ORIGIN AND SPREAD OF THE GERMAN COCKROACH, BLATTELLA GERMANICA TANG QIAN (B.Sc. (Hons), Wuhan University, China) A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF BIOLOGICAL SCIENCES NATIONAL UNIVERSITY OF SINGAPORE 2015 Declaration Declaration I hereby declare that this thesis is my original work and it has been written by me in its entirety. I have duly acknowledged all the sources of information which have been used in the thesis. This thesis has also not been submitted for any degree in any university previously. Tang Qian 31 Dec 2015 i Acknowledgement Acknowledgement My Ph.D. was supported by the NUS Research Scholarship from the Singapore Ministry of Education. The research project was funded by the Lee Hiok Kwee Endowed Fund of the Department of Biological Sciences, the National University of Singapore to Associate Professor Theodore Evans. I would like to thank the Singapore Ministry of Education and the National University of Singapore for providing me such opportunity to enter the academic world. This thesis could not be finished without the effort of my supervisors: Associate Professor Theodore Evans and Assistant Professor Frank Rheindt. Associate Prof. Evans initiated this ambitious research project with confidence and insights. Assistant Prof. Rheindt supported this project with professional advice and knowledge in the field of population genetics. This project requires much effort to collect samples. Associate Prof. Evans and Assistant Prof. Rheindt always offered me their advice and time. There are many people involved in my Ph.D. project, so I would like to cite their contribution by chapter: For chapter one, I would like to thank those who spent days in museums retrieving German cockroach specimens for my review. My supervisor Theodore Evan collected specimen information from British Museum of Natural History, London and the Smithsonian Natural History Museum in Washington D. C. My colleague Thomas Bourguignon collected specimen information from the Museum of Natural History in Paris. Curator Luc Willenmse provided information his collection in Naturalis, Leiden, Netherlands. Curator Eliane De Coninck provided information for her collection in the Royal Museum of Central Africa, Tervuren, Belgium. Curator Lars Lundqvist and his assistant Ellen Rehnberg ii Acknowledgement provided information for their collection in Museum of Zoology in Lund University. For Chapter Two, I thank Dr. Thomas Bourguinon in School of Biological Sciences, University of Sydney, Australia, former postdoctoral researcher in the Applied Entomology Lab in NUS. He directed my commencement of research in the field of molecular phylogeny. Without his patient guidance, there would be no Chapter Two in this thesis. I would also like to thank people devoted to sample collection of different cockroach species. Most of the cockroach samples were provided by Mr. Kyle Kandilian, owner of Roach Crossing, a lovely pet cockroach store. I thank him for his efforts preparing and shipping his cockroach collections to me as research materials. I would also like to thank my lab mates Aloysius Teo, Maria Lee and Naeem Iqbal, who helped me collect Blattella species in Singapore and Pakistan. For Chapter Three, I thank Prof. Ed Vargo, Professor and Endowed Chair in Urban and Structural Entomology, Department of Entomology, Texas A&M University, USA. He is one of the pioneers of studying population genetics of the German cockroach; his studies inspired my whole Ph.D. project. Prof. Vargo also contributed half of the samples for Chapter Three to provide this project with comprehensive coverage of worldwide German cockroaches. Moreover, I would like to thank Dr. Alison Wee Kim Shan and Dr. Annika Noreen for teaching me the ABCs of population genetics. I would like to thank people who sent me samples from all over the world: Dr. Lee Dong-kyu and his graduate student Dong- min in Kosin University provided Korean samples; Dr. Jiang Hong in Wuhan University provided Chinese samples; Dr. Noraishah Mydin Abd Aziz in Malaya University provided Malaysian samples; Dr. Intan Ahmad in Bandung Institute of Technology provided Indonesian samples; Dr. Tan Eng Kooi in Bayer Singapore provided samples from Holland, Germany and Brazil; Ms. Zuzana Varadinova in Charles University of Prague provided iii Acknowledgement samples from the Czech Republic, Morocco and Ghana; Dr. Melaku Wale and Mr. Fasil Adugna in Bahir Dar University provided Ethiopian samples. Last but not least, I would like to thank Dr. How Yee Fatt in Bentz Jaz, Singapore, Dr. Thomas Bourguinon in Sydney University and Mr. Nega Tassie helping me build connections with people who provided samples. For Chapter Four, I would like to give special thanks to Associate Professor Jiang Hong and his lab crews in Department of Medical Parasitology, School of Basic Medicine Science, Wuhan University, China. Associate Prof. Jiang helped me build connections to PCOs all around China; he also collected and stored all the samples from Chinese PCOs until I did DNA extractions. Special thanks also to my undergraduate mentor Professor Li Yangsheng and his lab members in Department of Genetics, College of Life Sciences, Wuhan University, China. Prof. Li provided very convenient lab environment for me to perform DNA extractions for the cockroach samples from China. I thank Prof. Li’s lab members, Yang Jing and Hu Daoheng who provided help and accompany during those hard-working days and nights in Wuhan. I would like to thank all the PCOs that collected samples from different cities in China; I thank Mr. Wu Xifeng, Shenyang; Mr. Nan Zengliang, Sinopest Co. Ltd, Beijing; Mr. Wang Jiansheng, Best Pest Control Co. Ltd., Nanjing; Mr. Mo Yuan, Chongqing; Mr. Pan Qiang, Huaguan Pest Control Co. Ltd., Guangzhou. I would like to thank all principal investigators and lab members in S14 level 3 labs. I appreciate your selfless help and I enjoy the time we spent together. I also thank the department administrative staff who provide high-quality services to assist me with issues associated with my research project and Ph.D. life. Thanks to everyone in the Department of Biological Sciences, whose professionalism and friendliness create a great environment iv Acknowledgement for me to accomplish my Ph.D. project. At last, I would like to thank my family. As the only son, I thank my parents’ constant understanding and encouragement to me of this career I choose. I thank Yan Yanyun, the love of my life. She took care of my life and stood with me for all these years facing challenges and an unknown future. v Table of Contnets Table of Contents Declaration………………………….……………………………………..…….....……...i Acknowlegement………………………..…………………………………….....…….…ii Table of Contents…………………..…………………………………………….....……vi Summary...………………………………………………………………...…….….…...vii List of tables………...…………………………………………………………...…...….i x List of figures…...………………………………………………………...………..……..x Chapter 1: Global spread of the German cockroach ................................................................... 1 1.1 Pest cockroach species ........................................................................................... 2 1.2 Origin and spread of the German cockroach ......................................................... 4 1.3 Oher pest Blattella species ..................................................................................... 6 1.4 Spread of the German cockroach........................................................................... 7 1.5 Factors limiting spread of the German cockroach ............................................... 12 1.6 Tracing spread of the German cockroach ............................................................ 13 1.7 Hypotheses and aims ........................................................................................... 15 Chapter 2: Phylogenetic reconstruction of order Blattodea and genus Blattella with complete mitochondrial genomes ............................................................................................ 18 2.1 Introduction ......................................................................................................... 18 2.2 Material and Methods .......................................................................................... 23 2.3 Results ................................................................................................................. 31 2.4 Discussion ............................................................................................................ 36 2.5 Conclusion ........................................................................................................... 44 Chapter 3: Global population structure of the German cockroach ............................................ 45 3.1 Introduction ......................................................................................................... 45 3.2 Materials and methods ......................................................................................... 53 3.3 Results ................................................................................................................. 57 3.4 Discussion ............................................................................................................ 67 3.5 Conclusion ........................................................................................................... 72 Chapter 4: Population structure of the German cockroach, in China ....................................... 73 4.1 Introduction ........................................................................................................