Acclimatization of tissue cultured citrus plantlets is a slow and difficult process. An efficient acclimatization protocol was developed from a series of experiments conducted on a wide range of citrus species. The protocol involved direct culturing of 4-week-old proliferated in vitro shoots in sterile soilrite over sterile farmyard manure. The shoots rooted in 7-15 days, depending upon the species. These plantlets were rooted in the different carriers in sterile tissue culture bottles.