ISHS Acta Horticulturae 411: VII International Workshop on Fire Blight
TRANSFORMATION OF GALA APPLE WITH LYTIC PROTEIN GENES |
| Authors: | H.S. Aldwinckle, J.L. Norelli, J.Z. Mills, S.K. Brown |
| Abstract:
Agrobacterium-mediated transformations of Royal Gala apple were conducted with eight plasmid binary vectors containing different lytic protein gene constructs, plus the plasmid binary vectors pBl121 and p35sGUS-INT. A transformation protocol was established for Royal Gala using media developed to maximize proliferation, regeneration, and recovery and elongation of regenerant meristems. One hundred and twenty-four transgenic Royal Gala lines have been recovered to date, and are being multiplied and histochemically assayed. The transgenic lines include representatives of four lytic protein genes, SB-37, Shiva-1, Attacin E, and hen eggwhite lysozyme. The transgenic recovery rate was 0.6% to 9.0% transformants per inoculated leaf. Eighty-six of the Royal Gala transgenic lines have been histochemically assayed for the localization of GUS activity in leaves and shoot cross sections to determine whether the line is composed only of transformed cells (solid) or is a mixture of transformed and untransformed cells (chimeric). Chimeric lines are being regenerated on a high kanamycin medium to select solid lines. Twenty-two transgenic lines now have been established as "solid" lines, and 18 of them have been micro-grafted to seedling or M.9 rootstock for greenhouse and field evaluation. Plants of three of these lines (total 78 plants) were planted in a field trial in June, 1995. | |
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