University of Montana ScholarWorks at University of Montana Graduate Student Theses, Dissertations, & Professional Papers Graduate School 1972 Characterization of two strains of tobacco mosaic virus Jacqueline Fletcher Perryman The University of Montana Follow this and additional works at: https://scholarworks.umt.edu/etd Let us know how access to this document benefits ou.y Recommended Citation Perryman, Jacqueline Fletcher, "Characterization of two strains of tobacco mosaic virus" (1972). Graduate Student Theses, Dissertations, & Professional Papers. 6656. https://scholarworks.umt.edu/etd/6656 This Thesis is brought to you for free and open access by the Graduate School at ScholarWorks at University of Montana. It has been accepted for inclusion in Graduate Student Theses, Dissertations, & Professional Papers by an authorized administrator of ScholarWorks at University of Montana. For more information, please contact [email protected]. CHARACTERIZATION OF TWO STRAINS OF TOBACCO MOSAIC VIRUS By Jacqueline F. Perryman B.S., Emory University, 1970 Presented in partial fulfillment of the requirements for the degree of Master of Science University of Montana 1972 Approved by: Chairman, Board of Examiners De^, G r S ch oo 1 n 7 z . Date ÿ f / 7 3 88 4 61 UMl Number: EP37457 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. UMT Oisssrtation PuMi«h*ng UMl EP37457 Published by ProQuest LLC (2013). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code Pro Quest ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 2- 2 !'7 2 ACKNOWLEDGEMENTS I wish to express my sincere thanks to Dr. Meyer Chessin for his encouragement and guidance during this investigation. I also appreciate the assistance, especially during the preparation of this paper, of Drs. R. A. Sol berg, R. N. Ushijima, and C. C. Gordon, as members of my graduate committee. Special thanks are due to Mr. Lallan Giri for his patient help and advice during the study, and to Mr, Klaus Lackschewitz for his care of the experimental plants in the greenhouse. 11 TABLE OF CONTENTS CHAPTER PAGE I. INTRODUCTION ........................................................................................................ 1 Statement of Problem ............................................................................................. 1 Review of Background Material .......................................................................... 2 Host Range Studies .......................................................................................... 4 Effects of Ultraviolet Light on Plant Viruses .................................. 5 II. MATERIALS AND METHODS ..................................................................................... 12 Host Range Study of the Virus Found in Ribgrass ...................................... 12 Source of Virus and Method of Inoculation .......................................... 12 Selection of Host Plants and Greenhouse Conditions ....................... 12 UV and Photoreactivation Experiments .......................................................... 13 Choice of the Virus and the Assay Plants ............................................ 13 Preparation of Virus Solutions ................................................................. 13 Inoculation ..................................... 15 Photoreactivation ................................................ 16 I I I . RESULTS ................................................................................................................... 18 Host Range of Ribgrass Virus ................................................ 18 UV and PR Experiments .......................................................................................... 25 CaVX 196 tests .................................................. 25 Ribgrass virus tests ........................................................................................ 27 Combined Results of UV and PR Experiments ................................................ 35 CaVX 196 ................................................................................................................ 35 Ribgrass Virus ................................................................................................... 35 i i i CHAPTER PAGE IV. DISCUSSION ............................................................................................................. 40 Host Range of HRV-M .............................................................................................. 40 UV and PR Experiments With CaVX 196 .......................................... 41 UV and PR Experiments With HRV-M ................................................................... 43 V. SUMMARY ...................................................................................... 47 BIBLIOGRAPHY ..................................................................................................................... 49 IV LIS T OF TABLES TABLE PAGE 1. Host range of the Montana ribgrass virus and comparisons with symptoms produced by Holmes' ribgrass virus, HRV-Y, and classical TMV ...................................................................... 19 2. Results of some back-inoculations from species inoculated with the Montana ribgrass virus ............................................... 25 3. Effect of UV treatment and post-UV illumination on virus survival ................................................................................ 28 LIS T OF FIGURES FIGURE PAGE 1. Nicotiana sylvestris infected with Cactus Virus X 196 showing chi orotic and necrotic lesions .................................................... 11 2. UV absorption spectrum of Montana ribgrass virus from recording spectrophotometer ........................................................................... 17 3. Montana ribgrass-infected Chenopodium quinoa. Left: uninoculated leaf with chlorosis Right: inoculated leaf with necrotic lesions ...................................... 22 4. Cheopodium quinoa. Left: healthy leaf. Right: uninoculated leaf showing chlorosis ............................................ 22 5. Plantago lanceolata. HRV-M inoculated leaves with many small local lesions .................................................................................. 24 6. Plantago lanceolata. HRV-M inoculated leaves with a few large necrotic lesions ........................................................ 24 7. Survival-dose curve of clarified juice of CaVX 196 after UV irradiation and inoculation into _N. glutinosa plants which were then exposed to visible light or placed in darkness for 24 hours ................................................................... 32 8. Survival-dose curve of clarified juice of CaVX 196 after UV irradiation and inoculation into _N. sylvestris plants which were then exposed to visible light or placed in darkness for 24 hours .................................................................................. 33 VI FIGURE page 9. Survival-dose curve of partially purified HRV-M after UV irradiation and inoculation into _N. gl utinosa plants which were then exposed to visible light or placed in darkness for 24 hours .......................................................................................... 34 10. Most accurage straight line plots of data combined from experiments using irradiated purified and unpurified solutions of CaVX 196 after inoculation into qlutinosa plants which were then exposed to visible light or placed in darkness for 24 hours ............................. 36 11. Most accurate straight line plots of data combined from all experiments using CaVX 196, comparing survival of UV irradiated virus in 1^. gl utinosa plants exposed to visible light to that in plants placed in darkness for 24 hours.............................................................................................. 37 12. Survival-dose curves of purified and partially purified solutions of HRV-M after UV irradiation and inoculation into glutinosa plants which were then exposed to visible light or placed in darkness for 24 hours ............................. 38 13. Most accurate straight line plots of data combined from experiments using HRV-M, comparing survival of irradiated virus in gl utinosa plants exposed to light to that in plants placed in darkness for 24 hours ........................................................ 39 VI 1 FIGURE PAGE 14. A comparison of the susceptibilities to UV inactivation of CaVX 196 and HRV-M with several other TMV strains studied by Siegel and Wildman, by survival-dose curves .......... 46 VI 1 1 Chapter I INTRODUCTION Statement of Problem My thesis research has been based on the characterization of two plant viruses using a variety of methods. During the winter of 1971, I became interested in studying a virus found commonly in the weedy plantain or ribgrass, Plantago media, on the lawns of the University of Montana. A plant virus isolated from a different species of ribgrass, P_. lanceolata, had been reported in 1940 by Holmes, and since then, other strains of the virus have been reported, I fe lt it would be interesting to