Vacuolar invertase knockout enhances drought tolerance in potato plants
Authors: Roitman, M., Teper-Bamnolker, P., Doron-Faigenboim, A., Sikron, N., Fait, A., Vrobel, O., Tarkowski, P., Moshelion, M., Bocobza, S., Eshel, D.
CRISPR/Cas9 knockout of the vacuolar invertase gene (StVInv) in potato enhanced drought resilience, with mutants maintaining higher stomatal conductance, transpiration, and photosynthetic efficiency, leading to improved agronomic water-use efficiency and biomass under water limitation. Metabolomic profiling showed accumulation of galactinol and raffinose, while ABA levels were reduced, indicating altered osmoprotective and hormonal responses that support sustained growth during drought.
The study engineered Tobacco rattle virus vectors incorporating distinct RNA secondary structures as mobility factors to improve guide RNA delivery to plant meristems. Using Nicotiana benthamiana plants expressing Cas9, optimal virus constructs were identified that generated both somatic and heritable edits, and these constructs were successfully applied to edit the emerging oilseed crop pennycress (Thlaspi arvense).
The study used comparative transcriptomics of dorsal and ventral petals across development, alongside expression profiling in floral symmetry mutants, to identify genes linked to dorsal (AmCYC-dependent) and ventral (AmDIV-dependent) identities in Antirrhinum majus. In situ hybridisation validated axis‑specific and boundary‑localized expression patterns, revealing that a conserved NGATHA‑LIKE1‑BRASSINAZOLE‑RESISTANT1‑miR164 module has been co‑opted to regulate AmDIV targets and shape the corolla. These findings delineate regulatory modules coordinating dorsoventral and proximal‑distal patterning in zygomorphic flowers.
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 sequenced genomes of ericoid mycorrhiza‑forming liverworts and experimentally reconstituted the symbiosis, revealing a nutrient‑regulated state that supports intracellular colonization. Comparative transcriptomics identified an ancestral gene module governing intracellular symbiosis, and functional validation in Marchantia paleacea through genetic manipulation, phylogenetics, and transactivation assays confirmed its essential role. The findings suggest plants have retained and independently recruited this ancestral module for diverse intracellular symbioses.
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 investigates the wheat Pm3 NLR allelic series, revealing that near-identical Pm3d and Pm3e alleles confer broad-spectrum resistance by recognizing multiple, structurally diverse powdery mildew effectors. Using chimeric NLR constructs, the authors pinpoint specificity-determining polymorphisms and demonstrate that engineered combinations of Pm3d and Pm3e further expand effector recognition, showcasing the potential for durable wheat protection through NLR engineering.
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 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.