Abstract: In this paper, Catopsis subulata was selected as the outgroup, and trnL-F genes of 31 Tillandsia species were analyzed and compared with each other to construct the phylogenetic trees by use of Clusta1 X1.81 and MEGA3.1 softwares. The results showed that the length of trnL-F genes in 31 Tillandsia plants were in the range of 806 bp to 835 bp, and there were 22 variable sites and 14 informative sites among them. On the other hand, the average genetic distance of 31 trnL-F gene sequences was 0.007, and at the points of 301-322 bp,there were 21 deoxyribonucleotides inserted in trnL-F gene sequences from T.fasciculata and T.tenuifolia,and a polyT sequence appeared at 826-830 bp.The phylogenetic trees built by ML method illustrated that 31 species could be clustered together into three groups. The clustered results were consistent with the morphologic identification. This indicated that trnL-F gene could be used for the Tillandsia relationship analysis and as the basis for the identification of the specific species.