The aim was to lay the foundation for the application of somatic hybridization in peanut breeding. The protoplasts isolated from leaves of Krapts and A. stenosperma were fused by PEG method, and then cultured in the modified liquid MSB5 medium (Murashige and Skoog salts plus B5 vitamins) with 2 mg/L picloram (Pic), 0.1 mg/L thidiazuron (TDZ), 2% coconut cream, 5 g/L polyvinyl pyrrolidone (PVP) and 0.1% 2-(N-morpholino) ethanesulfonic acid (MES). After 5 weeks culture, the formed small calli were transferred to the solid MSB5 medium supplemented with 3 mg/L zeatin, 0.2 mg/L BAP and 0.1 mg/L NAA for callus proliferation. First cell division was observed within 4 days after culture in the liquid medium. And the formed colonies grew up to 300 μm in diameter after 2 weeks culture and up to 2-3 mm after 5 weeks culture. When the small calli were transferred to the solid MSB5 medium, they grew rapidly. Genomic DNA of calli was extracted and detected by PCR, some of the bands from both parents and the new bands were amplified in some calli. The results indicated those calli were derived from fused cells.