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Transgene suppression in plants by foliar application of in vitro-synthesized small interfering RNAs

  • Applied genetics and molecular biotechnology
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Abstract

Recent research has shown that plants can uptake long dsRNAs and dsRNA-derived siRNAs that target important genes of infecting fungi or viruses when applied on the surface of plant leaves. The external RNAs were capable of local and systemic movement inducing plant resistance against the pathogens. Few studies have been made for plant gene regulation by foliar application of RNAs. In this study, several types of ssRNA and siRNA duplexes targeting the neomycin phosphotransferase II (NPTII) transgene were in vitro-synthesized and externally applied to the leaf surface of 4-week-old transgenic Arabidopsis thaliana plants. External application of the synthetic NPTII-encoding siRNAs down-regulated NPTII transcript levels in transgenic A. thaliana 1 and 7 days post-treatment with a higher and more consistent effect being observed for siRNAs methylated at 3′ ends. We also analyzed the effects of external NPTII-encoding dsRNA precursors and a dsRNA-derived heterogenous siRNA mix. Digestion of the NPTII-dsRNA to the heterogeneous siRNAs did not improve efficiency of the transgene suppression effect.

Key Points• Foliar application of siRNAs down-regulated a commonly used transgene in Arabidopsis.

• A more consistent effect was observed for methylated siRNAs.

• The findings are important for development of plant gene regulation approaches.

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Funding

This work was supported by a grant from the Russian Science Foundation (grant number 19-74-10023).

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Authors and Affiliations

  1. Laboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok, 690022, Russia

    Alexandra S. Dubrovina, Olga A. Aleynova, Andrey R. Suprun, Zlata V. Ogneva & Konstantin V. Kiselev

  2. The School of Natural Sciences, Far Eastern Federal University, Vladivostok, 690090, Russia

    Andrey R. Suprun & Konstantin V. Kiselev

Authors
  1. Alexandra S. Dubrovina
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  2. Olga A. Aleynova
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  3. Andrey R. Suprun
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  4. Zlata V. Ogneva
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  5. Konstantin V. Kiselev
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Contributions

ASD and KVK performed research design, data analysis, paper preparation, and experimental process. OAA, ARS, and ZVO performed plant management and qRT-PCRs.

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Correspondence to Alexandra S. Dubrovina.

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Dubrovina, A.S., Aleynova, O.A., Suprun, A.R. et al. Transgene suppression in plants by foliar application of in vitro-synthesized small interfering RNAs. Appl Microbiol Biotechnol 104, 2125–2135 (2020). https://doi.org/10.1007/s00253-020-10355-y

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