Recognizing the economic value of biofuels and due to the current growing interest in the non-toxic Jatropha plant's 
potential as a biofuel plant for the Egyptians future, Leaf culture of non-toxic Jatropha curcas Ecuador and Madagascar lines has been conducted at Fac. of Environ. Agric. Scie., Arish University during the period from 2019-2021 to identify an effective, simple, and rapid protocol for improvement the frequency regeneration of Jatropha curcas plantlets, depending on modification of various media, including MS, B5, and WPM. MS modified with 0.5 mgl-1 N-phenyl-N'-l,2,3-thidiazol-5-yl urea (TDZ) + 0.5 mgl-1 BA + 0.1 mgl-1 IBA resulted in the highest induction of adventitious shoot buds (5.3) in Ecuador line and (4.7) in Madagascar line from leaf culture. In in vitro rooting stage of different Jatropha, stable elongated shoots were then planted on half-strength MS, B5, and WPM medium, all of which contained varying concentrations of IBA. The MS medium modified with 0.5 mgl-1 IBA provided the largest frequency of root induction. The highest number of roots/shoot (6.06) with (2.93 cm) root length for Ecuador line, therefore, the largest number of roots/shoot (5.76) with (2.76 cm) root length for Madagascar line, after 6 weeks of culture. In the greenhouse, the in vitro Jatropha plantlets were successfully acclimatized and survived to other study in the future and it could be concluded that, the modified MS medium with TDZ or IBA and Ecuador line were more effective for leaf culture technique.