Regeneration and Acclimatization of Regenerants in Long-term in vitro Culture of Japanese Blood Grass (Imperata cylindrica ‘Rubra’)

Regeneration and Acclimatization of Regenerants in Long-term in vitro Culture of Japanese Blood Grass (Imperata cylindrica ‘Rubra’)

Long-term culture of cell lines is an important issue in in vitro culture and in plant science. In this study, the regeneration ability and ex vitro acclimatization of regenerants were evaluated. The ploidy level of regenerants derived from long-term cultured cell lines was measured in Imperata cylindrica ‘Rubra’, Poaceae. Adventitious buds (shoots) were successfully induced from five-year-cultured calli on MS medium containing 0.1 mg/L BA or 2.0 mg/L TDZ, combined with 0.01 mg/L auxins (IAA, IBA, NAA and 2,4-D), respectively. Adventitious roots were also induced on MS medium containing 0.01 mg/L auxins (IBA, NAA and 2,4-D), respectively. Interestingly, regenerants with both red and green leaf were successfully obtained when regenerants were cultured on MS medium with 9% sucrose. Regenerants derived from long-term cultured calli were transferred to pots using an optimal acclimatization process and successfully adapted to both pot and soil conditions. Moreover, the ploidy level was measured using calli and regenerants that had been kept on MS medium containing various kinds of plant growth regulators (PGRs).