Micropropagation of woody species is a challenging task due to difficulty in establishment of contamination free cultures. The present investigation was conducted to evolve techniques for successful establishment of contamination free cultures of Jatropha curcas. Explants (Nodal segment and apical shoots) were collected from the Jatropha plants growing under glass house condition and open field condition. Explants were treated with mercury chloride and carbendazin at different concentration for different durations. Among the different sterilization treatments, maximum culture establishment was obtained on treatment combination with carbendazin ( 0.2%) for 45 minutes + HgCl₂ (0.1%) for 4 minutes for shoot apices and nodal explants collected from bothglass house and field grown plants. Treatment combination of carbendazin (0.2%) for one hour and HgCl₂ (0.1%) for 10 minutes responded better than other treatment combinations for culture establishment during winter season.