Abstract: Mlo is an important member of plant diseases resistance genes. Cloning and sequence analysis of Mlo gene in Hevea brasiliensis will lay the foundation for breeding for disease resistant varieties through the transgenic technology. In this study, the barley Mlo gene (GenBank accession number: Z83834) was served as a query to search in the EST and transcript databases of Hevea brasiliensis, and then the homologous EST were assembled. The CDS fragment of Hevea Mlo gene with the expected size (1668 bp) has been successfully cloned by RT-PCR. The Hevea Mlo encoded 555 amino acid residues, with a predicted molecular mass of 63.14 kDa and a 8.85 isolectric point. The structure analysis revealed that Hevea Mlo protein had a conserved Mlo domain and 8 transmembrane domains. Phylogenetic analysis showed that Hevea Mlo shared the highest similarity with the Mlo protein from Manihot esculenta and Ricinus communis, with 92.1% and 87.6% sequence identities. In the genome of Arabidopsis thaliana, Hevea Mlo shared the highest similarity with AtMlo8 with 67% identical residues, which also contained 8 transmembrane domains. Therefore, the Hevea Mlo cloned in this study was designed as HbMlo8. Cloning and sequence analysis of HbMlo8 gene suggested that it belonged to Mlo gene family, these results provide a basis for further verify its function in Hevea brasiliensis.