Plant Growth Regulation

, Volume 72, Issue 3, pp 249–255 | Cite as

Effect of sucrose concentration and seed maturity on in vitro germination of Dendrobium nobile hybrids

  • Waraporn Udomdee
  • Pei-Jung Wen
  • Chen-Yu Lee
  • Shih-Wen Chin
  • Fure-Chyi ChenEmail author
Original paper


Orchid is a major floral crop around the world and Dendrobium hybrids are considered to be one of the most popular orchids. In vitro germination of hybrid seeds is a common practice among orchid growers, however, in many cross pollinations the embryos may not develop to maturity, leading to poor seed germination. The effect of seed maturity and sucrose concentration were investigated via asymbiotic germination of nobile Dendrobium hybrids. Capsules were harvested from two hybrids (Den. Lucky Girl × Den. Second Love ‘Kirameki’ and Den. Lucky Girl × Den. Hamana Lake ‘Kumi’) and one selfing of Den. Second Love ‘Kirameki’ at 2, 3, 4, and 5 months after pollination and immature seeds were taken. Immature seeds from 3- to 5-month old capsules could be successfully germinated on Hyponex based medium. Immature seeds from 4-month old capsules showed greatest germination rate of tested treatments, whereas 3-month old immature seeds showed the least germination. After 6 weeks of in vitro culture, protocorms derived from embryos developed on every concentration of sucrose, but germination was greater at lower concentrations. Greater concentration of sucrose decreased normal-developed protocorms.


Nobile Dendrobium Asymbiotic seed germination Capsule maturity Embryo rescue 



Activated charcoal


Coconut water


Murashige and Skoog medium


Photosynthetic photon flux



The first author thanks National Pingtung University of Science and Technology for providing a scholarship during her PhD study. We also thank members of the laboratory of plant biotechnology, Department of plant industry, National Pingtung University of Science and Technology for assistance. This work was supported by grants supported by the Council of Agriculture, Taiwan with grant numbers 100AS-5.3.1-ST-aB and 101AS-5.3.1-ST-aB.


  1. Arditti J, Ernst R (1993) Micropropagation of orchids. Wiley, New York, p 682Google Scholar
  2. Ávila-Díaz I, Oyama K, Gómez-Alonso C, Salgado-Garciglia R (2009) In vitro propagation of the endangered orchid Laelia speciosa. Plant Cell Tissue Organ Cult 99:335–343CrossRefGoogle Scholar
  3. Bridgen MP (1994) A review of plant embryo culture. HortScience 29:1243–1246Google Scholar
  4. Deb CR, Pongener A (2011) Asymbiotic seed germination and in vitro seedling development of Cymbidium aloifolium (L.) Sw.: a multipurpose orchid. J Plant Biochem Biotechnol 20:90–95CrossRefGoogle Scholar
  5. Deb CR, Sungkumlong (2008) Effects of different factors on immature embryo culture, PLBs differentiation and rapid mass multiplication of Coelogyne suaveolens (Lindl.) Hook. Indian J Exp Biol 46:243–248PubMedGoogle Scholar
  6. Deng Y, Chen S, Chen F, Cheng X, Zhang F (2011) The embryo rescue derived intergeneric hybrid between chrysanthemum and Ajania przewalskii shows enhanced cold tolerance. Plant Cell Rep 30:2177–2186PubMedCrossRefGoogle Scholar
  7. Hsu CC, Chen FC (2003) Plant regeneration from protocorm-like bodies induced in etiolated leave of Phalaenopsis aphrodite Rchb. f. J Chinese Soc Hortic Sci 49(4):335–342 (In Chinese with English abstract.)Google Scholar
  8. Ichihashi S (1978) Studies on the media for orchid seed germination II. The effects of anionic and cationic combinations relevant to seeding populations and culture periods on the growth of Bletilla striata seedlings. J Jpn Soc Hortic Sci 46:521–529CrossRefGoogle Scholar
  9. Islam MO, Ichihashi S (1999) Effects of sucrose, maltose and sorbitol on callus growth and plantlet regeneration in Phalaenopsis, Coritaenopsis and Neofinetia. J Japan Soc Hortic Sci 68:1124–1131CrossRefGoogle Scholar
  10. Johnson TR, Kane ME, Perez HE (2011) Examining the interaction of light, nutrients and carbohydrates on seed germination and early seedling development of Bletia purpurea (Orchidaceae). Plant Growth Regul 63:89–99CrossRefGoogle Scholar
  11. Kamemoto H, Amore TD, Kuehnle AR (1999) Breeding Dendrobium Orchids in Hawaii. University of Hawaii Press, Hononolu, pp 55–61Google Scholar
  12. Kananont N, Pichyangkura R, Chanprame S, Chadchawan S, Limpanavech P (2010) Chitosan specificity for the in vitro seed germination of two Dendrobium orchids (Asparagales: Orchidaceae). Sci Hortic 124:239–247CrossRefGoogle Scholar
  13. Knudson L (1946) A new nutrient solution for germination of orchid seed. Am Orchid Soc Bull 15:214–217Google Scholar
  14. Lee YI (2003) Growth periodicity, changes of endogenous abscisic acid during embryogenesis, and in vitro propagation of Cypripedium formosanum Hay. PhD Dissertation. National Taiwan University, Taipei, TaiwanGoogle Scholar
  15. Lee YI (2011) In vitro culture and germination of terrestrial Asian orchid seeds. In: Thorpe TA, Yeung EC (eds) Plant embryo culture: methods and protocols. Humana Press, New York, pp 53–62CrossRefGoogle Scholar
  16. Lee YI, Yeung EC (2010) Embryo development and in vitro seed germination of Bulbophyllum fascinator. Acta Hortic 878:243–250Google Scholar
  17. Lee YI, Yeung EC, Cheng MC (2005) Embryo development of Cypripedium formosanum in relation to seed germination in vitro. J Am Soc Hortic Sci 130:747–753Google Scholar
  18. Lee YI, Lu EF, Chung MC, Yeung EC, Lee N (2007) Developmental changes in endogenous abscisic acid concentrations and asymbiotic seed germination of a terrestrial orchid, Calanthe tricarinata Lindl. J Am Soc Hortic Sci 132:246–252Google Scholar
  19. Lee YI, Yeung EC, Lee N, Chung MC (2008) Embryology of Phalaenopsis amabilis var. formosa: embryo development. Bot Stud 49:139–146Google Scholar
  20. Lee SJ, Liao FS, Cheng SH (2010) Effects of culture media and capsule maturity on seed germination of intraspecific and interspecific crosses in the genus Calanthe. Acta Hortic 878:231–235Google Scholar
  21. Lo SF, Nalawade SM, Kuo CL, Chen CL, Tsay HS (2004) Asymbiotic germination of immature seeds, plantlet development and ex vitro establishment of plants of Dendrobium tosaense Makino—a medicinally important orchid. In Vitro Cell. Dev Biol Plant 40:528–535CrossRefGoogle Scholar
  22. Martin KP, Madassery J (2006) Rapid in vitro propagation of Dendrobium hybrids through direct shoot formation from foliar explants, and protocorm-like bodies. Sci Hortic 108:95–99CrossRefGoogle Scholar
  23. Mweetwa AM, Welbaum GE, Tay D (2008) Effects of development, temperature, and calcium hypochlorite treatment on in vitro germinability of Phalaenopsis seeds. Sci Hortic 117:257–262CrossRefGoogle Scholar
  24. Ngampanya B, Homla-aor W (2010) Simple media for Dendrobium orchid seed germination and protocorm development. Acta Hortic 878:219–223Google Scholar
  25. Niimoto DH, Sagawa NY (1961) Ovule development in Dendrobium. Am Orchid Soc Bull 30:813–819Google Scholar
  26. Pierik RLM (1988) In vitro culture of higher plants as a tool in the propagation of horticultural crops. Acta Hortic 226:25–40Google Scholar
  27. Roy AR, Patel PS, Patel VV, Sajeev S, Deka BC (2011) Asymbiotic seed germination, mass propagation and seedling development of Vanda coerulea Griff ex. Lindl. (Blue Vanda): an in vitro protocol for an endangered orchid. Sci Hortic 128:325–331CrossRefGoogle Scholar
  28. Sharrock S, Lusty C (2000) Nutritive value of banana. In: INIBAP annual report 1999. INIBAP: Montpellier (FRA), pp 28–31Google Scholar
  29. Stewart SL, Kane ME (2006) Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue Organ Cult 86:147–158CrossRefGoogle Scholar
  30. Tan TK, Loon WS, Khor E, Loh CS (1998) Infection of Spathoglottis plicata (Orchidaceae) seeds by mycorrhizal fungus. Plant Cell Rep 18:14–19CrossRefGoogle Scholar
  31. Thongpukdee A, Thepsithar C, Rojanawong T (2010) Optimum conditions for seed germination of Phalaenopsis Silky Moon. Acta Hortic 878:237–242Google Scholar
  32. Tsai CC, Chiang YC, Huang SC, Liu WL, Chou CH (2009) Intergeneric hybridization, embryo rescue and molecular detection for intergeneric hybrids between Ascocenda and Phalaenopsis. Acta Hortic 829:413–416Google Scholar
  33. Vasudevan R, Staden JV (2010a) In vitro asymbiotic seed germination and seedling growth of Ansellia africana Lindl. Sci Hortic 123:496–504CrossRefGoogle Scholar
  34. Vasudevan R, Staden JV (2010b) Fruit harvesting time and corresponding morphological changes of seed integuments influence in vitro seed germination of Dendrobium nobile Lindl. Plant Growth Regul 60:237–246CrossRefGoogle Scholar
  35. Vendrame WA, Carvalho VS, Dias JMM (2007) In vitro germination and seedling development of cryopreserved Dendrobium hybrid mature seeds. Sci Hortic 114:188–193CrossRefGoogle Scholar
  36. Vijayakumar S, Rajalkshmi G, Kalimuthu K (2012) Propagation of Dendrobium aggregatum through the culture of immature seeds from green capsules. Lankesteriana 12:131–1135Google Scholar
  37. Vyas S, Guha S, Bhattacharya M (2009) Rapid regeneration of plants of Dendrobium lituiflorum Lindl. (Orchidaceae) by using banana extract. Sci Hortic 121:32–37CrossRefGoogle Scholar
  38. Wotavová-Novotná K, Vejsadová H, Kindlmann P (2007) Effects of sugars and growth regulators on in vitro growth of Dactylorhiza species. Biol Plant 51:198–200CrossRefGoogle Scholar
  39. 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
  40. Zha XQ, Luo JP, Jiang ST, Wang JH (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 Biochem 42:344–351CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Waraporn Udomdee
    • 1
  • Pei-Jung Wen
    • 2
  • Chen-Yu Lee
    • 2
  • Shih-Wen Chin
    • 2
  • Fure-Chyi Chen
    • 2
    Email author
  1. 1.Department of Tropical Agriculture and International CooperationNational Pingtung University of Science and TechnologyPingtungTaiwan
  2. 2.Department of Plat IndustryNational Pingtung University of Science and TechnologyPingtungTaiwan

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