Genetius

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 constitutively active MLO (faNTA) can rescue the fer-4 root‑hair bursting and polarity defects, restoring tip‑focused cytosolic Ca2+ oscillations and ROS accumulation, highlighting a FERONIA‑MLO signaling module that governs Ca2+ influx and ROS production during root‑hair tip growth. Genetic analysis of mlo15-4 further confirms MLO15 as a key regulator of these Ca2+ and ROS dynamics. The findings suggest MLO proteins act downstream of FER to coordinate calcium and ROS signals essential for root‑hair integrity.

The study reveals that reactive oxygen species (ROS) have distinct temporal roles in Arabidopsis thaliana stigma papillae development, with superoxide promoting early growth and hydrogen peroxide marking mature, pollen‑receptive papillae. Pharmacological reduction of superoxide or transgenic over‑expression of superoxide dismutase under an early stigma promoter impairs papillae growth, highlighting ROS homeostasis as essential for proper papillae differentiation and successful pollination.

Loss‑of‑function mutations in the drought‑induced genes GASA3 and AFP1 confer enhanced drought tolerance in Arabidopsis thaliana, primarily through smaller stomatal apertures and increased ABA accumulation via hydrolysis of ABA‑GE. Constitutive overexpression of these genes heightens drought sensitivity, indicating that the AFP1/GASA3 module negatively regulates stomatal closure and ABA signaling.

The study identifies HISTONE DEACETYLASE COMPLEX 1 (HDC1) as a positive regulator of sepal size during maturation in Arabidopsis thaliana, showing that hdc1 mutants exhibit prolonged elongation due to delayed maturation. Integrated transcriptomic and proteomic analyses, together with genetic and chemical experiments, reveal that HDC1 promotes ethylene production, which in turn triggers ROS accumulation to terminate sepal growth. These findings elucidate a coordinated ethylene‑ROS signaling mechanism controlling organ size during plant development.

The study demonstrates that dihydroxy B‑ring flavonoids modulate the amplitude of the core circadian clock gene reporter TOC1:LUC in Arabidopsis by affecting cellular H2O2 levels, rather than auxin transport. Reducing reactive oxygen species restored normal TOC1:LUC amplitude in flavonoid‑deficient seedlings, and altered chloroplast Ca2+ levels suggest a retrograde signaling component.

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 identifies the scaffolding protein RACK1A as a cytoplasmic interaction partner of the antioxidant enzyme FSD1, revealing that RACK1A recruits FSD1 to cycloheximide-sensitive condensates that colocalize with stress granules during salt stress. Functional analyses show that this RACK1A‑FSD1 module modulates ROS levels, influences root hair tip growth, and determines salt‑stress resilience in Arabidopsis.

The study reveals that hydrogen sulfide (H₂S) and the cysteine desulfhydrase DES1 are essential for stomatal immunity, mediating flg22‑ and bacterial‑induced stomatal closure and influencing reactive oxygen species (ROS) dynamics. Loss‑of‑function des1 mutants show altered susceptibility to Pseudomonas syringae and reduced apoplastic and cytosolic H₂O₂ accumulation, while H₂S can induce ROS production independently of RBOHD.