
CONTROLLED ENVTRONMENT PROPAGATION AND CULTURE OF CASTILLEJA COCCINIIA (t. ) SPRENG., 4rNq4_O_NE PATENS VAR. WOLFGANGIANA 1ANSS. ) KOCH, AND CYPRIPEDIUM CALCEOLUS VAR. PARVIFLORUM (SNLTSE.) FERN. A Thesis Submitted to the Faculty of Graduate Studies fhe University of Manitoba By Judith Anne Zastre In Partial FuLfilLment oF the Requirements for the Degree of Master of Science Department of Plant Science Aprir r9B4 l.* :. / -.. l. ,,i:Í iì,1 i,t ï f 'ì CONTROLLED ENVIRONMENT PROPAGATION AND CULTURE OF CASTILLEJA clqcrNr4 (1. ) spnrNc., ANEMONE rATENS vAR I^IOLFGANGIAIIE (BESS. ) NOCH , AND cypnrpnortn't iar,cnolus vAR. PARVTFLoRUM (sAl,rs¡. ) rnnx BY JUDITH ANNE ZASTRE A thesis submitted to the Faculty of Graduate Studies of the university of Ma¡ritoba in partial fulfill¡nent of the requirenrents of the degree of MASTER OF SCIENCE o ,1985 Per¡nission has bee' granted to ttre LIBRARy oF THE UNIVER- SITY OF MANITOBA to lend or sell copies of this thesis. to the NATIONAL LIBRARY OF CANADA to microfilnr this thesis ard to lerd or sell copies of the film, and UNIVERSITY MICROFILMS to publish a¡r abstract of this thesis. The author reserves other publicatiorl rights, a¡rd neither the thesis nor extensive extracts fronl it may be printed or other- wise reproduced without the author's writtelr pernrission. l-I ACKNOWLEDGEMENTS The author wishes to gratefully acknowledge the follow- ing for their assistance in tire preparation of this thesis: prof . L. Lenz for his advice and gr:.i'Jance tÌrroughout my studies. The National scienct: and Engineering Research council and the university of i"lanitoba for their financiaL assist- ance. Dr.G.PlatfordandMr.H.Skinnerfort]reirkind donation of their time and planL materials' FelLowgracluatestudents,staffandfacultymembersfor all their unstinting assistance' And foremost, to my husband Erroll, for alL his st-tpport and understanding. l_l_]. TABLE OF CONTENTS Page LIST OF TABLES vi LIST OF I¡IGURES vi ii ABSTRACT x l- FOREWARD xr INTRODUCTION I CHAP'IER I. LITERATURtr REVIEW 3 Seed ProPagation 3 Germination 3 /1 DormancY Light Requirement 5 High EnergY Reaction 7 PhotoPigments I Low EnergY Reaction Mechanisms 9 High EnergY Reaction Mechanisms 10 Castilteia Seed Gei:mination II Anemone Seed Germination L2 Cypripe<fium Seed Germination l3 Seed and Seedling DamPing-off r5 Vegetative ProPagation L7 Vegetative and Reproductive Growth 20 Me<lia TYPes 20 PIant Nutrition 23 Vernalization 26 Parasitic PLants 29 iv Page Plant Descri,otion and Habitat 32 Castilleia coccinea 32 Anemone patens var. wol fqanqiana 34 Cypripedium calceolus var. parviflorum 37 MATERIALS AND METHODS 4L PreliminarY Studies 4L General Methods 42 CIIAPTER II. CONTROLLED ENVIRONI{ENT PROPAGA'IION AND CULTURE O.b- CASTILLEJA COCCINEA (I.) SPRENG 43 Tntroduction 43 l"taterials and Methods 45 Seed CoLlect.ion ê'nc1 Storage 45 Seed Germination 45 Low Light IntensitY StudY 50 High Light IntensitY StudY 5I Host SpecificitY 52 Results and Discr:ss ion 54 Conclusions an<1 SummarY BO CHAPTER III. CONTROLLED ENVIRONMENT PROPAGATION AND CULTURE OF ANEMON E PATBNS VAT. IVOLFGANGIANA ( ASSS .) KocH 82 r ntroduttion a2 Materials and Methods B5 Seed Collection and Storage B5 Seed Germination B5 PreliminarY StudY 85 Ttrermogradient StudY a7 V Page Seed Fungicide Treatments 90 Seed Treatments and Media 9r After-ripening 92 Plant StudY 92 FertiLízer Recluirement SLudY 92 Dormancy Induction antf Termination 93 Results and Discussion 95 Conclusions and SummarY L2L CHAPTER IV. CONTROLLED ENVIRONMENT PROPAGATION AND CULTURE OF CYPRIPEDIUM CALCEOLUS VAT. PARVIFLORUM (snr.rss. ) FERN L23 Introduction L23 MateriaLs and Methods L27 Plant Collection and PreParation r27 Vernalization and Dormancy of Unpotted P lants L27 Vernalization of PotteC Plants L29 Propagation L29 Results and Discussion r33 Conclusions and SummarY I48 I,ITERATURE CITED r,50 APPENDIX r59 VI LIST OF TABLES TabLe Page t The effect of light and dark and various tem- perature regimes on the germinat.ion of Castitteia coccinea seed 55 a 2 The germination of U coccinea seecl on thermograclient Plate 59 .) seed under J Germination of Castilleia coccinea low light int ens it ies with varior:s tempera- tures, filters, exposure and inbibition tiines 60 4 The effect of increased exposure bime of low intensity light on the germination of Castille a coccinea seed 6I seed 5 The germination of CastiÌIe a coccinea under high light intens i ty wl- th various fi lters 63 67 6 Effect oE Castilleia. on host survival 6B 7 Effect of ttosts on Castilleja s urvival s o'f B The effect of various hosts on the ize Casti lle ia 75 grovrn 9 Ttre mean cross sectional area of hosts or with . coccinea orC cocc inea forma alone I 79 lutescens . 10. The effect of light condition s and various temperature regimes of the nean germination of Anemone Patens var. wolfg angrana in vitro 96 ll. The effect of various fungicides on the germination of Anemone P.alene var. wolfg angrana in vitro r00 L2" The effect of various media on the germination and survival of Anemone Patens var' wol f an aana seed IOI 13" The effect of various fungicides on the germination and seedling survival of Anemone patens var. wolfg anqrana r03 L4. The effect of short P eriods of aft er-ripening on Anemone patens var. woI fgangiana seed in vitro r03 vrt Table Page r5. The eff ects of various fr:rtiLizer treaLments on Anemone patens var. wolfgangr-ana 110 16. The effects of various fertilizer treatmeilts on the dry and fresÌ't weights of Anemone patens var. wolfg angr-ana 1r3 L7 Ttre effect of length o:E time of low tetnperature storage on the flowering of Anemone patgLq. var' wolfg anql-ana r17 r8. Tlre effect of length of low temPerature storage on the flowering of rL edium calceolus var parviflorum r36 r9. The effect of low temPerature storage time on the increase in crovln numbers of C. calceolus var. parviflorum r45 20. The effect oE previous Plant height on the increase of crown numlters of C. calceolus var. parviflorum L46 V]- II LIST OF FIGURES Figure Page 1. Outline map of Manitoba showing collection sites 46 2 Diagram of the experimental set-up used for thermogradient studies 49 3 Rate o:E germination of C. coccinea seed under various temperature regr-mes 57 4 The survival of g. coccinea and cocc r-nea forma lutescens over t rme 66 5 The effect of various hosts on the dry weight of C. coccinea 7L 6 The effect ot various hosts on the dry weight of C. coccinea forma lutescens . 73 7 The effect of Castillefa on the dry weights of the host plants 77 B Outline map of Manitoba showing collection s ites B6 9 Diagram of the experimental set-up used for thermogradient studies B9 10. The effect of temPerature on the germination of Anemone atens var. wolf an iana seed 9B ll. The effect of length of after-ripening on the germination of Anemone patens var. wolfg angl-ana seed r06 L2. The effect of after-ripening on the germination of Anemone patens var. wolfg angr-ana s eed lo8 13. The effect of length of Iow temperature storage on leaf num'oer of Anemone Patens var. woI fgangiana ll9 L4. Outline map of Manitoba showing collection site L2B 15. Root and crown structure of Cypripedium calceolus var. Parviflorum r31 r6. The effect of length of low temperature s torage on the mean height of C. calceolus var. oarvi- florum storecl unPotted 135 ].X Page F igure L7. The effect of lengLh of low temperature storage on the height of potted g calceolus var. parviflorum r40 18. The effect of length of low temperature storage on the daYs to flowering of C . calceolus var. rviflorum L42 x ABSTRACT Zastre, Juclith Anne. M.Sc., The University of Manitoba, April, f985. Control led Environment Propagation and Cul-ture of Castilleia coccinea (1. ) Spreng., Anemone patens var. wolfg anqr-ana (ness. ) Koch, and Cypripedium calceolus var. parviflorum (saLisbury. ) Fern. Major Professor: Louis M. Lenz. Tlree native plants were propagatecl and,cultured indoors f or poss ible f uture commercial use as cut.rllowers or pot plants. seed germination ëxperiments were conducted on Castille ja coccinea and Anemone Patens var. wolf an rana while Cypripedium calceolus var. parviflorum v/as propagated by plant division. T'he restrlts on Casti I I e a seed indicated that the germination percent increaserf witfr j-ncreased fight intensity" Germination v/as inhibite<1 by red filtered Iight' Seed germinated readily at temperatures between 3.8 and 20"C but was inhib-ited at 2t2'C. The results indicated that light was not required for Anemone seed germination. The germina- tion percent increased as temperature increase'1 from 2'2 to 2l'C. Extensive damping-off in vitro was controlled only by dry benomyl. Moist after-ripening of Anemone seed decreased the percent germination. Transplanting and dì-viding xl- cypripedium stimulated a;oproximate ty an 80 percent increase in plant numbers. culture studies were underLaken to determine the effects of various factors on the growth of the plaats ' Results indicate.l tÌrat the root parasitic Castille a coccl-nea and 9. coccinea forma lutescens can survive without Jrost contact' but diFferent hosts ap.Ðear to enhance planL growth. The resulcs indicated that stcrage at temperatures a1fot below 0oC for six to l6 weeks is essential for the termination of plant dormancy of Anemone. rl_ edium required storage at temperatures above ooc for a minimum of 10 weeks for verna- lization and terminaLion of plant dormancy' Devernalization appeart:d to occur after l7 weeks of storage for Cvpripedium. Media type and amounts of N, P and K were important for Anemone plant survival and appearance" combinat-ions of several media components result i'n improved .olant survival ' Increased amounts of the nutrients N, P and K: N increased Ieaf numbers and dry shoot weights, P decreased dry shoot and root weights and K decrease<l leaf numbers arril dry shoot weights.
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