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Leibman, D., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Wolf, D., Department of Vegetable Research, ARO the Volcani Center, Bet Dagan 50250, Israel
Saharan, V., Department of Molecular Biology and Biotechnology, Maharana Pratap University of Agriculture and Technology, Udaipur-313001, Rajasthan, India
Zelcer, A., Department of Vegetable Research, ARO the Volcani Center, Bet Dagan 50250, Israel
Arazi, T., Department of Ornamental Horticulture, ARO the Volcani Center, Israel
Yoel, S., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Gaba, V., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Gal-On, A., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Gene-silencing has been used to develop resistance against many plant viruses but little is known about the transgenic small-interfering RNA (t-siRNA) that confers this resistance. Transgenic cucumber and melon lines harboring a hairpin construct of the Zucchini yellow mosaic potyvirus (ZYMV) HC-Pro gene accumulated different levels of t-siRNA (6 to 44% of total siRNA) and exhibited resistance to systemic ZYMV infection. Resistance to Watermelon mosaic potyvirus and Papaya ring spot potyvirus-W was also observed in a cucumber line that accumulated high levels of t-siRNA (44% of total siRNA) and displayed significantly increased levels of RNA-dependent RNA (RDR)1 and Argonaute 1, as compared with the other transgenic and nontransformed plants. The majority of the t-siRNA sequences were 21 to 22 nucleotides in length and sense strand biased. The t-siRNA were not uniformly distributed throughout the transgene but concentrated in "hot spots" in a pattern resembling that of the viral siRNA peaks observed in ZYMV-infected cucumber and melon. Mutations in ZYMV at the loci associated with the siRNA peaks did not break this resistance, indicating that hot spot t-siRNA may not be essential for resistance. This study shows that resistance based on genesilencing can be effective against related viruses and is probably correlated with t-siRNA accumulation and increased expression of RDR1. © 2011 The American Phytopathological Society.
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A high level of transgenic viral small RNA is associated with broad potyvirus resistance in cucurbits
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Leibman, D., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Wolf, D., Department of Vegetable Research, ARO the Volcani Center, Bet Dagan 50250, Israel
Saharan, V., Department of Molecular Biology and Biotechnology, Maharana Pratap University of Agriculture and Technology, Udaipur-313001, Rajasthan, India
Zelcer, A., Department of Vegetable Research, ARO the Volcani Center, Bet Dagan 50250, Israel
Arazi, T., Department of Ornamental Horticulture, ARO the Volcani Center, Israel
Yoel, S., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Gaba, V., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
Gal-On, A., Department of Plant Pathology and Weed Science, ARO the Volcani Center, Bet Dagan 50250, Israel
A high level of transgenic viral small RNA is associated with broad potyvirus resistance in cucurbits
Gene-silencing has been used to develop resistance against many plant viruses but little is known about the transgenic small-interfering RNA (t-siRNA) that confers this resistance. Transgenic cucumber and melon lines harboring a hairpin construct of the Zucchini yellow mosaic potyvirus (ZYMV) HC-Pro gene accumulated different levels of t-siRNA (6 to 44% of total siRNA) and exhibited resistance to systemic ZYMV infection. Resistance to Watermelon mosaic potyvirus and Papaya ring spot potyvirus-W was also observed in a cucumber line that accumulated high levels of t-siRNA (44% of total siRNA) and displayed significantly increased levels of RNA-dependent RNA (RDR)1 and Argonaute 1, as compared with the other transgenic and nontransformed plants. The majority of the t-siRNA sequences were 21 to 22 nucleotides in length and sense strand biased. The t-siRNA were not uniformly distributed throughout the transgene but concentrated in "hot spots" in a pattern resembling that of the viral siRNA peaks observed in ZYMV-infected cucumber and melon. Mutations in ZYMV at the loci associated with the siRNA peaks did not break this resistance, indicating that hot spot t-siRNA may not be essential for resistance. This study shows that resistance based on genesilencing can be effective against related viruses and is probably correlated with t-siRNA accumulation and increased expression of RDR1. © 2011 The American Phytopathological Society.
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