Roses - Patents granted to Florigene Europe B.V.

A method for producing a somatic rose embryo, which expresses an exogenous DNA sequence including a selectable marker gene, said method comprising:

A) culturing tissue from a rose plant on a callus induction medium containing nutrients, an energy source, an auxin, and a cytokinin in amounts effective to induce formation of embryogenic callus, wherein the tissue is cultured until a friable, granular embryogenic callus is produced;
B) combining cells from the embryogenic callus of step (A) with Agrobacterium cells carrying the exogenous DNA sequence in a co-cultivation medium containing nutrients, an energy source, and an induction compound under conditions which allow the Agrobacterium cells to infect the embryogenic callus cells and transfer the exogenous DNA sequence to the embryogenic callus cell chromosomes;
C) culturing embryogenic callus cells from step (B) in a selection medium containing nutrients, an energy source, an auxin, a cytokinin, and an agent which inhibits the growth of embryogenic callus cells which do not express the selectable marker gene; and
D) culturing the cells selected in step (C) in a maintenance medium containing nutrients, an energy source, an antibacterial agent, and a growth regulator, other than an auxin or a cytokinin, present in amounts effective to produce viable somatic embryos capable of being regenerated into transformed plantlets.

A method for genetically transforming callus cells from a rose plant, said method comprising: A) incubating friable, granular callus cells with Agrobacterium cells carrying an exogenous DNA sequence; and
B) selecting callus cells which express at least a portion of the exogenous DNA sequence.

A method for genetically transforming a rose plant, said method comprising: A) culturing tissue from the rose plant under conditions selected to produce a friable, granular callus;
B) incubating cells from the callus of step (A) with Agrobacterium cells carrying an exogenous DNA sequence;
C) selecting callus cells from step (B) which express at least a portion of the DNA sequence; and
D) producing transformed plantlets from the selected callus cells of step (C).

A somatic rose embryo produced by the method comprising: A) culturing tissue from a rose plant on a callus induction medium containing nutrients, an energy source, an auxin, and a cytokinin in amounts effective to induce callus formation;
B) combining cells from the callus of step (A) with Agrobacterium cells carrying the exogenous DNA sequence in a co-cultivation medium containing nutrients, an energy source, and an induction compound under conditions which allow the Agrobacterium cells to infect the callus cells and transfer the exogenous DNA sequence to the callus cell chromosomes;
C) culturing callus cells from step (B) in a selection medium containing nutrients, an energy source, an auxin, a cytokinin, and an agent which inhibits the growth of callus cells which do not express the selectable marker gene; and
D) culturing the cells selected in step (C) in a maintenance medium containing nutrients, an energy source, an antibacterial agent, and a growth regulator, other than an auxin or a cytokinin, present in amounts effective to produce somatic embryos.