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Ecology and Management of Pythium Species in Float Greenhouse Tobacco Transplant Production

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Ecology and Management of Pythium Species in Float Greenhouse Tobacco Transplant Production Ecology and Management of Pythium species in Float Greenhouse Tobacco Transplant Production Xuemei Zhang Dissertation submitted to the faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Plant Pathology, Physiology and Weed Science Charles S. Johnson, Chair Anton Baudoin Chuanxue Hong T. David Reed December 17, 2020 Blacksburg, Virginia Keywords: Pythium, diversity, distribution, interactions, virulence, growth stages, disease management, tobacco seedlings, hydroponic, float-bed greenhouses Copyright © 2020, Xuemei Zhang Ecology and Management of Pythium species in Float Greenhouse Tobacco Transplant Production Xuemei Zhang ABSTRACT Pythium diseases are common in the greenhouse production of tobacco transplants and can cause up to 70% seedling loss in hydroponic (float-bed) greenhouses. However, the symptoms and consequences of Pythium diseases are often variable among these greenhouses. A tobacco transplant greenhouse survey was conducted in 2017 in order to investigate the sources of this variability, especially the composition and distribution of Pythium communities within greenhouses. The survey revealed twelve Pythium species. Approximately 80% of the surveyed greenhouses harbored Pythium in at least one of four sites within the greenhouse, including the center walkway, weeds, but especially bay water and tobacco seedlings. Pythium dissotocum, followed by P. myriotylum, were the most common species. Pythium myriotylum, P. coloratum, and P. dissotocum were aggressive pathogens that suppressed seed germination and caused root rot, stunting, foliar chlorosis, and death of tobacco seedlings. Pythium aristosporum, P. porphyrae, P. torulosum, P. inflatum, P. irregulare, P. catenulatum, and a different isolate of P. dissotocum, were weak pathogens, causing root symptoms without affecting the upper part of tobacco seedlings. Pythium adhaerens, P. attrantheridium, and P. pectinolyticum did not affect tobacco seeds or seedlings. The consequences of Pythium infection were more likely to be severe when they occurred during seed germination than at seedling emergence, or after plant stem elongation when seedling roots had started to grow into underlying nutrient solutions, depending on the species of Pythium. High and low variation was observed among isolates of P. dissotocum and P. myriotylum, respectively. Pythium myriotylum co-existed with multiple other Pythium or oomycete species in the same environments within tobacco greenhouses, and significant in vitro and/or in vivo interactions between P. myriotylum and some naturally co-existing species were revealed. Pythium porphyrae may have the potential to protect tobacco seeds and seedlings from P. myriotylum infection. Greenhouse Pythium control trials identified ethaboxam, mefenoxam, and copper ionization as potentially promising alternatives to etridiazole for Pythium disease management in tobacco transplant production. The outcomes of this project provide useful new information to better understand the composition, distribution, and diversity of Pythium communities in tobacco transplant greenhouses and to improve Pythium disease management for tobacco transplant production. Ecology and Management of Pythium species in Float Greenhouse Tobacco Transplant Production Xuemei Zhang GENERAL AUDIENCE ABSTRACT Pythium diseases are common in tobacco transplant production and can cause up to 70% seedling losses in hydroponic (float-bed) tobacco transplant greenhouses. However, little is known about the composition and distribution of Pythium communities in tobacco transplant greenhouses. This project began with a tobacco transplant greenhouse survey, in which 12 Pythium species were recovered from center walkways, weeds, greenhouse bay water, and tobacco seedlings. Pythium dissotocum and P. myriotylum were the two types (species) of Pythium most commonly found in the survey. Pythium myriotylum, P. coloratum, and P. dissotocum were aggressive pathogens that suppressed seed germination and caused root rot, stunting, foliar chlorosis, and death of tobacco seedlings. Pythium aristosporum, P. porphyrae, P. torulosum, P. inflatum, P. irregulare, P. catenulatum, and an isolate of P. dissotocum, were weak pathogens causing root symptoms without affecting the upper part of tobacco seedlings. Pythium adhaerens, P. attrantheridium, and P. pectinolyticum did not affect tobacco seeds or seedlings. The symptoms caused by infection by Pythium species differed among host (tobacco) growth stages, except for the most aggressive species, P. myriotylum. High levels of variation were observed among isolates of P. dissotocum, in terms of vegetative growth rate (on V8 agar media) and aggressiveness on tobacco seed and seedlings. Pythium myriotylum was found to co- exist with multiple other Pythium or oomycete species (neighbor isolates) in the same environments within tobacco greenhouses. Significant interactions between P. myriotylum and some neighbor isolates were revealed, and these interactions significantly affect the consequences of P. myriotylum infection of tobacco seeds. Greenhouse Pythium control trials identified two chemical water treatments (ethaboxam and mefenoxam), and a non-chemical water treatment (copper ionization) as potentially promising alternatives to the current standard Pythium control (etridiazole) for Pythium disease management in tobacco transplant production. The outcomes of this project provide useful new information to both better understand the composition, distribution, and diversity of Pythium communities in tobacco transplant greenhouses and to improve Pythium disease management for tobacco transplant production. Dedications I am dedicating this work to my parents (Gui and Ying) and my brother (Song), who are always kind and supportive. Without their love and support, I would not be able to pursue my dream. This dissertation is also dedicated to the love of my life, Howard, and our lovely son Nick, who are the sunshine of my life. I am grateful for the joy they bring to me. vi Acknowledgements I would like to gratefully acknowledge many people who have been guiding and supporting me during my PhD study, without their help I would not have made it this far. I am extremely thankful for my advisor Dr. Charles Johnson, who has been a wonderful mentor to me. All the research and life experience that he has shared with me have guided me through many, many difficult times. I could not imagine where I would be if he were not there guiding and supporting me. I deeply appreciate this opportunity of working with him and learning from him. I would also like to express my gratitude to my advisory committee members: Dr. David Reed, Dr. Anton Baudoin, and Dr. Chuanxue Hong, who have been incredibly generous and supportive, providing advice, space, and experimental materials for me whenever I needed them. My heartfelt gratitude extends also to Mary Ann Hansen and Elizabeth Bush at the plant disease clinic on campus, who generously provided tremendous support in my research and personal life. I also would like to thank Stephen Barts, Taylor Clarke, and other colleagues who helped me collect survey samples. My sincere gratitude also goes to Spencer, Noah, Tyler, Laura, Molly, and my other laboratory mates, without whose help I would not be able to conduct so many trials at the Southern Piedmont Agricultural Research and Extension Center (SPAREC). I also would like to thank Dr. Carol Wilkinson, Brad, Melody, Margaret, Grace, and other folks in the SPAREC family; they have been so kind and supportive to me in every aspect. Last but not least, my special “thanks” goes to my dear Pythium friends. I am so thankful that they have been so patient with me and showed me a wonderful world that not everybody would get a chance to see. vii Contents Dedications .................................................................................................................................... vi Acknowledgements ....................................................................................................................... vii Contents ....................................................................................................................................... viii List of Figures ................................................................................................................................ xi List of Tables ................................................................................................................................ xv Chapter 1 ......................................................................................................................................... 1 Introduction ............................................................................................................................... 1 1.1 Background ................................................................................................................... 2 1.2 Objectives ..................................................................................................................... 3 1.3 Broader Impact ............................................................................................................. 4 Chapter 2 ......................................................................................................................................... 5 Review of Literature ................................................................................................................
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