Environmental and Experimental Botany

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

Atmospheric nitrogen (N) deposition is increasingly affecting global ecosystems, with nitrate contributing a growing proportion alongside ammonium. However, the interaction between N forms and leaf developmental stage in shaping physiological and metabolic strategies in Chinese fir remains poorly understood. In this study, a field experiment was conducted to explore the physiological and metabolic responses of young and old leaves to ammonium and nitrate N addition. Our findings showed that N addition enhanced photosynthetic performance in young leaves, with a stronger effect from nitrate. In contrast, old leaves exhibited limited photosynthetic response but accumulated higher non-structural carbohydrates and showed elevated N assimilation enzyme activities, particularly under nitrate addition. Phytohormone profiles varied between leaf ages, with young leaves having higher auxin levels while old leaves exhibiting increased abscisic and salicylic acid contents under N addition. Additionally, N addition induced differential reprogramming of amino acid metabolism, with age-dependent accumulation patterns. Metabolomic analysis identified key amino acids involved in coordinating carbon-nitrogen metabolism. These results highlighted the complementary metabolic strategies by young and old leaves of Chinese fir under contrasting N forms addition and emphasized the importance of considering both N form and leaf age in optimizing N management for sustainable plantation practices. HighlightsO_LINitrate enhanced photosynthesis in young Chinese fir leaves more effectively than ammonium. C_LIO_LIOld leaves prioritized C storage and N assimilation under N addition, especially nitrate. C_LIO_LIComplementary metabolic strategies between leaf ages optimized resource use under different N forms addition. C_LI

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.

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

Fluctuating extreme weather events, coupled with rising average temperatures, can severely impact grapevine physiology and yield. While biostimulants have been gaining acceptance as a short-terms tools to enhance grapevine resilience, their adoption is hindered by inconsistent efficacy, partly driven by unpredictable plant stress levels. Over two contrasting seasons, we integrated physiological, transcriptomic, and metabolomic analyses to investigate how a plant-based biostimulant modulates the sensibility of Vitis vinifera under varying intensities of heat, drought, and their combination. This panel of water status, ranging from -0.02 to -1.6 MPa, revealed that the physiological response induced by the biostimulant treatment alleviates water stress within a field-relevant hydraulic window located between -0.4 and -1.2 MPa. Moreover, moderate but constitutive reduction of growth parameters in biostimulant plants, suggests a trade-off between vegetative development and abiotic stress responses. Accordingly, gene expression analysis revealed an interaction between water availability and the plant response to the biostimulant, which suggest an activation of priming mechanisms. Metabolic profiling supported these findings, highlighting the central role of phenylpropanoid pathway modulation, together with adjustments in ROS dynamics and stress-related hormone responses, particularly abscisic acid. Overall, this work emphasizes the need for integrating detailed plant water status and leaf gas exchange to accurately evaluate biostimulant performances under abiotic stress.

Leaf senescence is a programmed plant developmental process that can also be regulated by environmental factors, like nutrient availability. Although phosphorus is an essential element determining plants growth and productivity, mechanisms underlying adaptation to phosphorus availability in plants are not well understood. In this study, we combined physiological, biochemical and molecular approaches to investigate the effect of phosphate supply on leaf senescence in rice. We show that short-term treatment of rice seedlings with low phosphate increases photosynthetic pigments content, confers tolerance to methyl viologen-induced oxidative stress in chloroplasts, and increases antioxidant enzyme activities. Leaves from low-Pi-treated plants also showed a reduction in membrane lipid peroxidation and electrolyte leakage. Opposite trends were observed in seedlings under high Pi supply, in which accelerated leaf senescence occurs. Further analyses indicated that CRISPR/Cas9-mediated editing of MIR827, and subsequent reduction in Pi content, promotes delayed leaf senescence, while Pi accumulation in MIR827 or MIR399 overexpressing plants accelerates senescence. These findings strongly support that short treatment with low phosphate delays rice leaf senescence. Transcriptomic analysis demonstrated multiple biological processes underlying adaptation of rice plants to low phosphate, including senescence-associated and metabolic processes. These findings provide novel insights into leaf senescence potentially contributing to sustainable rice production.

BackgroundNon-optimal temperatures have become a major constraint on plant development under rapidly changing climatic conditions. Both sub- and supra-optimal temperatures reduce physiological activity, alter plant morphology, lead to plant mortality, and ultimately decrease crop productivity. Temperature-tolerant plants employ physiological and morphological mechanisms to mitigate such stress. In this study, we aimed to identify the source of temperature tolerance in warm-climate adapted melon (Cucumis melo L.). ResultsSuboptimal temperature-tolerant accession (Ananas Yoqneam; AY) and susceptible accession (PI414723) were reciprocally grafted and grown under controlled temperature regimes of 16 {degrees}C, 25 {degrees}C, and 35 {degrees}C. Physiological and morphological traits were measured to characterize tolerance mechanisms and whole-plant responses. Temperature emerged as the dominant factor governing plant performance. Whereas non-grafted parental lines maintained consistent differences across all temperature regimes, reciprocal graft combinations diverged mainly under suboptimal (16 {degrees}C) conditions. Under these temperatures, scion identity strongly determined whole-plant performance through biochemical limitations. ConclusionThese results highlight the importance of scion-derived traits in abiotic stress tolerance and their downstream influence on root function.

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.

Cadmium is a very toxic heavy metal. We studied Cd-treated barley plants with especial focus on rare atypical plants with signs of chlorosis. Cd treatment decreased the maximal photochemical activities of both photosystems while the activity of photosystem I decreased more than activity of photosystem II. In photosystem II, Cd treatment inhibited non-photochemical quenching that increased portion of unquenched "closed" complexes of photosystem II. The latter effect increased balance of limitations between the acceptor side of photosystem II (qC) and the donor side of photosystem I (Y(ND)) and raised the ratio qC/Y(ND). All these effects were enhanced in the atypical more damaged plants. Cd treatment reduced K content in the first leaves; in atypical plants, K content decreased even more. Cd treatment changed a pattern of stomatal conductance possibly by means of reducing K content in leaves. The untreated barley plants kept different stomatal conductance at adaxial and abaxial sides of leaves and fulfilled a complicated diurnal dynamics with large ups and downs of stomatal conductance. The typical Cd-treated plants were less flexible and demonstrated medium values. Stomatal conductance in the untreated plants were higher or lower than in the typical Cd-treated plants depending on a particular time; average daytime stomatal conductance was equal in both variants. At 10.00, stomatal conductance in the atypical Cd-treated plants was smaller than in the typical ones. Levels of 13 chloroplast mRNAs remained unchanged, while psbD decreased in both types of Cd-treated plants. HighlightsO_LISeveral Cd effects were enhanced in more damaged (atypical) chlorotic plants C_LIO_LICd treatment decreased activity of photosystem I and non-photochemical quenching C_LIO_LIRatio of limitations between photosystems II and I [qC/Y(ND)] was rather constant C_LIO_LICd treatment reduced K content in the first leaves C_LIO_LICd treatment changed pattern of stomatal conductance C_LI

Aquaporins play a key role in plant responses to drought. Our previous work showed limited embolism in grapevine leaves under mild water stress and suggested that the outside-xylem water pathway plays a dominant role in reducing leaf hydraulic conductance (Kleaf) during dehydration. We used CRISPR-Cas9 to knockout the PIP2;1 aquaporin encoding gene in Vitis vinifera cv. Chardonnay to study how leaf function during dehydration is affected by this aquaporin isoform. We measured functional responses like stomatal and photosynthetic responses as well as Kleaf to compare wild-type and two independent PIP2;1 knockout lines. Under moderate drought, mutants maintained greater stomatal conductance (gs) and photosynthetic rates as {Psi}w declined. No significant differences were observed in mesophyll conductance (gm) across genotypes, however, mutants exhibited slightly higher values under moderate drought. Interestingly, all lines exhibited similar Kleaf vulnerabilities to drought. Our findings show that PIP2;1 induces earlier stomatal closure during dehydration while not modulating Kleaf responses across genotypes. This rapid response in WT plants would prevent further water loss that would lead to higher xylem tensions that can lead to embolism. These findings show that multiple mechanisms collectively limit leaf gas exchange and water loss during dehydration, enhancing our understanding of plant resilience to changing environments.

Chickpea is predominantly grown under rainfed conditions in regions where terminal drought limits yield, yet little is known about how this stress influences both vegetative allocation and reproductive dynamics leading to altered grain composition. We imposed a controlled terminal drought, with a rewatered treatment group, on three Desi cultivars (ICC4958, ICC1882 and CBA Captain) reported to have contrasting drought tolerance, quantifying vegetative biomass, reproductive node productivity across developmental regions and grain macronutrient composition. Under drought, vegetative responses reflected genotype-specific resource partitioning strategies particularly evident in severe root degradation and increase stem dry matter content that was only partially alleviated in rewatered plants. Reproductive outcomes were strongly influenced by developmental stage at the time of stress, with increased pod abortion observed particularly at nodes initiating seed development under drought treatment. Grain composition of seeds filled under drought was significantly altered by stress, with increased protein concentration and decreased starch content under both Drought and Recovery treatments independent of cultivar, likely due to water limitation at crucial filling stages. These findings demonstrate that the developmental timing of terminal drought interacts with cultivar growth strategy to influence pod production and grain nutritional quality in chickpea. HighlightThe developmental timing of terminal drought interacts with cultivar-dependent growth strategies to influence pod productivity and grain nutritional quality in chickpea.

C4 photosynthesis is a CO2-concentration mechanism that separates CO2 fixation between two cell types, thereby reducing photorespiration and making C4 plants more efficient than their C3 counterparts. While the C4 cycle has evolved multiple times across different genera, this study evaluates very closely related C3 and C4 species within the genus Flaveria. Apart from their carbon metabolism, C4 plants also possess adaptations in their mineral nutrition. One key nutrient which is also directly involved in photosynthesis is phosphorus. It is absorbed by the plant in the form of inorganic phosphate and is an essential component of DNA, ATP, lipids, and carbohydrates. In the Flaveria C4 species, but not in the C3 species, phosphate limitation was shown to affect the dark reactions of photosynthesis. This study investigates how phosphate deficiency impacts the light reactions in C3 and C4 Flaveria plants. We observed a differential response in the functionality of photosynthetic energy conversion between the two species. When exposed to a limited phosphate supply, the C3 species reduced its linear electron transport rate while dissipating excess energy through high-energy quenching, which was regulated by a higher pH gradient across the thylakoid membrane. In contrast, the C4 species did not regulate its photosynthetic light reaction under phosphate limitation. Instead, it exhibited increased stress levels, evidenced by a stronger biomass reduction and the induction of stress markers in the leaves. Additionally, this study uncovered an acceleration in NPQ relaxation during phosphate limitation, regardless of the photosynthesis type. HighlightPhosphate deficiency reduced linear electron transport rates and induced dissipation of excess energy through non-photochemical quenching in the C3 Flaveria species, while in the C4 species, despite elevated stress levels, the photosynthetic light reactions were unaffected.

Salinity and sodicity stresses adversely affect rice growth and yield. To overcome yield losses, suitable tolerant rice cultivars can be developed through a marker-assisted breeding (MAB) program. In the present study, genomic regions associated with sodicity stress tolerance at the reproductive stage were identified using a high-density 50kSNP array in a recombinant inbred line (RIL) population derived from the contrasting rice genotypes CSR11 and MI48. A total of 50 QTLs were detected for various yield-related traits; further, 19 QTLs with [≥]15% of phenotypic variance were selected for integrated (omics) analysis. RNA sequencing of leaves and panicles at the reproductive stage under sodic stress conditions was employed to find differentially expressed genes. A total of 1368 and 1410 SNPs; 104 and 144 indels were found for MI48 and CSR11, respectively, within the QTL regions from resequencing. At chromosomes 1 and 6, colocalized QTLs (qPH1-1/qGP1-1 and qGP6-2/qSSI6-2) were discovered. Differentially expressed genes (DEGs) were mapped over the QTL regions selected, and SNP variations and indels were screened for colocalized QTLs. Potential candidate genes, namely Os-pGlcT1 (Os01g0133400), OsHKT2;1 (Os06g0701600) and OsHKT2;4 (Os06g0701700), OsANTH12 (Os06g0699800), and OsPTR2 (Os06g0706400), were identified as being responsible for glucose transport, ion homeostasis, pollen germination, and nitrogen use efficiency, respectively, under salt stress. Finally, our study provides important insights into the genes and potential mechanisms affecting grain yield under sodic stress in rice, which will contribute to the development of molecular markers for rice breeding programs.

Climate change and increasing drought conditions significantly impedes citrus productivity in subtropical and tropical regions. This study explores the potential of combining arbuscular mycorrhizal fungi (AMF) Funneliformis mosseae and plant growth-promoting rhizobacteria (PGPR) Pseudomonas putida to mitigate drought resilience in Citrus reticulata (Red tangerine). AMF-mediated drought tolerance has been extensively documented; however, the collegial influence of PGPR and AMF on phytohormone signaling, photosynthetic efficiency, nutrient acquisition, and gene expression remains largely unexplored in citrus. We conducted a greenhouse experiment under both well water and drought stress conditions to assess the physiological and molecular responses to individual and co-inoculation with PGPR and AMF. Drought-stressed citrus plants, inoculated with AMF and PGPR, demonstrated significantly improved leaf water potential, stomatal conductance, carbon assimilation, and antioxidant defense. PGPR-AMF co-inoculation enhanced chlorophyll stability, osmotic adjustment, and nutrient uptake, while significantly reducing lipid peroxidation and ROS accumulation. The turquoise module emerged from transcriptomic and gene co-expression network analysis (WGCNA) as a potential key regulator of stress adaptation, revealed key regulatory transcription factors, e.g., CrMYB4, CrZFP8, CrSOS5, CrRGFR2, and CrQUA1, that were upregulated under combined inoculation, highlighting their potential role in stress adaptation. Our findings demonstrate that the synergistic PGPR-AMF interaction improves antioxidant enzyme activities and modulates gene expression to promote drought tolerance, providing new insights into the microbiomes role in plant resilience. These results offer a potential strategy to boost citrus growth and yield under water scarcity, with broad implications for agricultural resilience to climate change.

Sterol biosynthesis underlies significant physiological functions in plants, including the production of membrane structural sterols and hormones such as brassinosteroids and cytokinins. Inhibition of sterol biosynthesis has been shown to disrupt multiple aspects of Arabidopsis thaliana development. Here, the effects of lovastatin, an inhibitor of HMG-CoA reductase, on root development were investigated, focusing on auxin-cytokinin distribution and transport. Lovastatin inhibited primary root growth, especially cell elongation, in a dose-dependent manner. Additionally, lateral root density was considerably increased and lateral root primordia (LRP) emerged ectopically. In accordance to the above defects, auxin/cytokinin imbalance was recorded by the ectopic presence of the synthetic auxin marker DR5 and a significant decrease of cytokinins, as revealed by depletion of the TCS (two-component signaling) marker. Because auxin distribution appeared disturbed, auxin transport impairment was further examined. Plasma membrane localization of PIN auxin efflux carriers declined significantly, showing additional diffuse cytoplasmic localization in LRP cells. However, the cell-specific localization patterns of several PINs and their abundance at the transcript and protein level appeared unaffected or slightly increased. Fluorescence recovery after photobleaching (FRAP) analysis regarding membrane kinetics of PIN2 revealed altered PIN2 membrane dynamics and transmission electron microscopy (TEM) observations showed structural defects at the plasma membrane-cell wall interface. Together, these results support that sterol biosynthesis is essential for maintaining plasma membrane organization, which, in turn, is key factor for the distribution of hormones that control root development. HighlightsLovastatin treatment inhibits root growth and causes deregulated formation of lateral roots. Consistently, lovastatin causes altered patterns of auxin distribution relevant to PIN protein mis-localization and decreases cytokinin levels. These changes could be attributed to reduced structural sterols as exemplified from alteration in PIN2 membrane dynamics.

Acidic stress severely restricts crop growth by disrupting nutrient uptake, redox homeostasis, and membrane stability, yet mitigation strategies remain limited. Here, we investigated the role of melatonin (MT) in regulating growth, photosynthesis, oxidative stress, antioxidant defense and proton transport in peanut seedlings under controlled hydroponics acidic (pH 4.0) and near-optimal (pH 6.5) conditions, and validated these findings in naturally acidic field soil (pH 4.3-4.5). Acid stress markedly reduced biomass accumulation, chlorophyll content, and redox balance, while enhancing ROS (H2O2) and lipid peroxidation (MDA). Exogenous MT application, particularly at 50-100 {micro}M, significantly improved shoot and root biomass, restored chlorophyll pigments and reduced H2O2 and MDA accumulation, with more pronounced effects under pH 4.0 than pH 6.5. MT strongly activated antioxidant enzymes (SOD, CAT, APX), while POD activity declined, reflecting melatonins dual role as both a direct ROS scavenger and a regulator of enzymatic redox networks. Notably, MT induced strong, dose-dependent upregulation of HL-ATPase genes (AH1 and AH2) in both leaves and roots under acidic conditions, suggesting enhanced proton extrusion, intracellular pH homeostasis, and stress adaptation. The soil validation experiment confirmed the agronomic relevance of these findings, where MT dose-dependent concentrations improved germination, vegetative growth, chlorophyll fluorescence (Fv/Fm), and yield-related traits under natural acidic conditions. Although MT also conferred benefits at pH 6.5, responses were generally moderate compared with acid stress. Collectively, these results demonstrate that MT enhances peanut tolerance to acid stress across both controlled and natural field-relevant environments, highlighting its potential application for sustainable crop production on low-pH soils.

Non-structural carbohydrates (NSC) stored in the stem play a crucial role in supporting yield formation in rice. However, internode morphological determinants of NSC accumulation are unclear. This study aimed to clarify the relationship between internode morphology and NSC accumulation and to identify a robust morphological indicator for evaluating NSC accumulation capacity. Two years of field experiments were conducted using multiple cultivars. The NSC content was quantified for individual internodes and at the whole-plant culm level, and its relationships with internode morphological traits were analyzed. Since the upper internodes (UIN; first and second internodes) and lower internodes (LIN; third and subsequent internodes) exhibited contrasting roles in NSC accumulation, a novel index was introduced, the volume composition ratio (VCR) of UIN/LIN, which represents their relative volumetric contributions within a culm. The VCR of UIN/LIN showed the strongest correlation with culm NSC and high reproducibility across years, outperforming simple morphological traits. Manipulation of internode development using plant growth regulators demonstrated that altering VCR effectively modified culm NSC accumulation. Accordingly, the VCR of UIN/LIN serves as a robust morphological indicator of culm NSC accumulation capacity, providing a practical framework for improving NSC accumulation to achieve high and stable yield performance in rice. HighlightThis novel internode structural index robustly predicts the culm non-structural carbohydrate accumulation capacity, providing a practical morphological indicator for improving yield stability in rice.

Woody stems conduct both photosynthetic assimilation and respiration. The two processes work in concert, as stem photosynthesis helps refix CO2 released by stem respiration, thereby increasing carbon-use efficiency and generating a local pool of non-structural carbohydrates supporting cambial growth and stem hydraulic function. Despite its importance, little is known about seasonal variation in stem photosynthesis and the factors underlying its activity throughout the season. To fill this gap, we measured stem gas exchange together with growth activity, water status and photosynthetic pigment contents in two temperate species, Acer platanoides L. and Prunus avium L., over the season. In both species, gross photosynthetic rates (Pg) and dark respiration (Rd) changed significantly over the season in a similar pattern, indicating strong coordination between the two processes. Both Pg and Rd reached the highest values in May, during the period of rapid leaf expansion and secondary growth, and declined later in the growing season. At each measurement date, Rd exceeded Pg, resulting in a net CO2 efflux from the stems. The seasonal changes in Pg and Rd translated into seasonal variability in relative refixation of CO2, ranging from 3 to 59% and gradually decreasing towards the end of the season. Additionally, the Pg corresponded with the tissue hydration and increased significantly with increasing stem water potential. In contrast, total chlorophyll content showed less pronounced seasonal variation and thus explained substantially lower seasonal variability in Pg, except for the chlorophyll a/b ratio, which changed dynamically over the season and reached a minimum during the peak of the growing season. Overall, our results reveal that stem photosynthesis varies seasonally in accord with stem growth and water status, while the chlorophyll content has a lower impact on the seasonal changes. These findings are important for our understanding of the carbon relations of trees.

Glutathione peroxidases (GPXs) are widely recognized as key antioxidants that mitigate oxidative stress by detoxifying reactive oxygen species (ROS). However, GPXs are largely uncharacterized in citrus. Here, we demonstrated that Citrus sinensis contains four GPX proteins (CsGPX1-4). Unexpectedly, overexpression of CsGPX4, a homolog of AtGPX8 in Arabidopsis, in citrus resulted in typical oxidative stress phenotypes including severe growth inhibition, chlorosis, and elevated intracellular ROS accumulation. Transmission electron microscopy (TEM) analysis further revealed stress responses at cellular level. Whole genome shot gun sequencing analysis showed that T-DNA insertion occurs in the UTR of SWEET2 gene, which is unlikely to be responsible for the oxidative stress phenotypes. Immunoblotting revealed that CsGPX4 accumulates as a truncated protein in citrus, in contrast to the full-length version expressed in Nicotiana benthamiana. MALDI-TOF assays further confirmed the truncation of CsGPX4 in the transgenic line with the predicted cleavage site between L115-K117. This truncation was associated with altered subcellular localization, shifting from cytoplasmic and nuclear distribution in N. benthamiana to membrane association in citrus. Proteomic profiling further indicated extensive reprogramming of pathways involved in detoxification, cytoskeletal stability, hormone signaling, and cell wall modification. Our data suggests that de facto overexpression of truncated CsGPX4 may have dominant-negative effects on proteins interacting with CsGPX4, thus interfering with their normal functions. In conclusion, our study demonstrates CsGPX4 as a critical regulator of redox homeostasis and ROS homeostasis in citrus and reveals selective truncation of CsGPX4 as a unique proteolytic or regulatory strategies in such processes.

Photoperiod sensitivity (PS) is a key biological response in plants as they adapt to specific environments. Rice (Oryza sativa L.) exhibits a clear PS, as it implements critical phase transition decisions based on PS signals. In this study, we identified a novel PS gene, JMJ706, that is expected to deliver photoperiod-related signals to the flowering-time regulatory network in a day-length-dependent manner. The JMJ706 mutants exhibit early flowering under LD and later flowering under SD compared to WT plants. The gene encodes an H3K9me2 demethylase, and under long-day (LD) conditions, its demethylase activity facilitates the expression of Grain number, Plant height, and Heading-date7 (Ghd7). Since Ghd7 is a floral repressor in LD, it promotes the vegetative phase by delaying flowering. Under short-day conditions (SD), H3K9me2 demethylase activity facilitates Early heading-date 1 (Ehd1) expression, and it acts as a floral accelerator by inducing Heading date 3 (Hd3a) and RICE FLOWERING LOCUS T 1 (RFT1). Furthermore, we propose that the daylength-dependent promotion of target genes (Ghd7 and Ehd1) occurs through demethylation of specific promoter regions at a crucial time window. In addition, JMJ706 may play an important role in regulating plant architecture, including plant height. The natural variation in JMJ706 alleles shows high frequencies across major rice subpopulations, suggesting that JMJ706 could play an important role in the geographical distribution and adaptation of rice cultivars. Our results may add a new layer to the rice flowering-time regulatory pathway, supporting regional adaptation and potential for future breeding.