Planta Med 2010; 76(7): 743-750
DOI: 10.1055/s-0029-1240628
Biochemistry, Molecular Biology and Biotechnology
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Assessment of Cannabinoids Content in Micropropagated Plants of Cannabis sativa and Their Comparison with Conventionally Propagated Plants and Mother Plant during Developmental Stages of Growth

Suman Chandra1 , Hemant Lata1 , Zlatko Mehmedic1 , Ikhlas A. Khan1 , 2 , Mahmoud A. ElSohly1 , 3
  • 1National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, MS, USA
  • 2Department of Pharmacognosy, School of Pharmacy, University of Mississippi, University, MS, USA
  • 3Department of Pharmaceutics, School of Pharmacy, University of Mississippi, University, MS, USA
Further Information

Publication History

received May 18, 2009 revised Sept. 8, 2009

accepted October 27, 2009

Publication Date:
30 November 2009 (online)

Abstract

Gas chromatography-flame ionization detection (GC‐FID) was used to assess the chemical profile and quantification of cannabinoids to identify the differences, if existing, in the chemical constituents of in vitro propagated plants (IVP), conventionally grown plants (VP) and indoor grown mother plants (MP-Indoor) of a high THC yielding variety of Cannabis sativa L. during different developmental stages of growth. In general, THC content in all groups increased with plant age up to a highest level during the budding stage where the THC content reached a plateau before the onset of senescence. The pattern of changes observed in the concentration of other cannabinoids content with plants age has followed a similar trend in all groups of plants. Qualitatively, cannabinoids profiles obtained using GC‐FID, in MP-indoor, VP and IVP plants were found to be similar to each other and to that of the field grown mother plant (MP field) of C. sativa. Minor differences observed in cannabinoids concentration within and among the groups were not found to be statistically significant. Our results confirm the clonal fidelity of IVP plants of C. sativa and suggest that the biochemical mechanism used in this study to produce the micropropagated plants does not affect the metabolic content and can be used for the mass propagation of true to type plants of this species for commercial pharmaceutical use.

References

  • 1 Doyle E, Spence A A. Cannabis as a medicine?.  Br J Anaesth. 1995;  74 359-361
  • 2 Hammond C T, Mahlberg P G. Morphogenesis of capitate glandular hairs of Cannabis sativa (Cannabaceae).  Am J Bot. 1977;  64 1023-1031
  • 3 Mechoulam R, Ben-Shabat A. From gan-zi-gun-nu to anandamide and 2- arachidonoylglycerol: the ongoing story of Cannabis.  Nat Prod Rep. 1999;  16 131-143
  • 4 Sirikantaramas S, Taura F, Morimoto S, Shoyama Y. Recent advances in Cannabis sativa research: biosynthetic studies and its potential in biotechnology.  Curr Pharm Biotechnol. 2007;  8 237-243
  • 5 Brenneisen R, Egli A, ElSohly M A, Henn V, Spiess Y. The effect of orally and rectally administered Δ9-tetrahydrocannabinol on spasticity: a pilot study with 2 patients.  Int J Clin Pharmacol Ther. 1996;  34 446-452
  • 6 Formukong E A, Evans A T, Evans F. The medicinal uses of Cannabis and its constitutents.  Phytother Res. 1989;  3 219-231
  • 7 Grinspoon L, Bakalar J B. Marihuana, the forbidden medicine. New Haven; Yale University Press 1993
  • 8 Mattes R D, Shaw L M, Eding-Owens J, Egelman K, ElSohly M A. By passing the first pass effect for therapeutic use of cannabinoids.  Pharmacol Biochem Behav. 1993;  44 745-747
  • 9 Mattes R D, Egelman K, Shaw L M, ElSohly M A. Cannabinoids appetite stimulation.  Pharmacol Biochem Behav. 1994;  49 187-195
  • 10 Mechoulam R. The pharmacohistory of Cannabis sativa. Mechoulam R Cannabinoids as therapeutic agents. Boca Raton, Florida; CRC Press 1986: 1-19
  • 11 ElSohly M A, Ross S A, Mehmedic Z, Arafat R, Yi B, Banahan B F. Potency trends of Δ9-THC and other cannabinoids in confiscated marijuana from 1980–1997.  J Forensic Sci. 2000;  45 24-30
  • 12 Mehmedic Z, Chandra S, Slade D, Denham H, Foster S, Patel A S, Ross S A, Khan I A, ElSohly M A. Potency trends of Δ9-THC and other cannabinoids in confiscated Cannabis preparations from 1993–2008.  J Forensic Sci. 2010;  , accepted for publication
  • 13 Loh W H T, Hartsel S C, Robertson W. Tissue culture of Cannabis sativa L. and in vitro biotransformation of phenolics.  Z Pflanzenphysiol. 1983;  111 395-400
  • 14 Richez-Dumanois C, Braut-Boucher F, Cosson L, Paris M. Multiplication vegetative in vitro du chanvre (Cannabis sativa L.) application a la conservation des clones selections.  Agronomie. 1986;  6 487-495
  • 15 Mandolino G, Ranalli P. Advances in biotechnological approaches for hemp breeding and industry. Ranalli P Advances in hemp research. New York; Haworth Press 1999: 185-208
  • 16 Slusarkiewicz-Jarzina A, Ponitka A, Kaczmarek Z. Influence of cultivar, explant source and plant growth regulator on callus induction and plant regeneration of Cannabis sativa L.  Acta Biol Craco Series Bot. 2005;  47 145-151
  • 17 Bing X, Ning L, Jinfeng T, Nan G. Rapid tissue culture method of Cannabis sativa for industrial uses. CN Patent 1887043 A 20070103. 2007
  • 18 Lata H, Chandra S, Khan I, ElSohly M A. Propagation through alginate encapsulation of axillary buds of Cannabis sativa L. – an important medicinal plant.  Physiol Mol Biol. 2009;  15 79-86
  • 19 Lata H, Chandra S, Khan I, ElSohly M A. Thidiazuron induced high frequency direct shoot organogenesis of Cannabis sativa L.  In Vitro Cell Dev Biol Plant. 2009;  45 12-19
  • 20 Ross S A, Parker M, Arafat R, Lovett K, ElSohly M A. The analysis of confiscated marijuana samples for different cannabinoids using GC/FID.  Am Lab. 1996;  16 16-17
  • 21 Pierik R L M. Commercial aspects of micropropagation. Prakash J, Pierik RLM Horticulture – new technologies and applications. Dordrecht; Kluywer Academic Publisher 1991: 141-153
  • 22 Shenoy V B, Vasil I K. Biochemical and molecular analysis of plants derived from embryogenic cultures of nipper grass (Pennisetum purpureum K. Schum).  Theor Appl Genet. 1992;  83 940-955
  • 23 Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures.  Physiol Plant. 1962;  15 473-497
  • 24 Rani V, Parida P, Raina S N. Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh.  Plant Cell Rep. 1995;  14 459-462
  • 25 Wang X R, Szmidt A E, Nguyen H N. The phylogenetic position of the endemic flat-needle pine Pinus krempfii (Lec., Pinaceae) from Vietnam, based on PCR-RFLP analysis of chloroplast DNA.  Plant Syst Evol. 2000;  220 21-36
  • 26 Damasco O P, Graham G C, Henry R J, Adkins S W, Smith M K, Godwin I D. Random amplified polymorphic DNA (RAPD) detection of dwarf off types in micropropagated Cavendish (Musa spp. AAA) bananas.  Plant Cell Rep. 1996;  16 118-123
  • 27 Salvi N D, George L, Eapen S. Plant regeneration from leaf callus of turmeric and random amplified polymorphic DNA analysis of regenerated plants.  Plant Cell Tissue Organ Cult. 2001;  66 113-119
  • 28 Ma X, Gang D R. Metabolic profiling of in vitro micropropagated and conventionally greenhouse grown ginger (Zingiber officinale).  Phytochemistry. 2006;  2239-2255
  • 29 Lata H, Chandra S, Techen N, Khan I A, ElSohly M A. Assessment of the genetic stability of micropropagated plants of Cannabis sativa by ISSR markers.  Planta Med. 2009;  DOI: 10.1055/s-0029-1185945 , advance online publication
  • 30 Chandra S, Lata H, Khan I A, ElSohly M A. Photosynthetic response of Cannabis sativa L. to variations in photosynthetic photon flux densities, temperature and CO2 conditions.  Physiol Mol Biol Plants. 2008;  14 299-306

Ph.D. Suman Chandra

National Center for Natural Product Research
Research Institute of Pharmaceutical Sciences
School of Pharmacy
University of Mississippi

University, MS 38677

USA

Phone: + 1 66 29 15 69 54

Fax: + 1 66 29 15 55 87

Email: suman@olemiss.edu

>