臺灣博碩士論文加值系統

學號：M9922028
論文名稱: 苞舌蘭無菌播種及種子保存之研究
總頁數：147
學校名稱：國立屏東科技大學
系所別：熱帶農業暨國際合作系
畢業時間：101學年度第2學期 學位別：碩士學位
研究生：包瑞斯　　　　　　　　　　 指導教授：方中宜 博士

論文摘要內容：
台灣原生種紫苞舌蘭(Spathoglottis plicata)是一種栽培容易、花期長且花形和花色優美的植物。由於農業活動的過度發展，原生種之棲息環境受到破壞，因此急須發展出一套原生種遺傳資源的保存方法。本研究主要採用組織培養技術進行紫苞舌蘭的無菌播種、種苗生長、瓶苗馴化及種子保存的研究。種子採集自28天成熟的果莢並無菌播種在五種不同的培養基上，包括 ½ MS、蘭花播種培養基(OSSM)、BM-1陸生蘭培養基(BM-1)、Vacin and Went蘭花培養基(VW)及Knudson C蘭花培養基(KC)。結果顯示，種子在不同的培養基及光周處理下皆會發芽，而在黑暗下發芽速度比在光照下還快，但在光照的環境下，其萌發種子發育較快速(例如: stages 4-6)。在所有培養基中，種子播種於OSSM培養基黑暗及光照的環境處理及VW培養基光照的處理後11周有較好的發育情形(例如: stage 6)。在OSSM培養基中添加不同的細胞分裂素如6-benzylaminopurine (BAP) 和thidiazuron (TDZ) 及生長素如1-naphthaleneacetic acid (NAA) 和 2,4-dichlorophenoxyacetic acid (2,4-D) 組合，並觀察其種苗發育的情形，結果顯示種苗可在不含植物生長調節劑 (plant growth regulators; PGR)的條件下生長，而在含有PGR之培養基中其植株長度、葉片數、葉片重量、根數、根長及植株鮮重方面發現，以0.5 mg/L NAA的處理，其芽的長度、葉片重量、根
的數目及植株鮮重有最好的效果，而在0.5 mg/L NAA 以及BAP或TDZ的組合處理140天，能得到最多根的數目及最高的植株和葉片鮮重。將發育良好帶有葉片及根之植株移至介質為水苔及泥碳土的三吋盆中進行馴化，結果顯示實生苗可順利的在泥碳土介質中生長而水苔則否。將紫苞舌蘭之種子於低溫4和 -20˚C中儲藏，在低溫-20˚C中儲藏5周後其發芽率最高，而此處理可將種子保存2個月。最後，以不同的冷凍方法例如玻璃化冷凍法(vitrification)及密封脫水(encapsulation-dehydration)試圖長期保存苞舌蘭種子實驗中發現無法以液態氮來長期保存種子。但此研究可供未來紫苞舌蘭種源長期保存做為初步的參考。

Student ID: M9922028
Title of Thesis: Asymbiotic Seed Germination and Seed Conservation of Spathoglottis plicata
Total Page: 147
Name of Institute: Department of Tropical Agriculture and International Cooperation
Graduate Date: June, 2013 Degree Conferred: Master
Name of Student: Boris Bran Advisor: Fang Jong-Yi, Ph.D.
The Contents of Abstract in This Thesis:
Spathoglottis plicata, a native terrestrial orchid in Taiwan, is an attractive and easy to grow plant which can flower all year round in the garden. However, native orchid species are threatened as a consequence of environmental disturbances, succession of natural habitats, and overexploitation for horticultural purposes. Therefore, there is need to develop conservation procedures for preserving their genetic resources. The present research was conducted to study the in vitro asymbiotic seed germination, seedling growth, acclimatization and seed conservation of this orchid. Seeds of S. plicata from 28-day-old capsule were sown on five germination media, including half-strength Murashige and Skoog medium (½ MS), Orchid Seed Sowing Medium (OSSM), BM-1 Terrestrial Orchid Medium (BM-1), Vacin and Went Modified Orchid Medium (VW), and Knudson C Orchid Medium (KC). Germination occurred in all the media and photoperiods tested. Seeds subjected to darkness were found to germinate more rapidly than those exposed to light whereas seeds had a faster development to advanced seed development stages (i.e. Stages 4-6) when subjected to light.
IV
All the seeds grown on OSSM medium under light and dark regimes and those grown on VW medium under light condition proceeded to the advanced seed development stage (i.e. Stage 6) within 11 weeks of culture. In vitro seedling development were examined by subjecting the germinated seeds to OSSM medium supplemented with different combinations of cytokinins [i.e., 6-benzylaminopurine (BAP) and thidiazuron (TDZ)] and auxins [i.e., 1-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D)]. Results showed that seedlings could develop on medium devoid of plant growth regulators (PGR) in the same way as those cultivated with PGR(s) in terms of their shoot length, leaf number, leaf width, root number and length, and seedling fresh weight. The treatment which contained 0.5 mg/L NAA was found to provide the highest shoot length, leaf width, root number and seedling fresh weight; whereas 0.5 mg/L NAA in combination with BAP or TDZ yielded the highest number of roots, seedling fresh weight and leaf production after 140 days. Plantlets with well-developed leaves and roots were hardened in 3-inch pots filled with either sphagnum moss or peat moss. It was observed that the in vitro plantlets can be successfully acclimatized using peat moss, but not with sphagnum moss. The viability of S. plicata seeds was tested following storage under low temperature conditions of 4 and -20°C. Maximum germination of stored seeds was observed at week 5 under -20°C. Consequently, it can be used for the seed conservation of S. plicata which last a period of 2 months. Finally, different cryopreservation methods (i.e. vitrification and encapsulation-dehydration) were used to test their possibility for long-term conservating S. plicata seeds and protocorms. However, none of the stored explants survived liquid nitrogen storage. This study presents the first step towards the development of a long-term storage procedure for S. plicata genetic resources.

Chinese Abstract.................... ............................................................................ I
English Abstract............. ................................................................................. III
Acknowledgements .......................................................................................... V
Table of Contents ............................................................................................ VI
List of Tables ................................................................................................ VIII
List of Figures ................................................................................................. IX
1. Introduction ................................................................................................... 1
2. Literature Review .......................................................................................... 4
2.1 Spathoglottis plicata ................................................................................ 4
2.1.1 Taxonomy .......................................................................................... 4
2.1.2 Plant morphology ............................................................................... 4
2.1.3 Geographical distribution .................................................................. 5
2.1.4 Propagation of Spathoglottis plicata .................................................. 6
2.2 In vitro germination of native orchid seeds ............................................. 9
2.2.1 Symbiotic germination ....................................................................... 9
2.2.2 Asymbiotic germination .................................................................. 12
2.2.3 Factors affecting asymbiotic germination of native orchid seeds ... 15
2.3 Conservation of native orchid seeds...................................................... 23
2.3.1 Short-term seed conservation ........................................................... 23
2.3.2 Long-term seed conservation ........................................................... 25
3. Materials and Methods ................................................................................ 28
3.1 Plant materials ....................................................................................... 28 3.2 Estimation of seed number .................................................................... 28
3.3 Seed sterilization ................................................................................... 29
3.4 Effect of different culture media on asymbiotic seed germination ....... 29 3.5 Microscopic analysis of seed development ........................................... 32
3.6 Effect of different PGRs on seedling development .............................. 32 3.7 Acclimatization of in vitro plantlets ...................................................... 33 3.8 Low-temperature storage of seeds ......................................................... 34
VII
3.8.1 Low-temperature storage without any pretreatment ........................ 34 3.8.2 Low-temperature storage with lithium chloride pretreatment ......... 34 3.9 Cryopreservation of seeds ..................................................................... 35
3.10 Culture conditions and data analysis ................................................... 36
4. Results.................. ....................................................................................... 38
4.1 Estimation of seed number .................................................................... 38
4.2 Effect of different culture media on asymbiotic seed germination ....... 41
4.3 Microscopic analysis of seed development ........................................... 61
4.4 Effect of different PGRs on seedling development .............................. 65
4.5 Acclimatization of in vitro plantlets ...................................................... 86
4.6 Low-temperature storage of seeds ......................................................... 91
4.6.1 Low-temperature storage of seeds without pretreatment ................ 91
4.6.2 Low-temperature storage with lithium chloride pretreatment and cryopreservation procedures .................................................... 98
5. Discussion .................................................................................................. 100
6. Conclusions ............................................................................................... 113
7. References ................................................................................................. 114
Biosketch of Author ...................................................................................... 145

Alexander, B. 2006. An Analysis of Seed Production, Viability, Germination in situ, and Grazing Impacts on the Western Prairie Fringed Orchid (Platanthera praeclara Sheviak and Bowles). Dissertation, North Dakota State University, Fargo, USA.
Alvarez, M. R. 1969. Cytophotometric Study of Nuclear Proteins and Nucleic Acids in Parenchymatous Tissue of the Orchid Embryo. Experimental Cell Research 57: 179-184.
Anderson, A. B. 1996. The Reintroduction of Platanthera ciliaris in Canada. p. 73-76. In: Allen, C. (ed) North American Native Terrestrial Orchids Propagation and Production. North American Terrestrial Orchid Conference, Germantown, Maryland.
Arditti, J. 1967. Factors Affecting the Germination of Orchid Seeds. Botanical Review 33: 1-197.
Arditti, J. 1982. Orchid Seed Germination and Seedling Culture. p. 101-103. In: Arditti, J. (ed) Orchid Biology- Reviews and Perspectives III. Cornell University Press, New York, USA
Arditti, J., M. A. Clements, G. Fast, G. Hadley, G. Nishimura, and R. Ernst. 1982a. Orchid Seed Germination and Seedling Culture. p. 243-274. In: Arditti, J. (ed) Ordhic Biology: Reviews and Perspectives II. Cornell University Press, Ithaca, New York.
Arditti, J., M. A. Clements, and A. P. Oliva. 1982b. Practical Germination of North American and Related Orchids: Epipactis atrorubens, E. gigantean and E. hellborine. American Orchid Society Bulletin 51: 162-171.
115
Arditti, J., J. D. Michaud, and A. P. Oliva. 1981. Seed Germination of North American Orchids. I. Native California and Related species of Calypso, Epipactis, Goodyera, Piperia, and Platanthera. Botanical Gazette 142: 442-453.
Arditti, J. and R. Ernst. 1984. Physiology of Germinating Orchid Seeds. p. 58-60. In: Arditti, J. (ed) Orchid Biology: Reviews and Perspectives III. Cornell University Press, New York, USA. Arditti, J. and R. Ernst. 1993. Micropropagation of Orchids. p. 215. John Wiley and Sons Inc., New York, USA.
Arditti, J., R. Ernst, T. Wing Yam, and C. Glabe. 1990. The Contribution of Orchid Mycorrhizal Fungi to Seed Germination: A Speculative Review. Lindleyana 5: 249-255.
Arditti, J., J. D. Michaud, and A. P. Oliva. 1981. Seed Germination of North American Orchids. I. Native California and Related species of Calypso, Epipactis, Goodyera, Piperia, and Platanthera. Botanical Gazette 142: 442-453.
Athipunyakom, P., L. Manoch, C. Piluek, S. Artjariyasripong, and S. Tragulrung. 2004. Mycorhizal Fungi from Spathoglottis plicata and the Use of these Fungi to Geminate Seeds of S. plicata in vitro. Kasetsart Journal of Natural Science 37: 83-93.
Ávila-Díaz, E., K. Oyama, C. Gómez-Alonso, and R. Salgado-Garciglia. 2009. In vitro Propagation of the Endangered Orchid Laelia speciosa. Plant Cell Tissue and Organ Culture 99: 335-343.
Bajaj Y. P. S. 1995. Cryopreservation of Plant Cell, Tissue and Organ Culture for the Conservation of Germplasm and Biodiversity. p. 3-18. In: Bajaj Y. P. S. (ed) Biotechnology in Agriculture and Forestry Cryopreservation of Plant Germplasm I. Springer, New York. USA.
116
Ballard, W. W. 1987. Sterile Propagation of Cypripedium reginae from Seeds. American Orchid Society Bulletin 56:935-946.
Bapat, V. A. and S. Narayanaswamy. 1997. Rizhogenesis in a Tissue Culture of the Orchid Spathoglottis. Bulletin of Torrey Botany Club 104:2-4.
Barua, A. K. and S. K. Bhadra. 1999. In vitro Micropropagation of Cymbidium aloifolium (L.) Sw. and Spathoglottis plicata Bl. Plant Tissue Culture 9: 133-140.
Batty, A. L., K. W. Dixon, M. Brundrett, and K. Sivasithamparam. 2001. Constrains to Symbiotic Germination of Terrestrial Orchid Seed in a Meditterranean Bushland. New Phytologist 152: 511-520.
Batty, A. L., K. W. Dixon, M. C. Brundrett, and K. Sivasithamparam. 2002. Orchid Conservation and Mycorrrhizal Association. p. 195-226. In: Sivasithamparam, K., K. W. Dixon and R. L. Barrett (ed) Microorganisms in Plant Conservation and Biodiversity. Kluwer Academic Publishers.
Beechey, N. 1970. Propagation of Orchids from Serrial Roots. American Orchid Society Bulletin 39：1085-1088.
Beltrame, E. 2006. Spathoglottis-Inside and Out. Orchid Review 114: 68-71.
Bertolini, V., A. Damon, and A. N. R. Velázquez. 2011. Symbiotic Germination of Three Species of Epiphytic Orchids Susceptible to Genetic Erosion, from Soconusco (Chiapas, Mexico). European Journal of Environmental Sciences 1: 60-68.
Berjak. P. and N. W. Pammenter. 2001. Seed Recalcitrance - Current Perspectives. South African Journal of Botany 64: 79-89.
117
Berjak. P., J. M. Farrant, and N. W. Pammenter. 1989. The Basis of Recalcitrant Seed Behaviour. Cell Biology of the Homoiohydrous Seed Condition. p. 89-108. In: Taylorson, R. B. (ed) Recent Advances in the Development and Germination of Seeds. Plenum Press, New York.
Bewley, J. D. and M. Black. 1994. Seeds: Physiology of Development and Germination. Plenum Press, New York.
Beyrle, H. F. and S. E. Smith. 1993. Excessive Carbon Prevent Greening of Leaves in Mycorrhizal Seedlings of the Terrestrial Orchid Orchis morio. Lindleyana 89: 97-99.
Bidartondo, M. I. and D. J. Read. 2008. Fungal Specificity Bottlenecks during Orchid Germination and Development. Molecular Ecology 17: 3707-3716.
Bouman, H. and G. J. De Klerk. 1990. Cryopreservation of Lily Meristems. Acta Horticulturae 266: 331-337.
Borris, H. 1969. Samenvermehrung un Anzucht Europäi-scher Erdochideen. p. 10-13. In: Proceedings of the 2nd European Orchid Congress. Paris, France.
Breddy, N. C. 1953. Observations on the Raising of Orchids by Asymbiotic Cultures. American Orchid Society Bulletin 22: 12-17.
Brian, H. W., J. H. Wang, W. Q. Lin, N. Han, and M. Y. Zhu. 2002. Accumulation of Soluble Sugar, Heat Stable Proteins and Dehydrins in Cryopreservation of Protocorm-Like Bodies of Dendrobium candidum by air-drying method. Journal of Plant Physiology 159: 1139-1145.
118
Carlson, M. C. 1940. Formation of the Seed of Cypripedium parviflorum. Botanical Gazzette 102: 295-301.
Chang, D. C. N. 2008. Research and Application of Orchid Mycorrhiza in Taiwan. p. 299-305. In: Criley, R.A. (ed) Proceedings of the XXVII IHC-55 Ornamentals. Acta Horticulturae (ISHS).
Chang, C., Y. C. Chen, and H. F. Yen. 2005. Protocorm or Rhizome? The Morphology of Seed Germination in Cymbidium dayanum Reichb. Botanical Bulletin of Academia Sinica 46: 71-74.
Chang, D. C. N and L. C. Chou. 2001. Seed Germination of Haemaria discolor var. dawsoniana and the Use of Mycorrhizae. Symbiosis 30: 29-40.
Chang, J., X. Y. Ding, S. L. Bao, D. Y. Liu, J. He, F. Tang, and B. Z. Ding. 2004. Studies on Tissue Culture of Dendrobium lituiflorum. Zhongguo Zhong Yao Za Zhi 29: 313-317.
Chang, Y. B. M. Reed. 2001. Preculture Conditions Influence Cold Hardiness and Regrowth of Pyrus Meristems. In Vitro Cell Development and Biology 36: 1329-1333.
Chen, S. C. and A. Bell. 2009. Spathoglottis Blume, Bijdr. 400. 1825. Flora of China 25: 287-288.
Cheng, S. F., C. H. Yeh, C. H. Jan, C. N. Chang. 2012. Growth and Development of Phaius tankervilleae (Banks) Blume when Inoculated with Orchid Mycorrhizal Fungi. African Journal of Agricultural Research 7(42): 5644-5652.
Chennaveeraiah, M. S. and S. J. Patil. 1975. Morphogenesis in Seed Cultures of Spathoglottis plicata. Current Science 44: 68.
119
Chetia, S., P. C. Deka, and J. Devi. 1998. Germination of Fresh and Stored Encapsulated Protocorms of Orchids. Indian Journal of Experimental Biology 36: 108-111.
Ching, P. I. 2000. Spathoglottis plicata. p. 1031. In: Huang, T.-C. (ed), Flora of Taiwan 2nd edition Vol 5. Editorial Committee of Flora of Taiwan, National Taiwan University, Taiwan. Chou, L. C. and D. C. N. Chang. 2004. Asymbiotic and Symbiotic Seed Germination of Anoectochilus formosanus and Haemaria discolor and their F1 hybrids. Botanical Bulletin of Academia Sinica 45: 143-147.
Clements, M. A., H. Muir, P. J. Cribb. 1986. Preliminary Report on the Symbiotic Germination of European Orchids. Kew Bulletin 41: 437-445.
Comber, J. B. 2001. Spathoglottis plicata Blume. p. 294. In: Orchids of Sumatra. Natural History Publications, Malaysia.
Crafts, C. B. and C. O. Miller. 1974. Detection and Identification of Cytokinins Produced by Mychorrhizal Fungi. Plant Physiology 54: 586-588.
Currah, R. S. and C. Zelmer. 1992. A Key and Notes for the Genera of Fungi Mycorrhizal with Orchids and a New Species in the Genus Epulorhiza. Reports of the Tottori Mycological Institute 30: 43-59.
Curtis, J. T. 1936. The Germination of Native Orchid Seeds. American Orchid Society Bulletin 5: 42-47.
Curtis, J. T. 1947. Studies on the Nitrogen Nutrition of Orchid Embryos. I. Complex Nitrogen Sources. American Orchid Society Bulletin 16: 654-660.
120
Curtis, J. T and E. Spoerl. 1948. Studies on the Nitrogen Nutrition of Orchid Embryos II. Comparative Utilization of Nitrate and Ammonium Nitrogen. American Orchid Society Bulletin 17: 111-114.
Damon. A. 2011. Diversidad y Conservación de las Orquídeas del Corredor Biolóogico Tacaná-Boquerón. Final Report. FONDOS MIXTOS-CHIAPAS CHIS-2006-C06-45802.2007-2010.
De Pauw, M. A. and W. R. Remphrey. 1993. In vitro Germination of Three Cypripedium species in Relation to Time of Seed Collection, Media and Cold treatment. Canadian Journal of Botany 71: 879-885.
De Pauw, M. A., W. R. Remphrey, and C. E. Palmer. 1995. The Cytokinin Preference for in vitro Germination and Protocorm Growth of Cypripedium candidum. Annals of Botany 75: 267-275.
Deb, C. R. and T. Imchen. 2010. An Efficient in vitro Hardening Technique of Tissue Culture Raised Plants. Biotechnology 1-5.
Deb, C. R. and A. Pongener. 2011. Asymbiotic Seed Germination and in vitro Seedling Development of Cymbidium aloifolium (L.) Sw.: a Multipurpose Orchid. Journal of Plant Biochemistry and Biotechnology 20: 90-95.
Diggle, P. K. 1995. Architectural Effects and the Interpretation of Patterns of Fruit and Seed Development. Annual Review of Ecology and Systematics 26:5331-552.
Dijk, E. and N. Eck. 1995. Anexic in vitro Nitrogen and Phosphorus Responses of Some Dutch Marsh Orchids. New Phytologist 131: 353-359.
Downie, D. G. 1941. Notes on the Germination of some British Orchids. Transactions of the Botanical Society of Edinburgh 33: 94-103.
121
Duncan, D. B. 1995. Multiple Range and Multiple F-tests. Biometrics 11:1-42.
Dumet, D., A. Grapin, C. Bailly, and N. Dorion. 2002. Revisiting Crucial Steps of an Encapsulation/Dessication Based Cryopreservation Process: Importance of Thawing Method in the Case of Pelargonium Meristems. Plant Science 163: 1121-1127.
Dussert, S., N. Chabrillange, F. Engelmann, and S. Hamon. 1999. Quantitative Estimation of Seed Desiccation Sensitivity using a Quantal Response Model: Application to Nine Species of the Genus Coffea L. Seed Science Research 9: 135-144. Dutra, D., T. R. Johnson, P. J. Kauth, S. L. Stewart, M. E. Kane, and L. Richardson. 2008. Asymbiotic Seed Germination, in vitro Seedling Development, and Greenhouse Acclimatization of the Threatened Terrestrial Orchid Bletia purpurea. Plant Cell Tissue and Organ Culture 94: 11-21. Dutra, D., M. E. Kane, and L. Richardson. 2009. Asymbiotic Seed Germination and in vitro Seedling Development of Cyrtopodium punctatum: a Propagation Protocol for an Endangered Florida Native Orchid. Plant Cell Tissue and Organ Culture 96: 235-243. Endres, L. and H. Mercier. 2001. Ammonium and Urea as Nitrogen Sources for Bromeliads. Journal of Plant Physiology 158(2): 205-212. Engelmann, F. 1997. In vitro Conservation Methods. p. 119-162. In: Ford-Lloyd, B. V. H. J. Newbury, and J. A. Callow (eds) Biotechnology and Plant Genetic Resources: Conservation and Use. CABI, UK. Engelmann, F. 2011. Use of Biotechnologies for the Conservation of Plant Biodiversity. In Vitro Celullar and Developmental Biology - Plant 47: 5-16.
122
Engelmann, F. and J. M. M. Engles. 2002. Technology and Strategies for Ex Situ Conservation. p. 89-104. In: Ramanatha Rao V., A. H. D. Brown, and M. T. Jackson (ed) Managing Plant Genetic Diversity, Wallingford, Rome, CAB International, IPGRI. Ernst, R. 1982. Orchid Seeed Germination and Seedling Culture - A Manual: Paphiopedilum. p. 350-353. In: Arditti, J (ed) Orchid Biology-Reviews and Perspectives, II. Cornell University Press, Ithaca, NY.
Ervin, G. N. and R. G. Wetzel. 2002. Effects of Sodium Hypochlorite Sterilization and Dry Cold Storage on Germination of Juncus effuses L. Wetlands 22: 191-195.
Fabre, J. and J. Dereuddre. 1990. Encapsulation-Dehydration: A New Approach to Cryopreservation of Solanum Shoot-Tips. CryoLetters 11: 413-426.
Flachsland, E., G. Terada, A. Schocchi, H. Rey, L. Mroginsky, and F. Engelmann. 2006. Cryopreservation of Seeds and in vitro-Cultured Protocorms of Oncidium bifolium sims. (Orchidaceae) by Encapsulation-Dehydration. CryoLetters 27: 235-242.
Gamborg, O. L., T. Murashige, T. A. Thorpe, and I. K. Vasil. 1976. Plant Tissue Culture Media. In Vitro 12: 473-478.
Godo, T., M. Komori, E. Nakaoki, T. Yukawa, and K. Miyoshi. 2010. Germination of Mature Seeds of Calanthe tricarinata Lindl., an Endangered Terrestrial Orchid, by Asymbiotic Culture in vitro. In Vitro Celullar and Developmental Biology - Plant 46: 323-328.
Gogoi, K., S. Kumaria, and P. Tandon. 2012. Ex situ Conservation of Cymbidium eburneum Lindl.: A Threatened and Vulnerable Orchid, by Asymbiotic Seed Germination. 3 Biotech 2: 337-374.
123
Gogoi, K., S. Kumaria, and P. Tandon. 2013. Cryopreservation of Cymbidium eburneum Lindl. and C. hookerianum Rchb.f., Two Threatened and Vulnerable Orchids via Encapsulation-Dehydration. In Vitro Celullar and Developmental Biology - Plant 49: 248-254.
Hanlon, E. A. 1993. Soil pH and Electrical Conductivity: A County Extension Soil Laboratory Manual. Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida.
Harvais, G. 1973. Growth Requirements and Development of Cypripedium reginae in Axenic Culture. Canadian Journal of Botany 51: 327-332.
Harvais, G. 1980. Scientific Notes on a Cypripedium reginae of North-Western Ontario. American Orchid Society Bulletin 49: 237-244.
Harvais, G. 1982. An Improved Culture Medium for Growing the Orchid Cypripedium reginae Axenically. Canadian Journal of Botany 60: 2547-2556.
Hasan, M. N., A. Ansari, M. S. Z. Shahriar, M. N. K. Azam, and S. M. A. Awal. 2011. In vitro Seed Germination and Plantlets Development of Silvius appendiculatus (Orchidaceae). International Journal of Sustainable Crop Production 6: 1-5.
Hay, F. R., D. J. Merritt, J. A. Soanes, K. W. Dixon. 2010. Comparative Longevity of Australian orchid (Orchidaceae) Seeds under Experimental and Low Temperature Storage Conditions. Botanical Journal of the Linnean Society 164: 26-41.
Hirai, D. and A. Sakai. 1999. Cryopreservation of in vitro-Grown Axillary Shoot-Tip Meristems of Mint (Mentha spicata L.) by Encapsulation Vitrification. Plant Cell Reports 19: 150-155.
124
Hirano, T., T. Godo, M. Mii, and K. Ishikawa. 2005. Cryopreservation of Immature Seeds of Bletilla striata by Vitrification. Plant Cell Reports 23: 534-539.
Hirano, T., T. Godo, K. Miyoshi, K. Ishikawa, M. Ishikawa, and M. Mii. 2009. Cryopreservation and Low-Temperature Storage of Seeds of Phaius tankervilleae. Plant Biotechnology Reports 3: 103-109.
Hirano, T., K. Ishikawa, M. Ishikawa, and M. Mii. 2006. Advances in Orchid Cryopreservation. Floriculture, Ornamental and Plant Biotechnology 2: 410-414.
Hosomi, S. T., C. C. Custódio, P. T. Seaton, T. R. Marks, and N. B. Machado-Neto. 2012. Improved Assessment of Viability and Germination of Cattleya (Orchidaceae) Seeds following Storage. In Vitro Cell Development and Biology – Plant 48: 127-136.
Hossain, M. M. and R. Dey. 2013. Multiple Regeneration Pathways in Spathoglottis plicata Blume - A Study in vitro. South African Journal of Botany 85: 56-62.
Integrated Taxonomic Information System (ITIS). 2012. Spathoglottis Blume. Retrieved on Sep. 7, 2012, from the World Wide Web: http://www.itis.gov/index.html
Ishikawa, M., A. J. Robertson, and L. V. Gusta. 1995. Comparison of Viability Tests for Assessing Cross-Adaptation to Freezing, Heat and Salt Stresses Induced by Abscisic Acid in Bromegrass (Bromus inermis leyss) Suspension Cultured Cells. Plant Science 107: 83-93.
Ishikawa, K., K. Harata, M. Mii, A. Sakai, K. Yoshimatsu, and K. Shimomura. 1997. Cryopreservation of Zygotic Embryos of a Japanese Terrestrial Orchid (Bletilla striata) by Vitrification. Plant Cell Reports 16: 754-757.
125
Islam, M. O. and S. Ichihashi. 1999. Effects of Sucrose, Maltose and Sorbitol on Callus Growth and Plantlet Regeneration in Phalaenopsis, Coritaenopsis and Neofinetia. Journal of Japan Society and Horticultural Science 68: 1124-1131.
Israel, H. W. 1963. Production of Dendrobium Seedlings by Aseptic Culture of Excised Ovularies. American Orchid Society Bulletin 32: 441-443.
Johnson, T. R., M. E. Kane, and H. E. Perez. 2011. Examining the Interaction of Light, Nutrients and Carbohydrates on Seed Germination and Early Seedling Development of Bletia purpurea (Orchidaceae). Plant Growth Regulators 63: 89-99.
Johnson, T. R., S. L. Stewart, D. Dutra, M. E. Kane, and L. Richardson. 2007. Asymbiotic and Symbiotic Seed Germination of Eulophia alta (Orchidaceae)-Preliminary Evidence for the Symbiotic Culture Advantage. Plant Cell Tissue and Organ Culture 90: 313-323.
Kananot, N., R. Pichyangkura, S. Chanprame, S. Chadchawan, and P. Limpanavech. 2010. Chitosan Specificity for the in vitro Seed Germination of Two Dendrobium orchids (Asparagales: Orchidaceae). Scientia Horticulturae 124: 239-247.
Kano, K. 1972. Seed Germination of Oriental Cymbidium and their Shoot Tip Culture. p. 133-142. In: Arditti, J. (ed) Proceedings of the 6th World Orchid Conference, Sydney.
Kauth, P. J. 2005. In vitro Seed Germination and Seedling Development of Calopogon tuberosus and Sacoila lanceolata var. lanceolata: Two Florida Native Terrestrial Orchids. Master Thesis, University of Florida, Florida, U.S.A.
126
Kauth, P. J., W. A. Vendrame, and M. E. Kane. 2006. In vitro Seed Culture and Seedling Development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture 85: 91-102.
Kauth, P. J., T. R. Johnson, S. L. Stewart, and M. E. Kane. 2008. A Classroom Exercise in Hand Pollination and in vitro Asymbiotic Orchid Seed Germination. Plant Cell Tissue and Organ Culture 93: 223-230.
Kaviani, B. 2011. Conservation of Plant Genetic Resources by Cryopreservation. Australian Journal of Crop Science 5: 778-800.
Keller, E. R. and A. Senula. 2010. Cryopreservation of Plant Germplasm. p. 131-151. In: Davey M. R. and P. Anthony (ed) Plant Cell Culture Essential Methods. John Wiley and Son Inc., New York, USA.
Keel, B. G., L. W. Zettler, and B. A. Kaplin. 2011. Seed Germination of Habenaria repens (Orchidaceae) in situ Beyond its Range, and its Potential for Assisted Migration Imposed by Climate Change. Castanea 76(1): 43-54.
King, M. W. and E. H. Roberts. 1979. The Storage of Recalcitrant Seeds: Achievements and Possible Approaches. International Board for Plant Genetic Resources, Rome.
Kitsaki, C. K., S. Zygouraki, M. Ziobora, and S. Kintzios. 2004. In vitro germination, protocorm formation and plantlet development of mature versus immature seeds from several Ophrys species (Orchidaceae). Plant Cell Reports 23: 284-290.
Kheawwongjun, J. and K. Thammasiri. 2008. Breeding Spathoglottis spp. for Commercial Potted Orchids. Acta Horticulturae 788: 47-52.
127
Kitsaki, C. K., S. Zygouraki, M. Ziobora, and S. Kintzios. 2004. In vitro Germination, Protocorm Formation and Plantlet development of Mature versus Immature Seeds from Several Ophrys species (Orchidaceae). Plant Cell Reports 23:284-290.
Knudson, L. 1922. Nonsymbiotic Germination of Orchid Seeds. Botanical Gazette 73: 1-25.
Knudson, L. 1946. A New Nutrient Solution for Orchid Seed Germination. American Orchid Society Bulletin 15: 214-217.
Krishna-Mohan, P. T., S. M. Jorapur. 1986. In vitro Seed Culture of Acampe praemorsa (Roxb.) Blatt. And McC. p. 437. In: Vij, S. P (ed) Biology, Conservation and Culture of Orchids. Affiliated East West Press, New Delhi, India.
Kull, T. 1998. Fruit-Set and Recuitment in Population of Cypripedium calceolus in Estonia. Botanical Journal of the Linnean Society 126: 27-38.
Lavanya, M., B. Venkateshwarlu, and B. P. Devi. 2009. Acclimatization of Neem Microshoots Adaptable to Semi-Sterile Conditions. Indian Journal of Biotechnology 8: 218-222.
Laws, N. 2002. Orchid Commerce around the World. FloraCulture International 10: 28-29.
Le, B. V., N. T. Hang Phuong, L. T. Anh Hong, and K. T. Thanh Van. 1999. High Frequency Shoots Regeneration from Rhynchostylis gigantean (Orchidaceae) using Thin Cell Layers. Plant Growth Regulation 28:179-185.
Lee, N. 1987. Comparative Studies in Seed Germination and Seedling Growth of Pleione formosana and Phalaenopsis White hybrid. p. 117-
128
128. In: Proc. World Orchid Hiroshima Symposium, Hiroshima, Japan.
Lee, Y. L. 2003. Growth Periodicity, Changes of Abscisic Acid during Embryogenesis, and in vitro Propagation of Cypripedium fomosanum Hay. PhD. Thesis, National Taiwan University, Taiwan, R.O.C.
Lee, Y. L. 2011. In vitro Culture and Germination of Terrestrial Asian Orchid Seeds. p. 53-62. In: Thorpe T. A. and E. C. Yeung (ed) Plant Embryo Culture: Methods and Protocols, Methods in Molecular Biology.Vol. 710: DOI 10.1007/978-1-61737-988-8_5.
Lee, Y. L. and N. Lee. 2005. Embryo Development of Cypripedium formosanum in Relation to Seed Germination in vitro. Journal of American Society for Horticultural Science 130: 747-753.
Lee, Y. L., C. F. Lu, M. C. Chung. 2007a. Developmental Changes in Endogenous Abscisic Acid Concentrations and Asymbiotic Seed Germination of a Terrestrial Orchid, Calanthe tricarinata Lindl. Journal of American Society for Horticultural Science 132: 246-252.
Lee, Y. L., C. F. Lu, M. C. Chung, E. C. Yeung, and N. Lee. 2007b. Developmental Changes in Endogenous Abscisic Acid Concentrations and Asymbiotic Seed Germination of a Terrestrial Orchid, Calanthe tricarinata Lindl. Jounal of American Society for Horticultural Science 132: 246-252.
Lee, Y. L., N. Lee, E. C. Yeung, and M. C. Chung. 2005. Embryo Development of Cypripedium formosanum in Relation to Seed Germination in vitro. Journal of American Society for Horticultural Science 130: 747-753.
129
Lee, Y. L., N. Lee, E. C. Yeung, and M. C. Chung. 2008. Embryology of Phalaenopsis amabilis var. Formosa: Embryo Development. Botanical Studies 49: 139-146.
Lee, Y. I. and E. C. Yeung. 2010. Embryo Development and in vitro Seed Germination of Bulbophyllum fascinator. Acta Horticulturae 878: 243-250.
Lee, Y. I., E. C. Yeung, and M. C. Chung. 2007c. Embryo Development of Orchids. p. 23-43. In: Chen, W. H. and H. H. Chen (ed) Orchid Biotechnology. World Scientific Publishing, Singapore.
Leroux, G., D. Barabé, and J. Vieth. 1997. Morphogenesis of the Protocorm of Cypripedium acaule (Orchidaceae). Plant Systematics and Evolution 205: 53-72.
Light, M. H. S. and M. MacConaill. 1998. Factors Affecting Germinable Seed Yield in Cypripedium calceolus var. pubescens (Wild.) Correll and Epipactis helleborine (L.) Crantz (Orchidaceae). Botanical Journal of the Linnean Society 126:3-26.
Lo, S. -F., S. M. Nalawade, C. L. Kuo, C. L. Chen, and H. S. Tsay. 2004. Asymbiotic Germination of Immature Seeds, Plantlet Development and Ex Vitro Establishment of Plants of Dendrobium tosaense Makino - A Medicinally Important Orchid. In Vitro Cellular and Developmental Biology - Plant 40: 528-535.
Long, B., A. X. Niemiera, Z. Y. Cheng, and C. L. Long. 2010. In vitro Propagation of Four Threatened Paphiopedilum species (Orchidaceae). Plant Cell, Tissue and Organ Culture 101: 151-162.
Lurswijidjarus, W. and K. Thammasiri. 2004. Cryopreservation of Shoot Tips of Dendrobium Walter Oumae by Encapsulation/Dehydration. ScienceAsia 30: 293-299.
130
Mahendran, G. and V. Narmatha Bai. 2009. Mass Propagation of Satyrium nepalense D.Don. – A Medicinal Orchid via Seed Culture. Scientia Horticulturae 119: 203-207.
Maladabi, R. B. and K. Nataraja. 2006. Cryopreservation and Plant Regeneration via Somatic Embryogenesis using Shoot Apical Domes of Mature Pinus roxburghii Sarg. Trees. In Vitro Cellular and Developmental Biology - Plant 42: 152-159.
Malenmnganba. H., B. K. Ray, S. Bhattacharya, and D. C. Deka. 1996. Regeneration of Encapsulated Protocorms of Phaius tankervilliae Stored at Low Temperature. Indian Journal of Experimental Biology 34: 801-805.
Malmgren. S. 1996. Orchid Propagation: Theory and Practice. p. 63-71. In: Allen, C. (ed) North American Native Terrestrial Orchids Propagation and Production. North American Terrestrial Orchid Conference, Germantown, Maryland.
Mandal, J., S. Pattnaik, and P. K. Chand. 2000. Alginate Encapsulation of Axillary Buds of Ocimum americanum L. (hoary basil), O. basilicum L. (sweet basil), O. gratissimum L. (shrubby basil, and O. sactum L. (sacred basil). In Vitro Cellular and Developmental Biology - Plant 36: 287-292.
Maneerattanarungroj, P., S. Bunnag, and M. Monthatong. 2007. In vitro Conservation of Cleisostoma areitinum (Rchb.f.) Garay, Rare Thai Orchid Species by an Encapsulation-Dehydration Method. Asian Journal of Plant Science 6: 1235-1240.
Marzalina, M. and B. Krishnapillay. 1999. Recalcitrant Seed Biotechnology Applications to Rain Forest Conservation. p. 265-274. In: Benson, E. E. (ed) Plant Conservation Biotechnology. Taylor &; Francis Inc., Philadelphia, USA.
131
Masuhara, G. and K. Katsuya. 1992. Mycorrhizal Differences between Genuine Roots and Tuberous Roots of Adult Plants of Spiranthes sinensis var. amoena (Orchidaceae). Journal of Plant Research 105: 453-460.
Masuhara, G., S. Kimura, and K. Katsuya. 1988. Seasonal Changes in the Mycorrhiza of Bletilla striata (Orchidaceae). Transactions of the Mycological Society of Japan 29.
McKendrick, S. 2000. In vitro Germination of Orchids: A Manual. Ceiba Foundation for Tropical Conservation. Retrieved on Jul. 25, 2012, from the World Wide Web:
http://www.ceiba.org/documents/CFTCpropman.pdf
McKinley, T. C. and N. D. Camper. 1997. Action Spectra of in vitro Symbiotic Germination of Goodyera repens var ophioides. Lindleyana 12: 303-333.
Mead, J. W. and C. Bulard. 1975. Effects of Vitamins and Nitrogen Sources on Asymbiotic Germination and Development of Orchis laxiflora and Ophrys sphegodes. New Phytologist 74: 33-40.
Meeyot, W. and H. Kamemoto. 1969. Studies on Storage of Orchid Pollen. American Orchid Society Bulletin 38: 388-393.
Mendel, R. R. and R. Hänsch. 2002. Molybdoenzymes and Molybdenum Cofactor in Plants. Journal of Experimental Botany 53: 1689-1698.
Minea, M., C. Piluek, A. Menakanit and S. Tantiwiwat. 2004. A Study on Seed Germination and Seedling Development of Spathoglottis Bl. Orchids. Kasetsart Journal of Natural Science 38: 141-156.
Miyoshi, K. and M. Mii. 1995a. Enhancement of Seed Germination and Protocorm Formation in Calanthe discolor (Orchidaceae) by NaOCl
132
and Polyphenol Absorbent Treatments. Plant Tissue Culture Letters 12: 267-272.
Miyoshi, K. and M. Mii. 1995b. Phytohormone Pre-Treatment for the Enhancement of Seed Germination and Protocorm Formation by the Terrestrial Orchid, Calanthe discolor (Orchidaceae), in Asymbiotic Culture. Scientia Horticulturae 63: 263-267.
Miyoshi, K. and M. Mii. 1998. Stimulatory Effects of Sodium and Calcium Hypochlorite, Pre-Chilling and Cytokinins on the Germination of Cypripedium macranthos Seed in vitro. Plysiologia Plantarum 102: 481-486.
Mohanty, P., M. C. Das, S. Kumaria, and P. Tandon. 2012. High-Efficiency Cryopreservation of the Medicinal Orchid Dendrobium nobile Lindl. Plant Cell, Tissue and Organ Culture 109: 297-305.
Moraes, L. M., R. T. Faria, and F. L. Cuquel. 2005. Activated Charcoal for in vitro Propagation of Brazilian Orchids. Acta Horticulturae 683: 282-390.
Morel, G. M. 1965. Clonal Propagation of Orchids by Meristem Culture. Cymbidium Society News 20: 3-11.
Morel, G. M. 1972. Morphogenesis of Stem Apical Meristem Cultivated in vitro: Application to Clonal Propagation. Phytomorphology 22: 265-277.
Mweetwa, A. M., G. E. Welbaum, and D. Tay. 2008. Effects of Development, Temperature, and Calcium Hypochlorite Treatment on invitro Germinability of Phalaenopsis seeds. Scientia Horticulturae 117: 257-262.
133
Muir, H. J. 1989. Germination and Mycorrhizal Fungus Compatibility in European Orchids. p. 39-56. In: Pritchard, H. W. (ed) The Role of Physiology Ecology and Management. Cambridge University Press: Cambridge.
Murashige, T. and F. Skoog. 1962. A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Journal of Plant Physiology 122: 211-215.
Nagaraju, V., S. P. Das, P. C. Bhutia, and R. C. Upadhyaya. 2003. Response of Cymbidium lunavian Atlas Protocorms to Media and Benzylaminopurine. Indian Journal of Horticulture 60(1): 98-103.
Nayak, N. R., S. P. Rath, and S. Patnaik. 1998. High Frequency Plant Regeneration from Alginate Encapsulated Protocorm-Like Bodies of Spathoglottis plicata Bl., A terrestrial Orchid. Phytomorphology 48: 179-186.
Norstog, K. 1973. New Synthetic Medium for the Culture of Premature Barley Embryos. In Vitro 8: 307-308.
Oliva, A. P. and J. Arditti. 1984. Seed Germination of North American Orchids. II. Native California and Related Species of Aplectrum, Cypripedium and Spirenthes. Botanical Gazette 145: 495-501.
Ortega-Loeza, M. M., R. Salgado-Garciglia, C. Gómez-Alonzo, and I. Ávila-Díaz. Acclimatization of the Endangered Mexican Epiphytic Orchid, Laelia speciosa (H.B.K) Schltr. European Journal of Environmental Sciences 1: 48-54.
Otero, J. T., J. D. Ackerman, and P. Bayman. 2002. Diversity and Host Specificity of Endopytic Rhyzoctonia-Like Fungi from Tropical Orchids. American Journal of Botany 89: 1852-1858.
134
Paek, K. Y., E. J. Hahn, and S. Y. Park. 2011. Micropropagation of Phalaenopsis Orchids via Protocorms and Protocrom-Like Bodies. p. 293-306. In: Plant Embryo Culture. Humana Press.
Pacek-Bieniek, A., M. Dyduch-Sieminska, and M. Rudas. 2010. Influence of Activated Charcoal on Seed Germination and Seedling Development by the Asymbiotic Method in Zygostates grandiflora Lindl. Mansf. (Orchidaceae). Folia Horticulturae 22: 45-50.
Pammenter, N. W. and P. Berjak. 1999. A Review of Recalcitrant Seed Physiology in Relation to Dessication-Tolerance Mechanisms. Seed Science Research 9: 13-37.
Patil, J. G., M. L. Ahire, and T. D. Nikam. 2012. Influence of Plant Growth Regulators on in vitro Seed Germination and Seedling Development of Digitalis purpurea L. The Asian and Australasian Journal of Plant Science and Biotechnology 6: 12-18.
Paunescu, A. 2005. Conservation of the Endemic Plant Cerastium transsilvanicum Schur of Piatra Craiului Massif using in vitro Techniques. p. 99-103. In: Researches in Piatra Craiului National Park (ed). Phonix, Brasov.
Paunescu, A. 2009. Biotechnology for Endangered Plant Conservation: A Critical Overview. Romanian Biotechnological Letters 14(1): 4095-4103.
Pierik, R. L. M., P. A. Sprenkels, B. Vanderharst, and Q. G. Vandermeys. 1988. Seed Germination and Further Development of Plantlets of Paphiopedilum ciliolare Pfitz in vitro. Scientia Horticulturae 34: 139-153.
Pornuchi, W. and K. Thammasiri. 2008. Cryopreservation of Protocorms of Dendrobium cariniferum Rchb.f. Acta Horticulturae 788: 63-68.
135
Prakash, L., C. L. Lee, C. S. Loh, and C. J. Goh. In vitro Propagation of Commercial Orchids; An Assessment of Current Methodologies and Development of a Novel Approach- Thin Cross-Section Culture. p. 42-47. In: Islam, A. S. (ed) Plant Tissue Culture. Oxford and IBS Publishing Co. Pvt Ltd, New Delhi, Calcutta. Pritchard, H. W., A. L. C. Poynter, and P. T. Seaton. 1999. Interspecific Variation in Orchid Seed Longevity in Relation to Ultra-Dry Storage and Cryopreservation. Lindleyana 14: 92-101. Raghavan, V. 1964. Effects of Certain Organic Nitrogen Compounds on Growth in vitro of Seedling of Cattleya. Botanical Gazette 125: 260-267. Raghavan, V. and J.G. Torrey. 1964. Inorganic Nitrogen Nutririon of the Seedling of the Orchid, Cattleya. American Journal of Botany 51: 264-274. Ramanatha Rao, V. and R. Riley. 1994. The Use of Biotechnology for Conservation and Utilization of Plant Genetic Resources. Plant Genetic Resources Newsletter 97: 3-20. Rao, A. N. 1977. Tissue Culture in the Orchid Industry. p. 7-69. In: Reinert, J. and Bajaj, Y. P. S. (ed) Applied and Fundamental Aspects of Plant Cell, Tissue and Organ Cultures. Berlin Heidelberg, New York, Springer. Rao, N. K. 2004. Plant Genetic Resources: Advancing Conservation and Use through Biotechnology. African Journal of Biotechnology 3: 136-145.
Rasmussen, H. N. 1995. Terrestrial Orchid from Seed to Mycotrophic Plant. Cambridge University Press, New York, USA.
136
Rasmussen, H. N., T. F. Andersen, and B. Johansen. 1990. Temperature Sensitivity of in vitro Germination and Seedling Development of Dactylorhiza majalis (Orchidaceae) with and without Mycorrhizal Fungus. Plant Cell and Environment 13: 171-177.
Rasmussen, H. N., B. Johansen, and T. F. Andersen. 1991. Symbiotic in vitro Culture of Immature Embryos and Seeds from Listera ovata. Lindleyana 6: 134-139.
Redenbaugh, K., D. Slade, P. Viss, and M. Kossler. 1988. Artificial Seeds: Encapsulation of Somatic Embryos. p. 400-419. In: Valentine, F. A. (ed) Forest and Crop Biotechnology, Progress and Prospects. Springer-Verlag, New York, USA.
Richards, L. A. 1954. Diagnosis and Improvement of Saline and Alkali Soils. USDA Agriculture Handbook 60, Washington D. C.
Rhoades, J. D. and J. Loveday. 1990. Salinity in Irrigated Agriculture and Irrigation of Agricultural Crops. Agronomy Monograph 30: 1089: 1142.
Saiprasad, G. V. S. and R. Polisetty. 2003. Propagation of Three Orchid Genera using Encapsulated Protocorm-Like Bodies. In Vitro Cellular and Developmental Biology – Plant 3: 42-48.
Sakai, A., S. Koboyashi, and I. Oiyama. 1990. Cryopreservation of Nucellar Cells of Navel Orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by Vitrification. Plant Cell Reports 9: 30-33.
Sakai, A., S. Koboyashi, and I. Oiyama. 1991. Survival by Vitrification of Nucellar Cells of Navel Orange (Citrus sinensis Osb. var. brasiliensis) cooled to 196°C. Journal of Plant Physiology 137: 465-470.
137
Sakai, A. 1995. Biotechnology in Agriculture and Forestry. p. 53-69. In: Bajaj, Y. P. S. (ed) Cryopreservation of Plant Germplasm. Springer-Verlag, Berlin, Heidelberg, New York.
Salifah, H. A. B., M. Muskhazli, G. Rusea, P. Nithiyaa, and S. N. Nurul. 2009. Mychorrizae of Tiger Orchids (grammatophyllum spp.) and Symbiotic Germination of Seeds of Grammatophyllum stapeliiflorum (Teijsm. &; Mann.) S.S. Smith. Malaysian Applied Biology 38(2): 23-28.
Sarasan, V., R. Cripps, M. M. Ramsay, C. Atherton, M. McMichen, G. Prendergast, and J.K. Rowntree. 2006. Conservation of in vitro Threatened Plants – Progress in the Past Decade. In Vitro Cellular and Developmental Biology - Plant 42: 206-214.
SAS Institute Inc. 2002. SAS Version 8.2. SAS Institute, North Carolina.
Schafer-Menuhr, A., H. M. Schumacher, and G. M. Mix-Wagner. 1997. Long-Term Storage of Old Potato Varieties by Cryopreservation of Shoot-tips in Liquid Nitrogen. Plant Genetic Resources Newsletter 111: 19-24.
Seaton, P. T. 2007. Establishing a Global Network of Orchid Seed Banks. Lankesteriana 7 (1-2): 371-375.
Seaton, P. T. and N. S. J. Hailes. 1989. Effect of Temperature and Moisture Content on the Viability of Cattleya aurantiaca Seed. p. 17-30. In: Pritchard, H. D. (ed) Modern Methods in Orchid Conservation: The Role of Physiology, Ecology and Management. Cambridge University Press, New York.
Seaton, P. T. and H. W. Pritchard. 2003. Orchid Germplasm Collection, Storage and Exchange. P. 227-258. In: Dixon K. W. P., Cribb, S. Kell,
138
and R. Barrett (ed) Orchid Conservation. Natural History Publycations (Borneo), Kota Kinabalu, Sabah.
Seaton, P. T. and H. W. Pritchard. 2008. Life in the Freezer. Orchids 77: 762-773.
Seaton, P. T. and H. W. Pritchard. 2011. Orchid Seed Stores for Sustainable Use: A Model for Future Seed-Banking Activities. Lankesteriana 11: 349-353.
Seeni, S. and P. G. Latha. 2000. In Vitro Multiplication and Ecorehabilitation of the Endangered Blue Vanda. Plant Cell, Tissue and Organ Culture 61:1-8.
Sharma, A. and P. Tandon. 1986. Influence of Growth Regulators on Asymbiotic Germination and Early Seedling development of Coelogyne punctulata Lindl. p. 441-471. In: Vij, S. P (ed) Biology, Conservation and Culture of Orchids. Affiliated East West Press, New Delhi, India.
Sharma, A., P. Tandon, and A. Kumar. 1992. Regeneration of Plantlets from Encapsulated Dendrobium Protocorms. Indian Journal of Experimental Biology 30: 744-748.
Shiau, Y. J., S. M. Nalawade, C. N. Hsai, and H. S. Tsay. 2005. Propagation of Haemaria discolor via in vitro Seed Germination. Biologia Plantarum 49: 341-346.
Smith, S. E. 1973. Asymbiotic Germination of Orchid Seeds on Carbohydrates of Fungal Origin. New Phytologist 72: 497-499.
Silva Júnior, J. M., M. Rodrigues, E. Mauro de Castro, S. K. Vilela Bertolucci, and M. Pasqual. 2013. Changes in Anatomy and
139
Chlorophyll Synthesis in Orchids Propagated in vitro in the Presence of Urea. Acta Scientiarum 35: 65-72.
Singh, F. 1991. Encapsulation of Spathoglottis plicata protocorms. Lindleyana 6: 61-63.
Singh, S. K., M. K. Rai, P. Asthana, and L. Sahoo. 2010. Alginate-Encapsulation of Nodal Segments for Propagation, Short-Term Conservation and Germplasm Exchange and Distribution of Eclipta alba (L.). Acta Physiologiae Plantarum 32: 607-610.
Sinha, P., M. L. Hakim, and M. F. Alam. 2009. In vitro Mass Clonal Propagation of Spathoglottis plicata Blume. Plant Tissue Culture and Biotechnology 19: 151-160.
Stewart, S. L. and M. E. Kane. 2006. Asymbiotic Seed Germination and in vitro Seedling Development of Habenaria macroceratitis (Orchidaceae), a Rare Florida Terrestrial Orchid. Plant Cell, Tissue and Organ Culture 86: 147-158.
Stewart, S. L. and M. E. Kane. 2007. Symbiotic Germination and Evidence for in vitro Mycobiont Specificity in Spiranthes brevilabris (Orchidaceae), and its Implications for Species-Level Conservation. In vitro Cellular and Developmental Biology - Plant 43: 175-186.
Stoutamire, W. P. 1974. Terrestrial Orchids Seedlings. In: Withner, D. L. (ed) The Orchids: Scientific Studies. Wiley and Sons, New York, USA.
Straus, M. S. and D. M. Reisinger. 1976. Effects of NAA on Seed Germination. American Orchid Society Bulletin 45: 722- 723.
Sungkumlong. and C. R. Deb. 2008. Effects of Different Factors on Immature Embryo Culture, PLBs Differentiation and Rapid Mass
140
Multiplication of Coelogyne suaveolens (Lindl.) Hook. Indian Journal of Experimental Biology 46: 243-248.
Swarts, N. D. and K. W. Dixon. 2009. Terrestrial Orchid Conservation in the Age of Extinction. Annals of Botany 104: 543-556.
Taiz, L. and E. Zeiger. 2009. Fisiología Vegetal 4th ed. Porto Alegre: Artmed.
Takahashi, K., I. Ogiwara, and N. Hakoda. 2001. Inoculation of Habenaria (Pecteilis) radiata with Two Isolates of Orchid Mycorrhizal Fungi: Germination and Growth Response to Separate, Stimulateneous, and Consecutive Inoculation. Lindleyana 16: 102-108.
Tan, T. K., W. S. Loon, E. Khor, and C. S. Loh. 1998. Infection of Spathoglottis plicata (Orchidaceae) Seeds by Mycorrhizal Fungus. Plant Cell Reports 18: 14-19.
Teng, W. L., L. Nicholson, and M. C. Teng. 1997. Micropropagation of Spathoglottis plicata. Plant Cell Reports 16: 831-835.
Teixteira da Silva, J. A. 2013. Orchids: Advances in Tissue Culture, Genetics, Phytochemistry and Transgenic Biotechnology. Floriculture and Ornamental Biotechnology 7: 1-52.
Thakur, U. and N. Dongarwar. 2012. Artificial Pollination and in vitro Asymbiotic Seed Germination in Garden Orchid Spathoglottis plicata Blume (Orchidaceae). Recent Research in Science and Technology 4: 13-18.
Thammasiti, K. 2000. Cryopreservation of Seeds of a Thai Orchid (Doritis Pulcherrima Lindl.) by Vitrification. CryoLetters 21: 237-244.
141
Thammasiti, K. and L. Soamkul. 2007. Cryopreservation of Vanda coerulea Griff. Ex Lindl Seeds by Vitrification. ScienceAsia 33: 223-227.
Thompson, D. I., T. J. Edwards, and J. V. Staden. 2006. Evaluating Asymbiotic Seed Culture Methods and Establibshing Disa (Orchidaceae) Germability in vitro: Relationships, Requirements and First-Time Reports. Plant Growth Regulators 49: 269-284.
Tomita, M. 1995. Symbiotic Seed Germination and Development of Goodyera schlechtendaliana in vitro. Plant Tissue Culture Letters 12: 151-155.
Torrés, J., L. Laskowsky, and M. E. Sanabria. 2006. Efecto del Ambiente de Desarrollo sobre la Anatomía de la Epidermis Foliar de Cattleya jenmanii Rolfe. Bioagro 18: 93-99.
Utetake, Y. and R. L. Peterson. 1998. Association between Microtubles and Symbiotic Fungal Hyphae in Protocorm Cells of the Orchid Species, Spiranthes sinensis. New Phytologist 140: 715-722.
Vacin, E. H. and F. W. Went. 1949. Some pH Changes in Nutrient Solution. Botanical Gazette 110: 605-613.
Van Waes, J. C. and P. C. Debergh. 1986. In vitro Germination of some Western European Orchids. Physiologia Plantarum 67: 253-261.
Valmayor, H. L., M. L. Pimental, and M. T. Martinez. 1986. Callus Formation and Plantlet Morphogenesis in Vanda. Malayan Orchid Review 20:22-30.
Vasudevan, R. and J. Van Staden. 2010. In vitro Asymbiotic Seed Germination and Seedling Growth of Ansellia Africana Lindl. Scientia Horticulturae 123: 496-504.
142
Vij, S. P., A. Sood, and K. K. Plaha. 1981. In vitro Seed Germination of some Epiphytic Orchids. p. 473-481. In: Verma, S. C (ed) Contemporary Trends in Plant Science. Kalyani Publishers, New Delhi, India.
Vijayakumar, S., G. Rajalkshmi, and K. Kalimuthu. 2012. Propagation of Dendrobium aggregatum through the Culture of Immature Seeds from Green Capsules. Lankesteriana 12: 131-115.
Víllora, G., D. A. Moreno, and L. Romero. 2002. Response of Eggplant to Nitrogen Supply: Molybdenum-Nitrate Relationships. Biologia Plantarum 45: 621-623.
Vogel, I. N. and A. F. Macedo. 2011. Influence of IAA, TDZ, and Light Quality on Asymbiotic Germination, Protocorm Formation, and Plantlet Development of Cyrtopodium glutiniferum Raddi., A Medicinal Orchid. Plant Cell, Tissue and Organ Culture 104: 147-155.
Wang, J. H., J. G. Ge, F. Liu, H. W. Bian, and C. N. Huang. 1998. Cryopreservation of Seeds and Protocorms of Dendrobium candidum. CryoLetters 19: 123-128.
Wang, T. Y., C. F. Lin, and Y. T. Lin. 2004. Effect of Orchid Mycorrhizal Fungi on Seed Germination of Dendrobium. p. 60. In: Abstract the 8th Asia Pacific Orchid Conference in Taiwan, 6-8 March.
Warcup, J. H. 1973. Symbiotic Germination of some Australian Terrestrial Orchids. New Phytologist 72: 387-392.
Warcup, J. H. 1981. The Mycorrhizal Relationships of Australian Orchids. New Phytologist 87: 371-381.
Williams, B. S. 1894. Sphatoglottis plicata. p. 717. In: The Orchid-Grower’s Manual 7th edition: Containing Descriptions of the Best
143
Species and Varieties of Orchidaceous Plants. Victoria and Paradise Nurseries, London.
Withner, C. L. 1951. Effect of Plant Hormones and other Compounds on Growth of Orchids. American Orchid Society Bulletin 20: 276-278.
Withner, C. L. 1955. Ovule Culture and Growth of Vanilla Seedlings. American Orchid Society Bulletin 51:380-392.
Wochok, Z. 1981. The Role of Tissue Culture in Preserving Threatened and Endangered Plant Species. Biology Conservation 20: 83-89.
Wotavová-Novotná, K., H. Vejsadová, and P. Kindlmann. 2007. Effects of Sugars and Growth Regulators on in vitro Growth of Dactylorhiza species. Biologia Plantarum 51: 198-200.
Yam, T. W. and M. A. Weatherhead. 1988. Germination and Seedling Development of some Hong Kong Orchids I. Lindleyana 3: 156-160.
Yamazaki, J. and K. Miyoshi. 2006. In vitro Asymbiotic Germination of Immature Seed and Formation of Protocorm by Cephalanthera falcate (Orchidaceae). Annals of Botany 98(6): 1197-1206.
Yin, M. and S. Hong. 2009. Cryopreservation of Dendrobium candidum Wall. Ex Lindl. Protocorm-like Bodies by Encapsulation Vitrification. Plant Cell, Tissue and Organ Culture 98: 179-195.
Zeng, S., Z. Chen, K. Wu, J. Zhang, C. Bai, J. A. Texteira da Silva, and J. Duan. 2011. Asymbiotic Seed Germination, Induction of Calli and Protocorm-Like Bodies, and in vitro Seedling Development of the Rare and Endangered Nothodoritis zhejiangensis Chinese Orchid. Horticultural Science 46(3): 460-465.
144
Zettler, L. W., J. C. Burkhead, and J. A. Marshall. 1999. Use of Mycorrhizal Fungus from Epidendrum conopseum to Propagate Encyclia tampensis from Seed in vitro. Lindleyana 14(2): 102-105.
Zettler, L. W. and C. J. Hofer. 1997. Sensitivity of Spiranthes odorata seeds to Light during in vitro Symbiotic Seed Germination. Lindleyana 12: 26-29.
Zettler, L. W., S. B. Poulter, and K. I. McDonald. 2007. Conservation-driven Propagation of an Epiphytic Orchid (Epidendrum nocturnum) with a Mycorrhizal Fungus. Horticultural Science 42(1): 135-139.
Zha, X. Q., J. P. Luo, S. T. Jiang, J. H. Wang. 2007. Enhancement of Polysaccharide Production in Suspension Cultures of Protocorm-Like Bodies from Dendrobium huoshanense by Optimization of Medium Compositions and Feeding of Sucrose. Process Biochemistry 42: 344-351.