Abstract: In order to build the multiple SSR-PCR system for sugar beet, the 11 pairs of sugar beet SSR core primer were introduced to build 2 fold to 5 fold SSR-PCR reaction system, based on the different fragment size of SSR amplification products. The results showed as follows: on the basis of single SSR-PCR, the 2 fold to 3 fold SSR-PCR system for sugar beet showed: each one fold increase in SSR, there was only quantity increase in corresponding primer and quantity reduced in deionized water. As to the 2 fold to 3 fold SSR-PCR system for sugar beet, on the basis of single PCR, 0.5 fold increased in DNTPs content quantity and corresponding primer quantity, meanwhile correspondingly reduce or increase 0.5 fold quantity for individual primer according to the different amplification efficiency of individual primer, then we could successfully build 16 4-fold PCR and 9 5-fold PCR. The polymorphism produced by multiple PCR reaction was identical with that from single PCR, but there showed a 2 fold to 5 fold higher efficiency than the single PCR. The establishment of multiple PCR system for sugar beet would greatly accelerate the speed for identifying the variety purity and authenticity of sugar beet, and further promote the development of other areas in sugar beet molecular biology.