Study of Chondrostereum Purpureum and Its Role in the Decline of White Birch in Thunder Bay
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Lakehead University Knowledge Commons,http://knowledgecommons.lakeheadu.ca Electronic Theses and Dissertations Retrospective theses 1990 Study of Chondrostereum purpureum and its role in the decline of white birch in Thunder Bay McLaughlin, John A. http://knowledgecommons.lakeheadu.ca/handle/2453/967 Downloaded from Lakehead University, KnowledgeCommons A STUDY OF Chondrostereum purpureum AND ITS ROLE IN THE DECLINE OF WHITE BIRCH IN THUNDER BAY by John A. McLaughlin© A Graduate Thesis Submitted In Partial Fulfillment of the Requirements for the Degree of Master of Science in Forestry School of Forestry Lakehead University May, 1991 ProQuest Number: 10611832 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. 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ISBN 0-315-63304-6 Canada Abstract McLaughlin. J.A. 1991. A study of Chondrostereum purpureum and its role in the decline of white birch in Thunder Bay, 105 pp. + appendices. Major Advisor; Dr. Edson C. Setliff. Key Words: birch dieback and decline, crystalline secondary metabolite, fruiting, pathogenicity, sexuality, silver leaf. Various aspects of the biology of Chondrostereum purpureum (Pers.:Fr.) Pouz. and its role in dieback and decline of white birch (Betula papyrifera Marsh.) were studied. (1) Sixty-nine trees which exhibited symptoms of dieback and decline were sampled for C. purpureum infection in Thunder Bay, Ontario. Using a variety of sampling methods, including boring, and trunk and branch dissection, C, purpureum was isolated from 26 (38%) of 69 trees sampled. The increment borer sampling technique proved to be the most efficient method of isolating the pathogen, in terms of combining ease of execution and good success rate. (2) The pathogenicity of C. purpureum to birch seedlings was tested. All twenty—six seedlings infected with mycelium developed open wounds, 12 of which cankered, whereas the wounds in 26 controls closed within 4—6 weeks. Progress of the infection varied. Spread of the canker from the wound site (including stem girdling and dieback in two seedlings), spread through the xylem and formation of cankers further up the stem, and apparent containment of the infection at the wound site were observed. (3) The properties of white needle-like crystals which formed on heavy spore casts were investigated. The filamentous crystals were composed of a sesquiterpene compound (Cj^gH2gO, M^ 222, m.p. 136°C) similar to ( + )-torreyol . (4) The"^sexual ity of C. purpureum was investigated. The fungus is heterothal1ic, with tetrapolar sexual differentiation. Anomalies such as unilateral compatibility and unequal abundance of clamps and fruiting in compatible pairings were observed. (5) A method of inducing growth of C. purpureum basidiocarps and their storage was devised. IV Table of contents Page Abstract iii List of tables vii List of figures viii Acknowledgements x Introduction 1 Chapter 1. A Survey of Chondrostereum purpureum Associated With Birch Decline 3 Materials and methods 6 Tree selection 6 Sampling techniques 6 Isolation techniques 8 Results and Discussion 9 Sampling techniques 9 Isolation technique 11 Association with disease 14 Chapter 2. A Study of the Pathogenicity of Chondr'ostereum purpureum to White Birch. 19 Materials and methods 21 Experimental design 21 Stage 1 21 Inoculation technique 21 Stage 2 22 Response variables 23 Results 23 Field observations 23 Wounds and cankers 24 Progress of infection 24 Foliar symptoms 33 Height increment 35 Discussion 36 Inoculation 36 Wound closure and progress of infection 37 Foliar symptoms 39 Height increment 42 V Page Chapter 3. An Investigation of Crystals Produced by Chondrostereum purpureum 43 Materials and methods 45 Crystal production 45 Collecting the crystals 46 Crystal morphology 47 Instrumentation tests 47 Results 49 Crystal production 49 Crystal morphology 50 Instrumentation test results 50 Elemental analysis/X-ray microanalysis 50 Mass spectroscopy 50 XRD 51 FT/IR 51 FT/NMR 53 Melting point 53 Discussion 53 Origin of crystal compound 53 Inter-strain variation in crystal production 55 Secondary metabolites of Chondrostereum purpureum. 56 Identification of the crystal compound 57 Chapter 4. A Study of the Sexuality of Chondrostereum purpureum 60 Materials and methods 62 Results 63 Clamp formation 63 Fruiting 64 Macroscopic interactions 65 Discussion 70 Clamp formation 71 Fruiting 74 Macroscopic interactions 76 Chapter 5. Induced-Growth and Storage of Chondrostereum purpureum Basidiocarps 78 Materials and methods 80 Basidiocarp production 80 Basidiocarp storage 81 Results 83 Basidiocarp production 83 Basidiocarp storage 84 Discussion 86 Basidiocarp production 86 Basidiocarp storage 88 Effect of temperature on spore production 88 Freezing damage 89 Storage in glass vs paper 91 Conclusions and recommendations 91 vi Page Basidiocarp production 91 Basidiocarp storage 92 Summary and Recommendations 93 Literature cited 97 Appendices 106 Appendix I. Water demands of inoculated and non—inoculated seedlings under water stress 107 Appendix II. ANOVA of height increment Ill Appendix III. Instrumentation tests details 113 Appendix IV. A list of mating types and pairings... 116 Appendix V. Calculations of spore production 117 VI 1 List of tables Page Table 1.1. Chondrostereum purpureum isolations from symptomatic white birch 10 Table 2.1. Allocation of treatments in Stage 2 23 Table 2.2. Wound closure 24 Table 2,3. Water demands of inoculated and non- inoculated seedlings subjected to water stress over a period of 42 days 35 Table 2.4. Growth response of white birch seedlings to Chondrosteireum par-pui-eum inoculation and water stress 35 Table 3.1. Basidiocarp strains and crystal production 49 Table 4.1. Relative abundance of clamps and fruiting after 8 weeks 65 Table S.l. Basidiocarp production after two months... 83 Table 5.2. Effects of four storage treatments on spore release over 9 hours 85 VI 1 I List of figures Page Figure 1.1. A symptomatic tree among apparently healthy ones on the Lakehead University campus 9 Figure 1.2. Stages of limb dieback caused by C. purpureum-. death of the infected branch and dying foliage and twigs on the distal portions of the limb 12 Figure 1.3. Distinctive cultural appearance of C. purpureum with uneven margin and clock- wise rotation of leading hyphae 13 Figure 1.4. Typical C. purpureum basidiocarp as it forms in culture 14 Figure 1.5. Decline symptoms throughout a tree infected with C. purpureum 17 Figure 2.1. Dying tree infected with C. purpureum.... 25 Figure 2.2. Canker spreading down from infected branch 26 Figure 2.3. Column of discoloured wood extending from branch wound 27 Figure 2.4. Canker spreading after inoculation 28 Figure 2.5. Wound closure in non-inoculated seedling. 28 Figure 2.6. Canker continuing to spread during Stage 2 29 Figure 2.7. Spread of canker stopped during Stage 2.. 29 Figure 2.8. Canker formation further up stem 31 Figure 2.9. Dieback of an infected seedling 32 Figure 2.10. Vessels plugged by mycelium 32 IX Page Figure 2.11. Spread of the infection from the wound to the other side of the stem and the resultant extensive invasion of xylem.... 33 Figure 2.12. Leaf of seedling in advanced stage of disease 34 Figure 2.13. Healthy leaf of a non-inoculated seedling 34 Figure 3.1. Needle—like crystals arising from dense spore casts of C. purpureum 51 Figure 3.2. Needle-like crystals - various lengths and diameters 52 Figure 3.3. SEM micrograph showing filamentous bundle structure , 52 Figure 3.4. SEM micrograph showing striated, sheet -like structure of crystals 53 Figure 4.1. Mating reactions of single—spore isolates of C. purpureum 64 Figure 4.2. Macroscopic cultural interactions between paired single-spore isolates of C. purpureum after 2 wk. A. uncommon—AB X A2B2) B. common-A (A^B^ X A^B2> C. common-B (A^B^ X A2B^) D. common—B (A2B2 X A^B2> E. common-A (A2B2 X A2B^) F. uncommon—AB (A^B2 X A2B^) 67 Figure 4.2 (cont'd). G. common-AB (A2B2 X A2B2) H. common-AB (A^B2 X A^B2) I. & J. uncommon—AB (A^B^ X A2B2) after 5 wk, with and without fruiting, respectively K. & L. uncommon—AB (A^B2 X A2B^) after 5 wk, with and without fruiting, respectively 68 Figure 4.3.