DNA barcode is a technology which utilizes a short standardized DNA fragment to provide rapid DNA-based identification of unknown life samples. Its advantage is that the entire identification process would not be troubled by different life stages of the specimen, especially in fungi. The fungal identification is usually difficult because of their complex life cycle. In literature more studies have been conducted to screen a powerful suitable gene which could be used as the standard DNA barcode for fungi. The current review introduces the history of DNA barcode, DNA fragment screening, and the significance of DNA barcode to fungal taxonomy in China.