Verticillium Wilt of Cotton: Studies of Possible Seed Transmission
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VERTICILLIUM WILT OF COTTON: STUDIES OF POSSIBLE SEED TRANSMISSION Item Type text; Dissertation-Reproduction (electronic) Authors Allen, Ross Marvin, 1917- Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 05/10/2021 07:56:55 Link to Item http://hdl.handle.net/10150/289699 VERTICILLIUM WILT OF COTTON: STUDIES OF POSSIBLE SEED TRANSMISSION by Ross M. Allen A. Thesis submitted to the faculty of the Department of Plant Pathology in partial fulfillment of the requirements for the degree of DOCTOR OP PHILOSOPHY in the Graduate College, University of Arizona 1953 Approved!. ______________ , 20,1933 ^ Director of Thesis QDate This thesis has been submitted in partial fulfillment of requirements for an advanced degree at the University of Arizona and is deposited in the Library to be made available to borrowers under rules of the Library. Brief quotations from this thesis are allowable without special permission, provided that accurate aclmowledgment of source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department or the dean of the Graduate College when in their judgment the proposed use of the material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author. SIGNED:- 11 TABLE OP CONTENTS SECTION PAGE Introduction .................................. 1 Review of the I,i terature .......... 4 History . 4 Importance . ............... ...... 6 Distribution and Losses * .............. 6 Host Range . 10 Symptoms .. 10 On Mature Plants ......................... 10 On Seedling Plants ....................... 15 Variations of Expression 15 The Pathogen 17 Taxonomy ............ 17 Morphology .......... 17 Physiology ................... 21 Temperature Relations 21 Influence of p H ............ 26 Nitrogen Source ............ 26 Carbohydrate Source .......... 29 Enzyme and. Alkaloid Influence ............. 30 Effect of Dyes ................. 31 Cause of Wilt ..................... 31 Pathogenicity ........................... 33 Correlated with Morphology.......... 33 Temperature Influence 35 Cross-inoculation Experiments ............... .36 Mode of Attack .......... 40 Transmission and Spread . ... ... .. .. 41 By Seed ............... 41 By Root Contact ............... 44 By Vegetative Propagation ............. 45 By Plant Debris ........ I . 45 By Cultural Practices ................ 47 By Miscellaneous Means ........... 48 Environmental Factors Affecting the Disease . 48 Soil ............. 50 Soil Moisture ................ 52 Soil and Air Temperature . ..... ... 53 Soil Reaction. ............... 55 Soil Fertility .............. 56 Soil Microflora ................... 58 Crop History ............ 59 Vertical Distribution of Pathogen ....... 60 iv SECTION PAGE Control of the Disease ...................... 60 Resistant Varieties 60 Cultural Practices ........... 63 Soil Amendments .................... 65 Chemical Disinfection ........................... 65 Seed Treatment ...... 66 Legal Measures ........................... 67 Methods and Results . 68 Collection and Description of Cultures . 68 Preliminary Experiments with SxP Seeds .......71 Histological Studies of Verticillium-inoculated S e e d s .................................. 74 Pathogenicity Test TCsing Verticillium-infested Oats 76 Laboratory Studies ..................... ...... 79 Seed Inoculations, 1950 79 Seed Inoculations, 1951. ........... 85 Seed Inoculations, 1952 ............... .. 87, Field and Greenhouse Studies ..................... 92 Greenhouse Plantings- Inoculated Seeds, 1950 . , 92 Field Plantings- Inoculated Seeds, 1950 ..... 99 Field Plantings- Inoculated Seeds, 1951 102 Greenhouse Plantings- Inoculated Seeds, 1951 . 109 Field Plantings- Inoculated Seeds, 1952 ..... 112 Greenhouse Plantings- Inoculated Seeds, 1952 . 115 Field Inoculations, 1951 ............. 117 Greenhouse Inoculations, 1951 .......... 141 pathogenicity Test by Hypodermic Inoculation . 145 Culture of Young Bolls, from Verticillium- wilted Plants ......................... 149 Pathogenicity Studies of Isolates Using Tomato, Okra, and Eggplant . .......... 151 A Previously Undescribed Method of Isolating Verticillium ............... 152 Growth Comparisons of Isolates on Various Media . 156 Discussion and Conclusions ................... 161 Summary .............................. 174 Literature Cited ..... ........... ........ 177 Appendix LIST OF TABLES NUMBER PAGE 1, Additional plant hosts of Verticillium s p e c i e s ..................... 11 2. Temperature relations of Verticillium species ................... •••24 3^ pH range of Verticillium spp. in culture . 27 4* Results of pathogenicity test in artificially infested soil ......... •••••••••• 78 5. Inoculation of cotton seed by hypodermic Injection........ ......................... 80 6• Cultures of hypodermically-inoculated seeds, long-staple varieties •••••••... 82 7. Cultures of hypodermically-inoculated seeds, short-staple varieties .......... 83 8^ Drilled seed inoculated by Method B • • • • • 88 9^ Results of culturing seeds inoculated by Method C .................................. 91 10. Numbers of infected cotton plants in green house resulting from Verticillium-inocu- lated seeds ••••••••••«••••• 94 11. Numbers of infected cotton plants in green house resulting from Verticillium-inocu- lated seeds ••••....••.••.. 95 12. Numbers of infected cotton plants in green house resulting from Verticillium-inoeu- lated seeds ..•..••....•••.. 96 13. Numbers of infected cotton plants in green house resulting from Verticillium-inocu- lated seeds ....... .................. 97 14. Emergences occurring in field test, 1950 . 101 Vi WTJMBEÎR - FAGJil 15. Hills showing emergence 14 days after plant ing seeds inoculated by Method B .............105 16. Results of cultures of Method B-inocula ted seeds on potato-dextrose agar ....... 108 17. Results of planting seeds inoculated by Method B in sterile soil in the greenhouse, and of culturing 21-day old seedlings . Ill 18. Results of field planting, 1952 ....... 116 19. Results of planting unwounded, inoculated seeds in sterile soil in greenhouse .... 118 20. Culture of branches and branch-inoculated b o l l s ................. ............... .. 124 21. Culture of branches and branch-inoculated b o l l s ......... ........... .............. .. 125 22. Culture of bolls and seeds inoculated with Gliocladium throu^ boll pe d i c e l s ........... 134 23. Culture of bolls and seeds inoculated with Thomas strain of Verticillium through boll pedicels ............. 135 24. Culture of bolls and seeds inoculated with McNeal strain of Verticillium throu^ boll pedicels .......... ........ 136 25. Culture of bolls and seeds inoculated with Tipton strain of Verticillium throu^ boll pedicels ............... 137 26. Culture of bolls and seeds inoculated with CMI strain of Verticillium through boll pedicels ........... ............. 139 27. Location of Verticillium colonies growing from infectied seeds T .............. 142 28. Results of boll and pedicel inoculations of greenhouse plants .......... 146 vii LIST OF FIGURES (In Appendix) FUMBER 1. Photomicrograph of conidiophores and conidia of Gliocladium rossum. 2. Typical colonies of Verticillium strains and . Gliocladium on potato-dextrose agar. 3. Equipment used for inoculation of seeds by Method A, 4. Verticillium growing from wound-site and micropyle of SxP variety seeds, 5. Verticillium growing from germinated, inoculated seeds. 6. Photomicrograph of Verticillium conidiophore. 7. Photomicrograph of a portion of a longitudinal section of a Verticillium-infected seed, 8. Camera lueIda drawing of Verticillium hypha penetrating parenchyma cell-walis, 9. Ten-week old cotton plant showing typical foliar symptoms. 10, Verticillium growing from the wound-site in the cotyledon and from the fringe tissue inside the cast-off seed coat, 11, Seedlings showing Verticillium growing on coty ledons and cast-off seed coat. 12, Drilling apparatus used for preparation of seeds for inoculation by Method B. 13, Verticillium growing from seeds inoculated by Method C, 14, Verticillium colonies on potato-dextrose agar to which the fungus was carried from the in terior of seeds by seedlihgs. viii NUMBER 16. Growth, of Vertlcllli'um from the inoculation wound-rsite and from the chalazal end of the germinated seeds, 16. Cotyledons wounded in inoculation by Method B. Verticillium colonies at wound-sites, 17. Hypodermically-inoculated cotton stems showing vascular discoloration. IB. Modified kitchen strainer used for surface- sterilizing seeds. 19. Gliocladium cultured from pedicel and receptacle of a boll. 20. Locks of lint from bolls inoculated through the pedicel. 21. Photomicrographs of pseudosclerotia in and on lint fibers. 22. Photomicrographs of resting mycelium in and on lint fibers. 23. Gliocladium-invaded lint fiber. 24. Gliocladium on septa and placental column of cotton boll. 25. Wilted cotton plants 11 days after hypodermic inoculation. 26. Cotton plants 13 days after hypodermic inocu lation. 27. Growth of 4 isolates of Verticillium and 1 of Gliocladium on wort agar. 28. Growth of 4 isolates of Verticillium and 1