over the past 40 years, the regeneration ofCannabisvia in vitro propagation, has
been a challenge with very few reports available so far (Table13.1). Mostly, the
propagation ofCannabishas been achieved by two different routes of organo-
genesis i.e. direct and indirect organogenesis. The Murashige and Skoog
(MS) formulation is the most commonly used medium for in vitro propagation of
Cannabis genotypes (Murashige and Skoog 1962 ). However, the use of media such
as DARIA ind, Millers medium, B5 and MB medium has also been reported
(Feenay and Punja 2003 ; Plawuszewski et al. 2006 ; Wieglus et al. 2008 ).
13.2.3 Callus Production
The early work of growingCannabisin vitro was on callus cultures, particularly on
cell suspension cultures. Most of the studies were aimed at developing cell culture
system to obtain secondary metabolites, particularly the THC class of cannabinoids
those are specific to the genusCannabis(Turner et al. 1980 ). Different explants of
Cannabis sativa, including cotyledons, hypocotyls, epicotyls, leaves, petioles have
been used for the production of callus cultures by many researchers (Itokawa et al.
1975 ; John et al. 1978 ; Francoise and Vincent 1981 ; Fisse et al. 1981 ; Heitrich and
Binder 1982 ; Verzar-Petri et al. 1982 ; Loh et al. 1983 ; Braut-Boucher et al. 1985 ;
Fisse and Andres 1985 ). Using seed explants many different varieties of hemp have
also been studied to obtain callus cultures. Mandolino and Ranalli ( 1999 ) used
Carmagnola, Fibranova,UnikoandKompolti varieties,while, Feeney and Punja
( 2003 ) usedUniko-B,Kompolti AnkaandFelina-34.Slusarkiewicz-Jarzina et al.
( 2005 ) worked onSileia, Fibriman- 24 , Novosadska, Juso- 15 and Fedrina- 74 ,
whereas, Wielgus et al. ( 2008 ) have usedBeniko,SilesiaandBialobrzeskiefor the
callus production. Lata et al. (2009a,b, 2010 , 2012 ) have worked onMXvariety for
the propagation studies ofC. sativa.
In 1972, Veliky and Genest reported thefirst studies on Cannabis cell suspension
and investigated the accumulation of cannabinoids and phenolics in culture using
modified Gamborg’s medium (67-V) based on the research done in ( 1970 )by
Veliky and Martin. This was followed by research done by Itokawa et al. ( 1975 )on
the biosynthesis ofCannabiscallus cultures obtained from various explants like
hypocotyls, cotyledon, roots andfloral parts on MS medium supplemented with
0.1–0.01 ppm KIN and 1.0 ppm 2,4-D. Further, in 1977, Itokawa et al. studied the
biotransformation of cannabinoid precursors and alcohols using cell suspension
cultures ofC. sativa. In 1983, Loh et al. induced callus and suspension cultures
from various explants of embryo, leaf and stem explants using different combina-
tions of auxins [2,4-dichlorophenoxyacetic acid (2,4-D) and
2,4,5-trichlorophenoxyacetic acid (2,4,5-T)] and reported 2, 4, 5-T (3 mg/l) as the
best medium for calli growth using MS medium. Hartsel et al. ( 1983 ) also reported
the biotransformation of CBD to CBE in cell cultures ofC. sativagrown on MS
medium solidified with agar containing the vitamins of B5 medium, supplemented
288 H. Lata et al.