Ofere F. Emeriewen Institut für Züchtungsforschung an Obst Fire blight resistance of the wild apple species Malus fusca Dissertationen aus dem Julius Kühn-Institut Julius Kühn-Institut Bundesforschungsinstitut für Kulturpfl anzen Kontakt/Contact: Dr. Ofere F. Emeriewen Julius Kühn-Institut Federal Research Centre for Cultivated Plants Institute for Breeding Research on Fruit Crops Pillnitzer Platz 3a 01326 Dresden Germany Die Schriftenreihe ,,Dissertationen aus dem Julius Kühn-lnstitut" veröffentlicht Doktorarbeiten, die in enger Zusammenarbeit mit Universitäten an lnstituten des Julius Kühn-lnstituts entstanden sind. The publication series „Dissertationen aus dem Julius Kühn-lnstitut" publishes doctoral dissertations originating from research doctorates and completed at the Julius Kühn-Institut (JKI) either in close collaboration with universities or as an outstanding independent work in the JKI research fields. 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Bibliografische Information der Deutschen Nationalbibliothek Die Deutsche Nationalbibliothek verzeichnet diese Publikation In der Deutschen Nationalbibliografie: detaillierte bibliografische Daten sind im lnternet über http://dnb.d-nb.de abrufbar. Bibliographic information published by the Deutsche Nationalbibliothek (German National Library) The Deutsche Nationalbibliothek lists this publication in the Deutsche Nationalbibliografie; detailed bibliographic data are available in the Internet at http://dnb.dnb.de. ISBN 978-3-95547-066-1 DOI 10.5073/dissjki.2018.004 Herausgeber l Editor Julius Kühn-lnstitut, Bundesforschungsinstitut für Kulturpflanzen, Quedlinburg, Deutschland Julius Kühn-lnstitut, Federal Research Centre for Cultivated Plants, Quedlinburg, Germany Dieses Werk ist lizenziert unter einer Creative Commons – Namensnennung – Weitergabe unter gleichen Bedingungen – 4.0 Lizenz. This work is licensed under a Creative Commons – Attribution – ShareAlike – 4.0 license. - Fire blight resistance of the wild apple species Malus fusca EMERIEWEN, OFERE FRANCIS B.Agric, MSc. XXVII CYCLE Supervisors: Prof. Nicola Pecchioni (UNIMORE, Italy) Dr. Mickael Malnoy (IASMA, Italy) Dr. Andreas Peil (JKI, Germany) Dean of PhD School: Prof. Andrea Pulvirenti Date of thesis defense: 27th March 2015 Table of content………………………………………………………………………….........i Dedication…………………………………………………………...………………..............v Acknowledgements…………………………………………………………………..............vi Abstract………………………………………………………….…......................................vii Riassunto………………………...………………………………………..………………...viii Chapter 1: General Introduction………………………………….............…..……………….1 Origin and economic importance of apple (Malus × domestica Borgh.) .................................2 Fire blight - the most dreadful apple bacterial disease…..………….........................................5 Disease cycle of Erwinia amylovora - an uninvited guest.....................………….……...........7 Molecular mechanism of Erwinia amylovora pathogenicity and host resistance......................9 Fire blight resistance breeding in Malus ………...………………………..……..…………...12 Scope and aim of thesis…………………….............................................................................16 References…….........................................................................................................................18 Chapter 2: Identification of a major quantitative trait locus for resistance to fire blight in the wild apple species Malus fusca …………………...………………………………………….24 Abstract ………………………………………………………………………………………26 Introduction……...…………………………………………………………………………....27 Materials and methods…………….................................….....................................................30 Results…………………………….....………………………………………………………..35 Discussions…………………………………...……………….................................................47 Acknowledgements…………………………………………………………………...............54 References ……………………………………………………………………………………55 Chapter 3: The fire blight resistance QTL of Malus fusca (Mfu10) is affected but not broken down by the highly virulent Canadian Erwinia amylovora strain E2002A ………………….62 Abstract……………….….…...................................................................................................64 Background and findings.....................................................................................................65-73 Acknowledgement ……….............…………………………..…….……………...................74 References…………………………...……………..………………........................................75 Chapter 4: Fine mapping of the Malus fusca fire blight resistance locus (Mfu10)..……...…………………………………………………………………….................77 Abstract……………………………………………………………………………………….78 Introduction…………………………………………………………………………………...79 Materials and methods………………………………………………………………………..84 Results………………………………………………………………………………………...90 Discussions……………………………………………………………………………………97 References…………………………………………………………………………………...103 Chapter 5: General discussions and conclusions……………………………………………107 References…………………………………………………………………………………...113 Appendix…………………………………………………………………………………….115 Curriculum vitae ……………………………………………………………………………125 Publication activities………………………………………………………………………...126 i ii iii iv Dedicated to my lovely wife, Oluwaseun Atinuke Emeriewen; my parents, Rev Bernard Mevayabore and Kate Isioma Emeriewen; and my parents- in-law; Rev Bankole and Adefolake Ojumu ‘Although the fig tree shall not blossom, neither shall fruits be in the vines…Yet I will rejoice in the LORD…The LORD God is my strength….’ Habakkuk 3:17-19 v First and foremost, I am grateful to the Almighty God for giving me life with good health and a sound mind to be able to work and actualize dreams. His grace has been sufficient. I would like to thank Prof. Nicola Pecchioni for welcoming me into his group and for being my tutor at the University of Modena and Reggio Emilia. I want to place on record my gratitude to Prof. Andrea Pulvirenti and Dr. Luisa Antonella Volpelli of the PhD School of Agri-food Sciences, Technologies and Biotechnologies of UNIMORE for their kindness and assistance every single time I needed them. I also want to express my thanks to Dr. Mickael Malnoy of the Fondazione Edmund Mach – San Michele (Trentino), Italy for his supervision of the PhD project from that angle. I like to appreciate the technical support of Stefano Piazza of the Tissue Culture and Transformation Group, headed by Dr. Mickael Malnoy and also, every single member of this group which I was a part of. I want to specially thank Dr. Andreas Peil, of the Institute for Breeding Research on Fruit Crops – Julius Kühn Institute (JKI), Germany for his motivation, guidance, dedication and supervision of this PhD project. From the Germany angle, I also would appreciate the encouragements and pieces of advice of Prof. (Dr.) Viola Hanke (Head of JKI, Pillnitz, Dresden), and Dr. Henryk Flachowsky. I appreciate the technical support from members of staff of JKI. This PhD research was funded by the Genomics and Molecular Physiology of Fruits (GMPF) program of the Research and Innovation Centre (CRI) of the Instituto Agrario di San Michele all’ Adige (IASMA); Fondazione Edmund Mach (Trentino) Italy. Therefore, I would like to place on record my gratitude for sponsoring this research and selecting me as the scholarship candidate to carry out the research work. Last but by no means the least; I appreciate my beloved wife for the patience and encouragement throughout my PhD research. I am indebted to my family, immediate and extended, for the belief they showed in me and for their prayers, pieces of advice, encouragement, financial support and above all, for their unconditional love for me. It would have been absolutely impossible without the support of family and some friends whom have themselves become my family and too numerous to mention individually. I pray for you all that your dreams shall not die, your plans shall not fail and your destinies shall not be aborted, in the name that is above ALL names, JESUS CHRIST. vi Erwinia amylovora is the pathogen responsible for inciting fire blight - the most dreaded bacterial disease of apple (Malus × domestica) and other members of the Rosaceae family. The disease is very destructive as is difficult to control. Even though fire blight was first observed over two centuries ago in America, no sustainable control measure is known till date. Disease management practices such as pruning of affected tissues as well as the application of copper and antibiotics, for example, streptomycin, are to minimize the population of E. amylovora in an orchard. However, the use of streptomycin is not allowed in many European countries as a consequence of its environmental risks and the issue of raising antibiotic