Patent ID: 6455761
Filing Date: 2002-09-24
Classification: C12N

Abstract:
A method for transforming mature plants of turnip rape, comprising(i) excising an internode section of the inflorescence-carrying stem of a mature turnip rape plant, (ii) sterilizing said internode section, and cutting it in 4-8 mm segments to obtain an internode segment, (iii) placing an internode segment in a horizontal position on an agar pre-cultivation medium supplemented with 15-90 &mgr;M of silver nitrate and 2,4-dichlorophenoxyacetic acid (2,4-D) hormone, (iv) pre-cultivating the internode segment on said medium for 1 day, (v) immersing the internode segment in a MS solution inoculated with Agrobacterium tumefaciens bacteria carrying at least one gene heterologous to said turnip rape, (vi) placing the immersed internode segment in a horizontal position on an agar MS co-cultivation medium, (vii) co-cultivating the internode segment with Agrobacteria for 2 days, (viii) washing the internode segment to remove the Agrobacteria, (ix) placing the internode segment in a vertical position with the basal side down on MS agar medium for selection with an antibiotic, the medium being supplemented with cytokinine hormones and silver nitrate, to obtain an internode segment with regenerated primordia or embryonic green nodules, (x) placing the internode segment with regenerated primordia or embryogenic green nodules on a hormone-free regeneration medium, and (xi) recovering the transgenic shoots regenerated.