LONG-TERM STABLE EXPRESSION OF MULTIPLE TRANSGENES UNDER CONTROL OF THE SAME PROMOTER IN CYMBIDIUM ORCHID

M.R. Boase, N.W. Albert, H. Zhang, S. Arathoon, L. Wang, D.H. Lewis
A transformation protocol based on a particle-inflow-gun was successfully used to transfer multiple transgenes into ‘Jungfrau Dos Pueblos’, a commercial cultivar of Cymbidium orchid. Protocorm-like bodies (PLBs) were bombarded with three transgenes, a dihydroflavonol reductase (DFR) antisense construct, the green fluorescent protein reporter gene (GFP) and the neomycin phosphotransferase II (NPTII) selectable marker, all of which are under the control of the CaMV- 35S promoter. Transgenic plants were recovered from bombarded PLBs, which had been selected with geneticin. They were exflasked into a containment house and successfully grown to flowering. Seven years after bombardment, all three transgenes were shown to be expressed in floral tissue and no indication of gene silencing was observed. GFP fluorescence was monitored over this period and despite some masking by red autofluorescence in green tissue, was a useful visual marker in both vegetative and floral tissue. The antisense DFR (asDFR) construct was expressed in floral tissue but no obvious change in anthocyanin biosynthesis was observed initially. Induced anthocyanin accumulation in the flower lip was less in the asDFR flowers than in the non-transgenic flowers, possibly due to reduced DFR activity which is consistent with the action of this transgene.
Boase, M.R., Albert, N.W., Zhang, H., Arathoon, S., Wang, L. and Lewis, D.H. (2012). LONG-TERM STABLE EXPRESSION OF MULTIPLE TRANSGENES UNDER CONTROL OF THE SAME PROMOTER IN CYMBIDIUM ORCHID. Acta Hortic. 937, 597-604
DOI: 10.17660/ActaHortic.2012.937.72
https://doi.org/10.17660/ActaHortic.2012.937.72
transformation, anthocyanin, dihydroflavonol reductase, NPTII, green fluorescent protein
English

Acta Horticulturae