High efficiency transformation mediated agrobacteriun using cotton petiole explants.

摘要

A process for producing a transgenic cotton plant comprising the steps of: (a) obtaining petiole explants cotton, (b) exposing the petiole explants to a culture of Agrobacterium tumefaciens that harbors a vector comprising an exogenous gene and a selectable marker gene, the Agrobacterium being capable of effecting the stable transfer of the exogenous gene and selectable marker gene to the genome of the cells of the petiole explant, (c) culturing the petiole explants in medium containing 2,4-dichlorophenoxyacetic acid at a concentration ranging from more than 0 to 0.5 mg / l kinetin at a concentration ranging from more than 0 to 1 mg / l to induce callus formation, (d) selecting transformed callus that expresses the exogenous gene (e) culturing the selected callus in suspension culture in differentiation medium containing no plant hormone for less than 20 days to induce formation of embryoids, (f) germinating embryoids on a medium having a selected from the group nitrogen source consisting of asparagine, glutamine or both asparagine and glutamine and containing no plant hormone to produce transgenic cotton plants young, and (g) culturing the transgenic cotton plants young in a medium having 10 g / l glucose and 10 g / l sucrose as carbon source and containing no plant hormone to produce transgenic cotton plants, wherein the culture media used in steps (b) - (f) have glucose as the sole carbon source and wherein step (e) is performed at a pH between 5.8 and 7.5.