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In vitro propagation may play an important role in rapid propagation of lemon. Plants produced through in vitro propagation are true to type and are free from diseases. The present investigations were carried out to standardize surface sterilization of explants, suitable explants type for culture establishment, growth regulators for shoot multiplication and rooting, and to evaluate hardening process. The various concentrations of HgCl2 and NaOCl tried for surface disinfection. The explants viz. seeds, embryo and node were used as explants. It was observed that complete embryo gave the best results and emerged as appropriate explants for lemon culture establishment. Among different concentration of MS medium ½ MS medium gave early and maximum seeds germination. In all the combination of plant growth regulators, the highest number of shoots regeneration was observed in 1.0 mg/l 2,4D containing medium. Embryos germination were observed on 0.4 mg/l NAA & 1.5 mg/l BAP containing MS. Root Initiation was observed in combination MS + 0.5 mg/l BAP.