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 examined nitrogen use strategies in the model alga Chlamydomonas reinhardtii by comparing growth on ammonium, nitrate, and urea, finding similar molar nitrogen utilization efficiency under saturating conditions. Rapid nitrogen uptake and storage were demonstrated through pulse experiments, and source‑specific transcriptome analysis revealed distinct regulation of assimilation pathways and transporters, supporting a model of flexible nitrogen acquisition and storage.
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 investigates how maternal environmental conditions, specifically temperature and light intensity, influence seed longevity in eight Arabidopsis thaliana natural accessions. Seeds developed under higher temperature (27 °C) and high light showed increased longevity, with transcriptome analysis of the Bor-4 accession revealing dynamic changes in stored mRNAs, including upregulation of antioxidant defenses and raffinose family oligosaccharides. These findings highlight the genotype‑dependent modulation of seed traits by the maternal environment.
The study demonstrates that limonene, a natural essential‑oil component, strongly inhibits Fusarium oxysporum, the causal agent of potato dry rot, by impairing colony growth, hyphal morphology, spore viability, membrane integrity, and transcription/translation processes, as well as disrupting ion homeostasis. Combined treatments reveal additive effects with mancozeb and synergistic effects with hymexazol, highlighting limonene's potential as an eco‑friendly bio‑fungicide for potato disease management.
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 study investigates the evolutionary shift from archegonial to embryo‑sac reproduction by analyzing transcriptomes of Ginkgo reproductive organs and related species. It reveals that the angiosperm pollen‑tube guidance module MYB98‑CRP‑ECS is active in mature Ginkgo archegonia and that, while egg cell transcription is conserved, changes in the fate of other female gametophyte cells drove the transition, providing a molecular framework for this major reproductive evolution.
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.