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Isoflavonoids, the functional molecules of Fabaceae, are under clinical trials against cancer, osteoporosis and cardiovascular diseases. In this study, the efficacy of different plant growth regulators was evaluated for optimizing the production of isoflavonoids in Pueraria tuberosa. The cultures were maintained in Murashige and Skoog's medium containing 0.1 mg l -1 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) and 0.1 mg l -1 kinetin. The addition of 5.0 mg l -1 N 6 -(2-Isopentenyl) adenine (2iP) resulted in about *32-folds increase in production of isoflavonoids, while about *23-folds increase was recorded in the absence of kinetin in the maintenance medium. A maximum yield of isoflavonoids (*80 mg l -1 ; 82-folds increase) was obtained in cultures grown at 0.1 mg l -1 morphactin and 5.0 mg l -1 of 2iP. However, 2,4,5-T in combination with 2iP was ineffective for their production. Among different plant growth regulators tested, maximum yields of puerarin, genistin, daidzein and genistein were 17.4, 15.9, 69.0 and 0.04 mg l -1 , respectively. The study suggested that the presence of two cytokinins or 2iP with morphactin in the culture medium markedly enhanced the production of isoflavonoids in P. tuberosa.

The cell cultures of Pueraria tuberosa, a perennial leguminous lianas, were maintained in modified MS medium (KNO 3 475 mg l -1 , thiamine 1 mg l -1 , biotin 1 mg l -1 , calcium pantothenate 1 mg l -1 ) containing 0.1 mg l -1 2,4,5-trichloroacetic acid and 0.1 mg l -1 kinetin. Isoflavonoids (puerarin, genistin, daidzein, genistein) accumulation in cell suspension cultures was increased by 14-fold to *12 mg l -1 after 48 h of adding 100 lM ethrel. Ethrel inhibitors (silver nitrate and silver thiosulfate) completely inhibited this effect in the presence of ethrel and isoflavonoids were not detected in the spent medium. The increase was dose dependent and can be explored to trigger high yield of isoflavonoids production.

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