Genetius

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.

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 examined the pre‑activation state of the rice NLR pair Pik‑1 (sensor) and Pik‑2 (helper) when transiently expressed in Nicotiana benthamiana leaves. Both wild‑type and engineered Pik‑1 variants constitutively associate with Pik‑2 to form ~1 MDa hetero‑oligomeric complexes that localize to the plasma membrane in the absence of effector. These results reveal that some NLRs exist as pre‑assembled membrane‑associated complexes prior to pathogen perception.

The study demonstrates that subfamily I ethylene receptors form the core ethylene‑sensing module and act epistatically over subfamily II receptors, uniquely possessing Ca2+‑permeable channel activity that drives ethylene‑induced cytosolic calcium influx. This reveals a mechanistic link whereby subfamily I receptors integrate hormone perception with calcium signaling in plants.

The study uncovers how arbuscular mycorrhizal (AM) fungi induce lateral root formation in maize by activating ethylene‑responsive transcription factors (ERFs) that regulate pericycle cell division and reshape flavonoid metabolism, lowering inhibitory flavonols. It also shows that the rhizobacterium Massilia collaborates with AM fungi, degrading flavonoids and supplying auxin, thereby creating an integrated ethylene‑flavonoid‑microbe signaling network that can be harnessed to improve nutrient uptake and crop sustainability.

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 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.