To establish a highly efficient system for shoot regeneration in vegetable soybean (Glycine max (L.) Merrill), explants were obtained from six genotypes and adventitious shoots were regenerated from cotyledonary nodes cultured on medium supplemented with different concentrations of N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (thidiazuron TDZ) and 1-naphthylacetic acid (NAA). The most effective combination of plant growth regulators was selected first and then the effects of medium types, inoculation methods and genotypic differences on shoot regeneration were studied. The explants were inserted either vertically or horizontally into the medium and five basal media, including B5, ½ B5, MS, ½ MS, and MSB (MS salts and B5 organics), were tested. The shoot regeneration frequency of the six genotypes ranged from 53.5% to 88.9% and three of them reached 88.9%, 87.5% and 83.3% respectively on B5 medium supplemented with 1 mg•L-1 TDZ, 0.05 mg•L-1 NAA, and 5 mg•L-1 AgNO3. The shoot regeneration frequency of explants cultured on B5 medium was significantly higher than that of the other four basal media. The vertically inserted explants were found to yield a higher shoot regeneration frequency than that of horizontally inserted ones; the maximum difference of regenerating percentages between the two methods was 37.5%. In summary, TDZ was an efficient plant growth regulator for shoot induction. B5 medium and vertically inserted explants promoted shoot regeneration. We believe this highly efficient shoot regeneration system will provide foundation for the further transgenic studies in vegetable soybean.