Roses

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 controlled regeneration of a rose plantlet from a somatic embryo which comprises:

(a) providing a somatic embryo;
(b) culturing the somatic embryo on a maturation medium capable of inducing differentiation of the embryo to yield a differentiated embryo;
(c) germinating the differentiated embryo on germination medium to yield a germinated embryo; and
(d) propagating the germinated embryo on propagation medium to produce a mature plantlet capable of being transferred to soil conditions.

A method for obtaining at least one somatic embryo from mature somatic tissue of rose plant which comprises:

(a) culturing mature somatic tissue on callus induction medium comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin to obtain at least one induced callus; and
(b) culturing the induced callus in a regeneration media capable of inducing completion of the development of somatic embryos comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin in which the source of the auxin and cytokinin in the regeneration media differs from the source of the auxin and cytokinin in the callus induction medium to obtain at least one somatic embryo.

A method for obtaining a somatic embryo from a stamen filament of rose plant which comprises:

(a) culturing the stamen filament on callus induction medium comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin to obtain at least one induced callus; and
(b) culturing the induced callus in a regeneration media capable of inducing completion of the development of somatic embryos comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin in which the ratio of auxin to cytokinin is decreased by a factor of two to about 15 relative to the ratio of auxin to cytokinin in the callus induction medium to obtain a somatic embryo.

A method for obtaining a somatic embryo from a leaf explant of rose plant which comprises:

(a) culturing the leaf explant on a callus induction medium comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin to obtain at least one induced callus; and
(b) culturing the induced callus in a regeneration media capable of inducing completion of the development of somatic embryos comprising effective amounts of a nutrient medium, an energy source, an auxin and a cytokinin to which the source of the auxin and cytokinin in the regeneration media differs from the source of the auxin and cytokinin in the callus induction medium to obtain a somatic embryo.

• incubating the callus cells with Agrobacterium cells carrying an exogenous DNA sequence; and

• selecting callus cells which express at least a portion of the exogenous DNA sequence.

(a) culturing tissue from the rose plant under conditions selected to produce a 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) culturing tissue from a rose plant on a callus induction medium containing nutrients, an energy source, an auxin, and growth regulator, a cytokinin in amounts effective to induce callus formation wherein the tissue is derived from a plant part selected from the group consisting of stamen filaments, leaf explants, stem sections, shoot tips, petal, sepal, petiole, and peduncle;
(b) combining cells from the callus of step (a) with Agrobacterium cells carrying the exogenous DNA sequence in a cocultivation 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 regeneration medium containing nutrients, an energy source, an antibacterial agent, and a growth regulator selected from abscisic acid and giberellic acid, other than an auxin or a cytokinin, present in amounts effective to produce somatic embryos.

• incubating the callus cells with Agrobacterium cells carrying an exogenous DNA sequence; and

• selecting callus cells which express at least a portion of the exogenous DNA sequence.

(a) culturing tissue from the rose plant under conditions selected to produce a 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) 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 cocultivation 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.