Plant, Cell & Environment

Light wavelengths modulate plant growth, metabolism, and physiology. Amaranthus, a C4 underutilized climate resilient crop with promising nutritional properties remained unexplored in terms of metabolite enrichment under monochromatic light wavelengths of visible spectrum. In current study, two cultivars of Amaranthus tricolor (green and red) were exposed to seven light regimes of photosynthetically active radiation (PAR; 400-700 nm): deep blue, blue, green, amber, red, deep red, far red, and their metabolic responses were captured using Gas Chromatography-Mass Spectrometry. The metabolic analysis revealed wavelength-specific reprogramming in the levels of organic acids, sugars, amino acids, fatty acids as well as phenolics. In both the green and red Amaranthus, branched-chain amino acids and phenylalanine, which are nutritionally essential, were significantly elevated under far-red light. While the phenolics such as caffeic acid and ferulic acid were elevated under green and deep blue light respectively in green Amaranthus, amber light wavelengths enhanced these phenolics in red Amaranthus. The study highlighted cultivar-specific metabolic rewiring triggered by specific wavelengths. Altogether, these findings provides insights into metabolic adaptation and demonstrate the ability of light wavelength to specifically enrich the targeted metabolite of nutritional relevance in Amaranthus. It offers strategies to improve the nutritional value of crops in controlled agriculture systems. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=167 HEIGHT=200 SRC="FIGDIR/small/714947v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1a4477dorg.highwire.dtl.DTLVardef@518550org.highwire.dtl.DTLVardef@7682dorg.highwire.dtl.DTLVardef@4876e2_HPS_FORMAT_FIGEXP M_FIG C_FIG

Photosynthesis accounts for most of the final grain yield in rice, making improvements in radiation use efficiency (RUE) a key strategy for enhancing productivity. Agronomically, RUE is defined as the biomass produced per unit of total solar radiation or photosynthetically active radiation intercepted by the canopy. However, the interaction between carbon and nitrogen metabolism plays a critical role in determining plant growth and grain yield. Assimilated nitrogen is required for the synthesis of photosynthetic pigments and enzymes, while the reduction of nitrate (NOLL) and nitrite (NOLL), as well as the assimilation of ammonium (NHLL), depend on the reducing power and carbon skeletons generated by photosynthesis. In this study, two high-yielding rice (Oryza sativa) cultivars--an indica-type (El Paso 144) and a japonica-type (INIA Parao) were subjected to two nitrogen treatments (3 mM and 9 mM NOLL/NHLL) and two light intensities (850 and 1500 mol mL{superscript 2} sL{superscript 1}). A strong interaction between light intensity and nitrogen metabolism was observed, with contrasting responses between subspecies. These differences reflect a coordinated regulation of carbon assimilation and primary nitrogen metabolism. The results provide new insights into the metabolic strategies underlying nitrogen compound accumulation under variable irradiance. Such knowledge is essential for improving nitrogen fertilizer use efficiency and yield performance in elite rice genotypes cultivated under commercial field conditions.

Belowground, plants are exposed to a wide range of biotic stresses that vary in severity and nature, including tissue damage, disruption of vascular connectivity, and depletion of assimilates. How plants adapt their root systems to cope with different types of belowground biotic stresses is not well known. In this paper we compare above- and belowground plant adaptations to three nematode species with distinct tissue migration and feeding behaviours to study mechanisms underlying tolerance to different types of biotic stresses. We monitored both green canopy growth and changes in root system architecture of Arabidopsis inoculated with Pratylenchus penetrans, Heterodera schachtii, and Meloidogyne incognita. This revealed three distinct phases in aboveground plant responses: (i) initial growth inhibition associated with host invasion and tissue damage, (ii) persistent growth reduction associated with nematode sedentarism, and (iii) late growth stimulus in more advanced stages of infection. Specific adaptations in the root systems further revealed fundamentally different stress coping strategies. Tissue damage and intermittent feeding by P. penetrans in the root cortex did not induce significant changes in root system architecture. Tissue damage to the root cortex and prolonged feeding on host vascular cells by H. schachtii induced secondary root formation compensating for primary root growth inhibition. Prolonged feeding on host vascular cell by M. incognita alone did not induce secondary root formation, but was accompanied by typical local tissue swelling instead. Our data suggest that local secondary root formation and tissue swelling are two distinct compensatory mechanisms underlying tolerance to sedentarism by root-feeding nematodes. HighlightHow plants utilize root system plasticity to cope with different types of biotic stresses by root feeding nematodes remains largely unknown. Here, we report on specific adaptive growth responses in Arabidopsis roots to three nematode species, Pratylenchus penetrans, Heterodera schachtii, and Meloidogyne incognita, with fundamentally different strategies for host invasion, subsequent migration through host tissue, and feeding on host cells.

In cereal-legume intercrops, weed suppression is primarily driven by cereals, whose competitiveness is shaped by trait plasticity--morphological adjustments in response to the intercrop environment. However, how individual cereal traits respond plastically and contribute to system performance remains unclear, hampering improvements through breeding or system design. We combined field experiments with functional-structural plant modelling to quantify plastic responses of four cereal traits (tiller number, tiller angle, specific leaf area (SLA), and specific internode length (SIL)) and their effects on weed suppression and crop productivity. Field measurements revealed plasticity in tiller number, tiller angle, and SIL between sole crops and intercrops, while SLA showed minimal differences. Simulations showed that intermediate tiller numbers resulted in the strongest weed suppression and highest productivity, indicating an optimum, while more horizontal tillers suppressed weeds slightly better than vertical ones. Weed suppression increased with higher SLA values, while SIL showed a saturating response, increasing to intermediate SIL values and plateauing thereafter. In simulations with short-statured cereal phenotypes (low SIL), the reduction in cereal weed suppression was compensated by the legume component. This study demonstrates how FSP modelling can be used to investigate trait plasticity mechanisms and generate testable hypotheses about trait effects in complex intercrop systems. HighlightCereal trait plasticity shapes weed suppression in cereal-legume intercrops, with distinct response patterns per trait, while legumes can compensate for weakly competitive cereals, suggesting balanced competition over cereal dominance.

Malate and fumarate constitute a significant transient carbon stock that is dynamically synthesized during the photoperiod. These organic acids are diurnally stored and remobilised from the vacuole, and they have a key role in the cellular metabolic regulation. This function is well known in C4 and CAM plants. However, in C3 species that are the majority of terrestrial plants, the importance of the vacuolar accumulation/release and its influence on plant growth is still an open question. In Here we addressed this issue generating multiple knockout mutants in Arabidopsis thaliana lacking vacuolar anion channels of the Aluminium-Activated Malate Transporter (ALMT) family, to impair malate and fumarate transport to the vacuole. We show that in these mutants reducing vacuolar transport of malate and fumarate in mesophyll cells leads to a dramatic growth impairment. Metabolic and fluxomic analysis revealed that vacuolar malate and fumarate transport influences plant carbon and nitrogen metabolism as well as cellular pH and ionic homeostasis. In conclusion, our results show that the transport organic acids like malate and fumarate across the vacuolar membrane is essential for plant growth in a C3 plant too. These results establish the importance of the vacuolar pools of malate and fumarate in plant metabolism.

Previously, the chitin receptor-interacting protein kinase LIK1 (LysM receptor kinase 1/CERK1-interacting kinase) was shown to play an important role in regulating chitin signaling and plant defense. A limited proteolysis proteomics study revealed several LIK1-derived peptides that showed differential abundance between ATP-treated and mock-treated Arabidopsis samples, suggesting a possible involvement of LIK1 in extracellular ATP (eATP) signaling. To explore this possibility, LIK1 mutants were obtained and examined for their response to ATP. The results showed that mutations in LIK1 significantly reduced the expression of eATP-responsive genes. In addition, LIK1 was found to interact with the eATP receptor P2K1 and to be phosphorylated by it. The LIK1 protein was localized to the plasma membrane and its gene expression appeared to be ubiquitous. Collectively, these findings indicate that LIK1 not only contributes to chitin signaling but also participates in eATP signaling, highlighting its potential role as a shared component in multiple signaling pathways to regulate plant responses to diverse internal and external cues.

While most plant lineages are pigmented by anthocyanins, several families in the Caryophyllales represent a major exception by showing a replacement of anthocyanin pigmentation by betalain pigmentation. The mutual exclusion of anthocyanins and betalains at the family level has been well established for over 50 years and has been mechanistically explained. Chenopodiaceae are a betalain-pigmented lineage lacking a key anthocyanin biosynthesis gene and lacking the key activating transcription factor of the anthocyanin biosynthesis. A publication by Zhang et al., 2024 claims that anthocyanins would be responsible for the red pigmentation in leaves of Chenopodium quinoa. Here, we assessed this study and reanalyzed the RNA-seq datasets generated in this study to demonstrate that there is no evidence for anthocyanin biosynthesis, but activity of the betalain and carotenoid biosynthesis could explain the observed pigmentation of quinoa leaves.

BackgroundPlants are exposed to various environmental challenges. With ongoing climate change, droughts and insect outbreaks are expected to become more frequent. Thus, a better understanding is needed of how different plant species respond to such single and combined challenges. This study investigated common versus species-specific responses to environmental challenges in three perennial plant species of different growth forms and whether responses differ intraspecifically among accessions. Clones of different accessions of the herbaceous species Tanacetum vulgare, the woody vine Solanum dulcamara, and the tree Populus nigra were subjected to similar control, herbivory, drought, and combined (drought and herbivory) treatments for the same periods. After the exposure, concentrations of foliar phytohormones and various morphological traits were measured. ResultsAcross all species, several foliar phytohormones and one of ten morphological traits responded consistently to the environmental challenges. Jasmonoyl-isoleucine was induced by herbivory and the combined treatment, abscisic acid (ABA) by drought and the combined treatment, and indole acetic acid by the combined treatment in all species. Root mass remained unchanged in all species. However, structural equation models (SEMs) revealed a shared regulatory pathway across species in which ABA connected treatment and root mass, indicating a common hormonal response potentially linking challenges to growth responses. Despite these common patterns, species-specific responses were pronounced. In P. nigra, a unique induction of salicylic acid was found under the combined treatment, while aboveground mass and root-shoot ratio remained unaffected by any treatment, in contrast to the other two species. Species-specific SEMs further indicated distinct phytohormone-mediated pathways underlying morphological variation. Phenotypic plasticity reflected these species-specific patterns, with none of the phytohormones or morphological traits exhibiting uniform plasticity across species. Intraspecific variation further shaped responses, as phytohormone and morphological trait plasticity depended on accession, indicating substantial accession-specific plant responses. ConclusionsOur results indicate that some responses to comparable challenges may be conserved across species, while others are species-specific. The combined treatment elicited the most pronounced responses, and such complex responses may become more frequent under current global change. Our study highlights that comprehensive understanding of plant responses requires systematic comparisons at both interspecific and intraspecific scales.

Drought is a major constraint on the productivity of durum wheat across Mediterranean and North African regions. To elucidate the mechanisms underlying drought resilience, we employed a combination of scenario-controlled phenomics and flag leaf transcriptomics across ten durum wheat genotypes. These included the Tunisian landraces Chili and Mahmoudi, seven breeding lines, and the reference cultivar Svevo. The plants were grown to maturity under well-watered or long-term drought conditions in pots and rhizotrons, enabling a comprehensive assessment of growth, yield components, root architecture, physiological traits, and reaction norm plasticity. Drought markedly reduced performance, yet Chili and Mahmoudi consistently maintained superior biomass, grain number and intrinsic water use efficiency (iWUE). This was supported by balanced C/N allocation, strong osmotic adjustment, and the ability to sustain robust root systems under stress, albeit through partly divergent physiological strategies. Transcriptomic profiling revealed highly genotype specific responses, with drought tolerance unrelated to the number of differentially expressed genes. Instead, the landraces displayed distinct regulatory programs involving mainly photosynthesis protection, ABA-related transporters, osmotic adjustment pathways, and stress-responsive transcription factors. These mechanistic insights identify actionable physiological and molecular determinants of drought plasticity and provide high value targets for accelerating the breeding of climate resilient durum wheat. HighlightsIntegrated phenomics and transcriptomics revealed landrace-specific physiological and molecular mechanisms enabling superior drought resilience and identifying actionable targets for durum wheat improvement.

The chloroplast NADH dehydrogenase-like (NDH) complex mediates cyclic electron transport (CET) around photosystem I (PSI) and contributes to photosynthetic regulation and photoprotection under various environmental stresses. Although NDH function has been extensively characterized under controlled conditions, NDH-deficient mutants often show only subtle phenotypes in such environments, leaving its physiological importance under naturally fluctuating field conditions poorly understood. Here, we evaluated growth, yield, and photosynthetic performance of NDH-deficient rice cultivated under field conditions. Mutant plants exhibited reduced biomass accumulation and grain yield compared with wild type. Detailed physiological analyses revealed that NDH deficiency markedly decreased PSI electron transport and CO2 assimilation, particularly under low temperature and sub-saturating irradiance. At moderate and high temperatures, reductions in carbon fixation were largely confined to low-light conditions, whereas at low temperatures, impairment extended across nearly the entire light response range. Under repetitive fluctuating light regimes, NDH-deficient plants showed progressive declines in photosynthesis accompanied by a selective decrease in PSI photochemical capacity without changes in PSII maximum efficiency, indicating PSI-specific photoinhibition. These findings demonstrate that NDH-dependent CET plays a crucial role in sustaining photosynthetic efficiency and crop productivity in dynamic field environments by stabilizing PSI redox balance and maintaining long-term carbon gain. Summary StatementNDH-dependent cyclic electron transport supports photosynthesis and yield in field-grown rice by maintaining PSI function under fluctuating light, low temperature, and sub-saturating irradiance.

Improving photosynthesis is a promising approach to enhance sugar beet productivity. However, genetic variation in leaf photosynthesis and its relationship with disease resistance remain underexplored. We evaluated 98 sugar beet genotypes representing different breeding categories, including commercial F1 hybrids, seed-parent lines, and pollinator lines, in Hokkaido, northern Japan. Leaf gas exchange was measured during early growth under field conditions around the infection period of Cercospora leaf spot (CLS). To account for fluctuating irradiance during large-scale phenotyping, we applied a multilevel mixed-effects light-response model to estimate genotype-specific photosynthetic characteristics. Substantial genotypic variations in photosynthetic characteristics were detected. F1 hybrids exhibited higher photosynthetic capacity than breeding lines, whereas differences among breeding categories were unclear due to large within-category variation. Some breeding lines exhibited photosynthetic rates higher than those of hybrids, indicating exploitable genetic resources within the present genetic panel. We did not detect statistically significant trade-off between leaf photosynthesis and CLS resistance among 98 genotypes; in a subset of 19 genotypes analysed in detail, the relationship was even synergistic. Our results highlight the genetic diversity of leaf photosynthesis and its category-dependent structure, and suggest that selection for enhanced photosynthesis can proceed without substantial trade-off with CLS resistance. HighlightLeaf photosynthesis of 98 sugar beet genotypes showed significant genetic variation and dependence on breeding category. Active photosynthesis incurred minimal trade-off with Cercospora leaf spot resistance.

O_LIHydroxycinnamic acid amides (HCAAs) are phenylpropanoid-derived metabolites with known antimicrobial and structural roles in plant defence against pathogens. However, their contribution to mechanical wound responses remains unclear, especially in terms of tissue regeneration and signalling. C_LIO_LIHere, we used tomato transgenic plants overexpressing the tyramine hydroxycinnamoyl transferase (THT), the key biosynthetic enzyme for HCAA production, to investigate the role of HCAAs in wound-induced responses, combining targeted metabolite profiling, gene expression, confocal microscopy, antioxidant assays, and volatile analyses. C_LIO_LIWe show that THT overexpression enhances wound-induced accumulation of HCAAs, promoting vascular lignification, suberization, callose deposition, and increased regeneration capacity. Additionally, 35S::THT plants display a distinct VOC profile that modulates defence gene expression in neighbouring wild-type plants, even in the absence of injury. C_LIO_LIThese results identify THT as a key regulator of structural reinforcement and defence priming after mechanical damage. Our findings highlight a novel role for HCAAs in wound healing and interplant signalling, with potential applications for improving crop resilience to mechanical stress. C_LI

Napier grass (Cenchrus purpureus syn. Pennisetum purpureum), a perennial C4 forage and bioenergy crop, exhibits strong drought resilience, yet the integrative mechanisms underlying this tolerance remain incompletely understood. This study examined physiological, hydraulic, and metabolic responses of four Napier grass cultivars under PEG-induced osmotic stress and progressive soil water deficit. Drought significantly increased the root-to-shoot ratio, indicating preferential biomass allocation to roots, which supported maintenance of shoot growth and tissue water status. All cultivars showed an approximate twofold increase in water-use efficiency (WUE) under water deficit, with cv2 and cv7 displaying superior performance. Upregulation of aquaporin genes (PIP2;2 and PIP2;3) suggested active hydraulic regulation that sustained carbon assimilation under reduced transpiration. Metabolic profiling revealed pronounced root-centered osmotic adjustment, including accumulation of galactinol, myo-inositol, raffinose family oligosaccharides, proline, and several amino acids. Enhanced expression of the galactinol synthase gene confirmed activation of raffinose biosynthesis pathways. Genotypic variation highlighted cv2 as particularly drought resilient. Rapid post-stress regrowth further underscored the importance of perennial root persistence. In conclusion, drought tolerance in Napier grass arises from coordinated hydraulic resilience, osmotic adjustment, and C4 photosynthetic efficiency, supporting its suitability for forage and bioenergy production in water-limited environments. SignificantThis study shows drought tolerance in Napier grass relies on root-driven hydraulic and metabolic regulation with efficient water-use efficiency, rather than avoidance, and that PEG responses predict field performance.

Understanding the regulation of enzyme activity involved in photosynthesis is essential for engineering enhanced carbon fixation in crops. In C4 plants, the enzyme phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is one of the most abundant leaf enzymes and plays an essential role in photosynthetic carbon dioxide (CO2) fixation. The enzyme also plays a key role in central metabolism (e.g., providing intermediates to the citric acid cycle) and therefore must be highly regulated to coordinate its activity. The regulation of PEPC activity can occur allosterically by glucose 6-phosphate activation and malate inhibition, which is in part influenced by reversible phosphorylation. A specific light-dependent phosphorylation of PEPC at an N-terminal serine residue by the PEPC-protein kinase (PEPC-PK) can regulate its sensitivity to this allosteric regulation. However, the impact of this PEPC phosphorylation has not been tested in a C4 crop. Therefore, we created PEPC-PK mutant lines in Zea mays to assess the impact of PEPC phosphorylation on its allosteric regulation, photosynthesis, and growth. While the maximum PEPC activity was unchanged, PEPC in the PEPC-PK mutant plants was not phosphorylated under light and was more sensitive to malate inhibition. However, gas exchange, electron transport, and field biomass analyses showed no differences in the PEPC-PK mutant plants. These results demonstrate that in Z. mays PEPC phosphorylation affects enzyme sensitivity to malate in vitro but does not substantially alert photosynthetic performance or growth under field conditions suggesting additional regulation of PEPC activity in planta.

Photosynthetic electron transport is mediated by several protein supercomplexes that are spatially arranged in the thylakoid membranes of chloroplasts. The chloroplast NADH dehydrogenase-like (NDH) complex is part of the photosynthetic alternative electron transport (AET) chain, which reduces the plastoquinone (PQ) pool using reduced ferredoxin as a substrate. This NDH complex is associated with photosystem I (PSI) and mediates a portion of AET in stroma lamellae, whereas photosystem II (PSII) is concentrated in grana stacks. This study presents the findings regarding post-illumination chlorophyll fluorescence increase (PIFI), a protein crucial for regulating AET via the NDH pathway. A marked increase in NDH activity and a reduction in the PQ pool in the dark were observed in PIFI-deficient mutant strains (g-pifi) generated by genome editing. Blue native PAGE analysis indicated that PIFI was associated with the NDH-PSI supercomplex in the wild type, and the NDH complex was dissociated from PSI in the g-pifi mutants. Additionally, the g-pifi mutants exhibited a decrease in the maximum quantum yield of PSII (Fv/Fm). Notably, Fv/Fm was restored in a double mutant harboring both g-pifi and NDH-deficient pnsl1 mutations, demonstrating that deregulated NDH activity in g-pifi causes downregulation of PSII efficiency. However, the lower Fv/Fm was not observed in a mutant lacking thioredoxin m4 (trxm4), which showed deregulated NDH activity but maintained the NDH-PSI supercomplex. These data suggest that PIFI stabilizes the NDH-PSI supercomplex and maintains the spatial localization of PQ reduction via AET in thylakoid membranes, which is essential for the proper functioning of PSII.

Invasive plants can reshape ecosystems by altering soil biogeochemistry and microbial functioning under global change. Competitive interactions between the invasive Conyza bonariensis and the native Helminthotheca echioides were evaluated under warming, nitrogen enrichment, and elevated CO2, together with rhizosphere microbial function in solitary versus competitive growth. Plants were grown alone or in interspecific competition under elevated temperature (27 vs 29 {degrees}C), ammonium-nitrate fertilization versus no fertilization, and ambient versus elevated CO2 (400 vs 720 ppm). Plant traits and relative growth rate (RGR) were measured alongside potential extracellular enzyme activities (EEA) of -D-glucosidase (C acquisition) and N-acetyl-{beta}-D-glucosaminidase (NAGase; N acquisition) and functional gene abundances (nirS and bacterial amoA). To relate enzyme signals to plant demand and microbial biomass, we calculated a growth-normalized rhizosphere investment metric (Specific Rhizosphere Index; SRI) and a biomass-normalized investment metric (Specific Enzyme Activity; SEA). Competition effects were summarized as {Delta}SRI and {Delta}Tax (change from alone to competition) to quantify how competition altered growth- and biomass-normalized investment. Plant responses were driver- and context-dependent. Elevated CO2 produced the largest changes in growth traits, especially for the invasive species. Warming effects were modest in solitary plants but became apparent under competition, where elevated temperature reduced competitive suppression via increased invasive leaf production and reduced constraints on native leaf expansion. Fertilization caused comparatively small shifts in plant endpoints. Microbial responses depended strongly on soil conditioning history. Potential EEA showed limited shifts with warming and fertilization, whereas elevated CO2 enhanced NAGase mainly in invasive-conditioned soils and increased nirS across soils. Despite overlap in ecoenzymatic stoichiometry, SRI and {Delta}Tax revealed treatment- and legacy-dependent patterns in how competition re-scaled microbial C and N acquisition relative to plant growth and microbial biomass. Together, these results indicate that global change can decouple plant growth from enzymatic investment and reconfigure invasive-native interactions through shifts in above-belowground coupling.

Salt stress alters plant development, including the floral transition, but regulation of timing of flowering by salt is poorly understood at the molecular level. To identify genetic loci regulating the floral transition under high soil salinity, we performed a genome-wide association study (GWAS) in Arabidopsis thaliana and identified natural variation at the UGT74E1-UGT74E2-BT3 (UUB) locus that correlates with bolting time specifically in response to salt stress. Genetic analysis revealed BT3 as a novel repressor of the floral transition in control conditions. Similarly, the putative IBA glycosylases UGT74E1 & UGT74E2 delay the floral transition in control conditions. Furthermore, we identified that IBA homeostasis regulators TOB1 and ECH2/IBR10 play a key role in the floral transition, and that ECH2/IBR10 are required for the early flowering phenotype of the ugt74e1/ugt74e2 double mutant, indicating that UGT74E1 & UGT74E2 delay flowering by altering IBA homeostasis. A pangenome analysis of the UUB locus revealed variation in the occurrence of the DNA transposon SAUERKRAUT (SKRT). CRISPR-mediated SKRT deletion in Col-0 affected gene expression both within and outside the UUB locus and caused a salt-dependent delayed floral transition. The delayed bolting phenotype of the skrt-2 mutant also depends on ECH2/IBR10 function, indicating that SKRT accelerates the floral transition by altering IBA homeostasis. Finally, targeted demethylation of SKRT resulted in delayed floral transition under salt stress. Taken together, our data show a role for SKRT and its DNA methylation levels in the salt-dependent bolting time response in Arabidopsis, revealing a novel molecular mechanism to control flowering in adverse conditions.

During drought, numerous compounds accumulate in plant tissues, but their physiological roles remain unclear - they may function as osmolytes, osmoprotectants, or merely arise as by-products of stress-induced metabolic shifts. We developed an experimental approach to link accumulation patterns with specific functions, using Scots pine (Pinus sylvestris L.) saplings subjected to water deprivation and subsequent rewatering as a model system. We monitored changes in relative water content (RWC) and osmotic adjustment dynamics, employed untargeted primary metabolite profiling for preliminary screening of compounds correlated with water status, and performed quantitative GC-MS and LC-MS analyses of selected metabolites. Major inorganic cations (K, Ca{superscript 2}, Mg{superscript 2}) were also quantified to assess their potential roles. Our results revealed that tryptophan, valine, and lysine - though generally present in low abundance - exhibited selective accumulation under severely reduced RWC ([≤] 70%), suggesting their involvement as osmoprotectants. Major organic acids, particularly shikimic acid, showed trends consistent with osmotic adjustment. Notably, neither sucrose nor inorganic cations appeared to function as primary osmolytes in this context. The proposed approach offers a viable strategy for identifying compounds involved in plant adaptation to water deficit, with potential applications in breeding programs aimed at improving drought tolerance. HighlightsAn approach to identify osmolytes and osmoprotectants was implemented Accumulation of Trp, Val and Lys was consistent with their role in osmoprotection Osmotic adjustment relied predominantly on organic acids than on inorganic ions Monosaccharides but not sucrose correlates with changes in needle water status

Phenotypic plasticity is a key mechanism by which plants adjust their traits to environmental changes. These phenotypic adjustments are driven by plastic changes in gene expression regulated by gene regulatory networks. Drought, a major selective force in Mediterranean ecosystems, provides a powerful context to examine how genomic plasticity translates into phenotypic responses. Here, we used Dianthus inoxianus, a drought-tolerant Mediterranean carnation, in order to characterize the phenotypic and transcriptomic plasticity in response to drought stress combining ecophysiological measurements with RNA-seq, gene co-expression and gene regulatory network analyses. Most of the phenotypic traits exhibited low plasticity in response to drought, except water and osmotic potential. At transcriptome level, we identified 57 plastic genes, suggesting that drought tolerance in D. inoxianus relies predominantly on constitutive gene expression. These plastic genes were enriched in processes typically related to drought response, such as cell wall components and abscisic acid (ABA) signaling. Some plastic genes belonged to drought-responsive modules, while others were hubs in different modules acting as inter-modular connectors. Furthermore, the regulatory network revealed that these plastic genes were strongly regulated by multiple stress-responsive transcription factors, and that drought-associated modules were regulated through both ABA-dependent and ABA-independent pathways. In addition, we identified contrasting patterns of canalization and decanalization, with immune and post-transcriptional regulation remaining canalized under drought, whereas photosynthesis and amino acid metabolism became decanalized, potentially releasing cryptic genetic variation. Overall, our results emphasise that drought tolerance in D. inoxianus emerges from a strategy combining preadaptation with targeted plasticity in key molecular pathways.

The establishment of aphid-plant interaction involves the secretion of a salivary MIF protein. Morphological analyses revealed that aphid MpMIF1 prevents plant cell death, protects organelles from stress, and may promote plant cellular recovery. Co-expression of aphid MpMIF1 and the cell death inducer Npp1 revealed that MpMIF1 modulates autophagy-related genes ATG7/BECLIN1, impair plant senescence regulator ATAF1 and regulate apoptosis-like via Caspase-3-like activity. This effect on multiple-cell death pathways helps to maintain cellular homeostasis during aphid infection. Investigations on DNA Damage Response (DDR) signaling pathways demonstrated that aphid MpMIF1 reduces {gamma}H2A.X phosphorylation, maintains activity of the DNA repair protein RAD51 and stabilizes cell cycle checkpoint expression WEE1 under genotoxic stress. Therefore, MpMIF1 actively participates to the maintenance of a functional DDR. Finally, we showed that aphid MpMIF1 physically interacts with SOG1, a functional analog of animal p53 and central regulator of DDR, cell cycle arrest and programmed cell death in plants. These findings establish MpMIF1 as a key regulator of plant cell death during aphid-plant interactions and highlight its potential as a biotechnological tool for protecting major crops against aphid infection.