Plant Productions

Plant Productions

The Influence of Ultrasound and Growth Regulators on in vitro Micropropagation of Lilium Ledebourii Boiss

Document Type : Research Paper

Authors
Z. Azimzadeh 1
M. Mohebodini 2
E. CHamani 3
M. Erfani 4
1 Graduate M.Sc. of Horticulture Science, Department of Horticulture Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
2 Associate Professor, Department of Horticulture Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
3 Professor, Department of Horticulture Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
4 Ph.D. Student of Horticulture Science, Department of Horticulture Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran
10.22055/ppd.2018.13439
Abstract
Abstract
Background and Objectives
Lilium ledebourii is one of the wild species of Lilium genus, growing in northern Iran. This species has high potential for export. Plant tissue culture techniques are widely used in plant propagation, and using these methods can effectively provide micro-propagation of this plant in the large scale. High percentage of regeneration is necessary for plant protection, used in the breeding programs.
Materials and Methods
The experiment was factorial based on completely randomized design with four replications and was carried out in tissue culture Lab of Mohaghegh Ardabili University in 2015. For this purpose, segments of explant that were treated with ultrasound were cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. This experiment conducted Concentrations of NAA at four level Zero, 0.01, 0.1 and 1 mgl-1 and Concentrations of BAP at four levels Zero, 0.01, 0.1 and 1 mgl-1 and Ultrasound at four levels zero, 5, 10, 20 and 30 seconds. In order to remove possible contamination from the mediums, all media were autoclaved for 30 minutes at 121 °C. At the end of the experiment, the number of bulblet produced, root length, fresh weight of bulblet and etc were recorded. The cultures were kept at 20°C or 25°C under 16 h photoperiod or in darkness.
Results
The results showed the highest number of bulblets produced in 0, 0.01, 0.1 mg L-1 NAA and 5secend ultrasound, 0.1 mg L-1 NAA and 0, 5, 20 and 30 second ultrasound compared with control. The highest frequency of bulblet regeneration was produced in MS medium supplemented with 0, 0.01, 0.1 mg L-1 NAA and 5 second ultrasound. On the other hand, 0.1 mg L-1 NAA and 5 second ultrasound increased number and weight of bulblets. Different concentrations of NAA had also significant effects.
Discussion
On the other hand, ultrasound increased the number and weight of bulblets. Mechanical stress and micro streaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. Finally, it seems that ultrasound in combination with growth regulators have the potential to produce the highest average number of bulblets in the scale explant, using plant growth regulators in combination with an abiotic stress in in vitro. 
Keywords
Bulblet
Explant
Hormonal combination
In vitro
Scale
Subjects
References
Ananthakrishnan, G., Xia, X., Amutha, S., Singer, S., Muruganantham, M., Yablonsky, S., Fischer, E. and and Gaba, V. (2007). Ultrasonic treatment stimulates multiple shoot regeneration and explant enlargement in recalcitrant squash cotyledon explants in vitro. Plant Cell Reports, 26(3), 267-276.
Arzate-Fernandez, A. M., Miwa, M., Shimada, T., Yonekura, T. and Ogawa, K. (2007). In vitro propagation of Miyamasukashi-Yuri (Lilium maculatum Thunb. var. Bukosanense), an endangered plant species. Revista Fitotecnia Mexicana, 30(4), 373-379.
Azadi, P. and Khosh-Khui, M. (2007). Micropropagation of Lilium ledebourii (Baker) Boiss as affected by plant growth regulators, sucrose concentration, harvesting season and cold treatments. Electronic Journal of Biotechnology, 10(4), 582-591.
Azadi, P. and Mojtahedi, N. (2007). Effect of growth regulators, sucrose concentration and scale segment on micropropagation of Lilium ledebourii in winter harvested bulbs. Pajouhesh- va- Sazandegi in Agronomy & Horticulture Journal, 81, 154-159. [In Farsi]
Bakhshaie, M., Babalar, M., Mirmasoumi,  .Mand Khalighi, M. (2010). Somatic embryogenesis and plant regeneration of Lilium ledebourii (Baker) Boiss and endangered species. Plant Cell Tissue and Organ Culture, 102(2), 229-235.
Ekrami, T. (1980). Flower and ornamental. Tehran: University of Tehran Press. [In Farsi]
El-Naggar, H., Osman, A. and Sewedan, E. (2012). In vitro propagation and organogenesis of Lilium ‘Prato’. African Journal of Biotechnology, 11(82), 14771-14776.
Fahimi, H. (1998). Plant regulators. Tehran: University of Tehran Press. [In Farsi]
Farsam, H., Amanlou, M., Amin, G., Nezamivand-Chegini, G., Salehi- Surmaghi, M. H. and Shafiee, A. (2003). Anatomical and phytochemical study of Lilium ledebourii (Baker) Boiss; a rare endemic species in Iran. DARU Journal, 11(4), 164-170.
Gasemi Gahsareh, M. and Kafi, M. (2006). Scientific and practical floriculture (1st ed.). Isfahan: Golbon Press. [In Farsi]
Han, B. H., Yae, B. W., Yu, H. J. and Peak, K. Y. (2005). Improvement of in vitro micropropagation of Lilium oriental hybrid ‘Casablanca’ by the formation of shoot
with abnormally swollen basal plates. Scientia Horticulturae, 103(3), 351-359.
Jin, L., Zhang, Y., Yan, L., Guo, Y. and Niu, L. (2012). Phenolic compounds and antioxidant activity of bulb extracts of six Lilium species native to China. Molecules, 17(8), 9361-9378.
Kapoor, R., Kumar, S. and Kanwar, J. K. (2008). Bulblet regeneration from ex-vitro root explant in lily hybrids. Horticultural Science, 35(3), 107-112.
Kapoor, R., Kumar, S. and Kanwar, J. K. (2009). Bulblet production from node explant grown in vitro in hybrid lilies. International Journal of Plant Production, 3(4), 1-6.
Kumar, S., Awasthi, V. and Kanwar, J. K. (2007). Influence of growth regulators and nitrogenous compounds on in vitro bulblet formation and growth in oriental lily. Horticultural Science (Prague), 34(2), 77-83.
Kumar, S., Chaudhary, V. and Kanwar, J. K. (2008). Bulblet regeneration from in vitro roots of oriental lily hybrid. Journal of Fruit and Ornamental Plant Research, 16, 353-360.
Liu, Y., Takatsuki, H., Yoshikoshi, A., Wang, B. and Sakanishi, A. (2003). Effects of ultrasound on the growth and vascuolar H+-ATPase activity of Aloe arborescens callus cells. Colloids and Surfaces B: Biointerfaces, 32(2), 105-116.
Meamar-Moshrefi, M., Moieni, A. and Tavassolian, I. (2003). Effects of plant growth regulators NAA, BAP, different explants scale and photoperiod on tissue culture of Lilium ledebourii Boiss. Iranian Journal of Crop Sciences, 4(4), 253-261. [In Farsi]
Meamar-Moshrefi, M., Moieni, A and Tavassolian, I. (2005). Effects of Plant Growth Regulators and Scale on Propagation of Lilium ledebourii (Boiss). Iranian Journal of Agriculture Science, 35(4), 1033-1041. [In Farsi]
Miller, D. L. (1983). The botanical effects of ultrasound: A review. Environmental and Experimental Botany, 23(1), 1-27.
Mir, J. I., Ahmed, N., Itoo, H., Sheikh, M. A., Rashid, R. and Wani, S. H. (2012). In vitropropagation of Lilium (Lilium longiflorum). Indian Journal of Agricultural Sciences, 82(5), 455-558.
Niimi, Y. (1985). Factors affecting the regeneration and growth of bulblets in  bulb scale cultures of Lilium rubellum Baker. Journal of the Japanese Society for Horticultural  Science, 54(1), 82-86.
Padasht Dahkaei, M. N., Khalighi, A., Naderi, R. and Mousavi, A. (2006). Effect of temperature, propagation media and scale position on bulblet regeneration of Chelcheragh lily (Lilium ledebourii) by scaling method. Seed and Plant, 22(3), 383-397. [In Farsi]
Pelkonen, V.P. and Kauppi, A. (1999). The effect of light and auxins on the regeneration of lily (Lilium regale Wil.) cells by somatic embryogenesis and organogenesis. International Journal of Plant Sciences, 160(3), 483-490.
Tatari-Varnousefaderani, M., Fotouhi Ghazvini, R. and Hamidoghli Y. (2007). The effect of lily species, plant growth regulators and origin of explants on micropropagation of Chelcheragh (Lilium ledebourii) and Asiatic hybrid lilies. Iranian Journal of Agriculture Science, 38(1), 276-284. [In Farsi]
Wei, M., Yang, C. Y. and Wei, S. H. (2012). Enhancement of the differentiation of protocorm like bodies of Dendrobium officinale to shoots by ultrasound treatment. Journal of Plant Physiology, 169(8), 770-774.
Plant Productions
Volume 40, Issue 4
Winter 2018
Pages 11-20