Carnations - Patent application filed by Florigene Europe B.V.

A method for genetically transforming carnation plant material, said method comprising:

A) co-cultivating carnation plant material with Agrobacterium cells carrying an exogenous DNA sequence;
B) initiating callus formation in the plant material; and
C) selecting transformed plant cells.

A method for producing genetically altered carnation plants, said method comprising:

A) co-cultivation of carnation plant material with Agrobacterium cells carrying an exogenous DNA sequence including a selectable marker gene in a co-cultivation medium containing nutrients, an energy source, and an induction compound under conditions which allow the Agrobacterium cells to infect the plant material and transfer the exogenous DNA to the carnation chromosomes;
B) culturing plant material from step (A) in a callus initiation medium containing nutrients, an energy source, an auxin, a cytokinin, an anti-Agrobacterium antibiotic, and a plant selection agent which inhibits callus and shoot formation from plant material which does not express the selectable marker gene to produce transformed callus material; and
C) culturing transformed callus material in a regeneration medium containing nutrients, an energy source, an auxin, a cytokinin, an anti-Agrobacterium antibiotic, and the plant selection agent, present in amounts effective to produce transformed shoots.

Claim 23

A method for micropropagating shoots from carnation plant material, said method comprising:

A) culturing carnation plant material to produce a plurality of shoots; and
B) placing vitrified shoots from step (A) in a normalizing medium containing nutrients and an energy source but being substantially free from growth regulators, whereby new shoots are produced which are free from vitrification.

Claim 26

A method for micropropagating shoots from previously established carnation shoots, said method comprising:

A) separating individual shoots; and
B) culturing individual shoots in a multiplication medium comprising nutrients, an energy source, growth regulators and a solidifying agent for a time sufficient to produce at least about 50 shoots for each individual shoot cultured.

Claim 27

A method for regenerating carnation plants, said method comprising: culturing carnation plant material on a regeneration medium containing nutrients, an energy source, a solidifying agent, indole butyric acid at a concentration in the range from about 1 to 5 M, and thidiazuron at a concentration in the range from about 0.5 to 2 M, whereby shoots are produced at a regeneration frequency above about 20 percent.

Carnation callus material which expresses an exogenous DNA sequence.

A carnation plant having cells which express an exogenous DNA sequence.