The study presents an optimized Agrobacterium-mediated transformation toolkit for Sorghum bicolor that achieves up to 95.7% editing efficiency using CRISPR/Cas9 targeting the SbPDS gene, and demonstrates comparable performance with a PAM‑broadened SpRY variant. This platform enables multiplex genome editing and is positioned for integration of advanced tools such as prime and base editors to accelerate sorghum breeding.
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 integrated metabolomic and transcriptomic analyses of red clover (Trifolium pratense) roots infected with Fusarium oxysporum and Phoma medicaginis to identify candidate cytochrome P450 enzymes responsible for the methylenedioxy bridge formation in (-)-maackiain biosynthesis. Using co‑expression network analysis and phylogenetic screening, five P450 candidates were selected and screened in engineered Saccharomyces cerevisiae, revealing TpPbS/CYP76F319 as the enzyme catalyzing conversion of calycosin to pseudobaptigenin. This discovery enables reconstruction of the complete (-)-maackiain pathway for potential health and agricultural applications.
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 investigated how barley (Hordeum vulgare) adjusts mitochondrial respiration under salinity stress using physiological, biochemical, metabolomic and proteomic approaches. Salt treatment increased respiration and activated the canonical TCA cycle, while the GABA shunt remained largely inactive, contrasting with wheat responses.
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.
Unravelling the intraspecific variation in drought responses in seedlings of European black pine (Pinus nigra J.F. Arnold)
Authors: Ahmad, M., Hammerbacher, A., Priemer, C., Ciceu, A., Karolak, M., Mader, S., Olsson, S., Schinnerl, J., Seitner, S., Schoendorfer, S., Helfenbein, P., Jakub, J., Breuer, M., Espinosa, A., Caballero, T., Ganthaler, A., Mayr, S., Grosskinsky, D. K., Wienkoop, S., Schueler, S., Trujillo-Moya, C., van Loo, M.
The study examined drought tolerance across nine provenances of the conifer Pinus nigra using high‑throughput phenotyping combined with metabolomic and transcriptomic analyses under controlled soil‑drying conditions. Drought tolerance, measured by the decline in Fv/Fm, varied among provenances but was not linked to a climatic gradient and was independent of growth, with tolerant provenances showing distinct flavonoid and diterpene profiles and provenance‑specific gene expression patterns. Integrating phenotypic and molecular data revealed metabolic signatures underlying drought adaptation in this non‑model conifer.
The study systematically identified heterosis-associated genes and metabolites in rice, functionally validated three genes influencing seedling length, and integrated these molecules into network modules to explain heterosis variance. Predominant additive and partially dominant inheritance patterns were linked to parental genomic variants and were shown to affect 17 agronomic traits in rice, as well as yield heterosis in maize and biomass heterosis in Arabidopsis. The work highlights the quantitative contribution of transcriptomic and metabolomic variation, especially in phenylpropanoid biosynthesis, to hybrid vigor.