................................... 7 1.3.3 Physiological changes associated with diapause .......................................................... 8 1.4 Diapause in Dermestidae .................................................................................................. 10 1.5 Diapause in khapra beetle ................................................................................................ 13 1.5.1 Factors responsible for initiation of diapause ............................................................. 13 1.5.1.1 Role of larval density ............................................................................................ 13 1.5.1.2 Role of temperature............................................................................................... 14 1.5.1.3 Role of humidity .................................................................................................... 14 1.5.1.4 Role of fecal pellets ............................................................................................... 15 1.5.1.5 Role of diet quality ................................................................................................ 15 1.5.2 Factors responsible for terminating diapause ............................................................. 16 1.6 Cold and desiccation tolerance ........................................................................................ 16 1.7 Summary ............................................................................................................................ 19 1.8 Structure of thesis and goals of each chapter ................................................................. 20 CHAPTER 2: EFFECT OF DIET QUALITY AND AGE OF CULTURE ON DIAPAUSE TERMINATION IN KHAPRA BEETLE, Trogoderma granarium ....................................... 26 2.1 Abstract .............................................................................................................................. 26 2.2 Introduction ....................................................................................................................... 27 2.3 Materials and methods ..................................................................................................... 30 2.3.1 Insect’s origin and rearing .......................................................................................... 30 2.3.2 Experimental protocol ................................................................................................. 31 vi 2.3.3 Diet analysis ................................................................................................................ 32 2.3.4 Statistical analysis ....................................................................................................... 32 2.4 Results ................................................................................................................................ 33 2.4.1 Diet composition .......................................................................................................... 33 2.4.2 Diapause termination .................................................................................................. 33 2.4.3 Maintenance of diapause after 188 days ..................................................................... 34 2.5 Discussion ........................................................................................................................... 35 CHAPTER 3: CROSS TOLERANCE TO DESICCATION AND COLD IN KHAPRA BEETLE, Trogoderma granarium ............................................................................................. 44 3.1 Abstract .............................................................................................................................. 44 3.2 Introduction ....................................................................................................................... 44 3.3 Materials and Methods ..................................................................................................... 46 3.3.1 Insect’s origin and rearing .......................................................................................... 46 3.3.2 Desiccation .................................................................................................................. 47 3.3.3 Assessment of larval cold-tolerance ............................................................................ 48 3.4 Results ................................................................................................................................ 49 3.4.1 Cold-tolerance of larvae following desiccation ........................................................... 49 3.4.2 Diapause termination post cold exposure ..................................................................
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