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.
Characterization of a dominant SmNac-like gene as a candidate for photosensitivity in the fruit peel of eggplant
Authors: Gomis-Cebolla, J., Manrique, S., Arrones, A., Toledo-Tolgar, M. D., Luna, J., Baraja-Fonseca, V., Sanchez-Pascual, J., Gimeno-Paez, E., Plazas, M., Gramazio, P., Vilanova, S., Prohens, J.
The study identified that fruit photosensitivity in eggplant is governed by a single dominant gene, with QTLs clustering at the distal end of chromosome 10 (84.1-87.9 Mb). Bulked segregant analysis sequencing and RNA‑seq highlighted the SmNAC1‑like transcription factor as a likely regulator of anthocyanin accumulation, though no coding sequence mutations were detected, suggesting regulatory control at another level.
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 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.
Novel substrate affinity of FaCCR1 and FaCCR1/FaOCT4 expression control the content of medium-chain esters in strawberry fruit
Authors: Roldan-Guerra, F. J., Amorim-Silva, V., Jimenez, J., Mari-Albert, A., Torreblanca, R., Ruiz del Rio, J., Botella, M. A., Granell, A., Sanchez-Sevilla, J. F., Castillejo, C., Amaya, I.
The study identified a major QTL on chromosome 6A that accounts for 40% of variation in medium-chain ester (MCE) levels in strawberry fruit, pinpointing FaCCR1 and FaOCT4 as the causal genes. Functional validation through subcellular localization, transient overexpression, enzymatic assays, and molecular docking demonstrated that FaCCR1 also catalyzes MCE precursor reactions, and a KASP marker in FaOCT4 was developed for breeding fragrant cultivars.
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 identified seven adult plant resistance QTL for oat crown rust using two recombinant inbred line populations, with a major QTL (QPc_GS7_4A.2) on chromosome 4A closely linked to the Pc61 resistance gene. KASP markers targeting SNPs tightly linked to the four most significant QTL were developed, and genetic and haplotype analyses confirmed the association of QPc_GS7_4A.2 with both seedling and adult plant resistance, providing valuable tools for oat breeding.
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.