Abstract: The strains with better prebiotic characteristics were screened out from four Lactobacillus plantarum strains by in vitro prebiotic characteristics as well as safety evaluation tests,and the fermentation characteristics of selected strains were studied. The results showed that compared with other strains,the hydrophobic rate of p-xylene and cetane of L. plantarum B14 was more than 80%,the auto-aggregation ability was more than 85%,and the co-aggregation ability was more than 75%. At the same time,the adhesion ability of B14 to Caco-2 cells reached 30.756%and B14 was preliminarily determined as a safe strain. When L. plantarum B14 was added to the fermented milk as an auxiliary starter,the viscosity and titrated acid of the fermented milk were improved in 24 hours. It indicated that L. plantarum B14 would have potential application prospects as an auxiliary starter for fermented milk.