Using haploid technique in breeding of sugar beet as an allogamous plant will facilitate breeding efforts. In this method,a pure line with a desirable genotype can be achieved by unfertilized ovule culture. Therefore ovule culture was done from populations with numbers: 27621, 27624, 27626 and 27629. Young flowering stems were collected and ovules were isolated from the flower buds after sterilization and cultured in N6 medium containing 0.2 mg/l BA,0.5 mg/l NAA,6% sucrose ,0.1% activated charcoal and 0.8% agar and kept at 27ºC in dark condition for one month. Ovules with direct regeneration transferred to medium containing 0.5 mg/l BA, 0.2 mg/l NAA and 3% sucrose after four weeks. Calli were transferred to the regeneration medium. The percentage of cultured ovules response was between 1.6-4%. Then haploids micro shoots were cultured in basic medium containing 0.1% colchicines and 1.5% DMSO for 24 hours, and were subsequently transferred to basic medium with a low level hormone. Haploid micro shoots were cultured in basic medium containing 0.1% colchicines and 1.5% DMSO for 24 hours, and were subsequently transferred to basic medium with a low level hormone .Rooting of DH plants was done in vitro.