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Development of Microsatellites in Sycamore Maple (Acer Pseudoplatanus L.) and Their Application in Population Genetics

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Development of Microsatellites in Sycamore Maple (Acer Pseudoplatanus L.) and Their Application in Population Genetics Madhav Pandey Development of microsatellites in sycamore maple (Acer pseudoplatanus L.) and their application in population genetics Institute of Forest Genetics and Forest Tree Breeding Faculty of Forest Sciences and Forest Ecology Georg-August University of Göttingen Germany Development of microsatellites in sycamore maple (Acer pseudoplatanus L.) and their application in population genetics DISSERTATION Submitted in partial fulfilment of the requirements for the degree of Doctor of Forestry Science at the Faculty of Forest Sciences and Forest Ecology, Georg-August University of Göttingen, Germany by Madhav Pandey Born in Gulmi, Nepal Göttingen, 2005 D7 Supervisor: Prof. Dr. Hans H. Hattemer Co-supervisor: Prof. Dr. Reiner Finkeldey 1. Referee: Prof. Dr. Hans H. Hattemer 2. Referee: Prof. Dr. Reiner Finkeldey 3. Referee: Prof. Dr. Klaus von Gadow Date of oral examination: 1st August, 2005 This dissertation has been published through the library of Lower Saxony State and Georg-August University of Göttingen, Platz der Göttingen Sieben, 37073, Göttingen under the internet address: http://resolver.sub.uni-goettingen.de/purl/?webdoc-9 To my beloved family…… Acknowledgement A journey is easier when you travel together. Interdependence is certainly more valuable than independence. This dissertation is the result of three years of work whereby I have been accompanied and supported by many people. It is a pleasant aspect that I have now the opportunity to express my gratitude for all of them. My deepest thanks go to my supervisor Prof. Dr. Hans H. Hattemer for providing an opportunity to work in this project and for his excellent supervision, guidance, patience, and encouragement on this dissertation. My sincere thanks go to my co-supervisor Prof. Dr. Reiner Finkeldey, for his valuable supervision, guidance and suggestions to improve the dissertation. I am also thankful to him for his recommendation to join this project. I would like to thank Prof. Hattemer and Prof. Finkeldey also for their support in my daily life during the stay in Germany. I am also very thankful to Prof. Dr. Klaus von Gadow for consenting to be a co-referee and Prof. Dr. Dirk Hölscher for being a member of examination committee. My warm gratitude is extended to Dr. Oliver Gailing for his supervision and guidance in laboratory work and also for the constructive discussion. I extend my thanks to Dr. Dirk Fischer for his guidance and support during the microsatellite development work at the Institute of Plant Genetics-IPK, Gatersleben. I express my thanks to Prof. Dr. Martin Ziehe for his support and advice in diverse computer programme. My thanks also go to Dr. Ludger Leinemann and Dr. Barbara Vornam for fruitful discussions and providing valuable literature. I am grateful to Miss Olga Artes, Mr. Thomas Seliger, and Mr. Gerold Dinkel for their help in laboratory work. Likewise I am also thankful to Mr. August Capelle for his support in sample collection and mapping work. Friendly behaviour and fruitful discussion of Prof. Dr. Hans Rolf Gregorius, Dr. Elizabeth M. Gillet and Dr. Dirk Kownatzki is appreciated. I acknowledge the secretarial work of Ms. Marita Schwan. Many tanks to all my colleague at the Institute of Forest Genetics and Forest Breeding: Akindele Akinangbe, Taye Bekele, Cui Ping Cao, Alexandra Lucian Curtu, Dr. Natalia Decarli, Abayneh Derero, Nicolas-George Eliadess, Dr. Aki Höltken, Dr. Sapto Indrioko, Dr. Jürg Kleinschmit, Dr. Hong Truong Luu, Martin Motura, Sylvia Nascimento, Valdir Marcos Stefenon, Yanti Rachmayanti, and many others who could not be mentioned here for their friendship and valuable discussion. I would like to express my thanks to Mr. Ewarad Walter and Mrs. Margaretha Walter in Groß Lengden, Göttingen for their support in many aspects of my and my family’s daily life during the stay in Göttingen. I am also thankful to my all Nepalese colleague and their family in Göttingen for their friendship. I acknowledge the financial support of DFG (Deutsche Forschungsgemeinschaft) for this project. Last but not least my deepest and genuine gratitude is extended to my wife Manju for her countless support, love and patience during my study in Germany. I also thank my son Ashish for his understanding and obeying. One of the best experiences that we lived through in this period was the birth of our second son Amish, who provided an additional and joyful dimension to our life mission. I express special thanks to my parents and other family member in Nepal for their supporting and patience. Table of contents List of Figures……………………………………...……………………….……..… …….iv List of tables………………………………………………………………….….…......….viii 1. Introduction..........................................................................................................................1 1.1. Introduction into the species ...............................................................................................1 1.1.1. The genus Acer ..........................................................................................................1 1.1.2. Acer pseudoplatanus..................................................................................................1 1.1.2.1. Morphology.......................................................................................................1 1.1.2.2. Distribution .......................................................................................................2 1.1.2.3. Habitat and ecology ..........................................................................................3 1.1.2.4. Flower and sexual system .................................................................................4 1.1.2.5. Pollination biology............................................................................................5 1.1.2.6. Uses...................................................................................................................5 1.1.2.7. Propagation .......................................................................................................5 1.2. Polyploidy...........................................................................................................................6 1.2.1 Autopolyploids............................................................................................................6 1.2.2 Allopolyploids ............................................................................................................6 1.2.3. Genetic consequences of polyploidy .........................................................................7 1.3. Molecular markers ..............................................................................................................8 1.3.1. Microsatellites or Simple Sequence Repeats (SSRs).................................................8 1.3.2. Evolution of microsatellites (SSRs)...........................................................................9 1.3.3. Theoretical models of microsatellite mutation ........................................................10 1.3.4. Application of microsatellite markers......................................................................11 1.3.5. Advantages of microsatellite gene markers .............................................................12 1.3.6. Disadvantages ..........................................................................................................13 1.3.7. Development of microsatellite gene markers ..........................................................13 1.4. Genetic variation and genetic structure.............................................................................15 1.4.1. Genetic variation......................................................................................................15 1.4.2. Spatial genetic structure...........................................................................................16 1.5. Movement of genetic information ....................................................................................17 1.6. Genetic variation of Acer pseudoplatanus ........................................................................18 1.7. Objectives and research hypotheses..................................................................................19 2. Materials and Methods.....................................................................................................20 2.1. Plant material ....................................................................................................................20 2.1.1. Introduction to research areas ..................................................................................20 2.1.1.1. Södderich population ......................................................................................20 2.1.1.2 Weißwassertal population................................................................................20 2.1.2. Material collection ...................................................................................................24 2.1.2.1. Buds ................................................................................................................24 2.1.2.2. Leaves .............................................................................................................24 2.1.2.3. Seeds ...............................................................................................................27 2.2. Laboratory methods ..........................................................................................................27 2.2.1. Development of microsatellite (SSRs)
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