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Calli were initiated from the mature seeds of wheat (Triticum asetivum L.cv. Xuzhou 211), and suspension cultures were established. The protoplasts isolated from suspension cells were cultured in the modified MS medium solidified with 0.8% agarose. Regenerated cells divided and calli formed. Whole plants were regenerated from protoplast-derived calli. Colony formation was promoted when the medium with lower osmotic pressure was added after two weeks culture. The frequency of regenerated plants was increased with lower concentration of sucrose in the differentiation medium. Shoots were induced effectively with high concentration of cytokinins and calliferous shoots were avoided. The frequency of regenerated plants was affected when protoplast-derived calli were transferred onto the differentiation medium in different periods.