An efficient and reproducible protocol is described for shoot-bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of J. curcas. Treating the explants with high concentrations (5-120 mg/L) of TDZ for short durations (5-80 min) before inoculation culture increased significantly the regeneration frequency and improved the quality of the regenerated buds. The highest shoot-buds induction rate (87.35%) was achieved when petiole explants were treated with 20 mg/L TDZ solution for 20 min and inoculated on hormone-free MS medium for 30 days. Regenerated shoots of 0.5 cm or a little longer were isolated and grafted to seedling stocks of the same species, and then the grafted plantlets were planted on half-strength MS medium containing 0.1 mg/L IBA and 2 mg/L sodium nitroprusside (SNP). This grafting strategy was found to be very effective, to obtain that healthy grafted plantlets ready for acclimatization within 20 days. By the above mentioned protocol and with general Agrobacterium - mediated genetic transformation methods only 65 days were needed to obtain intact transgenic plants.

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An efficient method has been established to induce adventitious buds regeneration from 3 kinds of explants (hypocotyl, cotyledon leaf and leaf) of Jatropha curcas L. All the explants were treated by high concentrations (5-120 mg/L) of TDZ solution for different soak period (5-80 min) with the regeneration rate significantly enhanced and the quality of the regenerated buds dramatically improved. The best results of adventitious bud's regeneration were detected when explants were treated with 20 mg/L TDZ solution for 20 min before being inoculated on basic MS medium free of hormone for 30 days. What's more, addition of 0.4 mg/L GA3 into the elongation medium was the most conducive to the extension of adventitious buds. By applying this method described in our paper, regeneration effect of J. curcas was remarkably improved.