ISHS Acta Horticulturae 692: II International Symposium on Biotechnology of Tropical and Subtropical Species
TISSUE-SPECIFIC EXPRESSION OF LYTIC PEPTIDES IN TRANSGENIC GRAPEVINES VIA USE OF A NPTII/GFP FUSION MARKER |
| Authors: | D.J. Gray, Z.T. Li, S. Jayasankar |
| Keywords: | Vitis vinifera, grape, transformation, Pierce¿s disease |
| Abstract:
An NPTII/GFP fusion marker previously was used to optimize parameters for the stable transformation of grapevine (Vitis vinifera ‘Thompson Seedless’). Several hundred embryogenic culture lines were recovered, each presumably containing unique insertion(s) of the fusion marker. Subsequently, plants regenerated from the culture lines were evaluated for GFP expression pattern. It was discovered that certain plants exhibited enhanced GFP expression in their vascular systems. Furthermore, microscopic examination revealed that GFP expression was particularly high in both xylem and phloem tissue, as well as vascular cambium. Subsequently, a gene coding for the lytic peptide Shiva-1 was linked to the fusion marker and used to transform grapevine. ELISA analyses of regenerated plants revealed that lytic peptide was present in the same tissues that expressed GFP. These results suggest a method of selecting grapevines with enhanced antimicrobial gene expression in xylem tissue, since such genes can be linked to function in concert with the GFP/NPTII fusion marker. Expression of an antimicrobial transgene in the vascular system of grapevine might inhibit or suppress the xylem-limited bacterium (Xylella fastidiosa), which causes Pierce’s disease (PD). | |
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