Cannabinoids Production by Hairy Root Cultures of Cannabis sativa L.

Sayed Farag, Oliver Kayser^*

Technische Biochemie, Fachbereich Bio- und Chemieingenieurwesen, Technische Universität Dortmund, Dortmund, Germany.

DOI: 10.4236/ajps.2015.611188                

Abstract

Tetrahydrocannabinol (THC) derivatives are used clinically as analgesic, anti-inflammatory, appetite stimulant, anti-emetic and anti-tumor cannabinoids. THC and its related compounds are at present obtained by extraction from intact Cannabis plants or chemical synthesis, but plant cell cultures may be an alternative source of production. In the present study, hairy root cultures of C. sativa (Cannabaceae) were induced by incubation of aseptically grown callus culture with solid B5 medium supplemented with 4 mg/l naphthaleneacetic acid in darkness at 25°C. Hairy root growth profiles in shake flask, increased periodically during 35 days of growth cycle. The cannabinoid contents produced in minor levels and remained below 2.0 μg/g dry weight. The contents of can-nabinoid were analyzed by liquid chromatography and confirmed by mass spectrometry.

Keywords

Tetrahydrocannabinol, Cannabinoids, Callus, Hairy Root, Cannabis sativa

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Kim, E.S. and Mahlberg, P.G. (1997) Immunochemical Localization of Tetrahydrocannabinol (THC) in Cryofixed Glandular Trichomes of Cannabis (Cannabaceae). American Journal of Botany, 84, 336-342. http://dx.doi.org/10.2307/2446007
[2]	Happyana, N., Agnolet, S., Muntendam, R., Van Dam, A., Schneider, B. and Kayse, O. (2013) Analysis of Cannabinoids in Laser-Microdissected Trichomes of Medicinal Cannabis sativa Using LCMS and Cryogenic NMR. Phytochemistry, 87, 51-59. http://dx.doi.org/10.1016/j.phytochem.2012.11.001
[3]	Turner, C.E. and Elsohly, M.A. (1979) Constituents of Cannabis sativa L .16. Possible Decomposition Pathway of Delta-9-Tetrahydrocannabinol to Cannabinol. Journal of Heterocyclic Chemistry, 16, 1667-1668. http://dx.doi.org/10.1002/jhet.5570160834
[4]	Elsohly, M.A. and Slade, D. (2005) Chemical Constituents of Marijuana: The Complex Mixture of Natural Cannabinoids. Life Sciences, 78, 539-548. http://dx.doi.org/10.1016/j.lfs.2005.09.011
[5]	Elbatsh, M.M., Moklas, M.A., Marsden, C.A. and Kendall, D.A. (2012) Antidepressant-Like Effects of Delta (9)-Tetrahydrocannabinol and Rimon Abant in the Olfactory Bulbectomised Rat Model of Depression. Pharmacology Biochemistry and Behavior, 102, 357-365. http://dx.doi.org/10.1016/j.pbb.2012.05.009
[6]	Borgelt, L.M., Franson, K.L., Nussbaum, A.M. and Wang, G.S. (2013) The Pharmacologic and Clinical Effects of Medical Cannabis. Pharmacotherapy, 33, 195-209. http://dx.doi.org/10.1002/phar.1187
[7]	Waldman, M., Hochhauser, E., Fishbein, M., Aravot, D., Shainberg, A. and Sarne, Y. (2013) An Ultra-Low Dose of Tetrahydrocannabinol Provides Cardio Protection. Biochemical Pharmacology, 85, 1626-1633. http://dx.doi.org/10.1016/j.bcp.2013.03.014
[8]	Sirikantaramas, S., Taura, F., Tanaka, Y., Ishikawa, Y., Morimoto, S. and Shoyama, Y. (2005) Tetrahydrocannabinolic Acid Synthase, the Enzyme Controlling Marijuana Psycho Activity, Is Secreted into the Storage Cavity of the Glandular Trichomes. Plant and Cell Physiology, 46, 1578-1582. http://dx.doi.org/10.1093/pcp/pci166
[9]	Li, K., Feng, J.Y., Li, Y.Y., Yuece, B., Lin, X.H., Yu, L.Y., Li, Y.N., Feng, Y.J. and Storr, M. (2013) Anti-Inflammatory Role of Cannabidiol and o-1602 in Cerulein-Induced Acute Pancreatitis in Mice. Pancreas, 42, 123-129. http://dx.doi.org/10.1097/MPA.0b013e318259f6f0
[10]	Massi, P., Solinas, M., Cinquina, V. and Parolaro, D. (2013) Cannabidiol as Potential Anticancer Drug. British Journal of Clinical Pharmacology, 75, 303-312. http://dx.doi.org/10.1111/j.1365-2125.2012.04298.x
[11]	Slatkin, D.J., Doorenbo, N.J., Harris, L.S., Masoud, A.N., Quimby, M.W. and Schiff, P.L. (1971) Chemical Constituents of Cannabis sativa L. Root. Journal of Pharmaceutical Sciences, 60, 1891-1892. http://dx.doi.org/10.1002/jps.2600601232
[12]	Slatkin, D.J., Knapp, J.E., Schiff, P.L., Turner, C.E. and Mole, M.L. (1975) Steroids of Cannabis sativa Root. Phytochemistry, 14, 580-581. http://dx.doi.org/10.1016/0031-9422(75)85135-1
[13]	Latter, H.L., Abraham, D.J., Turner, C.E., Knapp, J.E., Schiff, P.L. and Slatkin, D.J. (1975) Cannabisativine, a New Alkaloid from Cannabis sativa L. Root. Tetrahedron Letters, 16, 2815-2818. http://dx.doi.org/10.1016/S0040-4039(00)75003-9
[14]	Grotenhermen, F. (2002) Review of Unwanted Actions of Cannabis and THC. Cannabis and Cannabinoids. Pharmacology, Toxicology, and Therapeutic Potential. The Haworth Press, Inc., New York.
[15]	Hillig, K.W. and Mahlberg, P.G. (2004) A Chemotaxonomic Analysis of Cannabinoid Variation in Cannabis (Cannabaceae). American Journal of Botany, 91, 966-975. http://dx.doi.org/10.3732/ajb.91.6.966
[16]	Hamill, J.D., Parr, A.J., Rhodes, M.J.C., Robins, R.J. and Walton, N.J. (1987) New Routes to Plant Secondary Products. Nature Biotechnology, 5, 800-804. http://dx.doi.org/10.1038/nbt0887-800
[17]	Flores, H.E. and Curtis, W.R. (1992) Approaches to Understanding and Manipulating the Biosynthetic Potential of Plant Roots. Annals of the New York Academy of Sciences, 665, 188-209. http://dx.doi.org/10.1111/j.1749-6632.1992.tb42584.x
[18]	Liu, C., Wang, Y., Guo, C., Ouyang, F., Ye, H. and Li, G. (1998) Enhanced Production of Artemisinin by Artemisia annua L. Hairy Root Cultures in a Modified Inner-Loop Airlift Bioreactor. Bioprocess Engineering, 19, 389-392. http://dx.doi.org/10.1007/pl00009026
[19]	Fisse, J., Braut, F., Cosson, L. and Paris, M. (1981) In Vitro Study of the Organogenetic Capacity of Cannabis sativa L. Tissues: Effect of Different Growth Substances. Plantes Médicinales et Phytothérapie, 15, 217-223.
[20]	Wahby, I., Arraez-Roman, D., Segura-Carretero, A., Ligero, F., Caba, J.M. and Fernandez-Gutierrez, A. (2006) Analysis of Choline and Atropine in Hairy Root Cultures of Cannabis sativa L. by Capillary Electrophoresis-Electrospray Mass Spectrometry. Electrophoresis, 27, 2208-2215. http://dx.doi.org/10.1002/elps.200500792
[21]	Wahby, I., Caba, J.M. and Ligero, F. (2013) Agrobacterium Infection of Hemp (Cannabis sativa L.): Establishment of Hairy Root Cultures. Journal of Plant Interactions, 8, 312-320. http://dx.doi.org/10.1080/17429145.2012.746399
[22]	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, 578-585. http://dx.doi.org/10.1079/IVP2003454
[23]	Sujatha, G., Zdravkovic-Korac, S., Calic, D., Flamini, G. and Kumari, B.D.R. (2013) High Efficiency Agrobacterium rhizogenes-Mediated Genetic, Transformation in Artemisia vulgaris: Hairy Root Production and Essential Oil Analysis. Industrial Crops and Products, 44, 643-652. http://dx.doi.org/10.1016/j.indcrop.2012.09.007
[24]	Sevon, N. and Oksman-Caldentey, K.M. (2002) Agrobacterium rhizogenes-Mediated Transformation: Root Cultures as a Source of Alkaloids. Planta Medica, 68, 859-868. http://dx.doi.org/10.1055/s-2002-34924
[25]	Gamborg, O.L., Miller, R.A. and Ojima, O. (1968) Nutrient Requirements of Suspension Cultures of Soybean Root Cells. Experimental Cell Research, 50, 151-158. http://dx.doi.org/10.1016/0014-4827(68)90403-5
[26]	Drozdowska, L. and Rogozinska, J. (1984) Effect of Growth Regulators and Light Conditions on the Morphogenesis of Excised Winter Rape Roots. Bulletin of the Polish Academy of Sciences. Biology, 32, 429-433.
[27]	Whitney, P.J. (1996) Hormone Independent Root Organ Cultures of Rye (Secale cereale). Plant Cell, Tissue and Organ Culture, 46, 109-115. http://dx.doi.org/10.1007/BF00034843
[28]	Bhadra, R. and Shanks, J.V. (1997) Transient Studies of Nutrient Uptake, Growth, and Indole Alkaloid Accumulation in Heterotrophic Cultures of Hairy Roots of Catharanthus roseus. Biotechnology and Bioengineering, 55, 527-534. http://dx.doi.org/10.1002/(SICI)1097-0290(19970805)55:3<527::AID-BIT9>3.0.CO;2-E
[29]	Datta, S., Kim, C.M., Pernas, M., Pires, N.D., Proust, H., Tam, T., Vijayakumar, P. and Dolan, L. (2011) Root Hairs: Development, Growth and Evolution at the Plant-Soil Interface. Plant and Soil, 346, 1-14. http://dx.doi.org/10.1007/s11104-011-0845-4
[30]	Liu, C.F., Zhu, J.Y., Liu, Z.L., Li, L., Pan, R.C. and Jin, L.H. (2002) Exogenous Auxin Effects on Growth and Phenotype of Normal and Hairy Roots of Pueraria lobata (Willd.) Ohwi. Plant Growth Regulation, 38, 37-43. http://dx.doi.org/10.1023/A:1020904528045