IiMYB34 was cloned (GenBank accession No. MF373610) from tenderleaves of Isatis indigotica with RTPCR method, and its bioinformatics and expression pattern analysis were performed in this study. The results indicated that: (1) the full length gDNA of IiMYB34 was 1 854 bp, including 3 exons and 2 introns; Its ORF was 951 bp in length, encoding 316 amino acid residues; The alpha helix structures and coiled coils were in higher proportions than those of the extension chain structures in the secondary structure of IiMYB34 protein, and the results were consistent with its tertiary structure; The functional structure domain analysis results showed that IiMYB34 protein was a typical R2R3MYB protein with two conserved MYB DNAbinding domains; The characteristic of the amino acid sequences of IiMYB34 was highly homologous with Brassica napus and Brassica oleracea. (2) The qRTPCR analysis results indicated that IiMYB34 exhibited a tissuesspecific expression and was highly expressed in leaf. Moreover, IiMYB34 could be induced significantly by MeJA, SA and Glu, but repressed by low temperature (4 ℃) and mechanical wounding. The obtained results laid the foundations for the further functional research on IiMYB34.