Document Type : Research paper


1 Agriculture Biotechnology Research Institute of Iran (ABRII) Seed and Plant Improvement Institutes Campus Mahdasht Road P. O. Box 31535-1897 Karaj, Iran

2 Agriculture Biotechnology Research Institute of Iran (ABRII), Seed and Plant Improvement Institutes Campus, Mahdasht Road, P. O. Box 31535-1897, Karaj, Iran


One of the most effective ways to propagate Lilium is by using tissue culture techniques, preferably through bulblet production. In addition to the surviving percentage of Lilium bulbs after transplantation, the small size of in vitro bulblets compared to commercial bulbs and bulblets‟ dormancy are the most important constraints to commercializing Lilium micropropagation. Different concentrations of sucrose or sorbitol as carbohydrate sources were investigated as maturation and enlargement factors on in vitro bulblets of Lilium longiflorum cv. „Ceb-dazzle‟. Five concentrations of gibberellic acid and 4°C cold treatment for 6, 7, 8, 9, 10 and 11 weeks in the greenhouse were studied in relation to the dormancy breaking of three in vitro Lilium bulblets cultivars: „Simplon‟, „Navona‟ and „Ceb-dazzle‟, which belong to the Oriental, Asiatic and LA hybrids. The results showed that 3% of sucrose and two weeks of culture significantly increased the bulblet maturation and enlargement of „Ceb-dazzle‟. One mg L-1 gibberellic acid for six weeks, or cold treatment for six to eight weeks, had significant positive effects on the dormancy breaking of in vitro bulblets of Lilium cultivars.


1. Aguttaz, P., A. Paffen, I. Delvallee, P. Van Der Linde, and G.J. De Klerk. 1990. The Development of Dormancy in Bulblets of Lilium speciosum Generated In Vitro. I. the Effects of Culture Conditions. Plant Cell Tiss. Org. Cult. 22: 167–172.
2. Azadi, P., and M. Khosh-Khui. 2007. Micropropagation of Lilium ledebourii (Baker) Boiss as Affected by Plant Growth Regulator, Sucrose Concentration, Harvesting Season and Cold Treatments. Electro. J. Biotech. 4: 582-591.
3. Bacchetta, L., P.C. Remotti, C. Bernardini, and F. Saccardo. 2003. Adventitious Shoot Regeneration from Leaf Explants and Stem Nodes of Lilium. Plant Cell Tiss. Org. Cult. 74: 37-44.
4. Bakhshaie, M., M. Babalar, M. Mirmasoumi, and A. Khalighi. 2010. Somatic Embryogenesis
and Plant Regeneration of Lilium ledebourii (Baker) Boiss., an Endangered Species. Plant Cell Tiss. Org. Cult. 102: 229-235.
5. Bong, H.H., B.W. Yae, H.J. Yu, and K.Y. Peak. 2005. Improvement of in Vitro Micropropagation of Lilium Oriental Hybrid 'Casablanca' by the Formation of Shoots with Abnormally Swollen Basal Plates. Sci. Hort. 103: 351-359.
6. Chang, C., C.-T. Chen, Y.-C. Tsai, and W.-C. Chang. 2000. A Tissue Culture Protocol for Propagation of a Rare Plant, Lilium speciosum Thunb. var. glorisoides Baker. Bot. Bull. Acad. Sinica 41: 139-142.
7. Dantu, P.K., and S.S. Bhojwani. 1995. In Vitro Corm Formation And Field Evaluation Of Corm-Derived Plants of Gladiolus. Sci. Hort. 61: 115-129.
8. De Klerk, G.J., I. Delvallée, and A. Paffen. 1992. Dormancy Release of Micropropagated 
Bulblets of Lilium speciosum after Long Culture in Soil. HortScience 27:147–148.
9. Delvallée, I., A. Paffen, and G.J. De Klerk. 1990. The Development of Dormancy in Bulblets of Lilium speciosum Generated In Vitro II. The Effect of Temperature. Physiol. Plant. 80: 431–436.
10. Dole, J.M. 2003. Research Approaches for Determining Cold Requirements for Forcing and Flowering of Geophytes. HortScience 38:341–346.
11. Finkelstein, R., W. Reeves, T. Ariizumi, and C. Steber. 2008. Molecular Aspects of Seed Dormancy. Annu. Rev. Plant Biol. 59:387-415.
12. Fuentes, S.R.L., M.B.P. Calheiros, J. Manetti-Filho, and L.G.E. Vieira. 2000. The Effect of Silver Nitrate and Different Carbohydrate Sources on Somatic Embryogenesis in Coffea canephora. Plant Cell Tiss. Org. Cult. 60:5-13.
13. Gerrits, M.,K.S., Kim, and G.J. De Klerk. 1992. Hormonal Control of Dormancy in Bulblets of Lilium speciosum Cultured in Vitro. Acta Hort. 352: 521-527.
14. Gua, C.H., M. Onjo, M. Hayashi, and C.H. Guo. 2000. Studies on Seed Tuber Production using Small Tubers and on the Breaking of Dormancy in Potatoes using GA. Jpn J. Trop. Agr. 44: 152–7.
15. Han, B.H., B.W. Yae, H.U. Yu, and K.Y. Peak. 2005. Improvement of in Vitro Micropropagation of Lilium Oriental Hybrid „Casablanca‟ by the Formation of Shoots with Abnormally Swollen Basal Plates. Sci. Hort. 103:351–359.
16. Ho, C.W., W.T. Jian, and H.C. Lai. 2006. Plant Regeneration Via Somatic Embryogenesis from Suspension Cell Cultures of Lilium ×formolongi Hort. using a Bioreactor System. In Vitro Cell. Devel. Biol.-Plant 42: 240–246.
17. Hobson, G.E., and J.N. Davies. 1978. Influence of the Extent and Duration of Cold Treatment on the Flowering Behavior, Composition and Metabolic Activity of Tulip Bulbs. Sci. Hort. 8: 279–287.
18. Ishimori, T., Y. Nimii, and D.S. Han. 2009. In Vitro Flowering of Lilium rubellum Baker. Sci. Hort. 120: 246-249.
19. Jeong, J.H. 1996. In Vitro Propagation of Bulb Scale Section of Several Korean Native Lilies. Acta Hort. 414: 269-276.
20. Kumar, S., M. Kashyap, and D.R. Sharma. 2005. In Vitro Regeneration and Bulblet Growth
from Lily Bulbscale Explants as Affected by Retardants, Sucrose and Irradiance. Biol. Plant. 49: 629-632.
21. Langens-Gerrits, M.M., S. Nashimoto, A.F. Croes, and G.J. De Klerk. 2001. Development of Dormancy in Different Lily Genotypes Regenerated in Vitro. Plant Growth Regul. 34: 215–222.
22. Langens-Gerrits, M.M., W.B.M. Miller, A.F. Croes, and G.J. De Klerk. 2003. Effect of Low Temperature on Dormancy Breaking and Growth after Planting in Lily Bulblets Regenerated in Vitro. Plant Growth Regul. 40: 267–275.
23. Lemos, E.E.P., and D.A. Baker. 1998. Shoot Regeneration in Response to Carbon Source on Internodal Explants of Annona muricata L. Plant Growth Regul. 25: 105-112.
24. Lian, M., D. Cjakrabarty, and K.Y. Paek. 2002. Growth and Uptake of Sucrose and Mineral Ions by Bulblets of Lilium Oriental Hybrid „Casablanca‟ During Bioreactor Culture. J. Hort. Sci. Biotech. 77: 253-257.
25. Lian, M., H.N. Murthy, and K.Y. Paek. 2003. Photoautotrophic Culture Conditions and Photosynthetic Photon Flux Influence Growth of Lilium Bulblets in Vitro. In vitro Cell. Dev. Biol.-Plant 39: 532-535.
26. Liu, H., P. Xu, Y. Yin, and H. Li. 2008. Research on the Growth of Oriental Lily and Lilium longiflorum Thunb. Bulblet in Vitro. J. Biotech.: S163 (abst.).
27. Memar Moshrefi, M. Moini, and I. Tavassolian. 2002. Effects of Plant Growth Regulators NAA , BAP , Different Explants Scale and Photoperiod on Tissue Culture of Lilium ledebourii Boiss. Iranian J. Crop Sci. 4: 253-664.
28. Mojtahedi, N., O. Dabirashrafi, A.A. Habashi, and P. Koobaz. 2010. Direct in Vitro Bulblet Production in Lilium longiflorum cv. Gironde. J. Hort. Sci. Tech. 11: 221-234.
29. Mori, S., Y. Adachi, S. Horimoto, S. Suzuki, and M. Nakano. 2005. Callus Formation and Plant Regeneration in Various Lilium Species and Cultivars. In vitro Cell. Dev. Biol.-Plant 41: 783–788.
30. Murashige, T., and F. Skoog. 1962. A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiol. Plant. 15: 473–497.
31. Nhut, D.T. 2003. The Control of in Vitro Direct Main Stem Formation of Lilium longiflorum Derived from Receptacle Culture and Rapid 
Propagation by using in Vitro Stem Nodes. Plant Growth Regul. 40:179-184.
32. Nhut, D.T., N.T.M. Hanh, P.Q. Tuan, LT.M. Nquyet, N.T.H. Tram, N.C. Chinh, N.H. Ngyen, and D.N. Vinh. 2006. Liquid Culture as a Positive Condition to Induce and Enhance Quality and Quantity of Somatic Embryogenesis of Lilium longiflorum. Sci. Hort. 110: 93–97.
33. Nowak, J., M. Saniewsky, and R.M. Rudnicki. 1974. Studies on the Physiology of Hyacinth Bulbs (Hyacinthus orientalis). I. Sugar Content and Metabolic Activities in Bulbs Exposed to Low Temperatures. J. Hort. Sci. 49: 383– 390.
34. Ohkawa, K. 1979. Effect of Gibberellins and Benzyladenin on Dormancy and Flowering of Lilium speciosum. Sci. Hort. 10: 255-260.
35. Padasht Dehkaei, M.N., A. Khalighi, R. Naderi, and A. Mosavi. 2008. Effect of Different Concentration of Benzyladenine (BA) and Naphthalene Acetic Acid (NAA) on Regeneration of Lilium ledebourii (Chelcheragh Lily) using Bulblets Microscales. Seed Plant 24: 321-332.
36. Rahman, M.H., M.S. Haque, MA. Karim, and M. Ahmed. 2006. Effects of Gibberellic Acid (GA3) on Breaking Dormancy in Garlic (Allium sativum L.). Int. J. Agr. Biol. 80: 63- 65.
37. Sharma, V.K., R. Hansh, R.R. Mendel, and J. Schulze. 2005. Seasonal Effect on Tissue Culture Response and Plant Regeneration Frequency from Nonbombarded and Bombarded Immature Scutella of Barley (Hordeum vulgare) Harvested from Controlled Environment. Plant Cell Tiss. Org. Cult. 81:19-26.
38. Sun, L., Z. Zhou, and K. Cheng. 2013. Plant Micropropagation from in Vitro Cultured Bulb
Scales of Lilium lancifolium. Life Sci. J.10:2689-2692.
39. Takayama, S., and M. Misawa. 1979. Differentiation in Lilium Bulb Scales Grown in Vitro. Effects of Various Cultural Conditions. Physiol. Plant. 46:184.
40. Terui, K., and N. Okagami. 1988. Loss of Short Day Requirement for Dormancy Breaking of Bulbils from Gibberellic Acid- Treated Plants in Sedum Bulbiferum. Plant Sci. 58:129- 134.
41. Tymoszuk, J., M. Saniewski, and R.M. Rudnicki. 1979. The Physiology of Hyacinth Bulbs. XV. the Effect of Gibberllic Acid and Silver Nitrate on Dormancy Release and Growth. Sci. Hort. 11:95-99.
42. Vashney, A., V. Dhawan, and P.S. Srivastava. 2000. A Protocol for in Vitro Mass Propagation of Asiatic Hybrids of Lily Through Liquid Stationary Culture. In vitro Cell. Dev. Biol.-Plant 36:383-391.
43. Wawrosch, C., P.R. Malla, and B. Kopp. 2001. Clonal Propagation of Lilium nepalense D. Don, a Threatened Medicinal Plant of Nepal. Plant Cell Rep. 20:285-288.
44. Xiao-Hual, L., D. Yi-Wei, Z. Ying-Jie, and L. Ying-Min. 2012. In vitro Micropropagation of Longiflorum-Asiatic (LA) Hybrids Lily (Lilium) Cultivar „Eyeliner‟. African J. Biotechnol. 11:13506-13517.
45. Xu R.Y., Y. Niimi, and D.S. Han. 2006. Changes in Endogenous Abscisic Acid and Soluble Sugars Levels during Dormancy-Release in Bulbs of Lilium rubellum. Sci. Hort. 111:68-72.