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Gaba, V., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Rosner, A., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Maslenin, L., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Leibman, D., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Singer, S., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Kukurt, E., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Shiboleth, Y.M., Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI 48109, United States
Gal-On, A., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Virus resistance can be effectively generated in transgenic plants by using the plant's silencing machinery. To study the specificity of gene-silencing-based resistance, homozygous tobacco (Nicotiana tabacum L.) plants containing a 597-nt hairpin RNA construct of the Potato Virus Y (PVY) replicase sequence were challenged with a variety of PVY strains. The transgene-carrying tobacco line was immune to five potato PVY strains with high sequence similarity (88.3-99.5%) to the transgene. Infection with more distant tomato and pepper PVY field strains (86-86.8% sequence similarity) caused delayed symptom appearance in the transgenic tobacco. Transgene production of small interfering (si) RNA was detected by northern blot and measured using a custom-designed microarray for the detection of small RNAs. siRNA accumulation peaks were observed throughout the inverted-repeat transgene. In the resistance-breaking tomato and pepper strains there were nucleotide differences in the sequences correlated to siRNA transgene accumulation, indicating the role of siRNA specificity in resistance breaking. The log of transgene siRNA signal intensity increased with probe GC content, indicating that the accumulating siRNA molecules were GC-rich. Sequence similarity of highly accumulating siRNAs with the target virus strain appears to be important for both resistance and resistancebreaking characteristics. © 2010 KNPV.
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Hairpin-based virus resistance depends on the sequence similarity between challenge virus and discrete, highly accumulating siRNA species
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Gaba, V., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Rosner, A., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Maslenin, L., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Leibman, D., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Singer, S., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Kukurt, E., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Shiboleth, Y.M., Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI 48109, United States
Gal-On, A., Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Hairpin-based virus resistance depends on the sequence similarity between challenge virus and discrete, highly accumulating siRNA species
Virus resistance can be effectively generated in transgenic plants by using the plant's silencing machinery. To study the specificity of gene-silencing-based resistance, homozygous tobacco (Nicotiana tabacum L.) plants containing a 597-nt hairpin RNA construct of the Potato Virus Y (PVY) replicase sequence were challenged with a variety of PVY strains. The transgene-carrying tobacco line was immune to five potato PVY strains with high sequence similarity (88.3-99.5%) to the transgene. Infection with more distant tomato and pepper PVY field strains (86-86.8% sequence similarity) caused delayed symptom appearance in the transgenic tobacco. Transgene production of small interfering (si) RNA was detected by northern blot and measured using a custom-designed microarray for the detection of small RNAs. siRNA accumulation peaks were observed throughout the inverted-repeat transgene. In the resistance-breaking tomato and pepper strains there were nucleotide differences in the sequences correlated to siRNA transgene accumulation, indicating the role of siRNA specificity in resistance breaking. The log of transgene siRNA signal intensity increased with probe GC content, indicating that the accumulating siRNA molecules were GC-rich. Sequence similarity of highly accumulating siRNAs with the target virus strain appears to be important for both resistance and resistancebreaking characteristics. © 2010 KNPV.
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