To study the technique of the callus induction from anther and plant regeneration of medicinal plants Liriope spicata var. prolifera.Callus was induced from anther of L. spicata var. prolifera on a MS medium supplemented with different hormones. The squash methods combined with a microscope were used to analyze chromosomes of regenerated plantlets.MS +2,4-D 1.0 mg x L(-1) + KT 2.0 mg x L(-1) gave the highest induction ratio which was 41.07%. MS +6-BA 1.5-2.0 mg x L(-1) + NAA 0.1-0.3 mg x L(-1) was suitable for the induction and proliferation of indefinite buds. The buds were transferred to 1/2 MS medium supplemented with NAA 0.1-0.3 mg x L(-1) for rooting. The shoots produced roots of culture and formed complete plantlets. The regenerated plantlets originated from somatic cells. At the same time, the effects of pretreatment of low temperature at 4 degrees C on the callus induction were studied and discussed.This paper sets up the method of tissue culture of anther somatic-cells and intermediate propagation of L. spicata var. prolifera.