Utilizing Potato Virus X to Monitor RNA Movement

Yu, Z., Cho, S.K., Zhang, P., Hong, Yiguo and Hannapel, D.J. (2020) Utilizing Potato Virus X to Monitor RNA Movement. In: RNA Tagging. Methods in Molecular Biology. Methods in Molecular Biology, 2166 (2166). Humana, New York, pp. 181-194. ISBN 9781071607114 (print) 9781-071607121 (online)

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.

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