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 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 performed a meta‑transcriptomic analysis of over twenty drought versus control experiments in Vitis vinifera and two hybrid rootstocks, identifying a core set of 4,617 drought‑responsive genes. Using transcription factor binding motif enrichment and random‑forest machine learning, gene regulatory networks were built, revealing key regulators such as ABF2, MYB30A, and a novel HMG‑box protein. These regulators and network hierarchies provide candidate targets for breeding and biotechnological improvement of grapevine drought tolerance.
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.
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.
The study reveals that in the liverwort Marchantia polymorpha, the UV‑B photoreceptor MpUVR8 forms homodimers that monomerize and accumulate in the nucleus upon UV‑B exposure, activating COP1‑dependent growth inhibition, gene expression reprogramming, and UV‑absorbing metabolite production. MpRUP promotes redimerization of MpUVR8, acting as a negative regulator, while MpSPA also negatively modulates UVR8 signaling, indicating lineage‑specific diversification of UV‑B signaling components that originated over 400 Myr ago.
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 reveals that the Arabidopsis O-GlcNAc transferase SEC is essential for timely ABA‑induced stomatal closure and drought tolerance, with sec-5 mutants showing delayed closure and increased water loss, while SEC overexpression enhances responsiveness. SEC influences guard‑cell microtubule remodeling, as loss of SEC impairs microtubule reorganization and SEC directly interacts with tubulin α‑4, suggesting tubulin as a target of O‑GlcNAcylation.