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Identification of the Causal Agent of Leaf and Crown Rot of Liriope In Louisiana State University LSU Digital Commons LSU Master's Theses Graduate School 2015 Identification of the Causal Agent of Leaf and Crown Rot of Liriope in Louisiana Carla Milena Proano Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_theses Recommended Citation Proano, Carla Milena, "Identification of the Causal Agent of Leaf and Crown Rot of Liriope in Louisiana" (2015). LSU Master's Theses. 70. https://digitalcommons.lsu.edu/gradschool_theses/70 This Thesis is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Master's Theses by an authorized graduate school editor of LSU Digital Commons. For more information, please contact [email protected]. IDENTIFICATION OF THE CAUSAL AGENT OF LEAF AND CROWN ROT OF LIRIOPE IN LOUISIANA A Thesis Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Master of Science in The School of Plant, Environmental and Soil Sciences By Carla Milena Proano Herrera B.S., Pan American School of Agriculture Zamorano, 2010 August 2015 ACKNOWLEDGEMENTS First of all I would like to thank God and my family for giving me the strength, support, love, and being the reason that encouraged me to keep pursuing and finish my degree. I would like to give my special thanks to Dr. Kuehny for giving me the opportunity to work with him, for his time, knowledge, guidance, patience and help during this process. Also, I would like to thank to my committee members Dr. Raghuwinder Singh, Dr. Edward Bush and Dr. Don Labonte for providing me their knowledge, help and guidance. I am also really thankful to the Botanic Gardens staff and student workers for being really helpful during my project and Timothy Burks for teaching me the laboratory methods needed for my research. My special gratitude to the LSU AgCenter and Dr. William Richardson for providing financial support which allowed me to study at Lousiana State University. Finally, I would like to thank my friends of ZAS (Zamorano Agricultural Society), for being my adopted family, for their unconditional support and friendship during all this time. My special gratitude to my friends that helped me any time I needed them: Fabián Díaz, Damir Torrico, José Brandao, to help me finish my project. This work would not have been possible without them. I am also thankful to Katherine Rubio and José Alonso for their unconditional support. ii TABLE OF CONTENTS ACKNOWLEDGEMENTS ............................................................................................................ ii LIST OF TABLES .......................................................................................................................... v LIST OF FIGURES ....................................................................................................................... vi ABSTRACT ................................................................................................................................. viii CHAPTER 1: INTRODUCTION ................................................................................................... 1 1.1 Ornamental industry in the United States and Louisiana ...................................................... 1 1.2 Ground Covers ...................................................................................................................... 1 1.3 Liriope in the United States and Louisiana ........................................................................... 2 1.4 Leaf and Crown Rot of Liriope ............................................................................................. 3 1.5 Objectives .............................................................................................................................. 6 CHAPTER 2: LITERATURE REVIEW ........................................................................................ 7 2.1 Liriope ................................................................................................................................... 7 2.1.1 Morphology or Phytography of Liriopogons.................................................................. 7 2.1.2 Culture ............................................................................................................................ 9 2.1.3 Insects and Diseases ..................................................................................................... 10 2.2 Phytophthora spp. ............................................................................................................... 12 2.2.1 Phytophthora palmivora: Biology, Ecology, Taxonomy ............................................. 16 2.2.2 Disease Cycle of Phytophthora Root Rot and Crown Rot ........................................... 17 2.2.3 Spread and Control ....................................................................................................... 18 2.2.3.1 Biological Control .................................................................................................. 19 2.2.3.2 Chemical Control ................................................................................................... 20 2.3 Fusarium spp. ...................................................................................................................... 20 2.3.1 Fusarium oxysporum: Biology, Ecology, Taxonomy .................................................. 23 2.3.2 Control .......................................................................................................................... 25 CHAPTER 3: IDENTIFICATION OF PHYTOPHTHORA PALMIVORA AND FUSARIUM OXYSPORUM ON SYMPTOMATIC LIRIOPE PLANTS .......................................................... 27 3.1 Introduction ......................................................................................................................... 27 3.2 Materials and Methods ........................................................................................................ 28 3.2.1 Isolation and Identification of Causal Organisms ........................................................ 28 3.2.1.1 Phytophthora spp. .................................................................................................. 28 3.2.1.2 Fusarium spp. ........................................................................................................ 29 3.2.2 Long Term Storage of Phytophthora and Fusarium .................................................... 30 3.2.3 DNA Extraction ............................................................................................................ 31 3.2.3.1 Phytophthora palmivora ........................................................................................ 31 3.2.3.2 Fusarium oxysporum ............................................................................................. 31 3.2.4 PCR (polymerase chain reaction) Amplification and Sequencing ............................... 32 3.3 Results ................................................................................................................................. 33 3.3.1 Phytophtora palmivora Sequence Analysis ................................................................. 34 3.3.2 Fusarium oxysporum Sequence Analysis ..................................................................... 36 iii 3.4 Discussion ........................................................................................................................... 36 CHAPTER 4: PATHOGENICITY TESTS ON LIRIOPE CULTIVARS .................................... 39 4.1 Introduction ......................................................................................................................... 39 4.2 Materials and Methods ........................................................................................................ 40 4.2.1 Plant Production and Experiment Design ..................................................................... 40 4.2.2 Experimental Design .................................................................................................... 41 4.2.3 Inoculum preparation .................................................................................................... 42 4.2.3.1 Phytophthora palmivora ........................................................................................ 42 4.2.3.2 Fusarium oxysporum ............................................................................................. 43 4.2.4 Inoculation Method....................................................................................................... 43 4.2.5 Data Analysis ................................................................................................................ 43 4.2.6 Re-isolation of Phytophthora palmivora and Fusarium oxysporum ............................ 44 4.2.7 Statistical Analysis ....................................................................................................... 44 4.3 Results ................................................................................................................................. 45 4.3.1 Leaf and Root Fresh and Dry Weight ........................................................................... 45 4.3.2 Percent of Disease Incidence ........................................................................................ 46 4.3.3 Root Rot Incidence
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