Protoplast isolation and plant regeneration of different genotypes of Petunia and Calibrachoa

Protoplasts were isolated from leaves of in vitro-grown shoot cultures of different Petunia and Calibrachoa genotypes by enzyme digestion with 0.6% macerozyme R-10 and 2.0% cellulase R-10. Shoot regeneration was achieved in five out of nine Calibrachoa and three out of four Petunia genotypes. Protoplast yield and frequency of shoot regeneration varied among experiments and genotypes. Among all regenerants, few morphological changes, such as chlorophyll deficiency, variegated leaves, and polyploidization, were observed.