Utilizing Potato Virus X to Monitor RNA Movement
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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 wordsPotato virus X RNA mobility assay FT BEL5
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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