In Vitro Micro Rhizome Induction

Species that normally produce such organs as bulbs, tubers, and corms can be induced to form these miniature propagules within in vitro cultures under appropriate environmental conditions. Plants that naturally produce tubers can be induced to produce miniature versions of the storage organs in a medium containing high cytokinin levels (George, 1993). Miniature storage organs have a great advantage as they can be readily removed from a culture flask in a dormant condition and stored ex vitro without precautions against sepsis. If they are produced in vitro from disease-free stocks, micro-tubers provide an ideal method for propagating and distributing disease-free planting material. When planted in soil, they behave as normal tubers.

In vitro induction of rhizomes and their germination in ginger has been reported by various workers (Sakamura et al., 1986; Sakamura and Suga, 1989; Bhat et al., 1994; Sharma and Singh, 1995; Nirmal Babu, 1997; Nirmal Babu et al., 2003). Bhat et al. (1994) reported in vitro induction of rhizomes in ginger at higher sucrose concentrations (9 to 12 percent). Quality analysis of in vitro—developed rhizomes indicated that they contain the same constituents as the original rhizome but with quantitative differences. The composition of basal medium seems to affect the composition of oil (Sakamura et al., 1986; Sakamura and Suga, 1989; Charlwood et al., 1988). Sharma and Singh (1995) reported microrhizomes with four to five buds weighing 73 to 459 mg that were induced on MS medium with 75 g/l sucrose. After storage in moist sand at room temperature for 2 months, 80 percent of the microrhizomes sprouted into plants.

Geetha (2002) and Peter et al. (2002) tried various combinations of sucrose and mannitol in different concentrations to induce microrhizomes in ginger (see Table 4.7).

Embryoids Suspension Culture

Figure 4.3 In vitro conservation and other biotechnological approaches in ginger: (a) cultures under medium-term conservation in minimal growth medium; (b) one-year-old culture under minimal growth; (c) synthetic seeds; (d) isolated protoplasts; (e) in vitro selection of embryoids against disease-causing organisms; (f) oil cells in cell suspension cultures; (g) transient expression of GUS in embryogenic calli.

Figure 4.3 In vitro conservation and other biotechnological approaches in ginger: (a) cultures under medium-term conservation in minimal growth medium; (b) one-year-old culture under minimal growth; (c) synthetic seeds; (d) isolated protoplasts; (e) in vitro selection of embryoids against disease-causing organisms; (f) oil cells in cell suspension cultures; (g) transient expression of GUS in embryogenic calli.

Microrhizomes of 0.05 to 15 g fresh weight per explant were induced in ginger tissue cultures in 1 to 12 months on MS basal medium supplemented with higher levels of carbon source. In 3 percent sucrose, microrhizome formation was observed in 30 to 40 percent cultures only after 12 months in culture. In the medium with 1 or 1.5 percent

Table 4.7 Effect of sucrose and mannitol on induction of microrhizomes in ginger on ms basal medium

Sucrose (%)

Mannitol (%)

Time period for induction of microrhizome (months)

Percentage responsea

Fresh weight (g)a

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