Genetius

The study reconstructed the evolutionary history of plant-specific GBF1-type ARF-GEFs by building phylogenetic trees and ortho‑synteny groups, identifying orthologs of AtGNOM and AtGNL1 across species. Functional analyses using transgenic Arabidopsis lines and yeast two‑hybrid assays revealed how duplication and loss events diversified GNOM paralogs, separating polar recycling from secretory trafficking functions.

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 demonstrates that N6‑methyladenosine (m6A) RNA methylation acts as a negative regulator of thermotolerance in Arabidopsis thaliana, with loss of m6A increasing heat‑responsive gene expression and mRNA stability. Heat shock triggers a transient reduction of m6A levels, which is linked to enrichment of the H3K4me3 histone mark at target loci, enhancing transcription of heat shock proteins. These findings reveal a coordinated interplay between RNA methylation and chromatin modifications that fine‑tunes the plant heat stress response.

The study examined how Arabidopsis thaliana integrates physiological, genetic, hormonal, and transcriptomic responses to combined water deficit, heat stress, and elevated CO2. Results show that stomatal aperture under these complex stress combinations is governed by a specific set of regulators, including nitric oxide, OPEN STOMATA 1, and the SLAH3 anion channel, distinct from those active under simpler stress conditions. This reveals a hierarchical stomatal stress code that could inform future research on plant resilience to global change.

The study examined how heat stress alters lipid droplet (LD) number and composition in Arabidopsis thaliana roots, revealing degradation of membrane lipids and accumulation of TAGs and LDs. Proteomic and lipidomic analyses of LDs from a specific Arabidopsis mutant identified novel LD-associated proteins, including triterpene biosynthetic enzymes, whose substrates and products also accumulate in LDs, indicating LDs function as both sinks and sources during stress‑induced membrane remodeling and specialized metabolism.

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.