The main factors in SSR on Vitis amurensis were optimized using the leaves of grape cultivar ‘Zuoshaner’ as material in this study.The concentrations of primer,DNA,dNTP and the dosage of TaqDNA polymerase were screened to optimize SSR amplification system.The results showed that the optimum concentrations of components in 20uL SSR reaction system were as follows: 1×PCR buffer,0.3umol/L primer,30ng DNA,0.2mmol/L dNTP,1.0U TaqDNA polymerase.The system developed was successfully applied in the amplification of 5 cultivars of Vitis amurensis with three pairs of primer.This indicated the suitability and steadiness of the system.