Abstract: The embryogenic calluses induced from the stolons of manilagrass [Zoysia matrella（L.）Merr.] were taken as materials and cultivated on three basic media（MS，N6 and N6AA）with different concentrations（0.5，1.0，1.5 and 2.0 mg · L-1）of 2,4-D for three successive generations to compare their effects. Growth rate，embryogenic maintenance and regeneration ability of the calluses，activities of anti-oxidant enzymes，contents of proline and ploidy of the regenerated plants were measured. The results showed that MS medium supplemented with 2.0 mg · L-1 2,4-D was optimal for callus growth and its embryogenic maintenance，and the rate of embryogenic callues reached 85.00%. However，there were no significant differences among the activities of SOD，POD，CAT and proline content in the regenerated plants acquired from the calluses that had been successively subcultured on three different basic media. The ploidy of the regenerated plants measured with flow cytometry showed unchanged. There were also no morphological differences appeared among individual plants regenerated from different treatments.

Key words: manilagrass, Zoysia matrella（L.）Merr．, medium, 4-D, ploidy