The study shows that silencing of NOR2 rRNA genes in Arabidopsis thaliana depends primarily on CHH-context cytosine methylation, particularly mediated by CMT2 and the chromatin remodeler DDM1, rather than CG or CHG methylation. Comparative promoter analysis revealed a prevalence of CHH sites in plant rDNA promoters, explaining why CHH methylation mutants disrupt NOR2 silencing more strongly, while NOR2 loci are hyper‑methylated and more condensed than NOR4.

The study demonstrates that RNA extracted from herbarium specimens can be used to generate high‑quality transcriptomes, comparable to those from fresh or silica‑dried samples. By assembling and comparing transcriptomes across specimen types, the authors validated a plant immune receptor synthesized from a 1956 collection, proving archival RNA’s utility for functional genomics. These findings challenge the prevailing view that herbarium RNA is unsuitable for transcriptomic analyses.