Genetius

The study applied a progressive, sublethal drought treatment to Arabidopsis thaliana, collecting time‑resolved phenotypic and transcriptomic data. Machine‑learning analysis revealed distinct drought stages driven by multiple overlapping transcriptional programs that intersect with plant aging, and identified high‑explanatory‑power transcripts as biomarkers rather than causal agents.

RNA‑Seq was used to compare Arabidopsis thaliana transcriptomes under drought, Meloidogyne incognita infection, and their combination, revealing a distinct set of genes uniquely regulated by the joint stress. Notably, AZI1, SAUR71, and DRN1 showed stress‑specific expression patterns, suggesting key roles in coordinating responses to simultaneous drought and nematode attack.

The study introduces a CRISPR/Cas9‑based restoration system (CiRBS) that reactivates a disabled luciferase reporter (LUC40Ins26bp) in transgenic Arabidopsis, enabling long‑term single‑cell bioluminescence monitoring. Restoration occurs within 24 h after particle‑bombardment‑mediated CRISPR delivery, with ~7 % of cells regaining luminescence and most restored cells carrying a single correctly edited chromosome, facilitating reliable analysis of cellular gene‑expression heterogeneity.

The study combined long‑read whole‑genome assembly, multi‑omics profiling (DNA methylation, mRNA, ribosome‑associated transcripts, tRNA abundance, and protein levels) in two Arabidopsis thaliana accessions to evaluate how genomic information propagates through the Central Dogma. Codon usage in gene sequences emerged as the strongest predictor of both mRNA and protein abundance, while methylation, tRNA levels, and ribosome‑associated transcripts contributed little additional information under stable conditions.

The study applied a CRISPR/Cas9 multiplex guide RNA strategy to delete entire open reading frames of four reproductive genes in Arabidopsis thaliana, achieving homozygous deletions already in the T1 generation with rates of 8.3–30%. Deletion efficiencies correlated with DeepSpCas9 prediction scores, and phenotypic analyses revealed unexpected effects of residual gene fragments on fertilization and seed development.

The study constructs a ~1‑million‑cell single‑nuclei transcriptome atlas of Arabidopsis leaves to reveal that drought stress accelerates transcriptional programs associated with maturation and aging, thereby limiting leaf growth in proportion to stress intensity. Targeted upregulation of FERRIC REDUCTION OXIDASE 6 in mesophyll cells partially rescues leaf growth under drought, demonstrating the functional relevance of these transcriptional changes.

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 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.