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Characterization and Management of Ralstonia Solanacearum Populations in South Asia

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Characterization and Management of Ralstonia Solanacearum Populations in South Asia Characterization and Management of Ralstonia solanacearum Populations in South Asia DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Nagendra Subedi Graduate Program in Plant Pathology The Ohio State University 2015 Dissertation Committee: Sally A. Miller, Advisor Christopher G. Taylor Pierce A. Paul Anne E. Dorrance Copyrighted by Nagendra Subedi 2015 Abstract Bacterial wilt caused by Ralstonia solanacearum (Smith) Yabuuchi is a major problem for tomato, eggplant and pepper production in South Asia. This disease is difficult to manage due to viability, adaptability, and diversity of the pathogen. To develop information regarding local pathogen population structure to inform disease management strategies, we characterized a collection of 100 R. solanacearum strains from South Asia using classical and recent molecular tools. All of the strains in this collection were race 1, phylotype I, and biovar III or IV. Based on phylogenetic analysis of endoglucanase gene sequences, we identified three sequevars (14, 47, and 48), and two putatively new sequevars among the South Asian strains. Cluster analysis of genomic fingerprinting profiles created by Rep-PCR divided strains into eight groups. Strains were not grouped based on geographic origin, host or biovar in either analysis. To identify resistant tomato, eggplant and pepper genotypes that can potentially be used in South Asia to manage this disease, we screened a worldwide collection of 37 tomato, eggplant, and pepper accessions against six selected South Asian R. solanacearum strains. Six tomato, nine eggplant, and three pepper accessions were highly resistant (≤10% wilt), and three tomato, two eggplant and one of the pepper accessions were moderately resistant (≤30% wilt). ii Biological control is a potentially economically feasible and environmentally safe and sustainable disease management practice. In this study we investigated 54 previously characterized bacterial biocontrol agents for their activity against diverse R. solanacearum strains from South Asia and evaluated the value of integration of these biocontrol agents with partial host resistance of tomato in management of bacterial wilt. Based on in vitro antagonism against 15 selected South Asian R. solanacearum strains, six biocontrol agents were selected to evaluate their biocontrol efficacy in a susceptible (L390) and a partially resistant (IRAT L3) tomato accessions. Biocontrol agents were almost four times more effective in suppressing bacterial wilt in IRAT L3 than in L390. Pseudomonas brassicacearum strains 93D8 and Wood 1R, and P. protegens strain Clinto 1 were the most effective strains with biocontrol efficacy of 67, 50 and 58% respectively, in IRAT L3. Bacterial wilt incidence was suppressed in all four experiments by Pseudomonas brassicacearum strain 93D8, and in three out of four experiments by P. vranovensis strain 15D11, P. protegens strain Clinto 1, and P. brassicacearum strain Wood 1R, in IRAT L3. However, in L390, the disease incidence was suppressed in only one experiment by P. protegens strains 15G2 and Clinto 1, and a mixture of all biocontrol agents. These results highlight the value of integration of biocontrol agents with host resistance in management of bacterial wilt. iii Dedication Dedicated to My Family iv Acknowledgments I wish to express my sincere thanks to my supervisor Professor Sally A Miller for her support and guidance throughout my research. I would like to thank the members of my student advisory committee, Dr. Christopher G Taylor, Dr. Pierce A. Paul and Dr. Anne E. Dorrance for their helpful comments and support. I am thankful to Dr. Brian McSpadden Gardener for helping me design and execute biocontrol experiments. My special thanks to Dr. Fulya Baysal Gurel, Dr. Melanie L. Ivey, Jony Mera, Bob James, Mafruha Afroz (Limi), Anna Testan, Claudio Vrisman, Nitika Khatri, Mynul Islam, Ferdous Elahi (Jabin), Xing Ma, Nick Rehm, Hugo Pantigoso and Angela Nanes for helping me carryout my experiments. I would like to thank Dr. Jaw-Fen Wang, AVRDC, Taiwan, Dr. Marie Christine Daunay, INRA, France, and Dr. Yousouf Mian, BARI, Bangladesh, for providing seeds for this research; and Bangladesh Agricultural Research Institute (BARI) and Nepal Agricultural Research Council (NARC) for providing laboratory facilities in Bangladesh and Nepal, respectively. I would like to acknowledge Integrated Pest Management Innovation Lab (IPM IL) and U.S. Agency for International Development (USAID) for financial support. v Vita 2001..................................................M. Sc. Botany, Tribhuvan University 2009..................................................M. S. Plant Pathology, The Ohio State University 2011 to present ................................Graduate Research Associate, Department of Plant Pathology, The Ohio State University Publications Miller, S. A., Mera, J. R., Pantigoso, H. A., Vrisman, C. M., and Subedi, N. 2015. Evaluation of fungicides for the control of foliar and fruit diseases of processing tomatoes, 2014. PDMR 9:V062. The American Phytopathological Society, St. Paul, MN. Subedi, N., Testen, A. L., Baysal-Gurel, F. and Miller, S. A.. 2014. First Report of Black Leaf Mold of Tomato Caused by Pseudocercospora fuligena in Ohio. Plant Disease 99:285. Subedi, N., Gilbertson, R. L., Osei, M. K., Cornelius, E. and Miller, S. A. 2014. First Report of Bacterial Wilt Caused by Ralstonia solanacearum in Ghana, West Africa. Plant Disease 98:840. vi Subedi, N., and Miller, S. A. 2014 Resistance of a worldwide collection of resistant tomato, eggplant and pepper lines to South Asian strains of Ralstonia solanacearum Phytopathology 104:S3. Baysal-Gurel, F., Subedi, N., Mamiro, D. and Miller, S. A. 2014. First Report of Anthracnose of Onion Caused by Colletotrichum coccodes in Ohio. Plant Disease 98:1271. Testen, A. L., Mamiro, D. P., Meulia, T., Subedi, N., Islam, M., Baysal-Gurel, F. and Miller, S. A. 2014. First Report of Leek yellow stripe virus in Garlic in Ohio. Plant Disease 98:574. Subedi, N., Baysal-Gurel, F., Hoitink, H., Ivey, M. and Miller, S. A. 2010. Regulation of genes involved in the interaction of tomato, Trichoderma hamatum 382 and Xanthomonas euvesicatoria. Phytopathology 100:S124 Miller S. A., Mera, J. R., Baysal-Gurel, F., and Subedi. N. 2009. Evaluation of fungicides to control Sclerotinia drop in Lettuce, 2008. PDMR 3:V010. The American Phytopathological Society, St. Paul, MN. Subedi, N. 2009. Use of biorational products for the control of diseases in high tunnel tomatoes and induction of certain defense genes in tomato by Trichoderma hamatum 382. MS Dissertation, The Ohio State University, Document no. osu1250602215. vii Fields of Study Major Field: Plant Pathology viii Table of Contents Abstract .......................................................................................................................... ii Dedication...........................................................................................................................iv Acknowledgments ...........................................................................................................v Vita ............................................................................................................................... vi List of Tables................................................................................................................. xi List of Figures ............................................................................................................. xiii Chapter 1: Introduction ...................................................................................................1 Chapter 2: Characterization of Ralstonia solanacearum populations and screening host resistance to manage bacterial wilt in South Asia ........................................................... 27 Introduction ............................................................................................................. 28 Materials and Method .............................................................................................. 31 Results ..................................................................................................................... 34 Discussion ............................................................................................................... 38 References....................................................................................................................45 Chapter 3: Combining partial host resistance with bacterial biocontrol strains improves outcomes for tomatoes infected with Ralstonia solanacearum ......................71 ix Introduction................................................................................................................72 Materials and Methods...............................................................................................75 Results..........................................................................................................................79 Discussion....................................................................................................................82 References....................................................................................................................88 Bibliography....................................................................................................................111 x List of Tables Table 2.1 Tomato,
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