Characterization of Evolvng Populations of Phytophthora
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CHARACTERIZATION OF EVOLVNG POPULATIONS OF PHYTOPHTHORA NFESTANS CAUSING LATE BLIGHT OF POTATO IN CANADA A Thesis Presented to The Faculty of Graduate Studies of The University of Guelph by RICK DANIEL PETERS In partial fulfilment of requirements for the degree of Doctor of Philosophy Apnl, 1998 O Rick Daniel Peters, 1998 National Library Bibliothèque nationale du Canada Acquisitions and Acquisitions et Bibliographie Services services bibliographiques The author has granted a non- L'auteur a accordé une licence non exclusive licence allowing the exclusive permettant à la National Library of Canada to Bibliothèque nationale du Canada de reproduce, loan, distribue or sell reproduire, prêter, distribuer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfiche/fiùs de reproduction sur papier ou sur format électronique. The author retaïns ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantid extracts fiom it Ni la thèse ni des extraits substantiels may be printed or otherwise de celle-ci ne doivent être imprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation. CHARACTERIZATION OF EVOLVING POPULATIONS OF PHYTOPHTHORA INFESTANS CAUSING LATE BLIGHT OF POTATO IN CANADA Rick Daniel Peters Advisors: University of Guelph, 1998 Dr. H.W. (Bud) Platt Dr. Robert HaU A collection of isolates of Phytophthora infstam (Mont.) de Bary was obtained From potato and tomato samples from across Canada in each of 1994, 1995, and 1996. Characterization of these isolates according to mating type (Al or A2), sensitivity to metalaxyl, growth in culture, and allozyme banding patterns, revealed eight distinct genotypes of the fungus. In 1994, the US- 1 genotype (A 1, metalaxyl-sensitive FIS]) was comrnody recovered in Canada (outside British Columbia). By 1996, the US- 1 genotype was no longer recovered fiom any samples, and outside British Columbia, the US-8 genotype (A2, rnetdaxyl-insensitive NI]) was the dominant genotype in Canada. In British Columbia, the g 1 1 genotype (Al, MI) became the dominant genotype recovered in 1995 and 1996. There was no corretation between recovery of MI strains and metalaxyl use. Allozyme banding patterns were strongly correlated to mating type. metalaxyl sensitivity, and cultural c haracten, revealing the clonal nature of populations. Most isolates of recently introduced genotypes were more aggressive (had greater fitness) on tuber tissue than isolates of the US-1 genotype. Variation also occurred within a genotype and an isolate of the US-8 genotype from New Brunswick was consistently Iess aggressive than other US-8 isolates. A total of 28 pathotypes were found among 80 isolates of P. infestans tested. There was a significant increase in the complexity of pathotypes fiom 1994 to 1996, which reflected the displacement of the US- 1 genotype (mean of 2.2 host differentials uifected) by the US-8 genotype (mean of 8.1 host differentials infected). Both mating types of the fungus were found in 7 fields and one sample was found to have oospores of P. infestans visibly present in plant tissues. Therefore, P. infestans cm reproduce sexually in nature in Canada. These results demonstrate that populations of P. infestam in Canada changed dramaticdly fiom 1994 to 1996 and the original US4 (Al, MS) genotype was rapidly displaced. Migration of new forms followed by genetic drift operating through founder effects and selection for fitter genotypes are postulated as the pnmary mechanisms responsible for the observed patterns of evolution. ACKNOWLEDGEMENTS 1 wodd like to express my sincere gratitude to my advisors, Dr. H.W. (Bud) Platt and Dr. R. Hall for their guidance and encouragement. 1 am also grateful to the other members of my advisory comrnitiee, Dr. G. Boland and Dr. T. Hsiang, for their help and constructive contribution to this thesis. 1wodd like to thank Agriculture & Agri-Food Canada (Charlottetown Research Centre) and al1 members of the potato industry fkom across Canada that contributed to Matchmg Investment Initiative Project #3026, for their financial and other contributions that made this research possible. 1 wouid also like to thank growers, potato industry professionals and govemment officiais fiom across Canada for sample submissions without which this research would not have been possible. There are many people at the Charlottetown Research Centre that deserve my thanks. Marco Medina, George Mahuku, Apnl Drisco 11, Sandy Jenkins, Erin Comors. Suzanne MacNeill, Anne MacPhail, Brian Matheson. Laurie McNally-Shanahan, Phi1 Maxwell, Richard Reddin and Velma MacLean al1 contributed in some way to the work presented here. Thanks also for your fiiendship, stimdating discussions and participation in many social engagements! A very special thanks to Bud and Shirley for their constant support and for being such good fiiends to Alison and me. You've made our stay on the Island a very memorable one! 1 would like to thank my parents, Henry and Mary Peters, for their unconditional support and love. I would also like to thank Bill and Phyllis Meadows for their support. Finaily, 1 would like to thank my wife Aiison for marrying me and for her patience, love, and unwavering support during the writing of this manuscript. It is to her that 1dedicate this thesis. TABLE OF CONTENTS AaOJOWLEDGEMENTS ...................................................................................................................i ... TABLE OF CONTENTS ........................................................................................................................ 111 LIST OF TABLES ...........................................................................................................................................ix LIST OF FIGURES ........................................................................................................................................?uv CHAPTER ONE Literature Review ............................................................................................................................................ The Host, Solanurn tuberosum L .............................................................................................. The Irish Potato Famine, 1845- 1849 ....................................................................................... The Pathogen. Phytophthora infistans (Mont.) de Bary ................................................ The Growth of P . hfestans in Culture .................................................................................... Symptoms of Disease ..................................................................................................................... The Disease Cycle .......................................................................................................................... Formation, Survival and Infectivity of Oospores ................................................ Primary Inoculum in Asexual Populations ........................................................ Polycyclic Disease Spread ............................................................................................ Infection of Tubers ........................................................................................................... Contro 1 Measures .............................................................................................................................. Culturai Control ................................................................................................................. Chernical Control ............................................................................................................. Disease Forecasting ......................................................................................................... ... 111 Hast Resistance and Host-Padiogen hteractions .............................................. 78 Global Migrations of P . infstam ..............................................................................................32 The A2 Mahg Type ........................................................................................................ 33 Metdaxyl Reskum......*................................................................................................. 34 Fitness of Introduced Genotypes .............................................................................. 35 Variability in P . infestans ............................................................................................................. 36 Detection of P . infemm ............................................................................................. 36 Mechanisms of Variability ........................................................................................... 37 Measurement of Variability .......................................................................................... 38 Research Objectives ........................................................................................................................................ 42 CHAPTER TWO Characterization of evolving populations of Phytophthora infestm in Canada using mating type and metalaxyl sensitivity markes .................................................................................. 43 Introduction ........................................................................................................................................