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 investigates the role of the chromatin regulator MpSWI3, a core subunit of the SWI/SNF complex, in the liverwort Marchantia polymorpha. A promoter mutation disrupts male gametangiophore development and spermiogenesis, causing enhanced vegetative propagation, and transcriptomic analysis reveals that MpSWI3 regulates genes controlling reproductive initiation, sperm function, and asexual reproduction, highlighting its ancient epigenetic role in balancing vegetative and reproductive phases.
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 investigates the role of two ATP-binding cassette transporters, MpABCG1 and MpABCG36, in the sequestration of specialized metabolites within oil bodies of the liverwort Marchantia polymorpha. Loss‑of‑function mutants displayed reduced accumulation of sesquiterpenes and, specifically for MpABCG1, decreased levels of bis‑bibenzyls, while oil‑body formation remained largely unaffected, indicating these transporters are essential for metabolite accumulation rather than organelle biogenesis.
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.
Discovery of tomato UDP-glucosyltransferases involved in bioactive jasmonate homeostasis using limited proteolysis-coupled mass spectrometry
Authors: Venegas-Molina, J., Mohnike, L., Selma Garcia, S., Janssens, H., Colembie, R., Kimpe, I., Jaramillo-Madrid, A. C., Lacchini, E., Winne, J. M., Van Damme, P., Feussner, I., Goossens, A., Sola, K.
The study applied limited proteolysis‑coupled mass spectrometry (LiP‑MS) to map JA‑protein interactions, validating known JA binders and uncovering novel candidates, including several UDP‑glucuronosyltransferases (UGTs). Functional omics, biochemical, enzymatic, and structural analyses demonstrated that two tomato UGTs glucosylate jasmonic acid, revealing a previously missing step in JA catabolism.
The study characterizes the single-copy S-nitrosoglutathione reductase 1 (MpGSNOR1) in the liverwort Marchantia polymorpha, showing that loss-of-function mutants generated via CRISPR/Cas9 exhibit marked morphological defects and compromised SNO homeostasis and immune responses. These findings indicate that GSNOR-mediated regulation of S‑nitrosylation is an ancient mechanism linking development and immunity in early land plants.