Plant Science

Iron (Fe) homeostasis is fundamental to plant growth and development and is centrally regulated by a well-conserved network of basic helix-loop-helix (bHLH) transcription factors. In Arabidopsis thaliana, bHLH121 regulates the expression of FER-like iron deficiency-induced transcription factor (FIT) and clade Ib bHLH38/39/100/101 genes, among others. However, its functional conservation remains unexplored, and some contradictory results have been reported regarding the effects of bHLH121 overexpression. Here, we investigated the evolutionary conservation of bHLH121 through complementary tests in the Arabidopsis bhlh121 mutant created by CRISPR/Cas9 gene editing. We also investigated the effects of ectopically expressing full-length, N-terminal, and C-terminal fragments of bHLH121 on plant responses to Fe deficiency in Arabidopsis. Transgenic plants overexpressing the full-length bHLH121 driven by the cauliflower mosaic virus 35S promoter (bHLH121 ox) resulted in shorter roots and more severe leaf chlorosis compared to wild-type plants under Fe limiting conditions. Moreover, overexpression of the N-terminal fragment (N121 ox) rendered plants highly sensitive to Fe deficiency, whereas overexpression of the C terminal fragment (C121 ox) did not. Consistent with the phenotypic observations, N121 ox plants accumulated lower Fe content, and the ferric chelate reductase activity was significantly reduced. In N121 ox plants, the induction of key Fe regulatory genes, especially FIT, was significantly inhibited under Fe deficiency. Confocal imaging showed that both N121-GFP and C121-GFP fusion proteins could localize to the nucleus. Electrophoretic mobility shift assay showed that both full-length bHLH121 and N121 could effectively bind the probes derived from the promoter of FIT. The present work reveals a conserved function of bHLH121 in Fe homeostasis across several dicot species and demonstrates that the N121 may possess a dominant nature in Fe deficiency responses.

Pepper fruit accumulated xanthophyll acyl esters during ripening. To characterize the genes involved, cDNAs for two putative xanthophyll acyltransferases named CaPYP1 and CaPYP1-like were cloned. The deduced amino acid sequences of CaPYP1 and CaPYP1-like had conserved hydrolase and acyltransferase domains. Phylogenetic analyses showed that both putative acyl transferases were conserved in Viridiplantae, suggesting important functions for both. CaPYP1 and CaPYP1-like GFP-fusion proteins were localized in the plastid when expressed in leaf tissue. Gene expression analyses indicated that both genes for CaPYP1 and CaPYP1-like were most expressed in ripening fruit (52 to 64 days after anthesis) and senescent leaf, CaPYP1 having relatively greater expression than CaPYP1-like. In virus-induced gene silencing experiments in pepper, xanthophyll esterification was greatly diminished when CaPYP1 was silenced with a simultaneous change in the ripening fruits color from dark red to bright red. In complementation tests, overexpression of CaPYP1 in a tomato mutant impaired for petal coloration and xanthophyll esterification, resulted in restoration of the petal color and the synthesis of both mono, diacyl and tri esters of xanthophylls. CaPYP1-like overexpression in the same genetic background resulted in the synthesis of relatively smaller amounts of xanthophyll monoesters. In an in vitro test, zeaxanthin was more sensitive to light than zeaxanthin dipalmitate but both were protected when triacylglycerol was mixed with it, suggesting that acyl moieties could improve xanthophyll stability. Together our results indicate that xanthophyll esterification during pepper fruit ripening is important for fruit color, xanthophyll accumulation and stability and is orchestrated by both CaPYP1 and CaPYP1-like with CaPYP1 playing a major role. HighlightRipening pepper fruit accumulates xanthophyll fatty acyl esters associated with nutritional quality and fruit color. The fruit expresses CaPYP1 and CaPYP1-like, two putative acyltransferases in their chromoplasts. In a tomato mutant impaired for xanthophyll esterification, transgenic expression of CaPYP1 promoted more xanthophyll esterification in ripe fruit than expressing CaPYP1-like.

Background Strigolactones (SLs) regulate diverse aspects of plant development and have been implicated in promoting leaf senescence. However, senescence phenotypes associated with SL deficiency have not been consistently observed across species, suggesting that this function may be species- or context-dependent. Moreover, the contribution of endogenous SL biosynthesis to senescence in leafy vegetable crops remains unclear. Here, we investigated the role of the SL biosynthetic gene MORE AXILLARY GROWTH1 (MAX1) in dark-induced leaf senescence in lettuce (Lactuca sativa). Results We found that endogenous SL biosynthesis plays a major role in dark-induced senescence in lettuce. SL pathway genes were induced during dark storage, while exogenous GR24 accelerated senescence and lettuce MAX1 (LsMAX1) complemented the delayed-senescence phenotype of the Arabidopsis max1 mutant. Consistent with these findings, CRISPR/Cas9-generated Lsmax1 mutants exhibited a pronounced stay-green phenotype during prolonged darkness, accompanied by strongly reduced induction of key senescence-associated genes. Despite this delayed visible senescence, Lsmax1 retained a substantial transcriptional response to dark storage. Strikingly, loss of LsMAX1 did not simply weaken the wild-type senescence program, but redirected part of the response toward a distinct stress-associated transcriptional state that was largely absent from wild type. Loss of LsMAX1 did not affect vegetative rosette architecture, although increased branching emerged after bolting. Conclusions Our findings establish MAX1-dependent SL biosynthesis as an important regulator of leaf senescence in lettuce and reveal a role that extends beyond controlling the rate of senescence. Rather than simply delaying the wild-type program, loss of LsMAX1 alters the transcriptional trajectory of senescence, favoring an alternative stress-associated state during prolonged darkness. The strong stay-green phenotype without detectable changes to vegetative rosette architecture further highlights SL biosynthesis as a potential target for extending postharvest longevity in lettuce and other leafy crops.

Drought stress severely limits tree growth and survival, a problem intensifying under climate change due to the rising frequency of extreme weather events. Identifying genetic variants that modulate drought stress responses and enhance resilience is therefore of great interest for developing climate-adapted trees. While the MADS-box transcription factor AGL16 has been implicated in drought responses and stomatal regulation in herbaceous species, the function of its homolog in woody perennials remains largely unexplored. Here, we investigate the role of PtaAGL16 in the poplar hybrid Populus tremula x P. alba. We generated CRISPR/Cas9 knockout lines of PtaAGL16 in the poplar clone INRA 717-1B4. Plants were subjected to a severe drought stress regime followed by a re-watering recovery phase. Morphological and physiological traits, including growth, stomatal conductance, shoot vitality, and stomatal density, together with stress-associated metabolites and drought-responsive gene expression, were analyzed. All agl16 knockout lines exhibited significantly reduced drought-induced damage compared to the wild-type. While the wild-type suffered extensive defoliation and persistent stem necrosis, the knockout lines retained higher shoot vitality and resumed growth after re-watering, including the formation of new leaves. Consequently, vitality in the knockout lines reached up to 86%, compared to only 34% in the wild-type. This improved resilience was accompanied by reduced persistent oxidative membrane damage, as indicated by lower MDA levels after recovery. These findings indicate that the disruption of PtaAGL16 is associated with enhanced drought resilience in a woody perennial species and highlight its potential as a genetic target for improving climate adaptation in fast-growing forest trees.

Groundnut (Arachis hypogaea) is an important oilseed crop, and its production is severely affected by drought worldwide. Improving the drought-tolerance of groundnut without compromising its agronomic traits is a major challenge for researchers. Here, a meta-transcriptome analysis identified tandemly duplicated G{beta} subunits (AhG{beta}1 and AhG{beta}3) as early signaling genes under water deficit conditions. Heterotrimeric G-protein {beta}-subunit is a major signaling element that can regulate diverse responses in plants. Heterologous complementation studies in Arabidopsis G{beta} mutant (agb1-2) demonstrated evolutionary conservation of AhG{beta} function, while the transgenic overexpression and RNAi knockdown in groundnut established AhG{beta}1 as a positive regulator of plant growth, yield, and drought tolerance. Integrated transcriptomic, transcription factor regulatory network, and metabolomic analyses demonstrated that AhG{beta}1 coordinates drought-responsive transcriptional and metabolic reprogramming characterized by suppression of cell wall-associated processes and activation of phenylpropanoid, flavonoid, and isoflavonoid biosynthesis. This reprogramming resulted in enhanced accumulation of antioxidant flavonoids, including quercetin and rutin, reduced ROS accumulation, and improved drought tolerance. Furthermore, exogenous application of these flavonoids enhanced drought recovery, highlighting their role as protective downstream metabolites. Collectively, our findings establish AhG{beta}1 as a conserved regulator linking early G-protein signaling with transcriptional and metabolic reprogramming to maintain redox homeostasis during drought stress and identify promising molecular and metabolic targets for improving drought resilience in groundnut.

Branched-chain amino acids (BCAAs) are essential amino acids involved in protein synthesis and energy metabolism. Because animals cannot synthesize BCAA de novo, plant-derived BCAAs are important to human nutrition. Although mungbean sprouts are widely consumed as functional plant-based food materials, changes in individual BCAA accumulation and their transcriptional regulation during mungbean germination remain poorly understood. In this study, amino acid contents and transcriptomic profiles were analyzed at three germination stages, 8H, 24H, and 72H. Total BCAA content increased during germination, whereas individual BCAAs exhibited distinct temporal accumulation patterns. Isoleucine and valine increased until 72H, while leucine increased during early germination and decreased after 24H. Transcriptome analysis revealed time-dependent expression changes in BCAA biosynthesis and degradation genes associated with the leucine decrease after 24H. These findings suggest that 24H represents an important transition point for BCAA accumulation and compositional change during mungbean germination. This study provides molecular evidence for the regulation of BCAA metabolism during mungbean germination and supports the potential use of germinated mungbean as a plant-based amino acid resource.

Amaranthus tricolor is a climate-resilient orphan crop whose betalain pigments contribute to nutritional quality and stress tolerance. However, breeding efforts have traditionally relied on field yield trials due to the limited availability of genomic information and genetic resources. To address this gap, we first used the Illumina and PacBio sequencing platforms to assemble a novel reference genome of A. tricolor, using the Japanese cultivar Bayam. By combining this reference genome with a core collection developed in our previous study, we aimed to identify genomic regions associated with pigmentation-related traits across winter and summer seasonal environments. Betalains are antioxidant phytonutrients that contribute to abiotic stress tolerance, making the regulation of their biosynthesis under heat stress particularly relevant for developing climate-resilient crops such as A. tricolor. As a result, ATR1.0CH03_5131103, which is associated with ATR1.0ch03g000565, was identified under the winter seasonal environment. Meanwhile, transcriptomic profiles in this study revealed that ATR1.0ch03g000565 was differentially expressed under both control and heat stress conditions, suggesting that it may be associated with the regulation of CYP76AD1 expression under optimal conditions. Under the elevated-temperature treatment, transcript levels of CYP76AD1 and 5GT decreased in Am087, coinciding with lower betacyanin-dominated absorbance indices. cDOPA-like homologs were upregulated; however, pigmentation-related absorbance remained reduced. Further molecular experiments, such as CRISPR-Cas9 knockouts or overexpression studies, are necessary to validate the regulatory mechanisms of CYP76AD1 and 5GT under heat stress.

Exposure to Ultraviolet-B (UV-B) light can adversely affect plant growth and cellular integrity by inducing oxidative stress and DNA damage. In this study, we investigated UV-B-induced DNA damage and repair responses in the aquatic monocotyledonous plant species Spirodela polyrhiza (duckweed). We exposed 13-day-old duckweed plantlets to broadband UV-B light for 1-10 min, followed by recovery periods of up to 24 h under normal growth conditions. We observed progressive chlorosis, wilting, and diminished plant vigor with longer durations of UV-B light exposure. Agarose gel electrophoresis demonstrated compromised genomic DNA integrity immediately after UV-B light treatment, with partial restoration of DNA quality during recovery. Immuno-slot blot assays established the accumulation of two major UV light-induced photoproducts, cyclobutane pyrimidine dimers (CPDs) and 6-4 pyrimidine-pyrimidone photoproducts [(6-4)PPs], in a dose-dependent manner following UV-B light exposure. Notably, the abundance of these DNA lesions declined substantially after recovery, indicating activation of endogenous DNA repair mechanisms. Staining with 3,3-diaminobenzidine revealed elevated accumulation of hydrogen peroxide immediately following UV-B exposure, suggesting enhanced oxidative stress. Collectively, these findings demonstrate that S. polyrhiza possesses efficient mechanisms for sensing, repairing, and mitigating DNA damage induced by oxidative stress resulting from UV-B light exposure. This study highlights the potential of duckweed as an effective model system for investigating DNA damage and repair pathways under UV-B light stress in plants.

Heat stress is one of the most damaging abiotic constraints on crop productivity, and its consequences are expected to intensify as extreme temperature events become more frequent and severe. Pepper (Capsicum annuum L.) is particularly vulnerable to sustained high temperatures, which can disrupt photosynthetic performance, cellular homeostasis, and redox regulation. However, the physiological and transcriptional dynamics underlying genotype-dependent responses to prolonged heat exposure remain insufficiently understood. We combined repeated physiological measurements with time-course RNA sequencing to compare GPC003240, previously identified as a candidate heat-tolerant accession, with two non-elite accessions, GPC010350 and GPC014930, which are phenotypically divergent from each other, under 40/30 {degrees}C Day/night temperatures for up to six days. GPC010350 maintained comparatively stable photosystem II performance and higher stomatal conductance, whereas GPC014930 showed progressive photochemical impairment and lower conductance; GPC003240 displayed a distinct, moderately responsive profile. Transcriptomic responses showed partial functional convergence during the early phase of stress exposure but diverged markedly after six days. When gene expression at day 6 was compared with the pre-treatment baseline separately within each genotype, 4,436 differentially expressed genes were detected in GPC010350, compared with 680 in GPC003240 and only 78 in GPC014930. The late response of GPC010350 was associated with enrichment of RNA- and ribosome-related, biosynthetic, DNA-repair, and genome-maintenance functions. By contrast, GPC014930 showed negative enrichment of photosynthesis, plastid organization, redox homeostasis, and translation-related processes. Global co-expression analysis identified a time-decreasing photosynthesis-associated module (ME5) and two time-increasing modules, ME12 and ME19, that were enriched in genes contributing to the late GPC010350 response. Integration of differential expressions, module membership, and functional annotation highlighted a heat shock transcription factor (Caz03g27980), HSP101 (Caz03g07770), and a dual-specificity phosphatase (Caz05g20970) as candidates for further investigation. Overall, the results suggest that genotype-dependent responses to prolonged heat exposure were associated not only with the magnitude of early transcriptional change, but also with differences in the temporal organization of stress-response, maintenance, and metabolic processes. The contrasting responses of the non-elite accessions GPC010350 and GPC014930 further highlight the value of phenotypically diverse germplasm for uncovering mechanisms relevant to future heat-tolerance breeding.

Stomata are the pores on plant surface, and these tiny pores are responsible for the flow of gas between plants and atmosphere. Currently, what effects of the broad and continuous increase in stomatal density achieved via genetic engineering on plant growth and development remain poorly understood. The 9 Arabidopsis transgenic lines with increased stomatal density were acquired through overexpressing FSTOMAGEN (the homologs of STOMAGEN, which are in Flaveria). The intermediate stomatal density (SD) lines exhibited increased trend in biomass. Compared with the lines with low SD, the biomass of Arabidopsis lines with intermediate SD (484 mm-2) significantly increased. There was a positive and significant correlation between biomass and relative water content. Across these transgenic lines, only during the earlier phase of growth, the leaf area exhibited a gradually increased trend as stomatal density increased, and there was both a significant linear relationship between SD and leaf growth rate and a strong linear relationship between SD and leaf area. In contrast, a clear relationship during the later phase wasnt observed. Under lower growth light intensity, there was an increased trend of biomass from other lines to the lines with intermediate SD, and the photosynthetic rate and stomatal conductance of the intermediate line were significantly increased. This study reveals plant-growth alterations that correspond to broad and near-continuous increases in stomatal density achieved via genetic engineering. Our study sheds light on the prerequisites for elevated stomatal density achieved via genetic engineering to promote plant growth.

Accelerating photoprotective regulation to improve carbon assimilation is a promising strategy to increase crop productivity. Although rapid non-photochemical quenching (NPQ) relaxation has been validated as a target through metabolic engineering, it remains unclear whether conventional breeding has improved this trait. Here, we investigated whether more than a century of soybean breeding enhanced NPQ relaxation alongside light-saturated carbon assimilation and seed traits. We evaluated a historical panel of 24 soybean genotypes across vegetative and reproductive developmental stages by integrating NPQ relaxation, gas exchange parameters, xanthophyll-cycle pigment profiles, expression of key photoprotective genes (VDE, PsbS, and ZEP), seed number and seed weight. NPQ relaxation parameters were not consistently associated with genotype release year, seed number, or seed weight at either developmental stage. The only exception was the amplitude of the rapidly relaxing NPQ component (AqE), which was negatively correlated with all three variables during the reproductive stage. In contrast, genotype release year was positively associated with maximum net CO2 assimilation rate (Amax), maximum carboxylation rate of Rubisco (Vcmax), maximum electron transport rate (Jmax), seed number, and seed weight, while Amax and Vcmax were positively correlated with seed number and seed weight. These findings indicate that the greater photosynthetic capacity of modern genotypes was not accompanied by faster photoprotective response. Thus, photoprotective regulation has not kept pace with gains in photosynthetic capacity under field conditions. We conclude that rapid NPQ relaxation remains an important target for synchronizing photoprotection with the high photosynthetic capacity of modern soybean lines.

High temperatures during grain filling stage are becoming increasingly frequent, compromising both grain and eating quality and thereby driving demand for heat-resilient cultivars. Such conditions are known to reduce the expression of granule-bound starch synthase I (GBSSI) and starch branching enzyme IIb (BEIIb), which are involved in starch biosynthesis, resulting in a decrease in amylose content and an increase in long-chain amylopectin. Thus, the present study introduced functional mutation in starch synthase IIa (SSIIa) that increases the proportion of short amylopectin chains to genetically compensate for the high-temperature-induced increase in amylopectin long chain. Rice lines carrying the ss2a mutation were grown at two locations with cooler (Akita) and warmer (Okayama) temperatures. Their grain traits, starch structure, and eating quality were compared. The ss2a mutant lines showed an increased proportion of short amylopectin chains as well as an increased apparent amylose content. Furthermore, these alterations in starch structure varied with the grain-filling temperature of the cultivation sites, ultimately affected eating quality. These results suggest that enriching short amylopectin chain via the ss2a mutation can counteract the increase in long amylopectin chain caused by high temperatures during grain filling, thereby maintaining a desirable starch structure and eating quality.

Plastidial phosphorylase (Pho1) regulates starch metabolism and also modulates photosystem I (PSI) activity. In rice, Pho1 contains an 80 amino acid intrinsically disordered region (IDR), L80, that acts as a negative regulator. While total removal of the IDR boosts seedling growth, flowering time, overall biomass, and grain yield and alters PSI redox properties, the precise underlying regulatory basis remains unclear. To pinpoint IDR regulatory elements, four Pho1 partial-IDR deletion variants ({delta}N, {delta}C, {delta}M, and {delta}N{delta}C) were evaluated. Lines carrying {delta}M and {delta}C exhibited enhanced starch metabolism, yielding larger, heavier grains, and flowered 5 to10 days earlier than wild type. Both d{delta}M and {delta}C share an acidic motif, (E/V)SEEV, identifying it as a primary negative regulator of starch synthesis and floral timing. By contrast, no partial deletion fully mirrored the PSI redox modifications of Pho1{delta}L80. These results demonstrate that starch biosynthesis and flowering are governed by one or more discrete sequence motifs within the middle and C-terminal regions, whereas PSI modulation likely requires the structural integrity of the intact IDR.

Cassava produces starch-filled tubers which are a crucial food source for many people, especially in Africa. This plant species is famous for drought tolerance and resilience, but changing conditions due to climate change require genetic improvements to ensure sustainable cassava cultivation. In this context, a comprehensive understanding of the genetic basis responsible for a variety of traits is beneficial. Here, we present insights into the anthocyanin metabolism, responsible for coloration and protection of cassava structures during stress. Intraspecific variation in the transcriptional control of the anthocyanin biosynthesis was characterized through comparative genomics and phylogenetic analyses. A member of the R2R3-MYB transcription factor family, orthologous to crucial activators of anthocyanin biosynthesis in other plant species, was identified as a genetic hotspot for pigment accumulation capacity between cultivars. Orthologs of this gene appeared absent from three cassava cultivars without pronounced anthocyanin pigmentation, while being present in cultivars known for anthocyanin formation. This discovery corroborates previous reports about the importance of transcriptional regulators in the intraspecific diversity of anthocyanin pigmentation.

Mitochondria are endosymbiont-derived organelles that play a central role in cellular metabolism, energy production and stress responses. While single mitochondria represent functional units, they continuously exchange their contents through fusion and fission, facing stress conditions as a dynamic population. To date, it remains largely unknown how stress alters mitochondrial dynamics in plants and how altered dynamics affect mitochondrial properties and plant stress resilience. Here, we investigate mitochondrial dynamics in response to oxidative stress in the non vascular model plant Physcomitrium patens. By creating mutants with impaired mitochondrial fission in different reporter lines for mitochondrial parameters, we additionally analyse effects of chronic changes to mitochondrial population dynamics. We found that Mito-Paraquat (MtPQ) treatment increased the glutathione redox potential EGSH in mitochondria, the cytosol and chloroplasts, as monitored via roGFP2-based genetically encoded biosensors. Mitochondria elongated within hours and showed a concomitant and heterogenous increase of matrix EOSred, that we propose as a marker for matrix protein damage. Mitochondrial fission mutants lacking PpELM1B (ELONGATED MITOCHONDRIA) displayed distinct changes of mitochondrial morphology parameters as determined by automated 3D-segmentation and feature mapping (MorphoMapper) of confocal z-stacks. Elongated mitochondria in Ppelm1bge lines showed an oxidative matrix EGSH shift and increased matrix EOSred while matrix mixing still occurred, albeit at the same slow rate as in wildtype, within days. Macroscopically, Ppelm1bge lines displayed reduced growth, decreased respiration, and a higher sensitivity to oxidative stress. Our results show that plant mitochondrial morphology and physiological parameters specifically shift in response to stress and impaired fission. Mitochondrial fission is vital to maintain a healthy mitochondrial population that sustains plant oxidative stress tolerance.

Plants make diverse metabolites to outcompete neighboring organisms for space and resources. Some of these toxic metabolites broadly disrupt conserved molecular mechanisms, such as protein biosynthesis. Nonproteogenic amino acids (NPAAs) are a structurally diverse class of metabolites that interfere with protein biosynthesis. The proline (Pro) analog azetidine-2-carboxylic acid (Aze) inhibits plant growth through misincorporation during protein biosynthesis. However, it is unknown if a cascade of downstream stress responses is triggered following Aze misincorporation. Here, we investigate the morphological and stress responses in Arabidopsis grown on Aze. Investigation of root morphological responses show not only reduced root growth, but increased root branching following growth on Aze. Altered root morphology is coupled with a reduced gravitropic response. Aboveground organs were also affected by Aze, including reduced chlorophyll content, reduced photosynthetic efficiency, and increased anthocyanin content. We then tested whether Aze induces reactive oxygen species (ROS) accumulation using multiple approaches and observed both immediate and sustained accumulation of general ROS and H2O2 following treatment with Aze. When plants were grown on Aze supplemented with Pro, ROS levels were restored to normal levels, suggesting that reducing misincorporation events results in less downstream stress responses. In summary, we find that following Aze treatment a cascade of downstream stress responses is induced that exacerbates the effects of toxic NPAAs. This study sheds light on the mechanism of action of NPAAs and provides information on the downstream consequences of translational errors.

Seed development and germination are two important transitions in a plant's life cycle. During such transitions, extensive transcriptional regulation reprogramming is performed to support major changes in cellular, tissue, and physiological levels. Transcription factors contain diverse interacting partners for precise transcriptional regulations. Different combinations of interactors would affect temporal and spatial transcriptional regulations. While a large number of transcription factors (TFs) are involved in these transcriptional networks, in vivo investigation on how TF interaction dynamics affect transcriptional regulation in seed-to-seedling transition is lacking. ABSCISIC ACID INSENSITIVE 3 (ABI3) has played a major role serving as the center hub of these transcriptional networks during seed development. ABI3 has diverse interacting partners, including ABI5. The presence of these interacting partners allows ABI3 to bind to diverse cis-regulatory elements (CRE) in a precise manner. One of the processes collectively governed by ABI3 and ABI5 is nutrient deposition, including lipid storage. ABI3 and ABI5 regulate lipid body (LB) related genes such as oleosins for controlling LB dynamics. Notwithstanding, not much is known on ABI5's significance on ABI3's binding towards CREs. Here, we showed that ABI5 impacts ABI3's binding strength towards G-Box motifs. The interaction of ABI3 and ABI5 strengthens ABI3's binding towards ABI3 induced genes. We further demonstrated that ABI3 and ABI5 regulates far more LB related genes during seed development and germination, suggesting that ABI3 and ABI5's have a more precise role in regulating LB dynamics. Our results had further consolidated ABI5's role in ABI3's binding preferences and strength and its significance in controlling LB during seed development and germination.

Legume sprouts contain diverse bioactive phytochemicals, yet species- and accession-level comparisons of antioxidant properties and secondary metabolites remain limited. Antioxidant capacity, total phenolic content (TPC), total flavonoid content (TFC), and 19 secondary metabolites were evaluated in sprouts of soybean (Glycine max (L.) Merr.), mungbean (Vigna radiata (L.) R. Wilczek), cowpea (Vigna unguiculata (L.) Walp.), and peanut (Arachis hypogaea L.), using ten accessions per species under standardized conditions. Mungbean and cowpea sprouts showed significantly higher antioxidant activity, TPC, and TFC than soybean and peanut; across accessions, ABTS and DPPH activities ranged from 13.67 to 49.33% and 7.91 to 55.16%, and TPC and TFC from 3.91 to 13.81 mg GAE/g and 0.05 to 1.04 mg QE/g, respectively. Metabolite profiling revealed species-specific patterns, including isoflavones in soybean, rutin in mungbean, coumestrol in cowpea and resveratrol in peanut. Both antioxidant and phytochemical profiles varied with species and accession, so both can be selected to obtain sprouts with targeted properties.

Although iron (Fe) is abundant in soils, its low solubility, particularly under alkaline conditions, often limits plant Fe acquisition. Under these conditions, coumarin-mediated Fe mobilization becomes increasingly important. Arabidopsis thaliana FE DEFICIENCY-INDUCED KELCH REPEAT PROTEIN 1 (FDK1, AT3G07720) is strongly induced by Fe deficiency and regulated by the central transcription factor FIT, but its function is unknown. We investigated FDK1 using genetic, physiological, metabolite, and transcriptomic analyses. Loss of FDK1 altered root growth, whereas this phenotype was restored in the FDK1-Ox overexpression line. Under alkaline Fe deficiency, growth medium from FDK1-Ox contained strongly increased levels of the Fe-mobilizing coumarins fraxetin and esculetin compared with wild type, whereas their induction was attenuated in fdk1-1. RNA-seq analysis linked FDK1 abundance to coumarin biosynthesis and iron-homeostasis gene expression. FIT and many FIT-associated genes were highly expressed in FDK1-Ox, whereas several upstream iron-homeostasis regulators, including bHLH Ib factors, were highest in fdk1-1. These findings identify FDK1 as a component of the alkaline Fe-deficiency response that promotes iron-mobilizing coumarin accumulation and is associated with coordinated regulation of iron-homeostasis genes.

Background: Weed control is one of the main challenges in agriculture today, particularly due to the increasing occurrence of herbicide-resistant populations. Among the most problematic species are Lolium multiflorum (L.) and Echinochloa crus-galli (L.) Beauv., for which glyphosate-resistant populations have been reported. In this context, glufosinate ammonium has emerged as an alternative for their control; however, its efficacy may vary depending on species and photosynthetic metabolism. Objective: The objective of this study was to evaluate the differential sensitivity of ryegrass (C3) and barnyardgrass (C4) to ammonium glufosinate by analyzing physiological responses associated with leaf senescence and photosystem II activity. Methods: Visual injury, chlorophyll fluorescence, and ammonium accumulation were assessed. Results: Results revealed a differential response between species. Barnyardgrass exhibited earlier symptom onset and a greater reduction in the quantum yield of photosystem II ({Phi}PSII), whereas ryegrass showed a slower senescence process. These differences indicate a higher sensitivity of barnyardgrass to glufosinate ammonium, possibly associated with its C4 photosynthetic metabolism. Conclusions: It is concluded that the effectiveness of glufosinate ammonium depends on the type of photosynthetic metabolism and on the ability of each species to cope with herbicide-induced oxidative stress. This information contributes to optimizing glufosinate ammonium use and to the development of management strategies aimed at delaying the evolution of herbicide resistance.