The authors introduce the ENABLE(R) Gene Editing in planta toolkit, a streamlined two‑step cloning system for creating CRISPR/Cas9 knockout vectors suitable for transient or stable transformation. Validation was performed in Oryza sativa protoplasts and Arabidopsis thaliana plants, and the toolkit includes low‑cost protocols aimed at facilitating adoption in the Global South.

The study examines how ectopic accumulation of methionine in Arabidopsis thaliana leaves, driven by a deregulated AtCGS transgene under a seed‑specific promoter, reshapes metabolism, gene expression, and DNA methylation. High‑methionine lines exhibit increased amino acids and sugars, activation of stress‑hormone pathways, and reduced expression of DNA methyltransferases, while low‑methionine lines show heightened non‑CG methylation without major transcriptional changes. Integrated transcriptomic and methylomic analyses reveal a feedback loop linking sulfur‑carbon metabolism, stress adaptation, and epigenetic regulation.

The authors created a fast‑cycling, isogenic barley line (GP‑rapid) by introgressing the wild‑type Ppd‑H1 allele from Igri into the Golden Promise cultivar and performing two backcrosses to limit the donor genome, achieving a 25% reduction in generation time under speed‑breeding conditions while retaining high transformation efficiency. CRISPR/Cas9‑mediated editing of Ppd‑H1 showed regeneration and transformation rates comparable to the original Golden Promise, establishing GP‑rapid as a rapid platform for transgenic and gene‑edited barley research.

The study applied CRISPR/Cas9 gene editing to Physalis peruviana to modify plant‑architecture genes and create a compact growth ideotype. This compact phenotype is intended to increase per‑plot yield and support future breeding efforts for this nutritionally valuable minor crop.

The study reveals that the cold‑repressed transcription factor MtCBF4 suppresses anthocyanin synthesis in Medicago truncatula by repressing MtLAP1 and modulating histone H3K27 trimethylation via activation of the demethylase MtJMJ13. MtJMJ13, in turn, demethylates the gene bodies of MtCBF4 and other negative regulators, creating a feedback loop that is inhibited under cold stress, thereby increasing anthocyanin accumulation and enhancing cold tolerance. This work identifies a transcription‑epigenetic regulatory module controlling anthocyanin production during cold stress.

The study used CRISPR/Cas9 to edit the downstream region of the Arabidopsis thaliana FLOWERING LOCUS T (FT) gene, identifying a 2.3‑kb segment containing the Block E enhancer as crucial for normal FT expression and flowering. Fine‑scale deletions pinpointed a 63‑bp core module with CCAAT‑ and G‑boxes, and revealed a cryptic CCAAT‑box that becomes active when repositioned, highlighting the importance of local chromatin context and motif arrangement for enhancer function.

The study optimized three wheat transformation methods—immature embryo, callus, and in planta injection—by systematically adjusting Agrobacterium strain, bacterial density, acetosyringone concentration, and incubation conditions, achieving transformation efficiencies up to 66.84%. Using these protocols, CRISPR/Cas9 knockout of the negative regulator TaARE1-D produced mutants with increased grain number, spike length, grain size, and a stay‑green phenotype, demonstrating the platform’s potential to accelerate yield and stress‑tolerance improvements in wheat.

The authors introduced a polycistronic tRNA‑gRNA array for CRISPR/Cas9 editing in Physcomitrium patens that doubled the frequency of large, targeted deletions compared with conventional single‑gRNA constructs. Using dual‑gRNA targeting, they achieved simultaneous deletion of two to four genes (katanin and TPX2 families) in a single transformation, reaching up to 42% efficiency per gene, though efficiency depended on gRNA pair design.

The study used CRISPR/Cas9 to generate rice snrk1 mutants and performed integrated phenotypic, transcriptomic, proteomic, and phosphoproteomic analyses under normal and starvation conditions, revealing SnRK1’s dual role in promoting growth and mediating stress responses. Findings indicate sub-functionalization of SnRK1 subunits and identify novel phosphorylation targets linked to membrane trafficking, ethylene signaling, and ion transport.

The study used RNA‑seq to compare early transcriptional responses to acute heat and cold stress in cultivated cranberry (Vaccinium macrocarpon) and its F1 hybrids with the cold‑adapted wild relative V. oxycoccos. Cold stress triggered differential expression in pathways such as photosynthesis, ribosomes, defense, and hormone signaling, with some hybrids showing transiently elevated cold‑responsive expression, suggesting potential cold‑tolerance introgression. The results highlight the utility of wild germplasm for breeding temperature‑resilient cranberries.