Moonshade™ carnation

Carnations with modified colour and sulfonylurea herbicide tolerance, produced by inserting two anthocyanin biosynthetic genes from Petunia hybrida, dihydroflavonol reductase (dfr) and Hf1 encoding flavonoid 3',5'hydroxylase (F3'5'H), whose expression results in a violet/mauve colouration. Tolerance to sulfonyl urea herbicides was produced through the introduction of a chlorsulfuron tolerant version of the acetolactate synthase (ALS) encoding gene from Nicotiana tabacum.

NOTE: This LMO was formerly referred to with the UID FLO-4Ø619-7.

A white carnation cultivar FE 123, belonging to Dianthus caryophyllus L., Dianthus, Caryophyllaceae, was used as the recipient organism.

pCGP1470

Gene expression
The T-DNA insertion contained three gene cassettes: Nicotiania tabacum acetolactate synthase (ALS; acetohydroxy acid synthase), Petunia hybrida flavonoid 3', 5' hydroxylase (F3'5'H) and P. hybrida dihydroflavonol-4-reductase (DFR).

Transcription of ALS is under control of the Cauliflower mosaic virus (CaMV) 35S promoter and the ALS terminator. A 5' untranslated leader from P. hybrida chlorophyll a/b-binding protein is also transcribed, but is not expected to be translated. The leader sequence augments the level of transcription (enhances gene expression) of ALS.

Transcription of F3'5'H is under control of the Antirrhinum majus chalcone synthase gene promoter and the P. hybrida D8 terminator.

Transcription of DFR is under control of the synthetic Mac-1 promoter and the Agrobacterium tumefaciens mannopine synthase gene terminator.


Note:
- The genetic element size of the ALS coding sequence includes the size of the terminator (3.76 kb = size of coding sequence + terminator)
- The Mac-1 promoter is a synthetic promoter compromised of A. tumefaciensmas promoter and CaMV enhancer sequences.

This transgenic carnation was developed using recombinant DNA techniques to produce flowers with a unique violet/mauve colour by introducing two genes from petunia (Petunia hybrida) that function together in the biosynthesis of the anthocyanin pigment delphinidin. The transgenic lines were derived from the parent cultivar 'White Unesco', which is a white coloured carnation that was selected for a mutation in the dihydroflavonol reductase (DFR) encoding gene that did not allow for expression of a functional enzyme, and thus did not produce the anthocyanin type pigments that give rise to blue and red coloured flowers. The two genes introduced into the transgenic carnation lines included a functional dihydroflavonol reductase encoding gene and a gene encoding the enzyme flavonoid 3', 5'-hydroxylase (F3', 5'H), a member of the NADPH-Cytochrome P450 reductase family. Expression of these genes allows for the production of blue coloured delphinidin anthocyanin pigments, which are not normally found in carnations.