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Spatial and Temporal Sensitivity of Alternaria Species Associated with Potato Foliar Diseases to Demethylation Inhibiting and Anilino-Pyrimidine Fungicides

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Spatial and Temporal Sensitivity of Alternaria Species Associated with Potato Foliar Diseases to Demethylation Inhibiting and Anilino-Pyrimidine Fungicides SPATIAL AND TEMPORAL SENSITIVITY OF ALTERNARIA SPECIES ASSOCIATED WITH POTATO FOLIAR DISEASES TO DEMETHYLATION INHIBITING AND ANILINO- PYRIMIDINE FUNGICIDES A Thesis Submitted to the Graduate Faculty of the North Dakota State University of Agriculture and Applied Science By Arthur Dimitri Lakshan Fonseka Gunawardena In Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE Major Department: Plant Pathology November 2015 Fargo, North Dakota North Dakota State University Graduate School Title SPATIAL AND TEMPORAL SENSITIVITY OF ALTERNARIA SPECIES ASSOCIATED WITH POTATO FOLIAR DISEASES TO DEMETHYLATION INHIBITING AND ANILINO-PYRIMIDINE FUNGICIDES By Arthur Dimitri Lakshan Fonseka Gunawardena The Supervisory Committee certifies that this disquisition complies with North Dakota State University’s regulations and meets the accepted standards for the degree of MASTER OF SCIENCE SUPERVISORY COMMITTEE: Dr. Neil C. Gudmestad Chair Dr. Gary A. Secor Dr. Julie S. Pasche Dr. Asunta L. Thompson Approved: November 17, 2015 Dr. Jack B. Rasmussen Date Department Chair ABSTRACT Early blight and brown spot, caused by Alternaria solani and Alternaria alternata, respectively, are important foliar diseases of potato, affecting both tuber yield and quality. Most of the commercial cultivars lack resistance; therefore, application of foliar fungicides remains a primary management strategy. Correlation coefficients comparing EC50 values for conidial germination and mycelial growth of A. solani and A. alternata in response to boscalid and fluopyram, respectively, were low, indicating that the association between these two in vitro assays was very weak. Baseline sensitivities of Alternaria spp. to difenoconazole, metconazole, and pyrimethanil using mycelial growth assays demonstrated high intrinsic activity against the two pathogens. Six out of 245 A. solani isolates exhibited reduced-sensitivity to pyrimethanil in in vitro assays and reduced-sensitive isolates were not controlled except at 100 µg/ml in greenhouse efficacy tests. The DMI chemistries and pyrimethanil remain valuable options for fungicide rotation programs in areas of high disease pressure. iii ACKNOWLEDGEMENTS I would like to extend my gratitude to my major professor, Dr. Neil C. Gudmestad for his guidance and advice during my graduate program. I am also grateful to Dr. Gudmestad for providing me an opportunity to conduct research on an interesting topic and providing valuable knowledge of plant pathology. I would also like to thank my committee members, Drs. Gary Secor, Julie Pasche, and Asunta Thompson, for their time and advice. A special thanks is extended Dr. Pasche for her guidance in the laboratory assays. Additionally, I thank Dr. Raymond Taylor, Christopher Johnson, Ipsita Mallik, Vanessa Johansen, Rachel Shjerve, Dean Peterson, Mitchell Bauske, Sarah Budde, Dr. Kishore Chittem, and Curt Doetkott for valuable suggestions and helping to conduct this research. The author is especially thankful to all the faculty, staff, and graduate students of the Department of Plant Pathology for their support and encouragement. Financial support from Bayer CropScience, Syngenta Crop Protection, and Valent U.S.A. Corporation is gratefully acknowledged. Finally, I would to like to thank my family members and friends for the unconditional support, encouragement, and understanding throughout the years. iv TABLE OF CONTENTS ABSTRACT ............................................................................................................................................... iii ACKNOWLEDGEMENTS .......................................................................................................... iv LIST OF TABLES ........................................................................................................................ vii LIST OF FIGURES .................................................................................................................... viii LIST OF APPENDIX TABLES ................................................................................................... ix LITERATURE REVIEW ................................................................................................................1 History of Potato .................................................................................................................1 Taxonomy and Nomenclature ..............................................................................................1 Biology of Alternaria solani and Alternaria alternata ............................................................ 6 Symptomatology ......................................................................................................................... 13 Epidemiology ............................................................................................................................... 14 Disease Management .................................................................................................................. 17 Fungicide Resistance .................................................................................................................. 28 MATERIALS AND METHODS ........................................................................................................... 40 Collection and Maintenance of Isolates ................................................................................... 40 Comparison of Conidial Germination Inhibition to Mycelial Growth Inhibition .............. 41 In Vitro Baseline Sensitivity of Alternaria solani and Alternaria alternata to Difenoconazole and Metconazole ........................................................................................ 46 In Vitro Baseline Sensitivity of Alternaria solani and Alternaria alternata to Pyrimethanil ............................................................................................................................ 47 Sensitivity of Non-Baseline Isolates of Alternaria solani to Difenoconazole, Metconazole, and Pyrimethanil ................................................................................................ 47 Effect of Reduced-Sensitivity of Alternaria solani to Pyrimethanil on Disease Control . 47 Statistical Analysis ...................................................................................................................... 49 RESULTS .................................................................................................................................................. 51 v Comparison of Conidial Germination Inhibition to Mycelial Growth Inhibition ............. 51 In vitro Baseline Sensitivity of Alternaria solani and Alternaria alternata to Difenoconazole and Metconazole ........................................................................................ 55 In Vitro Baseline Sensitivity of Alternaria solani and Alternaria alternata to Pyrimethanil .......................................................................................................................59 Sensitivity of Non-Baseline isolates of Alternaria solani to Difenoconazole, Metconazole, and Pyrimethanil ................................................................................................. 61 Effect of Reduced-Sensitivity of Alternaria solani to Pyrimethanil on Disease Control .. 64 DISCUSSION .......................................................................................................................................... 66 LITERATURE CITED ............................................................................................................................ 75 APPENDIX A. COLLECTION DATA OF ALTERNARIA SOLANI ISOLATES COLLECTED FROM FIELDS IN THE 2010, 2011, 2012, 2013, AND 2014 SEASONS FROM SEVERAL STATES .................................................................................................................. 92 APPENDIX B. COLLECTION DATA OF ALTERNARIA ALTERNATA ISOLATES COLLECTED FROM FIELDS IN THE 2011, 2013, AND 2014 SEASONS FROM SEVERAL STATES ................................................................................................................................ 99 APPENDIX C. CV-8 AGAR MEDIUM ............................................................................................ 102 APPENDIX D. L-ASPARAGINE (ASP-AGAR) MEDIUM .......................................................... 103 APPENDIX E. SUMMARY OF STATISTICAL ANALYSIS FOR THE IN VITRO COMPARISON OF CONIDIAL GERMINATION TO MYCELIAL GROWTH (FLUOPYRAM) ..................................................................................................................................... 104 APPENDIX F. SUMMARY OF STATISTICAL ANALYSIS FOR THE IN VITRO COMPARISON OF CONIDIAL GERMINATION TO MYCELIAL GROWTH (BOSCALID) ......................................................................................................................................... 105 APPENDIX G. SUMMARY OF STATISTICAL ANALYSIS FOR THE IN VITRO BASELINE SENSITIVITY TESTS OF ALTERNARIA SOLANI TO DMI FUNGICIDES ........ 106 APPENDIX H. SUMMARY OF STATISTICAL ANALYSIS FOR THE IN VITRO BASELINE SENSITIVITY TESTS OF ALTERNARIA ALTERNATA TO DMI FUNGICIDES ......................................................................................................................................... 107 APPENDIX I. SUMMARY OF STATISTICAL ANALYSIS FOR THE IN VITRO BASELINE SENSITIVITY TESTS OF PYRIMETHANIL............................................................ 108 APPENDIX J. SUMMARY OF STATISTICAL ANALYSIS FOR
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