Genetius

The authors created two Nicotiana benthamiana lines with CRISPR-mediated knockouts of two polyphenol oxidase genes, which exhibited slightly accelerated growth and retained normal transient GFP expression. These ppo-deficient plants produced leaf extracts that remained greener with markedly reduced enzymatic browning and protein crosslinking, leading to a nearly fourfold increase in yield and improved purity of a transiently expressed His‑tagged tomato protease. The study demonstrates that PPO depletion can enhance recombinant protein recovery from plant tissue.

The study demonstrates that calcium-dependent protein kinases NbCDPK4 and NbCDPK5 directly phosphorylate the NADPH oxidase NbRBOHB at Ser‑123, enhancing sustained ROS production during effector-triggered immunity in Nicotiana benthamiana. Constitutively active CDPKs also upregulate NbRBOHB transcription, and phosphorylation of Ser‑123 is amplified by Ca2+ influx triggered by an autoactive helper NLR (NRC4). These results define a NbCDPK‑NbRBOHB signaling module that links NLR activation to prolonged ROS bursts in ETI.

The study demonstrates that jasmonate (JA) enhances Arabidopsis thaliana responses to extracellular ATP (eATP) by upregulating the eATP receptor P2K1 and amplifying eATP‑induced cytosolic Ca²⁺ spikes and transcriptional reprogramming in a COI1‑dependent manner, whereas salicylic acid pretreatment suppresses these responses. These findings reveal a JA‑mediated priming mechanism that potentiates eATP signaling during stress.

The study establishes a transient Agrobacterium-mediated expression platform in Nicotiana benthamiana to produce glycosylated irregular monoterpenes by enhancing DMAPP biosynthesis through co‑expression of DXS, IDI, and HMGR. Engineering of plastidial and cytoplasmic pathways, including a bacterial cyclolavandulyl diphosphate synthase, led to the accumulation of six novel glucoside derivatives, reaching up to 6.6 µmol g⁻¹ fresh weight, the highest reported for plant‑based production.

The study characterizes a plasma membrane-localized, calcium‑permeable force‑gated channel named Rapid Mechanically Activated (RMA) in plants, using patch‑clamp and pressure‑clamp to elucidate its rapid activation, inactivation, and irreversible adaptation upon repeated mechanical stimulation. Kinetic modeling shows the channel functions as a pass‑band filter for frequencies between 10 Hz and 1 kHz, supporting its role in transducing high‑frequency mechano‑stimuli such as insect vibrations.

The authors present a protocol for the large‑scale isolation of RNA‑binding proteins cross‑linked to RNA, involving in vivo UV‑crosslinking, tissue lysis, fractionation, and organic‑solvent purification of RNA‑protein adducts for downstream proteomics analysis. Although developed with Nicotiana benthamiana leaves, the method can be adapted to other plant species and tissues.

The study visualizes subcellular dynamics following activation of the NRC4 resistosome, showing that NRC4 enrichment at the plasma membrane triggers calcium influx, followed by sequential disruption of mitochondria, plastids, endoplasmic reticulum, and cytoskeleton, culminating in plasma membrane rupture and cell death. These observations define a temporally ordered cascade of organelle and membrane events that execute plant immune cell death.

The study identified a suppressor mutation (sune42) in the Golgi-localized Ca2+ transporter CCX4 that alleviates the dominant‑negative root hair phenotype caused by the extensin‑less LRX1ΔE14 protein in Arabidopsis. Detailed Ca2+ imaging showed that LRX1ΔE14 disrupts tip‑focused cytoplasmic Ca2+ oscillations, a defect rescued by the sune42 mutation, highlighting the role of Golgi‑mediated Ca2+ homeostasis in root hair growth.

The study used Nicotiana benthamiana lines expressing fluorescent biosensors for PI4P and PI(4,5)P2 to visualize root colonisation by the pathogen Phytophthora palmivora and the mutualist fungus Funneliformis mosseae. Distinct phosphoinositide patterns distinguished the two interactions, and co‑colonisation induced recruitment of PI4P to pathogen haustoria, correlating with increased resistance, indicating dynamic remodeling of host membrane identity.

The study used transcriptomic and lipidomic profiling to investigate how chia (Salvia hispanica) leaves respond to short‑term (3 h) and prolonged (27 h) heat stress at 38 °C, revealing rapid activation of calcium‑signaling and heat‑shock pathways and reversible changes in triacylglycerol levels. Nearly all heat‑responsive genes returned to baseline expression after 24 h recovery, highlighting robust thermotolerance mechanisms that could inform improvement of other oilseed crops.