Somatic hybridization between Citrus reticulata and Citropsis gabunensis through electrofusion

Protoplasts from embryogenic calli of Citrus reticulata Blanco cv. Ponkan and Citropsis gabunensis (Engl.) Swing. & M. Kell (Cabon Cherry Orange), were isolated and fused using electric current. Maximum fusion frequency was obtained with AC at 75 kV/cm (1.0 MHz) for 15 s, followed by DC square-wave pulses at 1.25 kV/cm for 40 μs. Fusion-treated protoplasts were cultured on MT medium containing no growth regulators, solidified with 0.6% Bacto Difco agar. Protoplast-derived calli were proliferated on MT medium containing 1 mg/l zeatin and 0.9% agar. A total of 31 lines of somatic hybrid calli were obtained by screening on the basis of chromosome count and isozyme analysis. The somatic hybrids were tetraploid (2n=36). Plants were regenerated from the calli via somatic embryogenesis. The somatic hybrid plants exhibited morphological characteristics intermediate to the parental plants.