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.
Splicing regulation by RS2Z36 controls ovary patterning and fruit growth in tomato
Authors: Vraggalas, S., Rosenkranz, R. R., Keller, M., Perez-Perez, Y., Bachiri, S., Zehl, K., Bold, J., Simm, S., Ghatak, A., Weckwerth, W., Afjehi-Sadat, L., Chaturvedi, P., Testillano, P. S., Mueller-McNicoll, M., Zarnack, K., Fragkostefanakis, S.
The study identifies the serine/arginine-rich splicing factor RS2Z36 as a key regulator of ovary patterning and early fruit morphology in tomato, with loss‑of‑function mutants producing smaller, ellipsoid fruits and elongated pericarp cells. RNA‑seq and proteomic analyses reveal widespread alternative splicing and altered protein abundance, including novel splice‑variant peptides, while mutant pericarps show increased deposition of LM6‑detected arabinan and AGP epitopes.
The study functionally characterizes a conserved structured RNA motif (45ABC) in Arabidopsis RBP45 pre‑mRNAs, showing that its sequence and pairing elements mediate a negative auto‑ and cross‑regulatory feedback loop through alternative splicing that produces unproductive isoforms and reduces RBP45 expression. Transcriptome‑wide splicing analysis and phenotypic assessment of rbp45 mutants reveal that RBP45B plays a dominant role and that proper regulation of this motif is essential for root growth and flowering time.
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).
Integrating physiological, transcriptomic, and cellular analyses, the study shows that olive fruit abscission zones undergo lignification, alkalization, and extensive cell‑wall remodeling during natural maturation and after ethephon treatment. A set of 733 FAZ‑specific genes, including β‑1,3‑glucanases, pectate lyases, and pH‑regulating transporters, were identified, and increased glucanase activity together with reduced plasmodesmata callose suggest enhanced intercellular communication facilitates organ detachment in this non‑climacteric fruit.
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 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.
A large-scale proteomic study in Arabidopsis thaliana identified over 32,000 isoform-specific peptides, confirming that alternative splicing, particularly intron retention, produces translated protein isoforms. Integrated proteogenomic analysis, SUPPA classification, and AlphaFold modeling revealed structural impacts and a non-linear regulation of transcript and protein abundance, with mutant phenotypes linking splicing to growth, chlorophyll content, and anthocyanin accumulation.
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.