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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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Abstract

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 resistance-breaking characteristics.

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Acknowledgments

Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel, No. 511/08. The authors would like to thank the Israeli Ministry of Foreign Affairs for support to E. Kukurt, Dr. D. Shteinberg for statistical advice, Dr. T. Arazi for comments on the manuscript, and Dr. M. Lapidot for the tomato and pepper PVY strains. This work was supported in part by grants from the Chief Scientist's Office, Ministry of Agriculture, and the United States-Israel Binational Agricultural Research and Development Fund (BARD) (grant USA 3623-04).

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Author notes

  1. Yoel M. Shiboleth

    Present address: Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI, 48109, USA

Authors and Affiliations

  1. Department of Plant Pathology and Weed Science, ARO Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel

    Victor Gaba, Arie Rosner, Ludmilla Maslenin, Diana Leibman, Sima Singer, Emre Kukurt & Amit Gal-On

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  1. Victor Gaba
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Correspondence to Victor Gaba.

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Victor Gaba and Arie Rosner have contributed equally to this publication.

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Gaba, V., Rosner, A., Maslenin, L. et al. Hairpin-based virus resistance depends on the sequence similarity between challenge virus and discrete, highly accumulating siRNA species. Eur J Plant Pathol 128, 153–164 (2010). https://doi.org/10.1007/s10658-010-9654-5

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