Co-Authors:
Cohen, A., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Lipsky, A., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Arazi, T., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Ion, A., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Stav, R., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Sandler-Ziv, D., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Fintea, C., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Yedidia, I., Dept. of Ornamental Horticulture, ARO the Volcani Center, Bet Dagan 50250, Israel
Gollop, N., Dept. of Food Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Manulis, S., Dept. of Plant Pathology, ARO the Volcani Center, Bet Dagan 50250, Israel
Abstract:
Bacterial soft rot is one of the most destructive diseases affecting Ornithogalum, and affects many other bulbous crops, ornamental plants and vegetable crops, as well. This disease is caused by Perctobacterium carotovorum (Erwinia carotovora), which penetrates through wounds or stomatal openings, spreads through the apoplast secreting cell-wall degrading enzymes and, eventually, macerates the entire plant and spreads to neighboring plants. There is no known resistance or effective control measure for use against this disease, making utilization of transformation technologies the favored choice for the introduction of disease resistance into this crop. Tachyplesin, a small antimicrobial peptide isolated from hemocytes of the Japanese horseshoe crab, has been reported to inhibit the growth of both Gram-negative and Gram-positive bacteria. Liquid-grown cell clusters of O. dubium were bombarded with gold particles coated with a plasmid carrying the nptII gene, conferring kanamycin resistance, a GUS reporter gene, and the Tachyplesin 1 (TPN1) target gene under the control of either the polyubiquitin (UBQ3) or strawberry vein-banding virus ▲SVB) promoter. Following prolonged selection in a liquid medium supplemented with 80 mg/L kanamycin in darkness, the cultures were transferred to regeneration medium in the light, where hundreds of transgenic plantlets developed. The presence of the target gene was confirmed in all of the transgenic plants tested. Upon infection with a highly virulent bacterial isolate from calla lily (Zantedeschia aethiopica), the control plantlets were completely macerated within a week; whereas the transgenic plants showed varying levels of resistance. Half of the transgenic plantlets remained vital and flourished for months following the Erwinia challenge, despite the continued presence of the plant pathogen on and around the plants.
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