The article presents the results of a study on obtaining mutant doubled haploids of turnip rape. The culture medium for producing embryos in isolated microspore culture was optimized. The optimal medium formula was NLN + 13% S + 0.05 mg/L NAA + 0.05 mg/L BA. Embryos derived from isolated microspore culture were treated with EMS mutagen in three concentrations (4 mM, 8 mM, and 12 mM). Mutant doubled haploid plants and their seeds were obtained. Agronomic evaluation and analysis of fatty acid composition in mutant lines seeds showed that they had valuable traits in terms of seed oil quality and yield. Analysis of the fatty acid composition of the seeds of the obtained mutant doubled haploids showed an increase in the percentage of oleic acid (~11%–12%) in comparison with donor cultivars. Our results showed that mutagenesis of embryos from a culture of isolated microspores has potential for improving the qualitative traits of turnip rape.