The novel technique of identifying biological specimens using short DNA sequences from either nuclear or organelle genomes is called DNA barcoding. DNA barcoding not only helps in the identification of species but can also define species boundaries, flagging of new species and species delimitation [1,2]. The Consortium for the Barcode of Life Plant Working Group (CBOL) [3] evaluated seven chloroplast genomic regions across the plant kingdom and proposed a combination of matK and rbcL as plant barcodes. In closely related species, the discriminating ability of these two markers is low [4,5]. Therefore, the China Plant BOL Group [6] proposed the addition of nuclear ITS (Internal Transcibed Spacer) to the matK + rbcL combination as plant barcode in order to achieve maximum identification rates even in closely related species. Bromus L. with more than 151 species in the world comprises annual to perennial species. Bromus taxonomically is a complex genus with difficult nomenclatural history and many species are hard to distinguish due to their high degree of morphological similarity [7]. 17 species sequences beloning to Bromus sect. Bromus were aligned with Muscle program and adjusted manually. Phylogenetic reconstruction was perforemed using maximum parsimony method with heuristic search and TBR branch swapping as implemented in PAUP^* and Bayesian inference with GTR + G evolutionary model by MrBayes. We also constructed neighbour-joining trees based on the DNA sequences. Results from analysis of data showed that all species were recognized with nrDNA sequences and the sequences are an appropriate species marker for this genus.