Abstract: In the present investigation the effort was made to develop an efficient protocol for rooting and acclimatization of in vitro raised plantlets of guava cv. Allahabad safeda. In vitro plantlets were generated through nodal segment explant using 1.0 mg/l BA + 0.25 mg/l GA3 in MS medium. Maximum rooting (83.4%) and length of root/shoot (7.5 cm) and minimum days to root initiation (8.0 day) was observed in full strength of MS medium supplemented with 0.4 mg/l IBA. While, maximum length of shoots (8.1 cm) and maximum number of roots (5.52) were observed in treatment (Full MS + 0.2 mg/l IBA) followed by treatment ½ MS + 0.2 mg/l IBA. The plantlets grown in potting mixture containing vermicompost + soil (1:1v/v) showed better survival of plantlet (90.00 %). Survival of plantlets and growth of plantlets were significant influenced by different climate conditions. Maximum survival of plantlets (86.79 %) and length of shoot was maximum (9.33 cm) was reported in net house condition.