Abstract: In order to deeply understand the genetic background and the genetic diversity of Tibet walnut germplasm resources, FISHAFLP was conducted using 9 EcoRⅠ+3/MseⅠ+3 primer combinations to analyse 11 walnut (Juglans regia L.) advanced trees from Tibet at genomic DNA level. Totally 1 103 AFLP fragments were obtained, in which 1 076 were polymorphic and the average percentage of polymorphic bands (ppb) was 97.67%. The value of effective number of alleles (Ne) ranged from 1.00 to 1.99 with an average of 1.48±0.04; Neis gene diversity (H) ranged from 0.00 to 0.49 with an average of 0.29±0.02 and Shannons information index (I) ranged from 0.00 to 0.69 with an average of 0.44±0.02The genetic diversity and sibship among walnut advanced trees in Tibet were investigated and their fingerprintings were established in the research.