Allelopathic potential of Ophiopogon japonicus was investigated. The methanolic extract of O. japonicus roots strongly inhibited root and hypocotyls growth of lettuce. Sequential partitioning of the methanol extract with organic solvents showed that the diethyl ether and n-butanol extract possess strong plant growth inhibitory activities. The allelopathic constituents of the diethyl ether extract were isolated and identified as salicylic acid and p-hydroxybenzoic acid by NMR spectroscopy. Both of these phenolic acids were found in the aqueous extracts of leaves as well. The concentration of salicylic acid in roots and leaves were estimated as 0.011 and 0.02%, respectively, and it inhibited the root and shoot of tested plants by 50% even at less than 3 ppm. The p-hydroxybenzoic acid on the other hand was in less abundance (0.005%) and inhibited the plant growth to a lesser extent. The biological activity of commercially available O-methyl derivatives of these phenolic acids was also determined to establish structure–activity relationship. Among these, salicylic acid was found to be the most active one. These results suggest that Ophiopogon japonicus produces plant growth inhibitors, which are responsible for its potential allelopathic activity.