Recent Advances in Orchid Seed Germination and Micropropagation

  • Yuan-Yuan Li
  • Colin Chan
  • Corrine Stahl
  • Edward C. Yeung
Part of the Springer Protocols Handbooks book series (SPH)


This chapter provides recent information focusing on different seed germination and micropropagation methods for various species of orchids. The three primary methods, i.e., symbiotic seed germination, asymbiotic seed germination, and micropropagation, are emphasized. Additionally, some recent studies on artificial seeds for germination are included. The information is presented in a table form in order to provide a quick overview on recent publications at a glance.

Key words

Micropropagation Asymbiotic seed germination Symbiotic seed germination Artificial seeds 


  1. 1.
    Knudson L (1922) Nonsymbiotic germination of orchid seeds. Bot Gaz 73:1–25CrossRefGoogle Scholar
  2. 2.
    Arditti J (1990) Lewis Knudson (1994–1958): his science, his times, and his legacy. Lindleyana 5:1–79Google Scholar
  3. 3.
    Yam TW, Arditti J (2009) History of orchid propagation: a mirror of the history of biotechnology. Plant Biotechnol Rep 3:1–56CrossRefGoogle Scholar
  4. 4.
    Yam TW, Nair H, Hew CS, Arditti J (2002) Orchid seeds and their germination: an historical account. In: Kull T, Arditti J (eds) Orchid biology: review and perspectives, VIII. Kluwer, The Netherlands, pp 387–504CrossRefGoogle Scholar
  5. 5.
    Stoutamire WP (1974) Terrestrial orchid seedlings. In: Winther CL (ed) The orchids: scientific studies. Wiley, New York, NY, pp 101–128Google Scholar
  6. 6.
    Chugh S, Guha S, Rao IU (2009) Micropropagation of orchids: a review on the potential of different explants. Sci Hortic 122:507–520CrossRefGoogle Scholar
  7. 7.
    Teixeira da Silva JA (2013) Orchids: advances in tissue culture, genetics, phytochemistry and transgenic biotechnology. Floricult Ornament Biotechnol 7:1–52Google Scholar
  8. 8.
    Royal Botanical Gardens Kew (2016) Orchid research newsletter. Accessed 14 Jun 2017
  9. 9.
    Herman EB (2014) Recent advances in plant tissue culture XIX. Orchid tissue culture: 2007–2014. Agritech Consultants, Inc., Mohegan Lake, New YorkGoogle Scholar
  10. 10.
    Sebastián F, Vanesa S, Eduardo F, Graciela T, Silvana S (2013) Symbiotic seed germination and protocorm development of Aa achalensis Schltr., a terrestrial orchid endemic from Argentina. Mycorrhiza 24:35–43PubMedCrossRefGoogle Scholar
  11. 11.
    Jiang JH, Lee YI, Cubeta MA, Chen LC (2015) Characterization and colonization of endomycorrhizal Rhizoctonia fungi in the medicinal herb Anoectochilus formosanus (Orchidaceae). Mycorrhiza 25:431–445PubMedPubMedCentralCrossRefGoogle Scholar
  12. 12.
    Liu SS, Chen J, Li SC, Zeng X, Meng ZX, Guo SX (2015) Comparative transcriptome analysis of genes involved in GA-GID1-DELLA regulatory module in symbiotic and asymbiotic seed germination of Anoectochilus roxburghii (Wall.) Lindl. (Orchidaceae). Int J Mol Sci 16:30,190–30,203CrossRefGoogle Scholar
  13. 13.
    Herrera H, Valadares R, Contreras D, Bashan Y, Arriagada C (2017) Mycorrhizal compatibility and symbiotic seed germination of orchids from the Coastal Range and Andes in south central Chile. Mycorrhiza 27:175–188PubMedCrossRefGoogle Scholar
  14. 14.
    Yamamoto T, Miura C, Fuji M, Nagata S, Otani Y, Yagame T, Yamato M, Kaminaka H (2017) Quantitative evaluation of protocorm growth and fungal colonization in Bletilla striata (Orchidaceae) reveals less-productive symbiosis with a non-native symbiotic fungus. BMC Plant Biol 17:50PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Nikabadi S, Bunn E, Stevens J, Newman B, Turner SR, Dixon KW (2014) Germination responses of four native terrestrial orchids from south-west Western Australia to temperature and light treatments. Plant Cell Tiss Org Cult 118:559–569CrossRefGoogle Scholar
  16. 16.
    Bustam BM, Dixon KW, Bunn E (2014) In vitro propagation of temperate Australian terrestrial orchids: revisiting asymbiotic compared with symbiotic germination. Bot J Linnean Soc 176:556–566CrossRefGoogle Scholar
  17. 17.
    Fracchia S, Aranda-Rickert A, Rothen C, Sede S (2016) Associated fungi, symbiotic germination and in vitro seedling development of the rare Andean terrestrial orchid Chloraea riojana. Flora 224:106–111CrossRefGoogle Scholar
  18. 18.
    Sathiyadash K, Muthukumar T, Murugan SB, Sathishkumar R, Pandey RR (2014) In vitro symbiotic seed germination of South Indian endemic orchid Coelogyne nervosa. Mycoscience 55:183–189CrossRefGoogle Scholar
  19. 19.
    Yagame T, Funabiki E, Nagasawa E, Fukiharu T, Iwase K (2013) Identification and symbiotic ability of Psathyrellaceae fungi isolated from a photosynthetic orchid, Cremastra appendiculata (Orchidaceae). Amer J Bot 100:1823–1830CrossRefGoogle Scholar
  20. 20.
    Rafter M, Yokoya K, Schofield EJ, Zettler LW, Sarasan V (2016) Non-specific symbiotic germination of Cynorkis purpurea (Thouars) Kraezl., a habitat-specific terrestrial orchid from the Central Highlands of Madagascar. Mycorrhiza 26:541–552PubMedCrossRefGoogle Scholar
  21. 21.
    Pereira MCCAA, Rocha DI, Veloso TGR, Pereira OL, Francino DMT, Meira RMSA, Kasuya MCM (2015) Characterization of seed germination and protocorm development of Cyrtopodium glutiniferum (Orchidaceae) promoted by mycorrhizal fungi Epulorhiza spp. Acta Bot Brasilica 29:567–574CrossRefGoogle Scholar
  22. 22.
    Guimarães FAR, Pereira MC, Felício CDS, Torres DP, Oliveira SF, Veloso TGR, Kasuya MCM (2013) Symbiotic propagation of seedlings of Cyrtopodium glutiniferum Raddi (Orchidaceae). Acta Bot Brasilica 27:590–596CrossRefGoogle Scholar
  23. 23.
    Umata H, Ota Y, Yamada M, Watanabe Y, Gale SW (2013) Germination of the fully myco-heterotrophic orchid Cyrtosia septentrionalis is characterized by low fungal specificity and does not require direct seed-mycobiont contact. Mycoscience 54:343–352CrossRefGoogle Scholar
  24. 24.
    Swangmaneecharern P, Serivichyaswat P, Nontachaiyapoom S (2012) Promoting effect of orchid mycorrhizal fungi Epulorhiza isolates on seed germination of Dendrobium orchids. Sci Hortic 148:55–58CrossRefGoogle Scholar
  25. 25.
    Harshani HC, Senanayake S, Sandamali H (2014) Host tree specificity and seed germination of Dendrobium aphyllum (Roxb.) C.E.C. Fisch. in Sri Lanka. In: J Nat Sci Found, vol 41, pp 71–86Google Scholar
  26. 26.
    Zi XM, Sheng CL, Goodale UM, Shao SC, Gao JY (2014) In situ seed baiting to isolate germination-enhancing fungi for an epiphytic orchid, Dendrobium aphyllum (Orchidaceae). Mycorrhiza 24:487–499CrossRefPubMedGoogle Scholar
  27. 27.
    Hajong S, Kumaria S, Tandon P (2013) Compatible fungi, suitable medium, and appropriate developmental stage essential for stable association of Dendrobium chrysanthum. J Basic Microbiol 53:1025–1033PubMedCrossRefGoogle Scholar
  28. 28.
    Shao CS, Burgess KS, Cruse-Sanders JM, Liu Q, Fan XL, Huang H, Gao JY (2017) Using in situ symbiotic seed germination to restore over-collected medicinal orchids in Southwest China. Front Plant Sci 8:888PubMedPubMedCentralCrossRefGoogle Scholar
  29. 29.
    Khamchatra N-M, Dixon K, Chayamarit K, Apisitwanich S, Tantiwiwat S (2016) Using in situ seed baiting technique to isolate and identify endophytic and mycorrhizal fungi from seeds of a threatened epiphytic orchid, Dendrobium friedericksianum Rchb.f. (Orchidaceae). Agri Nat Res 50:8–13Google Scholar
  30. 30.
    Tsavkelova EA, Egorova MA, Leontieva MR, Malakho SG, Kolomeitseva GL, Netrusov AI (2016) Dendrobium nobile Lindl. Seed germination in co-cultures with diverse associated bacteria. Plant Growth Regul 80:79–91CrossRefGoogle Scholar
  31. 31.
    Tan XM, Wang CL, Chen XM, Zhou YQ, Wang YQ, Luo AX, Liu ZH, Guo SX (2014) In vitro seed germination and seedling growth of an endangered epiphytic orchid, Dendrobium officinale, endemic to China using mycorrhizal fungi (Tulasnella sp.) Sci Hortic 165:62–68CrossRefGoogle Scholar
  32. 32.
    Chen J, Wang H, Liu SS, Li YY, Guo SX (2014) Ultrastructure of symbiotic germination of the orchid Dendrobium officinale with its mycobiont, Sebacina sp. Aust J Bot 62:229–234CrossRefGoogle Scholar
  33. 33.
    Hoang NH, Kane ME, Radcliffe EN, Zettler LW, Richardson LW (2017) Comparative seed germination and seedling development of the ghost orchid, Dendrophylax lindenii (Orchidaceae), and molecular identification of its mycorrhizal fungus from South Florida. Ann Bot 119:379–393PubMedCrossRefGoogle Scholar
  34. 34.
    Zettler LW, Corey LL, Jacks AL, Gruender LT, Lopez AM (2013) Tulasnella irregularis (basidiomycota: tulasnellaceae) from roots of Encyclia tampensis in south Florida, and confirmation of its mycorrhizal significance through symbiotic seed germination. Lankesteriana 13:119–128Google Scholar
  35. 35.
    Park EJ, Lee WY (2013) In vitro symbiotic germination of myco-heterotrophic Gastrodia elata by Mycena species. Plant Biotech Rep 7:185–191CrossRefGoogle Scholar
  36. 36.
    Park EJ, Lee WY, Ahn JK (2012) In vitro propagation of myco-heterotrophic Gastrodia elata. Hort Environ Biotech 53:415–420CrossRefGoogle Scholar
  37. 37.
    Fracchia SCA, Aranda-Rickert A, Flachsland E, Terada G, Sede S (2014) Mycorrhizal compatibility and symbiotic reproduction of Gavilea australis, an endangered terrestrial orchid from south Patagonia. Mycorrhiza 24:627–634PubMedCrossRefGoogle Scholar
  38. 38.
    Long JRD, Swarts ND, Dixon KW, Egerton-Warburton LM (2012) Mycorrhizal preference promotes habitat invasion by a native Australian orchid: Microtis media. Ann Bot 111:409–418PubMedPubMedCentralCrossRefGoogle Scholar
  39. 39.
    Khamchatra N, Dixon K, Tantiwiwat S, Piapukiew J (2016) Symbiotic seed germination of an endangered epiphytic slipper orchid, Paphiopedilum villosum (Lindl.) Stein. from Thailand. S Afri J Bot 104:76–81CrossRefGoogle Scholar
  40. 40.
    Zhou X, Gao J (2016) Highly compatible Epa-01 strain promotes seed germination and protocorm development of Papilionanthe teres (Orchidaceae). Plant Cell Tiss Org Cult 125:479–493CrossRefGoogle Scholar
  41. 41.
    Cruz-Higareda JB, Luna-Rosales BS, Barba-Álvarez A (2015) A novel seed baiting technique for the epiphytic orchid Rhynchostele cervantesii, a means to acquire mycorrhizal fungi from protocorms. Lankesteriana 15:67–76CrossRefGoogle Scholar
  42. 42.
    Ercole E, Rodda M, Girlanda M, Perrotto S (2015) Establishment of a symbiotic in vitro system between a green meadow orchid and a Rhizoctonia-like fungus. Bio-protocol 5:1–7CrossRefGoogle Scholar
  43. 43.
    Aewsakul N, Maneesorn D, Serivichyaswat P, Taluengjit A, Nontachaiyapoom S (2013) Ex vitro symbiotic seed germination of Spathoglottis plicata Blume on common orchid cultivation substrates. Sci Hortic 160:238–242CrossRefGoogle Scholar
  44. 44.
    Sathiyadash K, Muthukumar T, Murugan SB, Sathishkumar R, Uma E, Jaison S, Priyadharsini P (2013) In vitro asymbiotic seed germination, mycorrhization and seedling development of Acampae praemorsa (Roxb.) Blatt. & Mc Cann, a common south Indian orchid. AsPac J Reprod 2:114–118Google Scholar
  45. 45.
    Hossein P, Promila P, Bhanwra RK (2013) Asymbiotic germination of immature embryos of a medicinally important epiphytic orchid Acampe papillosa (Lindl.) Lindl. Afr J Biotechnol 12:162–167CrossRefGoogle Scholar
  46. 46.
    Suja RM, Williams BC (2016) Micropropagation, phytochemical screening and antioxidant potential of a wild epiphytic orchid Acampe praemorsa (roxb) of Kanyakumari district, India. Euro J Pharm Med Res 3:572–576Google Scholar
  47. 47.
    Bhattacharjee B, Islam SMS (2014) Development of an efficient protocol for in vitro germination and enhancing protocorm-like body development in three indigenous orchid species in Bangladesh. AsPac J Mol Biol Biotechnol 22:209–218Google Scholar
  48. 48.
    Parab GV, Krishnam S (2012) Rapid in vitro mass multiplication of orchids Aerides maculosa Lindl. and Rhynchostylis retusa (L.) Bl. from immature seeds. Indian J Biotechnol 11:288–294Google Scholar
  49. 49.
    Verma J, Bhatti SK, Sembi JK (2015) Asymbiotic germination of immature seeds in an ornamentally important ‘Fox-tail’ Orchid, Aerides multiflora Roxb. Vegetos Int J Plant Res 28:162–168CrossRefGoogle Scholar
  50. 50.
    Wesley PS, Devi C, Shibu BS, Moin S (2013) In vitro propagation of Coelogyne breviscapa Lindl., Dendrobium aqueum Lindl., and Flickingeria nodosa (Dalz.) Seidenf. via asymbiotic seed germination. AsPac J Mol Biol Biotechnol 21:26–32Google Scholar
  51. 51.
    Kaur S, Bhutani KK (2014) In vitro conservation and asymbiotic propagation of Coelogyne flaccida (Lindl.): a threatened orchid. Plant Biosystems 148:935–944CrossRefGoogle Scholar
  52. 52.
    Parmar G, Pant B (2015) In vitro seed germination and seedling development of Coelogyne flaccida lindl. (Orchidaceae). Adv Forestry Sci 2:85–88Google Scholar
  53. 53.
    Koirala D, Pradhan S, Pant B (2013) Asymbiotic Seed germination and plantlet development of Coelogyne fuscescens Lindl., a medicinal orchid of Nepal. Sci World 11:97–100CrossRefGoogle Scholar
  54. 54.
    Gaurav P, Bijaya P (2016) In vitro seed germination and seedling development of the orchid Coelogyne stricta (D. Don) Schltr. Afr J Biotechnol 15:105–109CrossRefGoogle Scholar
  55. 55.
    Pradhan S, Regmi T, Parmar G, Pant B (2013) Effect of different media on in vitro seed germination and seedling development of Cymbidium aloifolium (L.) Sw. Nepal J Sci Technol 14:51–56CrossRefGoogle Scholar
  56. 56.
    Bhowmik TK, Rahman MM (2017) In vitro seed germination and flowering of Dendrobium palpebrae Lindl. Orchid. J Global Biosci 6:4748–4757Google Scholar
  57. 57.
    Mahendran G, Muniappan V, Ashwini M, Muthukumar T, Bai VN (2012) Asymbiotic seed germination of Cymbidium bicolor Lindl. (Orchidaceae) and the influence of mycorrhizal fungus on seedling development. Acta Physiol Plant 35:829–840CrossRefGoogle Scholar
  58. 58.
    Li F, Bao J, Sun Y, Liu C, Ma H, Zhang T, Chen X (2016) Embryo development and corresponding factors affecting in vitro germination of Cymbidium faberi × C. sinense hybrid seeds. Arch Biol Sci 68:541–550CrossRefGoogle Scholar
  59. 59.
    Deb CR, Pongener A (2013) A study on the use of low cost substrata against agar for non-symbiotic seed culture of Cymbidium iridioides D. Don. Aust J. Crop Sci 7:642–649Google Scholar
  60. 60.
    Wang Y, Li Z, Huang L, Su J (2013) In vitro mass scale propagation of wild Cymbidium lowianum with a rare and endangered plant. Amer. J Plant Sci 4:1500–1507CrossRefGoogle Scholar
  61. 61.
    Zhang Y, Lee YI, Deng L, Zhao S (2013) Asymbiotic germination of immature seeds and the seedling development of Cypripedium macranthos Sw., an endangered lady’s slipper orchid. Sci Hortic 164:130–136CrossRefGoogle Scholar
  62. 62.
    Huh YS, Lee JK, Nam SY, Paek KY, Suh GU (2016) Improvement of asymbiotic seed germination and seedling development of Cypripedium macranthos Sw. with organic additives. J Plant Biotechnol 43:138–145CrossRefGoogle Scholar
  63. 63.
    Huh YS, Lee JK, Nam SY, Hong EY, Paek KY, Son SW (2016) Effects of altering medium strength and sucrose concentration on in vitro germination and seedling growth of Cypripedium macranthos Sw. J Plant Biotechnol 43:132–137CrossRefGoogle Scholar
  64. 64.
    Vijayakumar S, Rajalkshmi G, Kalimuthu K (2013) Propagation of Dendrobium aggregatum by green capsule culture. Lankesteriana 12:131–135Google Scholar
  65. 65.
    Acharya R, Parmar G (2016) In vitro seed germination of endangered Nepalese orchid species: Dendrobium fimbriatum Hook. Adv J. Seed Sci Technol 3:72–74Google Scholar
  66. 66.
    Udomdee W, Wen PJ, Lee CY, Chin SW, Chen FC (2014) Effect of sucrose concentration and seed maturity on in vitro germination of Dendrobium nobile hybrids. Plant Growth Regul 72:249–255CrossRefGoogle Scholar
  67. 67.
    Zhao D, Hu G, Chen Z, Shi Y, Zheng L, Tang A, Long C (2013) Micropropagation and in vitro flowering of Dendrobium wangliangii: a critically endangered medicinal orchid. J Med Plant Res 7:2098–2110CrossRefGoogle Scholar
  68. 68.
    Bakar B, Latip MA, Gansau JA (2014) Asymbiotic germination and seedling development of Dimorphics lowii (Orchidaceae). Asian J Plant Biol 2:28–33Google Scholar
  69. 69.
    Paudel M, Pradhan S, Pant B (2013) In vitro seed germination and seedling development of Esmeralda clarkei Rchb.f. (Orchidaceae). Plant Tiss Cult Biotechnol 22:107–111Google Scholar
  70. 70.
    Suzuki RCAM, Moreira VC, Pescador R, Ferreira WDM (2012) Asymbiotic seed germination and in vitro seedling development of the threatened orchid Hoffmannseggella cinnabarina. In Vitro Cell Dev Biol Plant 48:500–511CrossRefGoogle Scholar
  71. 71.
    Arenmongla T, Deb CR (2012) Germination of immature embryos and multiplication of Malaxis acuminata D. Don, an endangered therapeutically important orchid, by asymbiotic culture in vitro. Indian J Biotechnol 11:464–469Google Scholar
  72. 72.
    Zhang YY, KL W, Zhang JX, Deng RF, Duan J, Teixeira da Silva JA, Huang WC, Zeng SJ (2015) Embryo development in association with asymbiotic seed germination in vitro of Paphiopedilum armeniacum S. C. Chen et F. Y. Liu. Sci Rep 5:1–15Google Scholar
  73. 73.
    Zeng S, Wang J, Wu K, Teixeira da Silva JA, Zhang J, Duan J (2013) In vitro propagation of Paphiopedilum hangianum Perner & Gruss. Sci Hortic 151:147–156CrossRefGoogle Scholar
  74. 74.
    Diengdoh RV, Kumaria S, Tandon P, Das MC (2017) Asymbiotic germination and seed storage of Paphiopedilum insigne, an endangered lady’s slipper orchid. S Afri J Bot 112:215–224CrossRefGoogle Scholar
  75. 75.
    Jiang N, Zhang JX, Teixeira da Silva JA, Duan J, Liu HT, Zeng SJ (2016) Stimulatory effects of sodium hypochlorite and ultrasonic treatments on tetrazolium staining and seed germination in vitro of Paphiopedilum SCBG Red Jewel. Seed Sci Technol 44:77–90CrossRefGoogle Scholar
  76. 76.
    Zeng S, Wu K, Teixeira da Silva JA, Zhang J, Chen Z, Xia N, Duan J (2012) Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh., an endangered terrestrial orchid. Sci Hortic 138:198–209CrossRefGoogle Scholar
  77. 77.
    Hossain MM, Dey R (2013) Multiple regeneration pathways in Spathoglottis plicata Blume—a study in vitro. S Afric J Bot 85:56–62CrossRefGoogle Scholar
  78. 78.
    Hrahsel L, Thangjam R (2015) Asymbiotic in vitro seed germination and regeneration of Vanda coerulea Giff. Ex. Lindl., an endangered orchid from Northeast India. J Plant Sci Res 2:1–5Google Scholar
  79. 79.
    Jualang AG, Devina D, Hartinie M, Sharon JS, Roslina J (2014) Asymbiotic seed germination and seedling development of Vanda dearei. Malays Appl Biol 43:25–33Google Scholar
  80. 80.
    Prakash B, Bais RT, Singh P, Khan S (2013) Effect of different pH on in vitro seed germination of Vanda tessellata (Roxb.) Hook.Ex.G an endangered medicinal orchid. Adv Life Sci Technol 8:4–7Google Scholar
  81. 81.
    Sagaya M, Divakar KM (2015) In vitro seed germination of Xenikophyton smeeanum (rchb.f.) Garay: a rare orchid of Western Ghats, Karnataka. Int Res J Eng Technol 2:280–286Google Scholar
  82. 82.
    Deepti S, Gayatri MC, Sitikantha S (2013) In vitro seed germination as an aid to conserve Aerides maculosum lindl., an endemic and endangered orchid of western Ghats. Indian Int J Pharm Bio Sci 4:478–486Google Scholar
  83. 83.
    Srivastava D, Gayatri MC, Sarangi SK (2015) In vitro seed germination and plant regeneration of an epiphytic orchid Aerides ringens (Lindl.) Fischer. Indian J Biotechnol 14:574–580Google Scholar
  84. 84.
    Papenfus HB, Naidoo D, Pošta M, Finnie JF, Staden JV (2016) The effects of smoke derivatives on in vitro seed germination and development of the leopard orchid Ansellia africana. Plant Biol 18:289–294PubMedCrossRefGoogle Scholar
  85. 85.
    Neto VBDP, Campos GDO, Boaretto AG, Zuffo MCR, Torrezan MDA, Benetão J (2013) In vitro behaviour of Aspasia variegata, an epiphytic orchid from the Brazilian Cerrado. Ciência Rural 43:2178–2184CrossRefGoogle Scholar
  86. 86.
    Hu W, Yang Y, Liaw S, Chang C (2013) Cryopreservation the seeds of a Taiwanese terrestrial orchid, Bletilla formosana (Hayata) Schltr. by vitrification. Bot Stu 54:33CrossRefGoogle Scholar
  87. 87.
    Santos SAD, Smidt EC, Padial AA, Ribas LLF (2016) Asymbiotic seed germination and in vitro propagation of Brasiliorchis picta. Afric J Biotech 15:134–144CrossRefGoogle Scholar
  88. 88.
    Hoque MM, Al-Forkan M (2016) Role of basal media, carbon sources and growth regulators in micropropagation of two valuable medicinal orchids of Bangladesh. Int J Sci Res 5:1022–1026Google Scholar
  89. 89.
    Bustam BM, Dixon K, Bunn E (2014) Proliferation and harvesting of secondary protocorms as a novel means for improving propagation of terrestrial orchids. Aust J Bot 62:614–621CrossRefGoogle Scholar
  90. 90.
    Schneider L, de Paula Araujo JS, Zaffari GR (2014) Seed germination of Cattleya intermedia and Cattleya warneri in alternative culture media. Amer Int J Contemp Res 4:60–66Google Scholar
  91. 91.
    Sasamori MH, Endres Júnior D, Droste A (2015) Asymbiotic culture of Cattleya intermedia Graham (Orchidaceae): the influence of macronutrient salts and sucrose concentrations on survival and development of plantlets. Acta Bot Brasilica 29:292–298CrossRefGoogle Scholar
  92. 92.
    Díaz-Álvarez EA, Torres-Galeano C, Rojas-Cortés ÁP, De La Barrera E (2015) In vitro germination and development of two endangered endemic Colombian orchids Cattleya mendelii and Cattleya quadricolor. Gayana Bot Concepcion 72:213–220CrossRefGoogle Scholar
  93. 93.
    Abraham S, Augustine J, Thomas TD (2012) Asymbiotic seed germination and in vitro conservation of Coelogyne nervosa A. Rich. an endemic orchid to Western Ghats. Physiol Mol Biol Plants 18:245–251PubMedPubMedCentralCrossRefGoogle Scholar
  94. 94.
    Gogoi K, Kumaria S, Tandon P (2012) Ex situ conservation of Cymbidium eburneum Lindl.: a threatened and vulnerable orchid, by asymbiotic seed germination. 3 Biotech 2:337–343PubMedCentralCrossRefGoogle Scholar
  95. 95.
    Mohanty P, Paul S, Das MC, Kumaria S, Tandon P (2012) A simple and efficient protocol for the mass propagation of Cymbidium mastersii: an ornamental orchid of Northeast India. AoB Plants 2012:pls023PubMedPubMedCentralCrossRefGoogle Scholar
  96. 96.
    Rodrigues LA, Neto VBDP, Boaretto AG, Oliveira JFD, Torrezan MDA, Lima SFD, Otoni WC (2015) In vitro propagation of Cyrtopodium saintlegerianum rchb. f. (Orchidaceae), a native orchid of the Brazilian savannah. Crop Breed App Biotechnol 15:10–17CrossRefGoogle Scholar
  97. 97.
    Hossain MM, Sharma M, Pathak P (2012) In vitro propagation of Dendrobium aphyllum (Orchidaceae)—seed germination to flowering. J Plant Biochem Biotechnol 22:157–167CrossRefGoogle Scholar
  98. 98.
    Parthibhan S, Benjamin JHF, Muthukumar M, Sherif NA, Kumar ST, Rao MV (2012) Influence of nutritional media and photoperiods on in vitro asymbiotic seed germination and seedling development of Dendrobium aqueum Lindley. Afric J Plant Sci 6:383–393CrossRefGoogle Scholar
  99. 99.
    Paul S, Kumaria S, Tandon P (2012) An effective nutrient medium for asymbiotic seed germination and large-scale in vitro regeneration of Dendrobium hookerianum, a threatened orchid of northeast India. AoB Plants 2012:plr023CrossRefGoogle Scholar
  100. 100.
    Utami ESW, Hariyanto S, Manuhara YSW (2017) In vitro propagation of the endangered medicinal orchid, Dendrobium lasianthera J.J. Sm through mature seed culture. Asian Pac J Trop Biomed 7:406–410CrossRefGoogle Scholar
  101. 101.
    Thejaswini R, Narasimhan S (2017) Undefined organic additives stimulates in vitro seed germination of Dendrobium ovatum (Willd.) Kraenzl, a medicinal orchid. Int J Pharma Med Biol Sci 6:29–31Google Scholar
  102. 102.
    Voges JG, Benevenuto RF, Fritsche Y, Guerra MP (2014) Protocorm development of Epidendrum fulgens (Orchidaceae) in response to different saline formulations and culture conditions. Acta Sci Biol Sci 36:287–292CrossRefGoogle Scholar
  103. 103.
    Jevšnik TCBE, Luthar Z (2015) Successful disinfection protocol for orchid seeds and influence of gelling agent on germination and growth. Acta Agri Slovenica 105:95–102CrossRefGoogle Scholar
  104. 104.
    Gangaprasad A (2013) Rapid in vitro multiplication of Eulophia cullenii (Wight) Blume-a rare, endemic and exquisite orchid of southern western Ghats. Int Agri Environ Biotechnol 6:721–727Google Scholar
  105. 105.
    Samala S, Te-chatob S, Yenchonb S, Thammasiri K (2014) Protocorm-like body proliferation of Grammatophyllum speciosum through asymbiotic seed germination. SciAsia 40:379–383Google Scholar
  106. 106.
    Giri L, Jugran A, Rawat S, Dhyani P, Andola H, Bhatt ID, Rawal RS, Dhar U (2012) In vitro propagation, genetic and phytochemical assessment of Habenaria edgeworthii: an important Astavarga plant. Acta Physiol Plant 34:869–875CrossRefGoogle Scholar
  107. 107.
    Vudala S, Ribas L (2017) Seed storage and asymbiotic germination of Hadrolaelia grandis (Orchidaceae). S Afr J Bot 108:1–7CrossRefGoogle Scholar
  108. 108.
    Magrini S, Vitis MD (2017) In vitro reproduction of three Limodorum species (Orchidaceae): impacts of scarification methods and nitrogen sources on mature seed germination and seedling development. Plant Biosyst 151:419–428CrossRefGoogle Scholar
  109. 109.
    Malabadi RB, Meti NT, Mulgund GS, Nataraja K, Kumar SV (2012) Smoke saturated water promoted in vitro seed germination of an epiphytic orchid Oberonia ensiformis (Rees) Lindl. Res Plant Biol 2:32–40Google Scholar
  110. 110.
    Thakur U, Dongarwar N (2013) A new report of in vitro flowering and multiple shooting in a wild epiphytic orchid Oberonia recurva Lindl. from asymbiotically germinated seedlings. Plant Knowledge J 2:113–118Google Scholar
  111. 111.
    Bektaş E, Cüce M, Sökmen A (2013) In vitro germination, protocorm formation, and plantlet development of Orchis coriophora (Orchidaceae), a naturally growing orchid species in Turkey. Turkish J Bot 37:336–342Google Scholar
  112. 112.
    Chen Y, Goodale UM, Fan XL, Gao JY (2015) Asymbiotic seed germination and in vitro seedling development of Paphiopedilum spicerianum: an orchid with an extremely small population in China. Global Ecol Conserv 3:367–378CrossRefGoogle Scholar
  113. 113.
    Lesar H, Čeranič N, Kastelec D, Luthar Z (2012) Asymbiotic seed germination of Phalaenopsis Blume orchids after hand pollination. Acta Agric Slovenica 99:5–11Google Scholar
  114. 114.
    Poff KE, Sharma J, Richards M (2016) Cold-moist stratification improves germination in a temperate terrestrial orchid. S Appalachian Bot Soc 81:292–301Google Scholar
  115. 115.
    Utami ESW, Purnobasuki H, Soedarti T, Hariyanto S (2015) Asymbiotic seed germination and in vitro seedling development of Paphiopedilum liemianum Fowlie, an endangered terrestrial orchid in northern Sumatra, Indonesia. J Plant Sci 10:25–34CrossRefGoogle Scholar
  116. 116.
    Kalita R, Das AK, Gogoi H, Veer V (2013) Effect of growth hormones on in vitro mass-propagation of orchid species Rhynchostylis retusa. J Cell Tiss Res 13:3711–3714Google Scholar
  117. 117.
    Cevdet G, Sebnem E (2012) Seed germination and development of Serapias vomeracea (Burm. fil.) Briq. ssp. orientalis Greuter in tissue culture. Res J Biotechnol 7:4–8CrossRefGoogle Scholar
  118. 118.
    Sebastinraj J, Muhirkuzhali S (2014) Asymbiotic seed germination and micropropagation of Spathoglottis plicata Blume. Int J Adv Phar Biol Chem 3:495–501Google Scholar
  119. 119.
    Singh M, Kumaria S, Tandon P (2014) In vitro multiplication of Thunia marshalliana Rcbh. f. through mature seeds and pseudonodes. Indian J Biotechnol 13:268–273Google Scholar
  120. 120.
    Bastin M, Jeyachandran R (2015) Ex situ conservation of Trichoglottis tenera (Lindl.) a threatened, and endangered orchid of western Ghats using asymbiotic seed germination technique. Int J Recent Sci Res 6:3488–2496Google Scholar
  121. 121.
    Sebastinraj J, Britto SJ, Kumar DV, Robinson JP, Thangavel P (2014) Rapid propagation of Vanda testacea (Lindl.) Rchb. F—a highly medicinal value epiphytic orchid of India. World J Agric Sci 10:223–230Google Scholar
  122. 122.
    Islam R, Kabir KR, Hossain S, Khalil I (2014) Efficient in vitro cultural techniques for seeds germination of Vanda roxburghii. World J Agric Sci 10:163–168Google Scholar
  123. 123.
    Zeng SJ, Chen ZL, KL W, Zhang JX, Bai CK, Teixeira da Silva JA, Duan J (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. HortScience 46:460–465Google Scholar
  124. 124.
    Linsmaier EM, Skoog L (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100–127CrossRefGoogle Scholar
  125. 125.
    Thomas TH, van Staden J (1996) Dormancy break of celery (Apium graveolens L.) seeds by plant derived smoke extract. Plant Growth Regul 17:195–198CrossRefGoogle Scholar
  126. 126.
    Malabadi RB, Teixeira da Silva JA, Mulgund GS (2008) Smoke-saturated water influence in vitro seed germination of Vanda parviflora Lindl. Seed Sci Biotechnol 2:65–69Google Scholar
  127. 127.
    McCown BH, Lloyd G (1981) Woody plant medium (WPM)—a mineral nutrient formation for microculture for woody plant species. Hort Sci 16:453Google Scholar
  128. 128.
    Yam TW, Arditti J (2017) Micropropagation of orchids, 3rd edn. Blackwell, CambridgeCrossRefGoogle Scholar
  129. 129.
    Devi HS, Devi SI, Singh TD (2013) High frequency plant regeneration system of Aerides odorata Lour. through foliar and shoot tip culture. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 41:169–176CrossRefGoogle Scholar
  130. 130.
    Mubbarakh SA, Rahmah S, Rahman ZA, Sah NNM, Subramaniam S (2014) Cryopreservation of Brassidium Shooting Star orchid using the PVS3 method supported with preliminary histological analysis. Appl Biochem Biotechnol 172:1131–1145PubMedCrossRefGoogle Scholar
  131. 131.
    Lin L, Xu Q, Ma J, Yuan B, Li W (2014) Cryopreservation of protocorm-like body-derived shoot tips of Calanthe davidii by droplet-vitrification. Cryo-Letters 35:129–137PubMedGoogle Scholar
  132. 132.
    Dewir YH, El-Mahrouk ME, Murthy HN, Paek KY (2015) Micropropagation of Cattleya: improved in vitro rooting and acclimatization. Hortic Environ Biotechnol 56:89–93CrossRefGoogle Scholar
  133. 133.
    Kaur S, Bhutani K (2013) In vitro mass propagation of ornamentally and medicinally important Coelogyne flaccida Lindl. through pseudobulb segments. Plant Tiss Cult Biotechnol 23:39–47Google Scholar
  134. 134.
    Deb CR, Pogener A (2013) In vitro regenerative competence of foliar explants of Cymbidium aloifolium (L.) and C. iridioides D. Don: two horticultural important orchids. Indian J Biotechnol 12:402–408Google Scholar
  135. 135.
    Nahar SJ, Haque SM, Kazuhiko S (2016) Application of chondroitin sulfate on organogenesis of two Cymbidium spp. under different sources of lights. Notulae Sci Biol 8:156–160Google Scholar
  136. 136.
    Roy AR, Sajeev S, Pattanayak A, Deka BC (2012) TDZ induced micropropagation in Cymbidium giganteum Wall. Ex Lindl. and assessment of genetic variation in the regenerated plants. Plant Growth Regul 68:435–445CrossRefGoogle Scholar
  137. 137.
    Vyas S, Guha S, Kapoor P, Rao IU (2010) Micropropagation of Cymbidium Sleeping Nymph through protocorm-like bodies production by thin cell layer culture. Sci Hortic 123:551–557CrossRefGoogle Scholar
  138. 138.
    Kaewjampa N, Shimasaki K, Nahar SJ (2012) Hyaluronic acid can be a new plant growth regulator for hybrid Cymbidium micropropagation. Plant Tiss Cult Biotechnol 22:59–64Google Scholar
  139. 139.
    Flachsland E, Terada G, Fernandez JM, Medina R, Shinini A, Rey H, Mroginski L (2011) Plant regeneration from root-tip culture of Cyrtopodium brandonianum Barb. Rodr. (Orchidaceae). Propag Ornam Plants 11:184–188Google Scholar
  140. 140.
    Nambiar N, Tee CS, Maziah M (2012) Effects of organic additives and different carbohydrate sources on proliferation of protocormlike bodies in Dendrobium Alya Pink. Plant Omics 5:10–18Google Scholar
  141. 141.
    Julkiflee AL, Uddain J, Subramaniam S (2014) Efficient micropropagation of Dendrobium sonia-28 for rapid PLBs proliferation. Emir J Food Agric 26:545–551CrossRefGoogle Scholar
  142. 142.
    Atichart P (2013) Polyploid induction by colchicine treatments and plant regeneration of Dendrobium chrysotoxum. Thai J Agric Sci 46:59–63Google Scholar
  143. 143.
    Bhattacharyya P, Kumaria S, Job N, Tandon P (2016) En-masse production of elite clones of Dendrobium crepidatum: a threatened, medicinal orchid used in traditional Chinese medicine (TCM). J App Res Med Aromatic Plant 3:168–176CrossRefGoogle Scholar
  144. 144.
    Kaewubon P, Hutadilok-Towatana N, Teixeira da Silva JA, Meesawat U (2014) Ultrastructural and biochemical alterations during browning of pigeon orchid (Dendrobium crumenatum Swartz) callus. Plant Cell Tiss Org Cult 121:53–69CrossRefGoogle Scholar
  145. 145.
    Pradhan S, Paudel YP, Pant B (2013) Efficient regeneration of plants from shoot tip explants of Dendrobium densiflorum Lindl., a medicinal orchid. Afr J Biotechnol 12:1378–1383Google Scholar
  146. 146.
    Lee PL, Chen JT (2014) Plant regeneration via callus culture and subsequent in vitro flowering of Dendrobium huoshanense. Acta Physiol Plant 36:2619–2625CrossRefGoogle Scholar
  147. 147.
    Bhattacharyya P, Kumaria S, Tandon P (2016) High frequency regeneration protocol for Dendrobium nobile: a model tissue culture approach for propagation of medicinally important orchid species. S Afr J Bot 104:232–243CrossRefGoogle Scholar
  148. 148.
    Qian X, Wang C, Ouyang T (2014) In vitro flowering and fruiting in culture of Dendrobium officinate kimura et migo. (Orchidaceae). Pakistan J Bot 46:1877–1882Google Scholar
  149. 149.
    Pant B, Thapa D (2012) In vitro mass propagation of an epiphytic orchid Dendrobium primulinum Lindl. through shoot tip culture. Afr J Biotechnol 11:9970–9974Google Scholar
  150. 150.
    Poobathy R, Sinniah UR, Rathinam X, Subramaniam S (2013) Histology and scanning electron microscopy observations of cryopreserved protocorm-like bodies of Dendrobium sonia-28. Turkish J Biol 37:191–198Google Scholar
  151. 151.
    Singh DR, Baskaran V, Abirami K, Simhachalam P (2016) In vitro micropropagation of Eulophia andamanensis—a tropical orchid. Int J Trop Agric 34:2173–2177Google Scholar
  152. 152.
    Panwar D, Ram KH, Shekhawat N (2012) In vitro propagation of Eulophia nuda Lindl., an endangered orchid. Sci Hortic 139:46–52CrossRefGoogle Scholar
  153. 153.
    Antony JJJ, Sundarasekar J, Rathinam X, Marimuthu K, Subramaniam S (2014) Microscopical analysis of in vitro Mokara Broga Giant orchid’s PLBs. Emir J Food Agric 26:73–81CrossRefGoogle Scholar
  154. 154.
    Chen J, Hong P (2012) Cellular origin and development of secondary somatic embryos in Oncidium leaf cultures. Biol Plant 56:215–220CrossRefGoogle Scholar
  155. 155.
    Baker A, Kaviani B, Nematzadeh G, Negahdar N (2014) Micropropagation of Orchis catasetum—a rare and endangered orchid. Acta Sci Pol Hortorum Cultus 13:197–205Google Scholar
  156. 156.
    Wu K, Zeng S, Teixeria da Silva JA, Chen Z, Zhang J, Yang Y, Duan J (2012) Efficient regeneration of Renanthera Tom Thumb ‘Qilin’ from leaf explants. Sci Hortic 135:194–201CrossRefGoogle Scholar
  157. 157.
    Jitsopakul N, Thammasiri K, Ishikawa K (2013) Efficient adventitious shoot regeneration from shoot tip culture of Vanda coerulea, a Thai orchid. Sci Asia 39:449–455CrossRefGoogle Scholar
  158. 158.
    Tan BC, Chin CF, Alderson P (2013) Effects of sodium nitroprusside on shoot multiplication and regeneration of Vanilla planifolia Andrews. In Vitro Cell Dev Biol Plant 49:626–630CrossRefGoogle Scholar
  159. 159.
    Saiprasad GVS (2001) Artificial seeds and their applications. Reson 6:39–47CrossRefGoogle Scholar
  160. 160.
    Sharma S, Shahzad A, Teixeira da Silva JA (2013) Synseed technology—a complete synthesis. Biotechnol Adv 31:186–207PubMedCrossRefGoogle Scholar
  161. 161.
    Gantait S, Sinniah R (2013) Storability, post-storage conversion and genetic stability assessment of alginate-encapsulated shoot tips of monopodial orchid hybrid Aranda Wan Chark Kuan ‘Blue’ X Vanda coerulea Grifft. ex. Lindl. Plant Biotechnol Rep 7:257–266CrossRefGoogle Scholar
  162. 162.
    Gantait S, Bustam S, Sinniah UR (2012) Alginate-encapsulation, short-term storage and plant regeneration from protocorm-like bodies of Aranda Wan Chark Kuan ‘Blue’ × Vanda coerulea Grifft. ex. Lindl. (Orchidaceae). Plant Growth Regul 68:303–311CrossRefGoogle Scholar
  163. 163.
    Maqsood M, Mujib A, Siddiqui ZH (2012) Synthetic seed development and conversion to plantlet in Catharanthus roseus (L.) G. Don. Biotechnology 11:37–43CrossRefGoogle Scholar
  164. 164.
    Pradhan S, Tiruwal BL, Subedee BR, Pant B (2014) Micropropagation of Cymbidium aloifolium (L.) SW., a medicinal orchid by artificial seed technology. J Nat Hist Museum 28:42–48CrossRefGoogle Scholar
  165. 165.
    Pradhan S, Tiruwal BL, Subedee BR, Pant B (2016) Efficient plant regeneration of Cymbidium aloifolium (L.) Sw., a threatened orchid of Nepal through artificial seed technology. Am J Plant Sci 7:1964–1974CrossRefGoogle Scholar
  166. 166.
    Mohanty P, Nongkling P, Das MC, Kumaria S, Tandon P (2012) Short-term storage of alginate-encapsulated protocorm-like bodies of Dendrobium nobile Lindl.: an endangered medicinal orchid from North-east India. 3 Biotech 3:235–239PubMedPubMedCentralCrossRefGoogle Scholar
  167. 167.
    Siew WL, Kwok MY, Ong YM, Liew HP, Yew BK (2014) Effective use of synthetic seed technology in the regeneration of Dendrobium White Fairy Orchid. J Ornam Plants 4:1–7CrossRefGoogle Scholar
  168. 168.
    Bustam S, Sinniah UR, Kadir MA, Zaman FQ, Subramaniam S (2012) Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on different media and short term storage of Dendrobium Shavin White. Plant Growth Regul 69:215–224CrossRefGoogle Scholar
  169. 169.
    Sharma V (2014) In vitro propagation of endangered orchid taxa using alginate-encapsulated protocorm-like bodies (PLBs). Nature Environ Pollut Technol 13:401–404Google Scholar
  170. 170.
    Sarmah DK, Borthakur M, Borua PK (2010) Artificial seed production from encapsulated PLBs regenerated from leaf base of Vanda coerulea Grifft. ex. Lindl.—an endangered orchid. Curr Sci 98:686–690Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yuan-Yuan Li
    • 1
  • Colin Chan
    • 2
  • Corrine Stahl
    • 2
  • Edward C. Yeung
    • 2
  1. 1.Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical CollegeBeijingPeople’s Republic of China
  2. 2.Department of Biological SciencesUniversity of CalgaryCalgaryCanada

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