In collaboration with Scientific Association of Iranian Medicinal Plants

Document Type : Research Paper

Authors

1 M.Sc. Student, Department of Plant Breeding, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Genetics and Agricultural Biotechnology Institute of Tabarestan, Agriculture and Natural Resources University of Sari, Iran

3 Department of Plant Breeding and Biotechnology, Science and Research, Islamic Azad University, Tehran, Iran

Abstract

The present study was carried out to investigate the possibility of micro-propagation and to determine the optimal medium composition and combination of Cannabis sativa L. growth regulators under in vitro conditions. Seeds were surface-sterilized and then cultured on MS basal medium. One month later, leaf and hypocotyl explants, obtained from the seedlings grown at in vitro condition, were used in MS culture medium containing NAA hormone (0.5, 1, 2 and 3 mg/l) either alone or in combination with 0.5mg/l BA; and 2,4-D (0.1, 0.2, 0.5 and 1 mg/l) alone or in combination with 0.5mg/l BA. Callus formation was the response of explants in most media. Direct shoot regeneration from explants was not observed but shoot induction from callus was seen only in 0.1 mg/L 2,4-D+ 0.5 mg/L BA. The highest volume of induced callus was formed on MS medium 0.1 mg/L 2,4-D+ 0.5 mg/L BA using leaf as explant. Root induction from some explants was observed in different treatments. The highest fresh weight of calli belonged to the leaf explant cultured on MS medium containing 1 mg/L 2,4-D+ 0.5 mg/L BA. Callus induction and rooting occurred easily and the explants did not respond well to regeneration.

Keywords

- Callaway, J.C. and Laakkonen, T.T., 1996. Cultivation of Cannabis oil seed varieties in Finland. Journal International Hemp Association, 3: 32-33.
- Chang, C., Chang-Tesrn, C., Tsai, Y.C. and Wei-Chin, C., 1999. A Tissue culture protocol for propagation of a rare plant, Lilium speciosum Thunb. var. glorisoides Baker. Botanical Bulletin of Academia Sinica, 41(2): 139-142.
- DeKlerk, G.J., 2006. Plant hormone in tissue culture. Plant Cell and Tissue Culture. Phytopathology Biochemical, 17-25.
- ElSohly, M.A., Holley, J.H., Lewis, G.S., Russell, M.H. and Turner, C.E., 1984. Constituents of Cannabis sativa L. XXIV: The potency of confiscated marijuana, hashish, and hash oil over a ten-year period. Journal of Forensic Sciences, 29(2): 500-514.
- Feeney, M. and Punja, Z.K., 2003. Tissue culture and Agrobacterium-mediated transformation of hemp (Cannabis sativa L.). In Vitro Cellular & Developmental Biology-Plant, 39(6): 578-585.
- Hendriks, H., Malingre, T. and Batterman, S., 1975. Mono and sesquiterpene hydrocarbons of the essential oil of Cannabis sativa L. Phytochemistry Journal, 14(3): 814-830.
- Kosiorek, P., Hryniewicz, A., Bialuk, L., Zawwadzka, A. and Winnicka, M.M., 2004. Cannabinoids alter recognition memory in rat. Journal Pharmacological Reviews, 55: 903-910.
- Lata, H., Chandra, S., Khan, I.A. and Elsohly, M.A., 2009. Propagation through alginate encapsulation of axillary buds of Cannabis sativa L. an important medicinal plant. Physiology and Molecular Biology of Plants, 15(1): 79-86.
- Lata, H., Chandra, S., Khan, I.A. and Elsohly, M.A., 2010. High frequency plant regeneration from leaf derived callus of high Δ9-tetrahydrocannabinol yielding Cannabis sativa. Planta Medica, 76(14): 1629-1633.
- Lusarkiewicz-Jarzina, A., Ponitka, A. and Kaczmarek, Z., 2005. Influence of cultivar, explant source and plant growth regulator on callus induction and plant regeneration of Cannabis sativa. Acta Biologica Cracoviensia-Series Botanica, 47(2): 145-151.
- Mand, Ch. and Mediavilla, V., 1998. Factor influencing the yield and quality of hemp (Cannabis sativa L.) essential oil. Journal of the International Hemp Association, 5: 16-20.
- Naik, P.K. and Nayak, S., 2005. Different modes of plant regeneration an factors affecting in vitro bulblet production in Ornithogalum virens. Science Asia, 31: 409-414.
- Nugent, G., Chandler, S.F., Whiteman, P. and Stevenson, T.W., 2001. Somatic embryogenesis in Eucalyptus globulus. Plant Cell, Tissue and Organ Culture, 67: 85-88.
- Raharjo, T.J., Eucharia, O., Chang, W.T. and Verpoorte, R., 2006. Callus induction and phytochemical characterization of Cannabis sativa cell suspension culture. Indonesian Journal of Chemistry, 6(1): 70-74.
- Small, E. and Cronquist, A., 1976. A practical and natural taxonomy for Cannabis. Taxon Journal, 25(4): 405-435.
- Vargas, T.E., Mejias, A. Oropeza, M. and De Garica, E., 2004. Plant regeneration of Anthurium andreanum CV Rubrun. Electionic Journal of Biotechnolgogy, 72: 285-289.
- Wang, X., Tang, C., Yang, X. and Gao, W., 2008. Characterization, amino acid composition and vitro digestibility of hemp (Cannabis sativa L) proteins. Food Chemistry, 107: 11-18.
- Wills, S., 1998. Cannabis use and abuse by man: a Historical perspective: 1-28. In: Brown, D.T. (Ed.), Cannabis: The Genus Cannabis, Harwood Academic Publishers, Amsterdam, 272p.
- Yan, M.M., Xu, C., Kim, C.H. and Um, Y.C., 2009. Effects of explant type, culture media and growth regulators on callus induction and plant regeneration of Chines jiaotou (Allium Chinense). Scientia Horticulturae, 123: 124-128.