Leaves of Pueraria thunbergiana possess allelopathic activity and the putative compounds causing this growth inhibitory effect were isolated from their aqueous methanol extract. The chemical structures of these growth inhibitors were determined by high-resolution MS and 1H NMR spectral data as cis,trans-xanthoxin and trans,trans-xanthoxin. cis,trans-Xanthoxin and trans,trans-xanthoxin inhibited the root growth of cress (Lepidium sativum L.) seedlings at concentrations greater than 0.3 and 3 microM, respectively. The doses required for 50% inhibition on the cress roots were 1.1 and 14 microM for cis,trans- and trans,trans-xanthoxin, respectively. The concentrations of cis,trans- and trans,trans-xanthoxin in P. thunbergiana leaves were 51.4 and 72.5 ng g(-1) fresh weight, respectively. The effectiveness of cis,trans- and trans,trans-xanthoxin on the growth inhibition and the occurrence of both xanthoxins in P. thunbergiana suggest that xanthoxins may contribute to the growth inhibitory effect of P. thunbergiana, and may play an important role in the allelopathy of P. thunbergiana after being released into the soil.