Microprojectile bombardment transformation of <i>Brassica </i>

摘要

The invention relates to the genetic manipulation of plants, particularly plants of the genus Brassica. Methods are provided for producing transgenic Brassica plants involving the introduction of a DNA construct by microprojectile bombardment into pre-incubated microspores, microspore-derived embryos and microspore-derived hypocotyls. The methods find use in the development of improved agricultural varieties of Brassica plants through the incorporation of desirable agronomic traits.

主权项

1. A method for producing a stably transformed Brassica plant, comprising:
(a) isolating a microspore from a donor Brassica plant; (b) culturing said isolated microspore for a period of about two to about ten days from the time of isolation to produce an isolated pre-incubated microspore; (c) prior to step (d), culturing under conditions of plasmolysis the isolated pre-incubated microspore; (d) introducing, under conditions of plasmolysis, a DNA construct by microprojectile bombardment into a pre-incubated microspore to produce a transformed pre-incubated microspore; (e) after step (d), culturing under conditions of plasmolysis the transformed pre-incubated microspore; (f) culturing the transformed pre-incubated microspore in or on a liquid selection medium to produce an embryo or tissue; and (g) regenerating a stably transformed Brassica plant from the embryo or tissue.