3% sucrose), supplemented with different combinations of BA and kinetin (Kin) as
cytokinins and indole-3-acetic acid (IAA), ANA and 2,4-dichlorophenoxyacetic
acid (2,4-D) as auxins, as follows: BA (1.0, 1.5, 2.0 and 4.0μgml−^1 ) in combi-
nation with 0.5μgml−^1 ANA; ANA (0, 0.025, 0.1, 0.2, 0.5, 1.0 and 3.0μgml−^1 )
in combination with 0.5μgml−^1 BA; Kin (0.5, 1.0 and 1.5μgml−^1 ) in combi-
nation with 0.05μgml−^1 ANA; 2,4-D (3.0 and 5.0μgml−^1 ) in combination with
0.1μgml−^1 BA; Kin (1.0μgml−^1 ) with IAA, ANA and 2,4-D (0.25μgml−^1
each). The rate of callus development on all these media for explants of the root line
used (an A4/CAN0221 derived root clone) was very high, ranging from 85%, on
media including 2,4-D to 100% on the others, and all these calli were friable in
texture. Other characteristics as callus growth, color or HRM response were more
dependent on the concentration and combination of plant growth regulators used.
(a)(b)(c)(d)(e)(f)(g)Fig. 14.3 Callusing responses ofCannabis sativahairy roots explants in different culture media
with defined concentrations and combinations (see text) of growth regulators. Horizontal bars
represent 0.5 cm
14 Hairy Root Culture as a Biotechnological Tool inC. sativa 311