Abstract:The antibacterial activity of Chinese herbal medicines was examined by agar-diffusing and filter-paper methods. Chinese herbal medicines were added into development and growth medium to observe the influence on growth and differentiation of calli from Lolium perenne, to select Chinese herbal medicines that possess significant antibacterial activity but do not show negative influence on the growth and differentiation of calli from Lolium perenne. The results showed that Scutellaria baicalensis Georgi, Taraxacum mongolicum, Forsythia suspensa and Coptis chinensis strongly inhibited the effects of A.tumefaciens cbf. A.tumefaciens 3301RT was pronouncedly inhibited by S.baicalensis Georgi and C.chinensis. The antibacterial effects on Agrobacterium tumefaciens by agar-diffusing method were more significant compared to filter-paper method. C.chinensis significantly suppressed the growth and differentiation of calli from L.perenne types. However, the other three Chinese herbal medicines didn’t affect calli’s development. S.baicalensis Georgi could be applied into plant gene engineering mediated by A.tumefaciens.