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Variabilin and Mechanism of Saskatoon Disease

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Variabilin and Mechanism of Saskatoon Disease VARIABILIN AND MECHANISM OF SASKATOON DISEASE RESPONSE TO EUTOMOSPORIUM MESPU A Thesis Submitted to the College of Graduate Studies and Research in Partial Futfilment of the Requirements for aie Degree of Doctor of Philosophy in the Department of Plant Sciences University of Saskatchewan Saskatoon, Saskatchewan, Canada b~ Philip Ronald Spring, 2001 Q Copyright Philip Ronald, 2001. Ali rights resenred. Aequiitions and Acquissions et Bibtiagraphic Services senrices bibliographiques The author has granted a non- L'auteur a accordé une licence non exclusive licence ailowing the exclusive permettant à la National Library of Canada to Bibliothèque nationale du Canada de reproduce, loan, distri'bute or seii reprodilire, prêter, distn'buer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. ia forme de microfiche/fIlm, de reproduction sur papier on sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni la thèse ni des substantieIs may be printed or otherwise de celle-ci ne doivent être imprimés reproduced withouî the author's ou autrement reproduits sans son permission. autorisation. VARlABlLlTY AND MECHANISM OF SASKATOON OlSEASE RESPONSE TO ENTOMOSPORIUM MESPIU -BY PHILIP RONALD A ttiesis submitted to the Faculty of Graduate Studies of the University of Saskatchewan in partial fulfilment of the requirements of the degree of DOCTOR OF PHILOSOPHY Permission has been granted to the LIBRARY OF THE UNIVERSITY OF SASKATCHEWAN to lend or seIl copies of this thesis, to the NATIONAL LIBRARY OF CANADA to microfilm this thesis and to lend or seIl copies of the film, and to UNIVERSITY MICROFILMS to publish an abstract of this thesis. The author reserves other publication rights, and neither the thesis nor extensive extracts from it may be pnnted or otherwise reproduced without the author's men permission. ABSTRACT Entomosporium leaf and beny spot (Entomosporium mespiii (DC.) Sacc., is a serious and economically important disease of saskatoon (Amelanchier ainifoIia Nutt.). The objectives of this study were to assess the variability of saskatoon disease response to E. mespiii, and to investigate the mechanisms of infection and host resistance for this pathogen. Cultivars of saskatoon were characterized for incidence and severity of Entomosporium leaf and berry spot. Detached leaves of 17 cultivars were evaluated for disease severity after inoculation with E. mespiii under controlled conditions. Leaves and fruit, sampled from plants of the same cultivars following natural infection, were evaluated for disease incidence and severity. Analysis of data from al1 experiments revealed significant differences (P 5 0.05) in disease response among saskatoon cultivars. Following natural infection in 1998 and 1999, cultivar means for leaf acervuli number ranged from 5.3 to 142-7 per leaf and from 6.8 to 98.8 per leaf, respectively. Parkhill, Regent and Success showed consistently lower values for the incidence and severity of fungal sporulation on leaves and fruit. Variation in leaf disease response to E. mespili was observeci among self- and cross-pollinated seedling populations derived from saskatoon cultivars. Seffing cultivars with partial resistance to Entomosporium leaf spot, praduced segregating progenies that were skewed tbward higher susceptibility. Mean leaf acervuli number ranged from 63.4 per leaf for seedlings of Honeywood, to 237.5 per leaf for seedlings of Smoky. Plants with resistance to leaf spot were identified among self- pollinated populations derived from saskatoon cultivars, as well as in three other species of Amelanchier. Population means for leaf acervuli number ranged frorn ** 11 2.0 per leaf for Amelanchisr intemedia to 134.8 per leaf for Amelanchier femaldii. Most seif-poilinated seedling progeny derived from a ptant of A. htemredia wmpletely nestricteci sporulation on leaves following infection by E mespili. Scanning electron microscopy was used to elucidate the infection process of E. mespili on leaves of two saskatoon cultivars, Buffalo and Success. Conidiospore germination occurred rapidly on leaves of both cultivars, usually by the extrusion of a single major germ tube from the larger of two body cells. Spore penetration of the leaf cuticle involved appressoria formed at germ tube termini or directly below the germinating spore. Sub-spore appressoria were most common and resulted in the early formation of leaf acenruli. At 2 days post-inoculation, 43% of genninating conidiospores had penetrated the leaf surface of Buffalo, compared with 12% on leaves of Success. Acervuli formed on leaves of al1 17 saskatoon cuitiiars, but cultivars dïfFered in acenrulus surface area and number of acenruli per leaf. Pre- and post-infection structural defense mechanisms were observed in inoculated leaves of saskatoon cultivars. Cuticle thickness was unifom across resistant and susceptible cultivars, but differences in leaf thicf ness and anatomy appeared to be important in reducing hyphal spread in leaves of resistant cultivars. Papiltae composed of suberin or callose were formed at sites along the adaxial epiderrnal layer in leaves of both resistant and susceptible cultivars. Hyphal encasements were rnost effective in resistant cultivars, Iikely due to their earlier formation. In leaves of resistant cuftivars, the formation of suberin deposits in palisade mesophyll cetls appeared to confine fungal activity to epidemal cells. When the pathogen accessed the leaf interior, the rapid and complete suberization of mesophyll celk at the periphery of the lesion sequestered hyphae in an isolated ... lll region and limited the nurnber of sporulation sites on the leaf surface. Characteriration of disease response to Entornosponurn leaf and beny spot among available saskatoon cultivars provides a valuable informational resource for individuals seeking to establish new orchards. This is especially true for growers interested in producing fruit organically, without synthetic fungicidal disease control. Planting less-susceptible saskatoon cultivars and implementing cultural disease controls, such as drip irrigation and the removal of suckers, may limit the amount of leaf and fruit damage caused by E. mespili. ACKNOWLEDOEMENTS The author is grateful to Drs. Richard St-Pierre, Jill Thomson, Geoff Hughes, Janet Taylor and Art Davis for their supervision and advice throughout the project. Dr. Piara Bains collected disease data at Edmonton in 1997 and provided constructive cnticism on the third chapter of this thesis. Dr. Bob Baker provided statistical advice on numerous occasions. In the winter of 1999,l was given the opportunity to spend a term of study in the Department of Plant Pathology at the University of Minnesota. Iwould like to acknowledge Dr. Bill Bushnell for supervising my study, as well as Gib Ahlstrand and Darryl Knieger for assistance with microscopy. Appreciation is extended to Valerie Catinot for valuable assistance with fieldlindoor studies and data collection. Dr. Campbell Davidson of Agriculture Canada in Morden, Manitoba, and Dr. Dirk Demeyre of the National Botanical Garden in Belgium provided seed samples of Amelanchier species. Appreciation is due to the faculty and students of the Department of Plant Sciences at the University of Saskatchewan. I have enjoyed my term of study at this institution and would recommend it to others. The friendships I have made and the mernories of the city will always be camed with me. Lastly, I would like to acknowledge the support of my farnily. My wife Karen has shown great patience and provided constant encouragement throughout the Wngprocess. She was also a great help in typesetting and proof-reading the thesis. My parents Wiibert and Sharon Ronald taught me to be diligent and persistent, qualities that have been instrumental in bringing me aius far in my education. A graduate scholarship from the College of Agriculture and grants hmthe Agriculture Development Fund funded this project TABLE OF CONTENTS COPYRIGHT CLAIMS............................................................................................. i ABSTRACT ............................................................................................................. ii ACKNOWLEDGMENTS .......................................................................................... v TABLE OF CONTENTS ......................................................................................... vi LIST OF TABLES ................~.....~.~..........................................................................ix LIST OF FIGURES ............................................................................................ xi 1. INTRODUCTION................................................................................................ 2 2.1 Plant Disease Response .................... ,, ............................................. 6 2 1 Concepts and terrninology ...................................................... 6 2.1.2 Assessrnent of disease response ................... .... ................. 7 2.1 -3 Measurement of disease response ........................................ 8 2.1.4 Genetic control of disease resistanœ ...................................10 2.1.5 Defence rnechanisms against disease ................................. 11 2.2 Entornospon'um Leaf and Berry Spot .............................................. 15 2.2.1 Causal
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