Crop landraces are an important component of crop genetic resources, which is of great significance to food security and people’s livelihood, thus becoming the main target of germplasm resource conservation and rural development. The Kam aromatic glutinous rice is one of the most important crops in Dong ethnic areas of China. It has become an important component of Dong ethnic traditional staple food crop for hundreds of years. At present, there are abundant germplasm resources of Kam aromatic glutinous rice in Qiandongnan Miao and Dong Autonomous Prefecture, Guizhou Province, China. Aimed at providing data for the conservation and utilization of Kam aromatic glutinous rice germplasm resources, the genetic diversity and similarity of Kam aromatic glutinous rice were explored at the molecular level in the area. In the study, an approach combining ethnobotany investigation and molecular biological experiment was used to evaluate the genetic diversity of the Kam aromatic glutinous rice landraces in Dong ethnic area of Qiandongnan Prefecture. The genomic DNA was extracted from young leaves of 95 Kam aromatic glutinous rice landraces collected from Dong villages following the CTAB method, and then AFLP (amplified fragment length polymorphism) molecular markers was adopted for enzyme digestion analysis, co??????nnection and pre-amplification and selective amplification, and 3730 XL sequencing facility was used for capillary electrophoresis. The polymorphic loci, percentage of polymorphic loci, genetic distance and genetic similarity were analyzed through PopGene1.32 and NTSYS pc-V. 2.1 software. The Kam aromatic glutinous rice landraces were clustered with UPGMA (unweighted pair group method analysis) method. A total of 707 clear bands were obtained, 619 of them were polymorphic bands and the percentage of polymorphic was 87.55%. Shannon’s information index was 0.3738. The genetic similarity coefficient of 8 populations of Kam aromatic glutinous rice is 0.9484-0.9860. The genetic distance was 0.0141-0.0529. The genetic similarity coefficient of Zhanli and Huanggang was the highest, while the genetic distance of Kengdong and Huanggang was the forest. There was no correlation between genetic distance and geographic distance. The genetic similarity coefficient of the 95 accessions was 0.7121-9958. It could be clustered into 2 big clusters (A and B) when the genetic similarity coefficient was 0.7546, with the addition of a special group of landraces. Branch A included a group of 88 landraces, and Branch B had a total of 6 landraces. The varieties were collected from the same and adjacent villages or the varieties with similar morphological traits that were not clustered completely together, there was no correlation between the genetic distance and the geographical location. The results show that there was higher genetic diversity in the landraces of Qiandongnan Prefecture, especially in the Dong villages where the genetic differentiation was high, and the genetic background was very complex in the Kam aromatic glutinous rice landraces. In addition, ethnic traditional classification has important reference value for understanding the diversity of rice landrace varieties. The formation of diversity was related to complex environment, ethnic traditional farming system and culture in the prefecture. The AFLP molecular system establishment and the reveals of diversity background of Kam aromatic glutinous rice provided a basis for the follow-up correlative study of the germplasm resources, which is conducive to further excavate and utilize the especial germplasm resources and provides an important basis for formulating relevant protection strategies.