Abstract:As an emerging organism identification method, DNA barcoding has been widely used in plants, animals and microorganisms for its advantage of higher sensitivity, accuracy, and objectivity. Even the nuclear ribosomal internal transcribed spacer (ITS) is used as a standard barcode in fungal identification frequently, nowadays, there are more and more new barcodes, such as the microcoding, ND6 and EF3. In this article we summarized the generation and developing history of DNA barcoding, also we present the advantage, shortcomings and the development trend based on fungal barcoding case studies. We indicated that DNA barcoding technique will be a good supplementary to the traditional morphology-based taxonomy, and towards a combination of natural evolutional relationships and morphological taxonomy in fungi.