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New protocol for the indirect regeneration of the Lilium ledebourii Bioss by using bulb explants
J Plant Biotechnol 2018;45:146-153
Published online June 30, 2018
© 2018 The Korean Society for Plant Biotechnology.

Sina Ghanbari1,3*, Barat Ali Fakheri1, Mohammad Reza Naghavi2,3, Nafiseh Mahdinezhad1

1Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Zabol, Zabol, Iran
2Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran
3Plant Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran
Correspondence to: S. Ghanbari (✉) e-mail:,
Received May 14, 2018; Revised June 11, 2018; Accepted June 11, 2018.
cc This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Lilium ledebourii Bioss is a wild species of Lilium, which grows naturally in some provinces of Iran. Previous studies on Lilium tissue culture have been linked to direct regeneration and a few studies have been conducted on indirect regeneration, which has been studied under bright conditions. In this study, for the first time in the world, all the stages of indirect regeneration (callus induction, shoot and root induction) have been studied under dark conditions. Callus formation and the regeneration levels of L. Ledebourii Bioss were examined for three replicates in an MS (Murashige and Skoog) medium with different hormonal compositions and by using a factorial experiment in the framework of a completely random plan. For callus initiation, 2,4-D and kinetin hormones were used in five and four levels, respectively, as auxin and cytokinin. Results showed that the highest percentage of the callus was found in 3 μM of 2,4-D and 0.5 μM of kinetin. In terms of callus wet weight, the highest amount was found in 3 μM of 2,4-D and 0.5 μM of kinetin. In addition, in terms of diameter, the highest amount was found in 3 μM of 2,4-D, and 0.5 μM of kinetin. In summary, the 2,4-D hormone had a major impact on the percentage of regeneration increase so that the best response was related to the composition of 3 μM of 2,4-D, and 0.1 μM of kinetin. This study contended that auxin and cytokinin can induce long shoots and roots through cell elongation in dark condition.
Keywords : Callus, Indirect regeneration, Hormone, in vitro, Lilium ledebourii Bioss

June 2018, 45 (2)
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