Abstract:Ganoderma leucocontextum is morphologically similar to certain species from its genus in nature. And the so-called distinguishing white flesh within its basidiocarp seemed non-unique according to Ganodermataceae sampling around Southern China. ITS region was reported a DNA barcode marker for macrofungi, anunique DNA markers are yet expected for species identification within genus or genus group. This article designed pair of specific PCR primers for G. leucocontextum identification. Mycelium samples of the 17 selected species phylogenetically closed to G. leucocontextum were harvested and their ITS were isolated and amplified, based on the universal primers. The alignment result showed multiplenon-conservative sites located at 120~140 bp and 450~470 bp. Two oligo sequences, BRLZ-F and BRLZ-R, were designed to match the 2 differentiate fragments as the forward and reverse primers for G. leucocontextum PCR identification. BRLZ-F and BRLZ-R bind efficiently to the DNA templates at 58 ℃ in annealing and only a 348 bp fragment was capable for amplification. Confirmatory experiment illustrates BRLZ-F and BRLZ-R is effective for G. leucocontextum identification within the Ganoderma genus.