Simplicillium Lanosoniveum, a Mycoparasite of Phakopsora Pachyrhizi and Its Use As a Biological Control Agent Nicole A
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Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2011 Simplicillium lanosoniveum, a mycoparasite of Phakopsora pachyrhizi and its use as a biological control agent Nicole A. Ward Louisiana State University and Agricultural and Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Plant Sciences Commons Recommended Citation Ward, Nicole A., "Simplicillium lanosoniveum, a mycoparasite of Phakopsora pachyrhizi and its use as a biological control agent" (2011). LSU Doctoral Dissertations. 3189. https://digitalcommons.lsu.edu/gradschool_dissertations/3189 This Dissertation 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 Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. SIMPLICILLIUM LANOSONIVEUM, A MYCOPARASITE OF PHAKOPSORA PACHYRHIZI AND ITS USE AS A BIOLOGICAL CONTROL AGENT A Dissertation 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 Doctor of Philosophy In The Department of Plant Pathology and Crop Physiology By Nicole A. Ward B.S., Louisiana State University, 2007 May 2011 ACKNOWLEDGEMENTS This project was funded by the Louisiana Soybean and Grain Research and Promotion Board, Southern Sustainable Agriculture Research and Education, Organic Crop Improvement Association, and the LSU Chapter of Sigma Xi. ii TABLE OF CONTENTS ACKNOWLEDGEMENTS ............................................................................................................ ii LIST OF TABLES ......................................................................................................................... vi LIST OF FIGURES ...................................................................................................................... vii ABSTRACT ................................................................................................................................... ix CHAPTER 1. INTRODUCTION .................................................................................................. 1 CHAPTER 2. COLONIZATION OF SOYBEAN RUST SORI BY SIMPLICILLIUM LANOSONIVEUM .......................................................................................................................... 9 2.1 Introduction ...................................................................................................................... 9 2.2 Methods .......................................................................................................................... 10 2.2.1 Identification of Colonists....................................................................................... 10 2.2.2 Verification of Colonization ................................................................................... 12 2.2.3 Morphological Characterization ............................................................................. 14 2.3 Results ............................................................................................................................ 14 2.3.1 Identification of Colonists....................................................................................... 14 2.3.2 Verification of Colonization ................................................................................... 15 2.4 Discussion ....................................................................................................................... 19 CHAPTER 3. SUPPRESSION OF PHAKOPSORA PACHYRHIZI, CAUSAL AGENT OF SOYBEAN RUST, BY THE MYCOPHILIC FUNGUS SIMPLICILLIUM LANOSONIVEUM 25 3.1 Introduction .................................................................................................................... 25 3.2 Methods .......................................................................................................................... 26 3.2.1 Co-inoculations of Disease-free Soybean Leaves................................................... 26 3.2.2 Inoculation of Infected Soybeans Leaves ............................................................... 29 3.2.3 Urediniospore Production and Germination ........................................................... 30 3.2.4 Sorus Age Study ..................................................................................................... 33 3.2.5 Statistical Analyses ................................................................................................. 34 3.3 Results ............................................................................................................................ 34 3.3.1 Co-inoculations of Disease-free Soybean Leaves................................................... 34 3.3.2 Inoculation of Infected Soybean Leaves ................................................................. 35 3.3.3 Urediniospore Production ....................................................................................... 36 3.3.4 Sorus Age Study ...................................................................................................... 41 3.4 Discussion ....................................................................................................................... 41 CHAPTER 4. EFFECTS OF FIELD INOCULATIONS OF SOYBEANS WITH THE MYCOPHILIC FUNGUS SIMPLICILLIUM LANOSONIVEUM ON PHAKOPSORA PACHYRHIZI AND SOYBEAN RUST ....................................................................................... 46 iii 4.1 Introduction .................................................................................................................... 46 4.2 Methods .......................................................................................................................... 47 4.2.1 Primer and Probe Development .............................................................................. 47 4.2.2 Field Studies............................................................................................................ 51 4.2.3 Field Experiments ................................................................................................... 52 4.2.4 DNA Quantification ................................................................................................ 55 4.4 Results ............................................................................................................................ 56 4.4 Discussion ....................................................................................................................... 63 CHAPTER 5. MYCOPARASITISM OF PHAKOPSORA PACHYRHIZI BY SIMPLICILLIUM LANOSONIVEUM: A MICROSCOPY STUDY ......................................................................... 69 5.1 Introduction .................................................................................................................... 69 5.2 Methods .......................................................................................................................... 70 5.2.1 Urediniospore Collection ........................................................................................ 70 5.2.2 Scanning Electron Microscopy ............................................................................... 71 5.2.3 Confocal Microscopy .............................................................................................. 72 5.2.4 Transmission Electron Microscopy ........................................................................ 74 5.3 Results ............................................................................................................................ 75 5.3.1 Scanning Electron Microscopy ............................................................................... 75 5.3.2 Confocal Microscopy .............................................................................................. 75 5.3.3 Transmission Electron Microscopy ........................................................................ 76 5.4 Discussion ....................................................................................................................... 80 CHAPTER 6. SURVEY OF ADDITIONAL RUSTS FOR SIMPLICILLIUM LANOSONIVEUM ........................................................................................................................ 88 6.1 Introduction .................................................................................................................... 88 6.2 Methods .......................................................................................................................... 89 6.3 Results ............................................................................................................................ 90 6.4 Discussion ....................................................................................................................... 90 CHAPTER 7. EFFECT OF EXPOSURE OF HYALINE AND BROWN UREDINIOSPORES OF PHAKOPSORA PACHYRHIZI TO SOLAR RADIATION ................................................... 96 7.1 Introduction .................................................................................................................... 96 7.2 Methods .......................................................................................................................... 97 7.3 Results ...........................................................................................................................