Cadmium (Cd) is a toxic heavy metal for plants, and although its mechanisms of toxicity are well characterized, the regulatory networks underlying plant responses remain incompletely understood. This study investigated membrane-associated molecular responses linked to RBOHC and RBOHF during Cd stress through a label-free proteomics of root microsomal membrane fractions from Arabidopsis thaliana wild-type (WT), rbohC and rbohF plants. Proteomic analysis revealed pronounced genotype-dependent responses, with most Cd-responsive differentially expressed proteins (DEPs) being genotype-specific. The rbohF mutant displayed broader proteomic reprogramming than rbohC, whereas rbohC exhibited a comparatively attenuated response. Functional analyses indicated that Cd stress altered proteins associated with detoxification, ion transport, membrane trafficking, redox regulation and stress signaling. WT plants showed coordinated accumulation of glutathione S-transferases (GSTs), MATE/DTX transporters, ABC transporters and Ca²⁺-related proteins, whereas rbohC lacked GST accumulation and showed reduced abundance of the IRT1-CIPK23 module. In contrast, rbohF exhibited enhanced accumulation of several GSTs together with altered abundance of proteins associated with redox regulation, Casparian strip organization and ion homeostasis, including SBP1, CASP1 and PER64. Physiological analyses further revealed increased superoxide accumulation in rbohC roots under Cd stress, while GST activity differed among genotypes, supporting distinct antioxidant responses. Together with previous physiological evidence, these findings identify membrane-associated proteins and pathways associated with the contrasting Cd responses of rbohC and rbohF mutants. Overall, this work provides new insights into membrane-associated molecular responses linked to RBOHC and RBOHF during Cd stress and identifies candidate proteins for future functional studies on Cd tolerance.

BACKGROUND: Reactive oxygen species (ROS) are key signaling molecules in plant responses against biotic stresses, however their regulation must be controlled to prevent cellular damage. WRKY transcription factors (TFs) play role in regulating biotic stresses via oxidative stress-related signaling molecules; however, specific functions of WRKY62 and WRKY63 in oxidative stress signaling remain poorly understood. The paper aims to investigate the physiological and molecular function of WRKY62 and WRKY63 TFs in Arabidopsis thaliana under biotic stress induced by oligogalacturonides (OGs). MATERIALS AND METHODS: In this study, photosynthetic performance, ROS accumulation, activities of antioxidants and selected defense-associated transcriptional responses were evaluated in wrky62 and wrky 63 mutants along with wild-type (WT) plants of A. thaliana following OGs treatment. RESULTS: In silico studies depicted conserved WRKYGQK motif; while phylogenetic analysis formed two clades of WRKY TFs with orthologs. Promoter analysis showed that they contain environment-related, hormone-related and light-responsive elements. Treatment of 200 μM OGs to WT and wrky mutant plants increased proline, ROS, malondialdehyde accumulation, and increased activities of enzymatic and non-enzymatic antioxidants, with lesser increase in wrky mutants. These results support with their predicted regulatory roles in stress signaling. Furthermore, expression analysis showed upregulation of wall-associated kinase 1 and respiratory burst oxidase homolog D genes in WT plants compared to wrky mutants. CONCLUSION: These findings suggest that WRKY62 and WRKY63 are strongly associated with OGs-induced redox homeostasis and antioxidant responses under the tested experimental conditions and could serve as possible targets for enhancing stress tolerance in plants.

Salt stress disrupts plant physiology and reprograms specialized metabolism. APETALA2/ethylene-responsive factor (AP2/ERF) transcription factors integrate stress and hormone signals, but their roles in linking salt tolerance with phenylpropanoid metabolism in non-model alpine plants remain poorly understood. Here, we performed a genome-wide survey of AP2/ERF genes and functionally characterized a salt-responsive ERF in Herpetospermum pedunculosum. We identified 147 HpAP2/ERF genes, whose promoters were enriched in hormone- and stress-responsive cis-elements. Salt treatment induced widespread expression changes of HpAP2/ERF genes in roots, with HpERF144 among the most rapidly and strongly up-regulated members. Virus-induced silencing and ZYMV-mediated overexpression in H. pedunculosum, together with heterologous overexpression in Nicotiana tabacum, showed that HpERF144 positively regulates salt tolerance by modulating oxidative stress-related physiological responses. Moreover, HpERF144 silencing reduced, whereas overexpression increased, total lignin and total lignan accumulation in roots under salt stress. DAP-seq and dual-luciferase assays further indicated that HpERF144 associates with and activates selected phenylpropanoid- and cell wall-related promoters, including 4CL- and CCR-like targets. These findings identify HpERF144 as a link between salt stress signaling and phenylpropanoid-associated metabolic regulation in a lignan-rich medicinal plant.

BACKGROUND: Drought stress severely restricts grape growth and yield. is widely involved in plant developmental processes as well as responses to biotic and abiotic stresses. Nevertheless, existing studies of the gene have primarily focused on cold and disease resistance, leaving its potential function in drought response largely unexplored. Therefore, investigating the role of under drought conditions is crucial for understanding the molecular mechanisms of stress tolerance in grape and for breeding drought-resistant cultivars. METHODS: A gene was cloned from drought-resistant cv. 'Thompson Seedless', followed by sequence analysis, subcellular localization assay, and expression pattern analysis. A dual-target gene editing vector of was subsequently constructed and transformed into embryogenic calli of 'Thompson Seedless' via -mediated genetic transformation. Gene-edited and wild type (WT) grapes were subjected to drought treatment. The biological function of under drought stress was determined by observing the plant growth status and stomatal aperture, measuring the proline and malondialdehyde (MDA) contents, antioxidant enzyme activities, and the expression levels of drought-related genes. In addition, proteins interacting with VvERF105 were screened and verified using yeast two-hybrid, bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation (Co-IP) assays. Their interaction was further confirmed using phosphorylation assays. RESULTS: is a stress-responsive gene localized in the nucleus. It responds to drought, cold, and high-temperature stresses and may act downstream of the ABA signaling pathway. mutant grapevine plants exhibited reduced resistance to drought stress. The edited lines exhibited smaller stomatal apertures, lower proline content, higher MDA content, and lower antioxidant enzyme activities compared to WT plants under drought stress. The expression levels of , , , , , and were also significantly downregulated. VvSnRK1 was identified as an interacting protein of VvERF105 and interacts with it in a phosphorylation-independent manner. CONCLUSIONS: is a nucleus-localized stress-responsive transcription factor that positively regulates grapevine drought stress responses. Its disruption significantly reduces drought resistance. Moreover, it interacts with the kinase VvSnRK1 in a phosphorylation-independent manner to mediate drought stress signaling in grapevines. The aforementioned results provide valuable genetic resources for molecular breeding of grapevines with enhanced drought resistance.

Drought stress inhibits the growth and development of , and WUSCHEL-related homeobox (WOX) transcription factors participate in plant growth and abiotic stress responses. However, the mechanism of the WOX-regulated drought response in remains to be elucidated. Here, we identified the drought-responsive WOX transcription factor ArWOX11. Overexpression of weakened drought resistance in and . ArWOX11 directly binds to the promoter (protein phosphatase 2C) and transcriptionally represses its expression. ArWOX11 interacts with itself and a novel regulator at the plasma membrane (NRPM) protein, ArNRPM3, in plants. Meanwhile, coexpression of ArWOX11 and ArNRPM3 enhanced the water loss rate and suppressed expression more than ArWOX11 alone, while abscisic acid (ABA) alleviated their inhibitory effect on . Overall, this study elucidates the function of the ArWOX11-ArNRPM3 module in the drought response, providing insights into the action mechanism of WOX in plant drought adaptation.

Sucrose non-fermenting-1 (SNF1)-related protein kinase 2 (SnRK2) members are central regulators in the abscisic acid signaling pathway and orchestrate plant drought stress responses. However, the functional diversity and mechanistic specificity among SnRK2 family members in wheat remain underexplored. This study reports TaSnRK2.8-5A, a nucleus-localized SnRK2 member in wheat (Triticum aestivum L.), whose transcription level was significantly upregulated under 12-h drought stress, with a 2.6-fold increase in leaves and 3.6-fold increase in roots compared with control conditions (P < 0.05), indicating its prominent role in stress signaling. Protein-protein interaction analysis revealed that TaSnRK2.8-5A interacts with TaPP2C53-1A and TabZIP23-6D, forming a novel ABA signaling module that distinguishes it from previously characterized SnRK2 members in wheat, which typically function through distinct or broader partners. Overexpression of TaSnRK2.8-5A or TabZIP23-6D alleviated drought-induced growth inhibition, whereas TaPP2C53-1A acted as a negative regulator. Under drought treatment, TaSnRK2.8-5A-OE, TabZIP23-6D-OE, and TaPP2C53-1A-KO lines displayed plant biomass increases of 32-42% compared with wild type plants. Physiological analyses demonstrated that these transgenic lines showed improved photosynthetic efficiency (net photosynthetic rate increased by 41%), enhanced osmolyte accumulation (proline increased by 14-25%; soluble sugars increased by 20-40%; soluble protein increased by 12-25%), and maintained ROS homeostasis (MDA reduced by 10-19%; SOD activity increased by 16.7-22%) under drought conditions (P < 0.05). These results indicated that the TaPP2C53-1A/TaSnRK2.8-5A/TabZIP23-6D module integrates multiple physiological processes to coordinately enhance drought tolerance in wheat. Mechanistically, yeast one-hybrid and transcriptional activation assays indicated that TabZIP23-6D binds to the promoters of TaPIN1, TaP5CS4, and TaPOD2, thereby activating their expression under drought stress. This regulation enhances root activity, proline biosynthesis, and ROS scavenging, respectively. Through the screening of a large wheat germplasm collection, the specific haplotype TaSnRK2.8-5A-Hap1 was identified as conferring superior drought tolerance. Collectively, these findings establish that a signal module centered on TaSnRK2.8-5A enhances the response of wheat to drought stress in coordination with physiological processes, providing both mechanistic insights into SnRK2-mediated drought adaptation and valuable gene resources for molecular breeding of drought-tolerant wheat cultivars.

Peanut ( L.) production faces persistent threats from various infectious diseases. Planting healthy varieties with robust botanical defense networks is critical for minimizing future costs. Non-expressor of pathogenesis-related (NPR) regulators are involved in immune activation and act as key targets for deeper stress adaptation, and are thus promising targets for genetic enhancement. In this study, we characterized the peanut NPR4B protein and demonstrated its local subcellular binding to the nucleus. Ectopic overexpression of in significantly enhanced resistance to the necrotrophic pathogen and enhanced cold tolerance, as supported by quantitative and statistical analyses ( < 0.05). As regards underlying molecular events, Y2H (Yeast 2-Hybrid) analysis revealed a binding in vitro physical relation of AhPR2-like to AhNPR4B. This binding was demonstrated in vivo through BiFC (Bimolecular Fluorescence Complementation). These results suggest that the AhNPR4B-AhPR2-like complex may act as a key regulatory module associated with biotic and abiotic stress signaling, potentially contributing to broad-spectrum stress resistance. These findings provide foundational insights into the functional roles of and its interaction with in regulating stress resistance and support its potential as a candidate target for future genetic improvements to enhance stress resilience in peanuts.