ISHS Acta Horticulturae 689: VII International Symposium on Grapevine Physiology and Biotechnology
TRANSGENIC GRAPEVINE PLANTS EXPRESSING GREEN FLUORESCENT PROTEINS TARGETED TO THE APOPLAST |
| Authors: | C.B. Aguero, A.M. Dandekar, C.P. Meredith |
| Keywords: | Vitis vinifera, genetic transformation, protein secretion |
| Abstract:
The green fluorescent protein gene (gfp) is a powerful tool for plant transformation and other types of genetic experiments because it permits the monitoring of gene expression in living tissues. Pre-embryogenic calli originating from anthers of Vitis vinifera L. cvs. ‘Thompson Seedless’ and ‘Chardonnay’ were transformed via Agrobacterium tumefaciens with three gene constructs to study gfp expression and protein secretion in grapevines. These constructs included the coding sequences of a synthetic GFP that was fused with the amino-terminal sequences obtained from the secreted protein trichosanthin (TCS; tcs-gfp) or the xylem specific protein XSP30 (xsp30-gfp), transcriptionally regulated by the CaMV 35S promoter. Strong fluorescence was found in embryos, roots, stems and leaves obtained from plants transformed with gfp and xsp30-gfp but the level of fluorescence observed in tcs-gfp transformants was very low. In all cases fluorescence was detected only inside the cells, indicating that either the signal peptides were not recognized by the grape secretory machinery or the physiochemical properties of gfp were not sufficient to permit its secretion. | |
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