The invention relates to an isolated culture method of an unfertilized ovary of lilium, belonging to the technical field of plant haploid culture. The method provided by the invention comprises the following steps: with a flower bud one day before bloom as a testing piece, sterilizing and taking the ovary; transversely cutting the testing piece to sheets; then, inoculating a BDS induction medium with the ovary sheets to induce embryogenesis so as to obtain the embryo rate with high frequency, wherein averagely each ovary contains 108-223 embryo ovules; and then, transferring the induced ovary sheets to the BDS regeneration medium to obtain a regeneration plant with other ploidy such as ploidy, diploid and aneuploid through the path of embryogenesis. The method provided by the invention has important meaning for accelerating lilium breeding, germplasm innovation and basic research of genetics.