Abstract: High quality DNA required for construction of AFLP analysis system was isolated with an improved method of CTAB from young leaves of Castanea mollissima. A silver-staining AFLP reaction system was constructed for genetic analysis of C. mollissima by optimizing DNA restriction-ligase reaction, pre-amplification and selective amplification.Resultsshowed that the system of restriction of DNA was better than restriction and link of DNA. The polymorphism bands were obtained from three primer pairs (E-AAC+M-CAA, E-AAC+M-CAT, E-AGT+M-CAT), but E-AGT+M-CAT could amplify more uniform and high sharp bands. Our study showed that the silver-staining AFLP was available for genetic diversification, variety identification, construction of molecular map, identification and classification.