[Objectives] This study was conducted to better understand the variation law of sugarcane clones during tissue culture process,and to provide a reference for rapid propagation and detection of healthy sugarcane seedlings.[Methods] The genetic stability of tissue culture clones of three sugarcane varieties was analyzed using the AFLP molecular marker technique.[Results]The average number of polymorphic loci was 19. 58 for each primer pair,and the percentage of polymorphic loci was 41. 74%. Compared with the donor varieties,all tissue culture materials were mutated. There were 3-16 missing bands,with an average of 5. 2 bands,and there were 0-17 increased bands,with an average of 3. 3 bands. The total number of missing and increased bands was 4-33,with an average of 8. 5. The band difference rates were in the range of 0. 009 4%-0. 077 6%,with an average of 0. 020 6%. The genetic similarity coefficients between materials ranged from 0. 685 6 to 0. 998 2,with an average of 0. 818 4. The three sugarcane varieties and their tissue culture clones were clustered into three groups.[Conclusions] Although variations occur in tissue culture,the variations are not too obvious,and the genetic stability is relatively high. It is recommended to minimize the number of subculture generations and cultivation time to reduce the occurrence of variation during tissue culture for rapid propagation of sugarcane seedlings.