The study examined how DNA methylation influences cold stress priming in Arabidopsis thaliana, revealing that primed plants exhibit distinct gene expression and methylation patterns compared to non-primed plants. DNA methylation mutants, especially met1 lacking CG methylation, showed altered cold memory and misregulation of the CBF gene cluster, indicating that methylation ensures transcriptional precision during stress recall.

The authors adapted OpenPlant kit CRISPR/Cas9 tools to enable multiplex gRNA expression from a single transcript using tRNA sequences in the liverwort Marchantia polymorpha, markedly enhancing editing efficiency and scalability. They coupled this vector system with a simplified, optimized thallus transformation protocol, providing a rapid and versatile platform for generating CRISPR/Cas9 mutants and advancing functional genomics in this model species.