Efficient Approaches to in vitro Multiplication of Lilium candidum L. with Consistent and Safe Access throughout Year and Acclimatization of Plant under Hot-Summer Mediterranean (Csa Type) Climate


  • Sheida DANESHVAR ROYANDAZAGH Tekirdağ Namık Kemal University, Faculty of Agriculture, Department of Agricultural Biotechnology, Değirmenaltı Campus 59030 Süleymanpaşa-Tekirdağ (TR)




in vitro; Lilium candidum; rooting; single scale bulb


Lilium candidum L. is one of the most fragrant plant species among Liliaceae. The indigenous and natural populations of L. candidum are significantly affected by fast increasing anthropological population pressure and increased carbon fuel pollution. There is a necessity for development of a micropropagation protocol to serve in effective manner for both commercial multiplication and protection of the plant. The study targeted to develop a strategy for effective in vitro plant propagation system using static liquid culture, seismomorphogenic treatments with mixed liquid culture and semi solid culture medium using single scale explants. The former two techniques failed to regenerate new bulblets effectively whereby the later technique induced 100% regeneration on all treatments. Maximum number of bulblets regenerated on 0.27 µM Thidiazuron + 1.08 µM Naphthalene acetic acid. The scale explant could be recultured and regenerated on the mother culture medium for six cycles. The daughter bulblets regenerated on Murashige and Skoog medium containing both 0.18 or 0.27 µM TDZ + 1.08 µM NAA were successfully rooted using 1/2 × MS medium containing 2.45 µM indole butyric acid. Regardless of rooting development, acclimatization was noted on all bulblets in the greenhouse. These plants are showing continuous flowering since last four years under hot-summer Mediterranean (CSa type) climate field conditions of Tekirdag province of Thrace region in Turkey.


Metrics Loading ...


Aasim M, Khawar KM, Özcan S (2009). Comparison of shoot regeneration on diferent concentrations of Thidiazuron from shoot tip explant of cowpea on gelrite and agar containing medium. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 37(1):89-93.

Arzate-Fernandez AM, Nakazaki T, Okumoto Y, Tanisaka T (1997). Efficient callus induction and plant regeneration from filaments with anther in lily (Lilium longiflorum Thunb.). Plant Cell Reports 16(12):836-840.

Asokan MP, O'Hair SK, Litz RE (1984). In vitro plant regeneration from corm callus of Amorphophallus rivieri Durieu. Scientia Horticulturae 24(3-4):251-256.

Azadi P, Kosh KIU (2007). Micropropagation of Lilium ledebori (Baker) as effected by plant growth regulators, sucrose concentrations, harvesting time and cold treatment. Electronic Journal of Biotechnology 10(4):587-591.

Bacchetta, Loretta, Remotti, Patrizio C, Bernardini C, Saccardo F (2003). Adventitious shoot regeneration from leaf explants and stem nodes of Lilium. Plant Cell, Tissue and Organ Culture 74(1):37-44.

Bakhshaie M, Khosravi S, Azadi P, Bagheri H, van Tuyl JM (2016). Biotechnological advances in Lilium. Plant Cell Reports 35(9):1799-1826.

Barazani O, Perevolotsky A, Hadas R (2008). A problem of the rich: Prioritizing local plant genetic resources for ex situ conservation in Israel. Biological Conservation 141(2):596-600.

Blamey M, Grey-Wilson C (1993). Illustrated flora of Brıtaın & Northern Europe. Ed. Hodder and Stroughton. Sevenoaks. Kent, UK.

Bouyoucous GJ (1951). A recalibration of hydrometer method for making mechanical analysis of soils. Agronomy Journal 43(9):434-438.

Burun B, Şahin O (2013). Micropropagation of Lilium candidum L.: a rare and native bulbous flower of Turkey. Bangladesh Journal of Botany 42(2):185-187.

Cardoso JC, Teixeira da Silva JA (2013). Gerbera micropropagation. Biotechnology Advances 31(8):1344-1357.

Chang C, Chang-Tesrn Chen, Tsai Yu-Ching, Chang Wei-Chin (2000).. A tissue culture protocol for propagation of a rare plant. Lilium speciosum Thunb. var. glorisoides Baker. Botanical Bulletin of Academia Sinica 41:39-142.

Comber HF (1949). A new classification of the genus Lilium. Lily Year-Book of RHS. UK, pp 86-105.

Cristiano G, Murillo-Amador B, De Lucia B (2016). Propagation techniques and agronomic requirements for the cultivation of Barbados Aloe (Aloe vera (L.) Burm. F.): A review. Frontiers in Plant Science 7:1410.

Curtis WR (2005). Application of bioreactor design principles to plant micropropagation. Plant Cell, Tissue and Organ Culture 81(3):255-264.

Daneshvar-Royandezagh S, Khawar KM, Ozcan S (2009). In vitro micropropagation of garden thyme (Thymbra spicata L. var. spicata L.) collected from Southeastern Turkey using cotyledon node. Biotechnology & Biotechnological Equipment 23(3):1319-1321.

Davis PH (1985). Flora of Turkey and the East Aegean Island. Edinburgh University, Edinburgh.

De la Vina G, Pliego-Alfaro F, Driscollb SP, Mitchellb VJ, Parryb MA, Lawlorb DW (1999). Effects of CO, and sugars on photosynthesis and composition of avocado leaves grown in vitro. Plant Physiology and Biochemistry 37(7-8):587-595.

Dostal J (1989). Nova kvetena CSSR [New Flora of CSSR]. Priroda, Praha.

Feinbrun Dothan N, Danin A (1991). Analytical flora of Eretz Israel. In Hebrew, Jerusalem.

Godo T, Kobayashi K, Tagami T, Matsui K, Kida T (1998). In vitro propagation utilizing suspension cultures of meristematic nodular cell clumps and chromosome stability of Lilium formolongi. Scientia Horticulturae 72(3-4):193-202.

Granados-Sánchez D, Castañeda-Pérez AD (1998). Sábila (Aloe barbadensis Mill.): Planta agroindustrial (Medicinal) del desierto. Universidad Autónoma Chapingo. Apoyos Académicos, Texcoco.

Greweling T, Peech M (1960). Chemical soil tests. Cornell University Agricultural Experiment Station, New York State College of Agriculture.

Gutterman Y, Chauser-Volfson E (2007). Secondary phenol metabolites (SPhMs), distribution and content of some Aloe species, originated from arid zones of South Africa review. American Journal of Food Technology 2:555-569.

Stocker TF, Field CB, Qin D, Barros V, Plattner GK, Tignor M, … Ebi KL (2010). Meeting report of the intergovernmental panel on climate change expert meeting on detection and attribution related to anthropogenic climate change. Ed. IPCC Working Group I Technical Support Unit, Switzerland.

Jackson ML (1985). Soil chemical analysis: advanced course. UW-Madison Libraries Parallel Press, Wisconssin, USA.

Jeong JH (1996). In vitro propagation of bulb scale section of several Korean native lilies. Acta Horticulturae 414:269-276.

Khan S, Naz S, Mariam R, Kausar N, Naseeb A, Saeed B (2010). Cultivation of Lilies (Lilium regale) for commercialization in Pakistan. Pakistan Journal of Botany 42(2):1103-1113.

Khawar KM, Cocu S, Parmaksiz I, Sarihan EO, Sancak C, Ozcan S (2005). Mass proliferation of Madona Lilly (Lilium candidum L.) under in vitro conditions. Pakistan Journal of Botany 37(2):243-248.

Krishnan PN, Sudha CG (1995). Rapid propagation through shoot tip culture of Trichopus zeylanicus Gaertn. a rare ethnomedicinal plant. Plant Cell Reports 14(11):708-711.

Kumar S, Sharma DR, Sharma YD, Pathania NS (2001). In vitro propagation of Asiatic hybrid lily from bulb scales. The Indian Journal of Agricultural Sciences 71(7):463-465.

Kumar S, Sharma DR, Sharma YD, Pathania NS (2007). Influence of growth regulators and nitrogenous compounds on in vitro bulblet formation and growth in oriental lily. Horticultural Science 34:77-83.

Kumar S, Kashyap M, Sharma DR (2005). In vitro regeneration and bulblet growth from lily bulb scale explants as affected by retardants, sucrose and irradiance. Biologia Plantarum 49(4):629-632.

Lai CC, Lin H M, Nalawade SM, Fang W, Tsay HS (2005). Hyperhydricity in shoot cultures of Scrophularia yoshimurae can be effectively reduced by ventilation of culture vessels. Journal of Plant Physiology 162(3):355-361.

Lian M, Chakrabarty D, Paek KY (2002). Growth and uptake of sucrose and mineral ions by bulblets of Lilium oriental hybrid ‘Casablanca’ during bioreactor culture. The Journal of Horticultural Science and Biotechnology 77(3):253-257.

Lian ML, Murthy HN, Paek KY (2003). Photoautotrophic culture conditions and photosynthetic photon flux influence growth of Lilium bulblets in vitro. Cellular and Development Biology-Plant 39(5):532-535.

Mabberley DJ (1990). The Plant-Book. Cambridge University Press, Cambridge.

Mouterde P (1966). La Nouvelle Flore du Liban et de la Syrie. Tome II Beyrouth Imprimerie Catholique, French.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum 15(3):473-497.

Nakano M, Sakakibara T, Suzuki S, Saito H (2000). Decrease in the regeneration potential of long-term cell suspension cultures of Lilium formosanum Wallace and its restoration by the auxin transport inhibitor, 2,3,5-triodobenzoic acid. Plant Science 158(1-2):129-137.

Nhut DT, Le BV, Fukai S, Tanaka M, Van TT (2001). Effects of activated charcoal, explant size, explant position and sucrose concentration on plant and shoot regeneration of L. longiflorum via young stem. Plant Growth Regulation 33(1):59-65.

Nhut DTH, Nguyen TD, Le Bui Van DS, Jaime T, Fukai S, Tanaka M (2002). The changes in shoot regeneration potential of protocorm-like bodies derived from Lilium Iongiflorum young stem explants exposed to medium volume, pH, light intensity and sucrose concentration pre-treatment. The Journal of Horticultural Science and Biotechnology 77(1):79-82.

Niimi Y, Nakano M, Goto M (1995). Comparison of seedling production among several embryo rescue techniques in Lilium formosanum Wallace. Plant Tissue Culture Letters 12(3):317-319.

Niimi Y, Onozawa T (1979). In vitro bulblet formation from leaf segments of lilies, especially Lilium rubellum Baker. Scientia Horticulturae 11(4):379-389.

Ocak A, Yıldırım H, Pirhan AF, Emecen AA (2014). Ak Zambak (Lilium candidum) Tür Koruma Eylem Planı. Orman ve Su İşleri Bakanlığı Doğa Koruma ve Milli Parklar Genel Müdürlüğü IV. Bölge Müdürlüğü - İzmir Şube Müdürlüğü, İzmir.

Overmyer K, Brosch M, Kangasjärvi J (2003). Reactive oxygen species and hormonal control of cell death. Trends in Plant Science 8(7):335-342.

Ozel CA, Khawar KM, Arslan O (2008a). A comparison of the gelling of isubgol, agar and gel rite on in vitro shoot regeneration and rooting of variety Samsun of tobacco (Nicotiana tabacum L.). Scientia Horticulturae 117(2):174-181.

Ozel CA, Khawar KM, Karaman S, Ates MA, Arslan O (2008b). Efficient in vitro multiplication in Ornithogalum ulophyllum Hand-Mazz from twin scale explants. Scientia Horticulturae 116(1):109-112.

Özen F, Temeltaş H, Aksoy Ö (2012). The anatomy and morphology of the medicinal plant. Lilium candidum L. (Liliaceae), distributed in Marmara Region of Turkey. Pakistan Journal of Botany 44(4):1185-1192.

Paek KY, Murthy HN (2002). High frequency of bulblet regeneration from bulb scale sections of Fritillaria thunbergii. Plant Cell Tissue And Organ Culture 68(3):247-252.

Palta MA (1982). Clonal propagation of Eucalyptus by tissue culture. Plant Science Letter 26(1):1-11.

Park SW, Jeon JH, Kim HS, Parlk YM, Swath CA, Joung H (2004). Effect of sealed and vented gaseous microenvironments on hyperhydricity of potato shoots in vitro. Scientia Horticulturae 99(2):199-205.

Parmaksiz İ, Khawar KM (2006). Plant regeneration by somatic embryogenesis from immature seeds of Sternbergia candida Mathew. et T. Baytop, an endangered endemic plant of Turkey. Propagation of Ornamental Plants 6(3):128-133.

Polunin O, Huxley A (1987). Pomegranate. Flowers of the Mediterranean. Hogarth Press, pp 54-57.

Preece JE, Sutter EG (1991). Acclimatization of micropropagated plants to the greenhouse and field. In: Kluwer Academic Publishers, Boston, pp 71-93.

Priyakumari I, Sheela VL (2005). Micropropagation of gladiolus cv. ‘Peach Blossom’ through enhanced release of axillary buds. Journal of Tropical Agriculture 43(1-2):47-50.

Rout GR, Mohapatra A, Mohan Jain S (2006). Tissue culture of ornamental pot plant. A critical review on present scenario and future prospects. Biotechnology Advances 24(6):531-560.

Saadon S, Zaccai M (2013). Lilium candidum bulblet and meristem development. In Vitro Cellular & Developmental Biology-Plant 49(3):313-319.

Sağlam M, Dengiz O (2012). Influence of selected land use types and soil texture interactions on some soil physical characteristics in an alluvial land. International Journal of Agronomy and Plant Production 3(11):508-513.

Sevimay CS, Khawar KM, Parmaksiz I, Cocu S, Sancak C, Sarihan EO, Özcan S (2005). Prolific in vitro bulblet formation from bulb scales of meadow lily (Lilium candidum L.). Periodicum Biologorum 107(1):107-111.

Sharma JR, Sharma A, Singh AK, Kumar S (1996). Economic potential and improved varieties of aromatic plants of India. Journal of Medicinal and Aromatic Plant Science 19:512-522.

Siljak-Yakovlev S, Peccenini S, Muratovic E, Zoldos V, Robin O, Valles J (2003). Chromosomal differentiation and genome size in three European mountain Lilium species. Plant Systematics and Evolution 236(3-4):165-173.

Simmonds J.A, Cumming BG (1976). Propagation of Lilium hybrids. II. Production of plantlets from bulb-scale callus cultures for increased propagation rates. Scientia Horticulturae 5(2):161-170.

Snedecor GW, Cochran WA (1976). Statistical methods Iowa State University Press, Ames. Statistical methods. 7th Edition.

Tanimoto S, Matsubara Y (1995). Stimulating effect of spermine on bulblet formation in bulb-scale segments of Lilium longiflorum. Plant Cell Reports 15(3-4):297-300.

Teixeira da Silva JA, Dobranszki J (2013). Plant thin cell layers: a 40-year celebration. Journal Plant Growth Regulation 32(4):922-943.

Temeltaş H (1999). Balıkesir Yöresinde Doğal Yayılış Gösteren Lilium candidum L. (Beyaz Zambak) un İç Morfolojisi. Dış Morfolojisi ve Ekolojisi. MSc thesis, Balıkesir Univ, Turkey.

Tribulato A, Remotti PC, Loffler HJM, Van Tuyl JM (1997). Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb. Plant Cell Reports 17(2):113-118.

Tubives (2017). Tubives. Retrieved 2017 February 12 from www.tubives.com.

Uysal E, Kaya E (2013). Farklı miktarlarda uygulanan azotun, bazı doğal çiçek soğanlarında (Lilium candidum L., Galanthus elwesii, Leucojum aestivum) soğan büyüklüğü üzerine etkileri. V. Süs Bitkileri Kongresi II 729-732.

Van Tuyl JM, Van Dien MP, Van Creij MGM, Van Kleinwee TCM, Franken J, Bino RJ (1991). Application of in vitro pollination, ovary culture, ovule culture, and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses. Plant Science 74(1):115-126.

Waters WE and Wilkins HF (1967). Influence of intensity, duration and date of light on growth and flowering of uncooled Easter lily, Lilium longiflorium Thunb and Georgia. Proceedings of the American Society for Horticultural Science 90:433-439.

Wawrosch C, Malla PR, Kopp B (2001). Clonal propagation of Lilium nepalense D. Don: a threatened medicinal plant of Nepal. Plant Cell Reports 20(4):285-288.

Yamagishi M (1998). Effect of culture temperature on the enlargement, sugar uptake, starch accumulation, and respiration of in vitro bulblets of Lilium japonicum Thunb. Scientia Horticulturae 73(4):239-247.

Ziv M (1989). Enhanced shoot and cormlet proliferation in liquid cultured gladiolus buds by growth retardants. Plant Cell Tissue and Organ Culture 17(2-3):101-110.




How to Cite

DANESHVAR ROYANDAZAGH, S. . (2019). Efficient Approaches to in vitro Multiplication of Lilium candidum L. with Consistent and Safe Access throughout Year and Acclimatization of Plant under Hot-Summer Mediterranean (Csa Type) Climate. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(3), 734–742. https://doi.org/10.15835/nbha47311486



Research Articles
DOI: 10.15835/nbha47311486