Induction of calli from young ears and immature embryos of Secale cereale CV. wintergrazer-70 and regeneration of plantlets

The segments of young ears and immature embryos from wintergrazer-70, a Secale cereale sub-species were inoculated on modified MS media supplemented with varied phytohorme combinations and calli that could be subcultured for many generations were obtained. On the differentiation medium, plantlets were regenerated from the calli. The induction frequency and quality of calli were significantly influenced by the concentrations of 2,4-D and 6-BA. Young ears and smaller immature embryos produced calli in higher frequency. The segments of young ears produced the highest induction frequency and better quality of calli on modified MS medium supplemented with 1.0 mg/L 2,4-D+200 mg/L glutamine. Immature embryos formed the highest induction frequency and much better quality of calli on modified MS medium supplemented with 2.0 mg/L 2,4-D+0.1 (mg/L) 6-BA +200 mg/L glutamine. On the medium supplemented with 1.0 mg/L 2,4-D, 0.1 mg/L TDZ and 200 mg/L glutamine, large amounts of plantlets differentiated from the calli that had been subcultured for several times. The system established in this study could be applied in genetic manipulation and cellular engineering for the breeding of wintergrazer-70.