Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Nov 7;25(12):5780–5809. doi: 10.1038/s44319-024-00304-5

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

PMC Copyright notice

(A) Schematic representation of the three EVD silencing steps. Upon EVD reactivation, ribosome stalling during translation of EVD shGAG transcript triggers PTGS. 21-nt siRNA produced through RDR6 and DCL4 and loaded into AGO1. With increasing EVD copies across generations, the excess of dsRNA produced by RDR6 is processed by DCL3 to generate 24-nt siRNAs. Loaded into AGO4, shGAG siRNAs trigger DNA methylation (5mC) through RDR6-RdDM at GAG coding sequences without silencing. At 40–50 copies per genome, TGS is installed through Pol IV-RdDM, coincidentally with the appearance of DNA methylation and 24-nt siRNAs on the LTR sequences. (B) Crossing scheme to generate rdr6 mutant lines with active EVD. F2 plants were genotyped to select homozygous WT and mutant RDR6 lines, propagated through selfing until the F6 generation. (C) EVD copy number analysis by qPCR in RDR6-EVD and rdr6-EVD lines at generations F2, F4, and F6 derived from two independent F1s (biological replicates), using the EVD-GAG sequence as target. (D) qPCR analysis of shGAG expression normalized to ACT2 in EVD-RDR6 and EVD-rdr6 lines at generations 2, 4, and 6 derived from two independent F1s (biological replicates). In (C) and (D), each biological replicate, consistent of bulks of 8–10 plants, are represented for each genotype at each generation, dots show technical replicates. (E) RNA blot analysis of EVD siRNAs against GAG and LTR in RDR6 and rdr6 lines with active EVD at generations F2, F4, and F6. tasiR255 probe is used as control for RDR6 mutation, miR171 and snoRNA U6 are shown as loading controls. WT Col-0 and rdr6 with no reactivated EVD are shown as negative control for EVD activity. Source data are available online for this figure.