Biological invasion, which has caused huge economic losses and severe environmental disruption, has become an international challenge. Traditional methods of customs inspection has several potential problems, ranging from inefficient sampling strategy, low accuracy, and a lack of specialists, all of which can create opportunities for biological invasion. The technology of identifying species based on DNA differences, the results of DNA barcoding may provide a useful addition to the inspection toolkit. Identification does not depend on morphology but on the quality of the isolated DNA; the process requires molecular biological techniques that are becoming more and more simple. Consequently, these methods will have a huge potential in customs inspection of invasive plants. Considering the complex evolution and variation in plants, the internal transcribed spacer (ITS) with great differences within or between species is suggested as the basis of barcoding supplemented by two chloroplast genes:matK and rbcL. The database of DNA barcode of alien invasive plants based on DNA barcoding, extended ultra-barcoding, and meta-barcoding combined with iFlora will be a good new method to the fast identification of invasive plants.