Planta Med 2010; 76(14): 1629-1633
DOI: 10.1055/s-0030-1249773
Biochemistry, Molecular Biology and Biotechnology
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

High Frequency Plant Regeneration from Leaf Derived Callus of High Δ 9-Tetrahydrocannabinol Yielding Cannabis sativa L.

Hemant Lata1 , Suman Chandra1 , Ikhlas A. Khan1 , 2 , Mahmoud A. ElSohly1 , 3
  • 1National Center for Natural Products 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 Nov. 2, 2009 revised January 14, 2010

accepted March 3, 2010

Publication Date:
30 March 2010 (online)


An efficient in vitro propagation protocol for rapidly producing Cannabis sativa plantlets from young leaf tissue was developed. Using gas chromatography-flame ionization detection (GC‐FID), high THC yielding elite female clone of a drug-type Cannabis variety (MX) was screened and its vegetatively propagated clones were used for micropropagation. Calli were induced from leaf explant on Murashige and Skoog medium supplemented with different concentrations (0.5, 1.0, 1.5, and 2.0 µM) of indole- 3-acetic acid (IAA), indole- 3- butyric acid (IBA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxy–acetic acid (2,4-D) in combination with 1.0 µM of thidiazuron (TDZ) for the production of callus. The optimum callus growth and maintenance was in 0.5 µM NAA plus 1.0 µM TDZ. The two-month-old calli were subcultured to MS media containing different concentrations of cytokinins (BAP, KN, TDZ). The rate of shoot induction and proliferation was highest in 0.5 µM TDZ. Of the various auxins (IAA, IBA, and NAA) tested, regenerated shoots rooted best on half strength MS medium (1/2 – MS) supplemented with 2.5 µM IBA. The rooted plantlets were successfully established in soil and grown to maturity with no gross variations in morphology and cannabinoids content at a survival rate of 95 % in the indoor growroom.


Ph.D. Hemant Lata

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

University, MS 38677


Phone: + 1 66 29 15 59 28

Fax: + 1 66 29 15 55 87