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RNA Tagging pp 181–194Cite as

Utilizing Potato Virus X to Monitor RNA Movement

Part of the Methods in Molecular Biology book series (MIMB,volume 2166)

Abstract

Mobility assays coupled with RNA profiling have revealed the presence of hundreds of full-length non-cell-autonomous messenger RNAs that move through the whole plant via the phloem cell system. Monitoring the movement of these RNA signals can be difficult and time consuming. Here we describe a simple, virus-based system for surveying RNA movement by replacing specific sequences within the viral RNA genome of potato virus X (PVX) that are critical for movement with other sequences that facilitate movement. PVX is a RNA virus dependent on three small proteins that facilitate cell-to-cell transport and a coat protein (CP) required for long-distance spread of PVX. Deletion of the CP blocks movement, whereas replacing the CP with phloem-mobile RNA sequences reinstates mobility. Two experimental models validating this assay system are discussed. One involves the movement of the flowering locus T RNA that regulates floral induction and the second involves movement of StBEL5, a long-distance RNA signal that regulates tuber formation in potato.

Key words

  • Potato virus X
  • RNA mobility assay
  • FT
  • BEL5

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Acknowledgments

This work was in part funded by Ministry of Science and Technology of China (National Key R&D Program 2017YFE0110900), Ministry of Agriculture of China (National Transgenic Program 2016ZX08009001-004), the National Natural Science Foundation of China (31872636, 31370180), Zhejiang Provincial Natural Science of Foundation (LY19C020002), Hangzhou Normal University (Sino-EU Plant RNA Signaling S&T Platform Initiative 9995C5021841101), and a NSF Plant Genome Research Program award no. DBI-0820659 to DH.

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Correspondence to Yiguo Hong or David J. Hannapel .

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Yu, Z., Cho, S.K., Zhang, P., Hong, Y., Hannapel, D.J. (2020). Utilizing Potato Virus X to Monitor RNA Movement. In: Heinlein, M. (eds) RNA Tagging. Methods in Molecular Biology, vol 2166. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0712-1_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0712-1_10

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  • Publisher Name: Humana, New York, NY

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