In order to develop effective approaches to obtain uncultured bacteria in soil，four culture media containing MS mineral salt and reduced organic nutrition named as MS+TSG，MS+TSA，MS+R2A and MS+SE respectively，were designed by adding 1/100×TSB，1/50×R2A or soil extraction(SE)into the MS basic medium with agar(A)or gellan gum(G)as coagulant. A content of 1.5 g/L sodium propionate was added into the media mention above to obtain a group of antioxidant culture media. Bacteria in the rhizosphere soil of healthy tomato plants were cultured on the eight media mentioned above，and the representative colonies selected randomly from different media were identified by 16S rRNA gene sequencing. The results indicated that，on the four media containing MS mineral salt plus reduced organic nutrition，more than 1×107cfu were obtained from per gram of soil after being cultured at 28℃ for 28 d. The order of colony account on media from high to low was MS+SE > MS + TSG > MS + R2A > MS + TSA. In the condition of mineral salt plus low level of organic nutrients，gellan gum could help to obtain more colonies compared to agar，but the supplement of sodium propionate dramatically reduced the diversity of colonies. Among the 84 representative colonies identified，a total of 52 species from 34 genera were included，of which 7(8.33% of detected colonies)are possible new species. The order of species diversity on media from high to low was MS + TSA>MS+R2A>MS+TSG>MS+SE. The results suggest that media consisting of MS mineral salt and low level of organic nutrients are effective to isolate uncultured bacteria and new antagonism resource to plant pathogens from soil.