Effects of light, methyl jasmonate(MJ), 6-benzylaminopurine(BA), thidiazuron(TDZ), and 2,4-dichlorophenoxyacetic acid(2,4-D) on the contents of ecdysteroids (β-ecdysone+polypodine B) in Polypodium vulgare L. and Achyranthes japonica Nakai were studied. When the plants of P. vulgare were cultured under light control, the ecdysteroids contents in both leaves and rhizomes decreased with increasing light intensity. The ecdysteroids contents in A. japonica were also lower when cultured under light than under dark. Among the tested plant growth regulators, MJ, BA, and TDZ increased the ecdysteroids contents in both P. vulgare and A. japonica.