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Conserved Sequences of Replicase Gene-Mediated Resistance to Potyvirus through RNA Silencing

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Abstract

Nuclear inclusion protein b (NIb) genes of three Potato virus Y isolates PVY-SD1 (O strain), PVY-SD4 (N strain), PVY-SD5 (NTN strain), and Tobacco etch virus isolate TEV-SD1 in Shandong Province were cloned and sequenced. Sequence analysis showed that the sequence homology of the entire NIb gene among the four viruses ranged from 65% to 95%. Hairpin RNA (hpRNA) constructs were designed based on five conserved regions derived from PVY-SD1 and introduced into tobacco plants. After asexual propagation, the transgenic plants were analyzed for resistance to PVY-SD1, PVY-SD4, PVY-SD5, and TEV-SD1. We obtained resistance ratios of 26.2%, 22.7%, 36.4%, 20.3%, and 21.7% to PVY-SD1. When inoculated with the PVY-SD5 virus, the transgenic plants had resistance ratios ranging from 2.4% to 15.9%, but no resistance at all to the other viruses, PVY-SD4 and TEV-SD1. No correlation was found between resistance of transgenic plants and the transgene copy numbers. Northern blot and small interfering RNA (siRNA) analysis demonstrated that the resistance was attributable to RNA silencing. Genetic analysis demonstrated that virus resistance was stably inherited in progeny T1 and T2. These results indicate that the siRNA molecules against conserved regions can confer virus resistance but are restricted to viruses with more than 90% sequence homology.

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Acknowledgements

This research was financially supported by the National Natural Science Foundation of China (Nos. 30771408 and 30500327).

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Authors and Affiliations

  1. College of Plant Protection, Shandong Agricultural University, Tai’an, Shandong, 271018, People’s Republic of China

    Li Xu

  2. State Key Laboratory of Crop Biology, Shandong Agricultural University, Daizong Street No 1, Tai’an, Shandong, 271018, People’s Republic of China

    YunZhi Song, XingQi Guo, ChangXiang Zhu & FuJiang Wen

  3. College of Biology and Technology, Beijing City University, Beijing, 100083, People’s Republic of China

    JunHua Zhu

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  1. Li Xu
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Correspondence to ChangXiang Zhu or FuJiang Wen.

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Li Xu and YunZhi Song have equal contribution.

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Xu, L., Song, Y., Zhu, J. et al. Conserved Sequences of Replicase Gene-Mediated Resistance to Potyvirus through RNA Silencing. J. Plant Biol. 52, 550–559 (2009). https://doi.org/10.1007/s12374-009-9071-5

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