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.
The study demonstrates that salinity stress induces a photomorphogenic‑like response in dark‑grown Arabidopsis thaliana seedlings, resulting in reduced apical hook curvature and impaired soil emergence. This phenotype is linked to disrupted asymmetric epidermal cell elongation, decreased auxin signaling and PIN3 abundance on the hook’s concave side, repression of BBX28 expression, and loss of a spatial COP1 gradient, highlighting spatial regulation as a key factor in stress‑affected seedling development.
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.
SPOROCYTELESS/NOZZLE acts together with MADS-domain transcription factors to regulate an auxin-dependent network controlling the Megaspore Mother Cell development
Authors: Cavalleri, A., Astori, C., Manrique, S., Bruzzaniti, G., Smaczniak, C., Mizzotti, C., Ruiu, A., Spano, M., Movilli, A., Gregis, V., Xu, X., Kaufmann, K., Colombo, L.
The study elucidates the SPL/NZZ‑dependent regulatory pathway governing megaspore mother cell (MMC) differentiation, revealing that SPL/NZZ directly targets genes and interacts with ovule‑identity MADS‑domain transcription factor complexes. Integration of multi‑omics data with genetic complementation and mutant analyses uncovers an auxin‑dependent downstream network that drives MMC formation.
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 characterizes the liverwort-specific NPR protein (MpNPR) in Marchantia polymorpha, demonstrating that it controls oil body formation and confers resistance to gastropod herbivory through interaction with the transcription factor MpERF13. Loss- or gain-of-function of MpNPR disrupts MpERF13‑dependent gene expression and compromises defense against snail feeding, revealing a lineage‑specific immune pathway distinct from tracheophyte NPR functions.
The study demonstrates that red and blue light have opposing effects on thallus growth orientation in Marchantia polymorpha, with red light promoting epinasty and blue light promoting hyponasty. Loss-of-function mutants in the respective photoreceptors and BBX transcription factors reveal antagonistic interactions that balance thallus flatness under white light. Time‑resolved transcriptomics identified rapid light‑induced genes, including all six MpBBX members, whose mutant phenotypes support this antagonistic model.
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 demonstrates that very long chain sphingolipids in the outer membrane leaflet interdigitate with inner‑leaflet phosphatidylserine, forming a vertical bridge that organizes PS nanodomains and enables auxin‑induced activation of the Rho‑GTPase ROP6. Disruption of sphingolipid biosynthesis disperses these nanodomains, impairing ROP6 signaling, cytoskeletal dynamics, and directional growth, highlighting interleaflet coupling as a key mechanism linking membrane asymmetry to plant signal transduction.
Nanoclustering of a plant transcription factor enables strong yet specific DNA binding
Authors: Arfman, K., Janssen, B. P. J., Romein, R., van den Boom, S., van der Woude, M., Jansen, L., Rademaker, M., Hernandez-Garcia, J., Ramalho, J. J., Dipp-Alvarez, M., Borst, J. W., Weijers, D., van Mierlo, C. P. M., Sprakel, J.
The study reveals that the Auxin Response Factor MpARF2 in Marchantia polymorpha forms nanoscopic clusters within the plant nucleus, representing a distinct mode of DNA binding distinct from monomeric/oligomeric binding and liquid phase-separated condensates. These nanoclusters provide high‑affinity, switch‑like, sequence‑specific DNA interaction, suggesting a novel mechanism for transcriptional regulation by TF nanoclustering.