The study performed a comprehensive computational analysis of the Arabidopsis thaliana proteome, classifying 48,359 proteins by melting temperature (Tm) and melting temperature index (TI) and linking thermal stability to amino acid composition, molecular mass, and codon usage. Machine‑learning and evolutionary analyses revealed that higher molecular mass and specific codon pairs correlate with higher Tm, and that gene duplication has driven the evolution of high‑Tm proteins, suggesting a genomic basis for stress resilience.

The study identifies four Arabidopsis REM transcription factors (VDD, VAL, REM12, REM13) that bind specific DNA sequences and, together with GDE1, recruit RNA polymerase IV to produce 24‑nt siRNAs that direct DNA methylation at designated loci. Loss of GDE1 causes Pol IV complexes to relocalize to sites bound by REM8, indicating that REM proteins provide sequence‑specific cues for epigenetic patterning.

The study reveals that a set of REPRODUCTIVE MERISTEM (REM) transcription factors, termed RIMs, are essential for directing RNA‑directed DNA methylation (RdDM) to CLSY3 targets in a sex‑specific manner in Arabidopsis reproductive tissues. Disruption of RIM DNA‑binding domains or their target motifs abolishes RdDM at these loci, demonstrating that genetic cues can guide de novo methylation patterns.

The study generated two allotriploid Brassica hybrids (ArAnCn) to investigate asymmetric subgenome dominance, finding that the Cn subgenome dominates despite the An subgenome showing highest expression levels. Increased density of accessible chromatin regions (ACRs) in the Cn subgenome correlates with dominant gene expression, while changes in CHH methylation and specific RNA‑directed DNA methylation pathway mutants affect subgenome bias.

The study generated de‑novo genome assemblies for the resistant wild relative Solanum dulcamara and the susceptible Solanum nigrum using a hybrid Oxford Nanopore and Illumina sequencing strategy. Comparative genomic analyses identified auxin‑transport genes and novel pattern recognition receptor orthogroups unique to resistant species, as well as differential gene‑body methylation that may underlie resistance to Ralstonia solanacearum.