Abstract: The CYP83B1 gene of Isatis indigotica Fort. was cloned and its expression patterns were analyzed. Results showed that the length of the IiCYP83B1 gene was 1652 bp, and included two exons and one intron. The full length cDNA of IiCYP83B1 was 1500 bp, encoding a protein of 499 amino acids. IiCYP83B1 was a hydrophobic protein located in the endoplasmic reticulum, without a transmembrane domain or signal peptide. Its secondary structure mainly included alpha helixes and irregular coils. Homologous comparison illustrated that IiCYP83B1 has close relationship with Raphanus sativus Linn., Brassica napus L., Brassica oleracea L., and Brassica rapa L. qRT-PCR analysis indicated that IiCYP83B1 was expressed in root, stem, flower, and fruit, and highly expressed in leaf. It was also highly expressed in the seedling, vegetative growth, and flowering stages, compared with the germination period. Moreover, IiCYP83B1 could be induced significantly by methyl jasmonate (MeJA) and glucose (Glu), but repressed by low temperature (4℃) and salicylic acid (SA). Results in this experiment provide reference for further functional study on IiCYP83B1.