Patent ID: 6020538
Filing Date: 2000-02-01
Classification: C12N

Abstract:
A method of genetically engineering an orchid with a DNA construct comprising an expression cassette, which construct comprises, in the 5' to 3' direction, a transcription initiation region and a peptide coding region positioned downstream from said transcription initiation region and under the transcriptional regulation thereof, said method comprising the steps of:(a) culturing primary protocorm-like bodies (PLBs) of the orchid in liquid medium to induce secondary PLBs;(b) placing the PLBs comprising both primary and secondary PLBs on a target surface;(c) physically accelerating at the PLBs microprojectiles which comprise copies of a foreign genetic construct comprising at least one foreign gene of interest and at least one selectable marker gene;(d) culturing the PLBs from (c) in liquid medium to form proliferative PLBs;(e) inducing the proliferative PLBs to form shoots in liquid medium containing at least one selection agent for which the at least one selectable marker gene confers resistance, imparts prototrophy or imparts a visible phenotype to the proliferative PLB in the presence of the selection agent to form selectable PLBs having shoots, said selectable PLBs being transformed plant tissue;(f) culturing selectable PLBs which have shoots; and(g) regenerating the shoots of the selectable PLBs into clonal transgenic orchid plants wherein, said orchid is a member of genus Cymbidium.