Method for regenerating viable and fertile citrus plants by tissue culture from explants

摘要

The invention provides a method for regenerating viable and fertile Citrus plants by tissue culture from explants of field-grown mature trees, said method comprising the steps of incubating the explant in agarified media to develop axillary buds, subculturing the explant alongwith sprouted axillary buds in agarified media, excising the meristem domes along with leaf primordia in another medium to generate shoots, subculturing the meristem-regenerated shoots on filter paper bridge employing a liquid medium in order to obtain healthy shoots of an average length of about 1.5 cm without intervening callusing within a period of 20-25 days, proliferating the meristem-regenerated shoots in agarified media to obtain well-developed shoots, rooting the isolated well-developed shoots in agarified media, transferring the freshly developed roots of Citrus sinensis to a different medium while allowing the shoots of C. aurantifolia to grow in the same medium for about 15 days to develop tap roots, hardening the rooted shoots ex vitro in a liquid medium, and allowing the shoots to grow in the same medium under incubation conditions, transferring the hardened shoot meristem-raised plants to soilrite in pro-trays for ex vitro growth for about 7 days to ensure 95% to 100% survival of the shoot meristem raised Citrus plants.