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A Cultural Study of Paceykytospora Tuberculosa (Dc

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A Cultural Study of Paceykytospora Tuberculosa (Dc A cultural study of Pachykytospora tuberculosa (DC. ex Fr.) Kotl. et Pouz. Authors Eboh, Daniel Okoye 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 26/09/2021 10:33:34 Link to Item http://hdl.handle.net/10150/318710 A CULTURAL STUDY OF PACEYKYTOSPORA TUBERCULOSA (DC. ex Fr.) Kotl. et Pouz. ■by- Daniel Okoye Eboh A Thesis Submitted to the Faculty of the DEPARTMENT OF PLANT PATHOLOGY In Partial Fulfillment of the Requirements For the Degree of MASTER OF SCIENCE In the Graduate College THE UNIVERSITY.OF ARIZONA 19 6 8 STATEMENT BY AUTHOR This thesis has been submitted in partial fulfillment of requirements for an advanced degree at The University of Arizona and is deposited in the University 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 acknowledgment of source is made. Requests for permission for extended quotation from or reproduction of this manu­ script in whole or in part may be granted by the head of the major department or the Dean of the Graduate College when in his judgment the proposed use of material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author. SIGNED: APPROVAL BY THESIS DIRECTOR This thesis has been approved on the date shown below: ROBERT"DnT2T?nm L.r GILBERTSONr<TTDTnmonM DatevtT+rr f Professor of Plant Pathology ACKNOWLEDGMENTS The author expresses sincere gratitude for the guidance and counsel of Dr. Robert L. Gilbertson, Pro­ fessor of Plant Pathology, University of Arizona, under whose inspiration and direction this study was conducted. Thanks are also extended to Dr. Alice M. Boyle of the Department of Plant Pathology, University of Arizona, for her motherly interest in my work. Appreciation is expressed to the United States Agency for - International Development for the USAID scholarship under which the graduate study program was conducted. TABLE OF CONTENTS Page LIST OF ILLUSTRATIONS..................... .. vi LIST OF TABLES ................ viii' ABSTRACT .... ... ............... ix INTRODUCTION . ............... 1 LITERATURE REVIEW ........ 3 Taxonomy ................ 3 Geographical Distribution ........... 5 Substratum Relationships . •.............. 6 MATERIALS AND METHODS .................. 7 Study of Basidiocarps .... ..... .. .... ." 7 Specimens examined . .............. 7 Macroscopic and microscopic study methods ........ 8 Study of Cultures . .. ... .’ 8 Isolation methods . .......... 8 Media u s e d .......... ............. 8 Oxidase tests . ...................... 9 Temperature study ........ ....... 9 Effect of light on mycelial growth . 10 Growth on different concentrations of tannic acid . ' . ....... 10 Attempts to induce fruiting in vitro •. 12 Chemical Monokaryotization .......... 13 Mo no kar y o t i z at i on by blending ....... 14 Utilization of carbon sources ....... 15 RESULTS AND DISCUSSION . ;. 19 Morphology of Basidiocarps . ............ - 19 • Macroscopic c h a r a c t e r s ......... 19 Microscopic characters . ... ... 20 Remarks on similar fungi .............. 25 iv V Page Cultural Morphology..... ................ 35 Growth characters 35 Microscopic characters ........ .... 35 Key patterns according to Davidson and Nobles .............. 36 Remarks on possible phylogeny ...... 36 Temperature Study . ...... 37 Effect of Light . ...................... 38 Monokaryotization- . ... .' . 4.0 Attempts to Induce Fruiting in Vitro . .... 43 Growth on Different Concentrations of Tannic Acid .................... 43 Carbon Utilization 44 SUMMARY ..............•.............. 45 LITERATURE CITED . 48 LIST OF 'ILLUSTRATIONS Figure ■ Page 1 Oak twig medium set. -up to induce in vitro fruiting of _P. tuberculosa (RLG 7712) . 11 2 Basidiocarp of _P. tuberculosa (RLG 7252) on the bark of an oak b r a n c h .............. 18 3 Microscopic characters of marginal hyphae of basidiocarps of P. tuberculosa (RLG 7252) a, thin-walled generative hyphae; b, thick- walled skeletal hyphae with apical and inter­ calary swellings . ';.................... 21 4 Context hyphae of basidiocarp of _P. tuber­ culosa (RLG 7252) a, thick-walled skeletal hypha; b, thin-walled generative hypha . 22 5 Hymenial structure of P. tuberculosa (RLG 7 2 5 2) a, cystidioles; b, basidiospores; c, basidia bearing basidiospores; d, basidia in different stages, of development showing globose form of young b a s i d i a ............ 23 6 Basidium bearing basidiospores of _P. tuber­ culosa (RLG 7 7IO) . • • . , 24 7 Basidiospores of _P. tuberculosa (RLG 7252) showing the • "verrucas11 that appear in more or less definite rows ........... 26 8 Culture of P. tuberculosa (RLG 7708) showing fan-pat tern type of growth ........ 27 9 Culture of J?. tuberculosa (RLG 7711) showing distinct mycelial clumps or puffs ..... 27 10 Culture of P. tuberculosa (RLG 7708) on tannic acid medium showing a strong positive oxidase reaction . ....... ; 28 vi vii Figure Page 11 A Petri dish culture of J?. tuberculosa (RLG- 7252) growing on malt extract agar medium with oak t w i g s ................ 29 12 A flask culture of _P. tuberculosa (RLG 7252) • growing on malt extract agar medium with oak t w i g s ................................. 29 13 Microscopic structures of advancing zone of a young culture of J?. tuberculosa (RLG 7709) a, thick-walled hyphae; b, thin-walled hyphae 3 c, crystals ................... 30 14 ' Distorted, thin-walled generative hyphae of _P. tuberculosa (RLG 7708) from a raised tuft of mycelium on a culture grown on malt extract agar and incubated at 25°C 31 15 Skeletal hyphae of ]?. tuberculosa (RLG 7706) from oak medium incubated at 2b°C. Note the terminal and intercalary swellings ........ 32 16 Hyphae of advancing zone of an old culture of P..tuberculosa (RLG 7708) showing chlamydospore-like cells with thin walls . 33 17 Microscopic features of P. tuberculosa (RLG.7 7 0 8) showing the intricately branched complexes of generative hyphae ....... 34 18 Generative hyphae of P. tuberculosa (RLG 7 7 0 8) showing thin-walled vesicle-like ' s t r u c t u r e ............................ 34 19 Growth of the Fungus at different tempera- • t u r e s .................... .. 39 20 Growth of the Fungus at.different concen­ trations of tannic acid •.............. 4l 21 Cultures of JP. tuberculosa (RLG 7708) grown on different concentrations of tannic acid . a) Culture grown on 0.04^ concentration of tannic acid b) Culture grown on 0.09^ con­ centration of tannic acid c) Culture grown on 0 .1 9 0 concentration of tannic acid. .. 42 LIST OF TABLES Table . Page I Temperature study using malt extract agar as the medium ...... „ 38 II 'Temperature study using malt extract agar •containing pieces of oak as the medium . 40 III ■ Growth of the fungus on three concentrations of tannic acid . ........................ 44 viii ABSTRACT Pachykytospora tuberculosa is a fungus that is known in the western hemisphere only from southern Arizona. It is associated with white rot of Arizona white oak (Quercus arizonica Sarg.). Although this plant is of no : economic importance for lumber, its recreational and watershed values should not be overlooked. In this research, morphological and physiological studies of this fungus were carried out. Ten isolates were cultured from the basidiocarps obtained from the field. Microscopic examinations revealed that two hyphal types are present in both basidiocarps and cultures, the thin-^walled generative hyphae with clamp connections, frequent branching and occasional swellings j and thick- walled skeletal hyphae with occasional branching, and swellings. The optimum temperature for growth is 25°C and growth is inhibited at 30°C and above. There was no success in the investigation on the carbon utilization since this fungus did not grow in any of the liquid media used. The fungus showed extreme variability and erratic growth in culture. Swollen and highly contorted hyphae are present in .some cultures. The fungus fruits" ' poorly In vitro, and It seems to require an additional metabolite, presumably tannin or tannin derivative, for good vegetative growth. It grows much better on malt extract agar medium containing oak twigs. INTRODUCTION 'Very little research has been done on the diseases and decays of the many hardwood species occurring in Arizona. Although these trees are not of great commercial importance for lumber production, they are extremely impor­ tant fOr watershed protection and for their aesthetic value in recreational areas. .This research concerns a fungus causing decay in Arizona white oak (Quercus. arizonica Sarg.). • Pachykytospora tuberculosa (DC. ex Fr.) Kotl. et Pouz., is a Bymenomycete in the family Polyporaceae. It has been known in Europe since 1815 but has been known ■ from North America only recently' (Gilbertson, 1 9 6 1) and has been found here only in southeastern Arizona. No further investigations on this interesting fungus have been carried out and its.geographical distribution and substratum relationships in North America, cultural characteristics, sexuality, physiology, and pathological significance are unknown. The objectives of this research are as follows: (1) to.determine the cultural morphology
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