Cell wall-degrading enzymes are viewed as extracellular virulence factors. However, their potential to act as immune elicitors and interact with intracellular immune receptors remains poorly defined. Here, we demonstrate that six conserved glycoside hydrolase 7 (GH7) family proteins from the fungal pathogen Verticillium dahliae have dual roles in virulence and immune activation. Deletion of each GH7 gene significantly impaired virulence without affecting fungal growth. The secretion of each GH7 protein was confirmed using yeast invertase and triphenyltetrazolium chloride assays. Live-cell imaging revealed that all GH7 proteins translocate into plant cells and localize to the cytoplasm, where they triggered canonical immune responses, including hypersensitive cell death, reactive oxygen species burst, and callose deposition, independent of their enzymatic activity. Genetic disruption of Nicotiana benthamiana Suppressor of G2 allele of Skp1 (SGT1) and Non-race specific Disease Resistance 1 (NDR1), but not Enhanced Disease Susceptibility 1 (EDS1), compromised GH7-triggered cell death, indicating that GH7-triggered cell death partially depends on a classical NLR-mediated signalling pathway. Thus, GH7s act as both virulence factors and unconventional intracellular effectors that activate effector-triggered immunity via the SGT1-NDR1 pathway. This reveals how fungal metabolic enzymes evolve effector-like properties and offers new insight into host-pathogen coevolution.

Seed dormancy is a vital adaptive trait that ensures germination occurs under favorable environmental conditions, significantly affecting crop yield and quality. However, the genetic regulation of dormancy remains poorly understood in peanut, limiting efforts to improve preharvest sprouting (PHS) resistance. In this study, we carried-out a genome-wide analysis of the DELAY OF GERMINATION 1 (DOG1)-domain-containing gene family in peanut and identified 35 members, comprising 12 DOG1-like and 23 TGA-related genes. Phylogenetic and structural analyses revealed clade-specific divergence, with AhDOG1-2 showing strong seed-preferential expression and responsiveness to abscisic acid (ABA). Subcellular localization assays showed that AhDOG1-2 localizes to both the nucleus and cytoplasm, and yeast transactivation assays confirmed its transcriptional activity. Functional validation showed that AhDOG1-2 overexpression in Arabidopsis enhances seed dormancy and increases ABA sensitivity. Furthermore, transgenic rice expressing AhDOG1-2 exhibited reduced PHS without significant agronomic penalties. Expression analysis revealed that these phenotypic changes were accompanied by reprogramming of genes involved in ABA and gibberellin (GA) pathways. These findings identify AhDOG1-2 as a conserved dormancy regulator with cross-species potential, providing the first functional characterization of DOG1 homologs in peanut and highlighting AhDOG1-2 as a promising molecular target for genetic improvement of dormancy and PHS resistance in crops.

Leaf intrinsic water use efficiency (iWUE) quantifies the trade-off between carbon assimilation and water loss in plants, and is constrained by leaf traits such as maximum carboxylation capacity (V) and stomatal conductance. Yet, the potential links of iWUE with leaf elementomes across different forest types remain unclear. Here, we analyzed iWUE (estimated by leaf carbon isotopes) variability and its associations with V, stomatal conductance (estimated by O enrichment in leaf dry matter above source water, ΔO), and leaf elementomes across 82 tree species from temperate, subtropical and tropical forests, and evaluated the effectiveness of leaf reflectance spectroscopy as an indicator of iWUE variability and trait-iWUE associations. Across species, V, ΔO, leaf mass per area (LMA) and leaf iron, nitrogen, sodium and manganese concentrations were the traits most strongly associated with cross-site iWUE variability. Furthermore, climatic factors (mean annual precipitation, mean annual temperature and climate moisture index) shaped trait-iWUE covariation by negatively linking leaf elements and positively with LMA, which affected iWUE more directly than indirectly via V and ΔO. Leaf reflectance spectroscopy accurately predicted iWUE (R = 0.83), and the trait-iWUE relationships derived from spectral modelling were consistent with those obtained through field measurements. These findings reveal strong linkages between the leaf elementomes and iWUE, and highlight the potential of reflectance spectroscopy for characterizing iWUE variability and trait-iWUE relationships, thereby improving process modelling of forest carbon and water cycles.

Unseasonal high temperatures caused by global warming adversely impact crop growth and yield. Post-translational modifications (PTMs) of proteins play critical roles in plant responses to environmental stresses. However, the PTM-based regulatory mechanisms of heat stress proteins remain poorly understood. In this study, we report that the histone deacetylase OsHDAC1 overexpression in rice enhances heat tolerance, whereas OsHDAC1 RNA interference (RNAi) lines exhibit increased heat sensitivity. OsHDAC1 physically interacts with OsHSA32, a plant-specific atypical heat-induced protein, while oshsa32 knockout mutants display reduced tolerance to heat. Mechanistically, OsHDAC1 stabilizes OsHSA32 via deacetylation, thereby promoting the heat tolerance of rice. Further analysis reveals that OsHDAC1-mediated deacetylation inhibits the interaction between OsHSA32 and the RING-type E3 ubiquitin ligase OsRFPH2-11, thereby preventing ubiquitin-proteasome-mediated degradation of OsHSA32. Collectively, our findings uncover a PTM-based regulatory mechanism by which deacetylation modulates the stability of a heat-induced protein to enhance thermotolerance in rice.

Transpiration restriction under high vapour pressure deficit (VPD), measured indoors with individual plants, increases water use efficiency (WUE). However, VPD is not the only factor driving transpiration, individual plants in a field rapidly become a canopy, and reports on the transpiration restriction versus WUE are scant. We analyzed the transpiration response to the evaporative demand (Penman-Monteith reference evapotranspiration, ET) and WUE in sorghum canopies outdoors. These responses showed no plateau at high ET in 47 genotypes. The slope of the resulting linear relationship over the whole range of ET showed a large genetic variability. Unexpectedly, this slope was positively correlated with WUE in experiments with high ET. Conversely, a (classical) negative correlation was observed under low ET. Genotypes with high WUE and response to ET allowed maximum light penetration into the canopy, via more erect leaf orientation. VPD in the canopy was also lower than in open air when the leaf area index reached 2.5-3. We interpret that higher WUE related to a larger proportion of plant photosynthesis being contributed by lower level leaves that received light and faced lower VPD than leaves exposed to air VPD. This study opens new opportunities, agronomic and genetic, to improve WUE.

The phyllosphere encompasses all above-ground plant parts, covering ~10 km and hosting as many as 10 microbial cells, yet its chemical ecology remains understudied compared to the rhizosphere. This review synthesizes recent advances in metabolite-mediated communication orchestrating phyllosphere microbiome assembly, function and host feedback. Leaf structural traits, host immune genes, developmental stage, and fluctuating environmental drivers create spatiotemporal chemical niches that filter incoming microbes. We then examine four major classes of plant-derived signals, including primary metabolites, secondary metabolites and phytohormones, with an emphasis on their dual functionality. Microbial feedback occurs through phytohormone synthesis/catabolism, volatile and soluble effectors and antimicrobial metabolites that collectively modulate plant immunity, growth and stress tolerance while structuring inter-microbial competition. These bidirectional exchanges form a dynamic network where plants and microbes continuously negotiate cooperation and conflict under diurnal and seasonal oscillations. We outline translational prospects, including probiotic foliar applications, metabolite priming and breeding for beneficial consortia, while identifying key challenges in signal attribution, microbiota stabilization and deciphering community-level crosstalk dynamics for sustainable crop protection.

Microbial fortification represents a promising approach for selenium biofortification in crops. Building on the previous discovery that Bacillus cereus SESY enhances selenium uptake in Brassica napus, this study employed an integrated multi-omics approach to investigate the mechanism by which B. cereus SESY enhances Se bioavailability in the Brassica napus rhizosphere. Inoculation with B. cereus SESY significantly increased selenium content in Brassica napus roots and shoots in calcareous soil by 42.9% and 21.5%, respectively, and increased the selenium content of shoots in yellow brown soil by 30.7%. B. cereus SESY promoted the transformation of residual Se into bioavailable forms and enriched bacterial taxa with high motility and Se-transforming capacity (e.g., Lysobacter, Rhodanobacter, Sphingomonas and Burkholderiaceae) in rhizosphere soil. Key genes of these bacteria involved in Se metabolism (e.g., trxA, narH, cysE, cysK, metB) and cell motility genes (e.g., FlgG, CheW, FliH) were up-regulated. Core rhizosphere metabolites such as N-formylmethionine and xanthine correlated strongly with enriched bacteria abundance and available Se. Joint application of these metabolites with enriched bacteria increased plant Se content by 144% and rhizosphere soil available Se by 13.4%. These results reveal a metabolite-mediated microbial network that enhances Se mobility and plant uptake, providing a novel strategy for microbiome-driven biofortification.

Lysine deficiency in staple crops like maize, rice, and wheat remains a major cause for global protein malnutrition, underscoring the urgent need for effective biofortification strategies. This review critically examines recent advances in enhancing lysine content, spanning conventional breeding and metabolic engineering to cutting-edge precision genome editing. While conventional breeding, exemplified by Quality Protein Maize, has improved lysine levels, it is often constrained by yield and quality trade-offs. Metabolic engineering strategies, including overexpression of lysine biosynthetic genes, suppression of catabolic genes, and modification of storage proteins, have achieved substantial lysine enrichment but face regulatory and consumer acceptance challenges due to their transgenic nature. The advent of CRISPR/Cas technology now enables precise, transgene-free editing of key enzymes such as DHDPS, AK, and LKR/SDH offering a powerful alternative, though concerns regarding off-target effects and pleiotropy remain. While integrating multi-omics with AI-driven predictive modelling can optimise metabolic flux for higher lysine yield, coupling next-generation genome editing with speed breeding offers a transformative route to develop high-lysine, high-yielding crops for sustainable nutritional security.

Cadmium (Cd) is a toxic metal that accumulates in plants to inhibit growth and enters the food chain to harm human health. Although Cd accumulation and tolerance in plants have been extensively analysed, their regulation is less understood. Here, we identify a stress-responsive receptor-like kinase (OsSRK) involved in rice Cd accumulation and tolerance. Our results show that OsSRK expression was strongly induced by Cd treatment. OsSRK overexpression decreased while its silencing or mutations increased both Cd accumulation and Cd-induced leaf chlorosis in rice. OsSRK is a close homologue of MULTIPLE SPOROCYTE 1 (MSP1), which controls sporogenic development with its TAPETUM DETERMINANT1 (TPD1)-LIKE 1 A (OsTDL1A) ligand. OsSRK interacts with OsTDL1B, an OsTDL1A homologue, in both yeast and plant cells. Like OsSRK, expression of OsTDL1B was induced by Cd treatment, and mutations of OsTDL1B enhanced both Cd accumulation and Cd-induced symptoms in rice. These results strongly support that OsTDL1B acts as a ligand for the OsSRK receptor kinase in Cd stress signalling. Comparative transcriptome and proteome profiling support that OsSRK plays a critical role in rice Cd accumulation and tolerance through the regulation of genes in Cd accumulation and oxidative stress responses.

Chilling is an important abiotic stressor that significantly affects cucumber production. Melatonin (MT) modulates chilling responses by interacting with multiple signalling molecules; however, the molecular link between MT and nitric oxide (NO) in cucumbers under chilling stress remains elusive. Herein, we found prolonged chilling stress induced the accumulation of endogenous NO, whereas overexpression of MT biosynthesis gene N-acetylserotonin methyltransferase (CsASMT), with higher endogenous MT content, significantly increased chilling tolerance of cucumbers with decreased accumulation of NO via upregulation of the relative expression of S-nitrosoglutathione reductase gene (CsGSNOR), accompanied by decreased membrane lipid peroxidation and reactive oxygen species (ROS) accumulation. Moreover, we identified a transcription factor zinc finger of Cucumis sativus 10 (CsZAT10), and found CsZAT10 could directly bind to the promoter of CsGSNOR. Furthermore, we found CsZAT10 overexpression enhanced cucumber chilling resistance by directly activating CsGSNOR expression to mediate NO homoeostasis, whereas the suppression of CsZAT10 obviously decreased the chilling tolerance and CsGSNOR expression in cucumber induced by MT. Overall, our results demonstrate that MT enhances chilling tolerance in cucumber by regulating the CsZAT10-CsGSNOR-NO module.

Flowering is essential for plants to reach the survival of species and flowering is influenced by many environmental factors. However, trithorax group (TrxG) mediated epigenetic modification mechanisms of Physalis grisea under low temperature on flowering remain largely unknown. Here, we report that TrxG core member ULTRAPETALA1 (PgULT1) inhibits flowering in P. grisea by interacting with Polycomb Group (PcG) member LIKE-HETEROCHROMATIN-PROTEIN 1 (PgLHP1) and transcription factor DNA-BINDING-ONE-FINGER 3.4 (PgDOF3.4) to regulate H3K4me3 and H3K27me3. PgULT1 overexpression delayed flowering, yet flowering was relatively promoted under low temperatures. Similarly, PgDOF3.4 confers delayed flowering by transcribing PgULT1, PgLHP1, and FLOWERING LOCUS C (PgFLC). Protein interaction assays indicated that PgULT1, PgDOF3.4 and PgLHP1 can interact with each other, enhance PgFLC transcription and suppress FLOWERING LOCUS T (PgFT) transcription. Genetic evidence demonstrated that PgULT1 and PgLHP1 inhibit flowering by depositing H3K4me3 and H3K27me3 at the PgFLC and PgFT transcription start sites, respectively. PgULT1, PgDOF3.4 and PgLHP1 expression are suppressed under low temperatures, leading to reduced H3K4me3 and H3K27me3 modifications on PgFLC and PgFT promoters, thereby promoting flowering. Collectively, the functional interactions between epigenetic modifiers and transcription factors reveal a cooperative mechanism between TrxG and PcG to respond to low temperatures and promote flowering in P. grisea.

As plants grow taller, increasing conductive pathlength imposes hydraulic resistance, challenging the maintenance of water transport to leaves. While tip-to-base conduit widening along the stem helps mitigate this resistance, theoretical models and empirical data suggest that stem widening alone is insufficient to fully compensate. Here, we explore whether leaf length could contribute to maintaining hydraulic conductance by influencing vessel diameters in the stem. Across a diverse set of angiosperm species, we found that leaf length strongly predicts vessel diameter at the petiole base, and that petiole vessel diameter, in turn, scales positively with vessel diameter at the twig tip. These relationships imply that longer leaves are associated with wider conduits in the stem, potentially boosting stem-wide permeability. Simple fluid dynamic models show that the steep rate of conduit widening in angiosperm leaves plausibly buffers the resistance costs of increased leaf length. Because vessel diameter scales with the fourth power of conductance, modest increases in leaf length, and thus stem conduits, could lower the resistance not buffered by conduit widening in the stem. Leaf length during height growth may serve as a key mechanism in maintaining hydraulic supply, complementing conduit widening in the stem.

OsAAI1 belongs to the HPS_like subfamily of the AAI_LTSS superfamily, yet the molecular mechanism by which it regulates root development under osmotic stress remains unclear. In this study, we found that overexpressing OsAAI1 significantly promoted rice root system growth. Specifically, the primary root length, lateral root number, lateral root density and adventitious root count in the overexpression line (OE19) markedly exceeded those in the wild type (ZH11) and the osaai1 mutant. Consistent with this phenotypic enhancement, the root IAA content in OE19 was substantially higher than in ZH11 and osaai1. We further demonstrated that exogenous IAA application compensated for the root growth defects in the osaai1 mutant. Under PEG-induced osmotic stress, OE19 exhibited the most extensive and densely distributed root system, and exogenous IAA also rescued the inhibited growth of the osaai1 mutant. Mechanistically, we identified an interaction between OsAAI1 and the MADS-box transcription factor OsMADS25. This interaction enhanced the transcriptional expression of two key osmotic stress tolerance genes, LAX1 and OsBAG4. Furthermore, it upregulated the auxin biosynthesis gene OsYUC4 while suppressing the auxin inhibitory factor OsIAA14. This coordinated gene regulation promotes the auxin signalling pathway, thereby stimulating root growth and enhancing osmotic stress tolerance. Collectively, our findings indicate that OsAAI1 and OsMADS25 fulfil critical functions in rice osmotic acclimation by orchestrating downstream gene expression and modulating the auxin pathway.

Tartary buckwheat, valued for its nutritious and medicinal quercetin. Following two independent domestication events, distinct quercetin accumulation patterns have emerged between the southwestern (SL) and northern (NL) landrace populations. However, the genetic mechanisms underlying these metabolic divergences remain elusive. Here, we identified the transcription factor FtNAC2 through genome-wide association study (GWAS) of quercetin content in 480 accessions of Tartary buckwheat. Haplotype analysis identified two single nucleotide polymorphisms (SNPs) in the FtNAC2 promoter that defined three major haplotypes, with higher promoter activity and gene expression observed in Hap2. Functional characterization revealed that FtNAC2 promotes quercetin accumulation in Tartary buckwheat hairy roots and potentially serves as a multifunctional regulator influencing both drought tolerance in buckwheat and seed size in Arabidopsis. Transcriptome co-clustering and pull-down mass spectrometry (MS) indicated FtNAC52 as a potential regulatory partner of FtNAC2. DNA affinity purification sequencing (DAP-seq) and quantitative reverse transcription PCR (qRT-PCR) analyses demonstrated that FtNAC2 promoted quercetin biosynthesis by upregulating FtF3'H and FtF3'5'H genes. Collectively, our results elucidated how FtNAC2 influences quercetin content variation in Tartary buckwheat, providing molecular insights into the differential quercetin accumulation between cultivated populations.

Leaf spot disease, caused by the fungal pathogen Alternaria alternata f. sp. mali, poses a severe threat to apple production. Pathogenesis-related (PR) genes are crucial for plant immunity, yet their regulatory networks remain poorly understood. Here, we report that MdDREB2A, a transcription factor known for its role in abiotic stress, negatively regulates apple resistance to A. alternata by suppressing the expression of MdPR10 genes. We demonstrated that MdDREB2A overexpression plants exhibited increased susceptibility to A. alternata infection, whereas its knockdown conferred enhanced resistance. Based on DAP-seq analysis, we identified three MdPR10 genes as direct targets of MdDREB2A. This direct repression was confirmed by ChIP-qPCR, EMSA, and dual-luciferase assays, which showed that MdDREB2A binds to the promoters of MdPR10s to inhibit their transcription upon pathogen infection. Furthermore, functional studies revealed that MdPR10 proteins possess antifungal activity, and their overexpression enhanced resistance in apple leaves. Consequently, in MdDREB2A overexpression plants, the suppression of MdPR10s leads to diminished antifungal resistance. This study establishes MdDREB2A as a negative regulator of defense against A. alternata in apple, which operates by repressing the expression of three pathogenesis-related genes, thereby proposing a new strategic direction for developing resistant apple cultivars.

Insect-derived molecular cues can prime plant defences against herbivore attack. The genes that are sensitive to priming, and how their expression changes on the scale of days, have not been fully resolved. Moreover, priming may affect interactions with insects that are not the source of the priming cue. We primed tall goldenrod (Solidago altissima) plants by exposure to the volatile emission of a specialist herbivore, the goldenrod gall fly (Eurosta solidaginis) then subjected the plants to 48 h of herbivory from an unrelated generalist, corn earworm (Helicoverpa zea). Using RNA sequencing, we identified transcriptome-wide gene expression patterns between exposed and unexposed plants. We identified biotic stress-associated genes that were more abundant during herbivory in primed plants, including defence-related transcription factors, thaumatin-like receptors and chitinases. We observed a surprising rise and fall in expression of hundreds of defence-related genes in a 48-h phase in primed damaged plants only. Our results support the hypothesis that primed defences are stronger than typical induced defences and suggest that primed defences target herbivores in the short term. We show that the threat cue from a specialist can affect plant defences against an unrelated herbivore.

Photoperiod regulates flowering time and maturity in soybean, thereby determining yield performance and latitudinal adaptation. However, the molecular network through which photoperiod regulates flowering remains incompletely elucidated. Here, we identify two BBX family transcription factors, BBX32a and BBX32b, that act as positively regulators flowering under long-day (LD) conditions in soybean. We demonstrate that BBX32a and BBX32b can form both homologous and heterologous dimers. The bbx32a and bbx32b mutants exhibit significantly delayed flowering compared to wild-type W82. However, the bbx32a bbx32b double mutants flower at a similar time to the single mutants, suggesting that the BBX32a-BBX32b heterodimer plays a central role in regulating soybean flowering. E3 and E4 upregulate the transcription of BBX32a and BBX32b, which repress E1 transcription to promote flowering under LD conditions. Genetic evidence demonstrates that BBX32a and BBX32b regulate flowering time, completely dependent on functional E3, E4 and E1 family genes. Four haplotypes of BBX32a were identified in 1295 soybean accessions; BBX32a exhibits significantly reduced nuclear accumulation relative to BBX32a. The BBX32a allele is predominantly fixed in cultivated soybeans, whereas BBX32a and BBX32a alleles remain largely unexploited. Collectively, our findings identify novel genetic targets for developing novel soybean cultivars adapted to high-latitude regions, thereby maximising yield potential.

The development of commercially available porometers has allowed for higher throughput measurement of stomatal conductance, but a body of evidence has suggested a persistent positive bias in their measurements relative to "reference" measurements from instrumentation based on infra-red gas analysis. We compiled a data set comprised of 25 angiosperm species, across a range of field conditions and found that the LI-COR LI-600, an open flow-through porometer, produced an exponentially increasing bias relative to the LI-COR LI-6800 infra-red gas analyser-based instrument in response to increasing stomatal conductance and decreasing relative humidity. This bias was minimal at lower stomatal conductance (below roughly 0.25 mol m s ), but was pronounced for larger values. We hypothesised that this bias is the result of the assumption of a constant air temperature throughout the flow stream used by the instrument software to estimate stomatal conductance from raw sensor measurements. We relaxed this assumption, and applied psychrometrics to augment the typical gas exchange equations with an additional energy balance constraint to solve for the temperature change throughout the air flow stream. We found that including this temperature difference corrects the computed transpiration and stomatal conductance values, and brings the porometer measurement into agreement with that of the infra-red gas analysis-based system. Software is provided to apply the correction to LI-600 output files. For future instrument design iterations, explicit measurement of temperature variation in the flow stream provides a potential opportunity for improvement in measurement accuracy at high stomatal conductance.