In Vitro Regeneration in Pomegrante (Punica Granatum L.) Cv. Bhagwa Using Double Nodal Segment

India is the second largest pomegranate producer in the world but the productivity is lower mainly due to non-availability of healthy and disease free planting material. Conventional methods of propagation of pomegranate through stem cutting is time consuming and tedious as it requires about one year to raise the saplings. The present study was carried out with the objective to standardize efficient and reproducible protocol for direct regeneration of plantlets in pomegranate “Bhagwa”. Different explants, duration of mercuric chloride treatment, different growth regulators on in vitro shoot multiplication and root regeneration were tried. The current season twigs containing shoot tip as well as different region internodes were collected from healthy mother plant. Among the various treatments, surface sterilization of double nodal explants containing third fourth nodes with HgCl2 0.10% for 3 minute resulted maximum establishment aseptic culture (55 %) and lowest contamination ((15%)) and lesser intensity of browning ( ) when incubated onto MS medium containing BAP 1 mg/l AgNO3 1 mg/l activated charcoal 2000 mg/l. Shoot proliferation was found superior, when aseptic cultures were transferred onto the MS medium containing ancymidol 0.02 mg/l AgNO3 1 mg/l activated charcoal 500 mg/l with 5 shoot/explants, 4.97 cm shoot length and 18.23 number of leaves. Finally regenerated microshoots were transferred onto the full and half strength MS medium supplemented with NAA and IBA, 1-5 mg /l along with activated charcoal 200 mg/l. Highest rooting was observed on half strength MS medium supplemented with IBA 2 mg/l AgNo3 1 mg/l activated charcoal 200 mg/l, which took 22 days for rooting with 48 % rooting, 5.50 primary root/shoot and 4.32 cm root length. Later the rooted plantlets were successfully hardened under polytunnel with 75 % survival rate in potting mixture containing sterilized coco pith, perlite, and sand (1:1:1 v/v).