In vitro studies were undertaken for culture establishment in pomegranate cv. G-137 using shoot tips and nodal segment collected from mature plants. Sealing of the cut ends of explants with pre-sterilized paraffin was found to minimize browning by preventing phenol exudation and giving high culture establishment (83.95%) as compared to control (17.55%). Fungicidal pre-treatment of explants with Bavistin® 0.1%(carbendazim)+ Ridomil® 0.1%(Syngenta) for 1 h gave the highest culture establishment (80.60%) with nodal segment and lowest microbial contamination (40.60%), with shoot tip as explant. Interaction between explant, sterilant and duration of treatment revealed that highest culture establishment (81.00%) was observed with 3 min. surface sterilization with 0.1% HgCl₂. The maximum culture establishment was obtained with treatment 2.0 mg/l BAP+ 0.1 mg/l NAA+ 0.5 mg/l GA, which also gave the earliest bud sprouting (5.67 days) as compared to control.