A sustainable improvement in jute productivity under less favourable environment can only be achieved with a constant flow of new genetic materials. The existing variability for constraints, like insect-pest and diseases, poor soil fertility, water stress, fiber quality, photo-insensivity etc. is a serious issue that needs to be addressed . It is evident that all the tissue culture techniques play a vital role in the enrichment of genetic variability. Among the phytohormone combination, MS+ 2 mg/l BAP + 0.5 mg/l IAA showed the highest shoot regeneration (60.66%). The variety CVE-3 response better than O-72 towards shoot regeneration at different pH level. Among the sucrose concentration 3% (30mg/l) showed the highest shoot regeneration (87.49%). Among the different concentration of vitamins x2 times (3.0mg/l) showed the highest shoot regeneration. Using different concentration of surfactant 0.1% surfactant showed the highest shoot regeneration (91.663%). In the NaCl experiment, among the three varieties CVE-3 was found highly responsive to shoot regeneration (97.22%) and 0.125, 0.25 and 0.50 (%) NaCl shows the better responsive to shoot regeneration.