Abstract: The embryogenic callus induction and shoot regeneration were studied systematically in elite clone GL9 ofEuca lyptus cultivated widely in south China. We investigated TDZ 0. 02 - 0. 05 mg ·L-1 was the p roperconcentration. The callus induction rate was 92. 9%—100%. When TDZ concentration was higher than 0. 1 mg·L-1, the axillary bud germination was comp letely inhibited. TDZ 0. 02 mg·L-1 with combinations of Cocl2 0. 125mg·L-1 could induct the embryogenic callus. The direct regeneration rate was 13. 39% ±1. 03%, and withcombinations ofNAA 0. 1mg·L-1 could not differentiate directly from callus, but higher regeneration rate (20. 2%±13. 3% ) could be obtained by transferring callus onto regeneration medium. The size of callus can increase to1. 4 fold of its original size in the first subculture in modified MS + TDZ 0. 02 mg·L-1 + NAA 0. 1 mg·L-1medium and the average number of deep2p ink2coloured masses of embryogenic cells on each calluswas 8. 4. In thesecond and third subculture, callus stopped growing further and the number of masses of embryogenic cellsdecreased gradually. Regeneration system could lay a good foundation for further transformation research.