Abstract: In order to direct the construction of plant germplasms by elucidating the relatives among plants at the level of gene, CYP86MF gene analogues from 11 species of 6 genera in Cuciferae were respectively obtained by PCR strategy using gene specific primers designed from conserved regions of CYP86MF gene reported. Sequence comparisonindicated that the similarities among the genes at nucleotide level were over 80%, and the similarities at amino acid level remained above 70%. The differences between the genes at nucleotide and amino acid level between species were 1.0% ~ 5.7% and 2.6% ~ 7.3% respectively, while those between genera 5.6% ~ 22.5% and 7.3% ~ 31.2%, respectively. Phylogenetic analysis showed that Brassica was closely related to Raphanus, followed by Rorippa Scop, Arabidopsis Heynh, Capsella Medic orderly, most distantly related to Orychophrogmus. It was concluded that CYP86MF gene was not applicable to specie and subspecie taxon but genus taxon because the differences of sequences in nucleotides and amino acids were lower between species than genera.