Leucoanthocyanidin reductase (LAR) is a key enzyme involved in the biosynthesis and accumulation of proanthocyanidins (PAs). Here, we conducted a genome-wide identification and systematic characterization of the LAR gene family in the legume forage species sainfoin (Onobrychis viciifolia Scop.). A total of 83 OvLAR genes were identified in the sainfoin genome, and they are unevenly distributed across 28 chromosomes. Phylogenetic analysis classified the 83 OvLAR proteins into five evolutionary clades. Furthermore, segmental duplication was the major driving force for the expansion of the OvLAR gene family. Cis-acting element profiling indicated that OvLAR gene expression is potentially regulated by both endogenous cues and environmental signals, implicating these genes in complex regulatory networks that coordinate secondary metabolism and adaptive development. Consistently, qRT-PCR results revealed tissue-specific expression patterns among OvLAR genes, as well as differential transcriptional responses to multiple abiotic stresses. Functional assays further demonstrated that overexpression of the key gene OvLAR71 in tobacco and sainfoin markedly promoted PA biosynthesis and accumulation in leaves and flowers, accompanied by distinct changes in floral pigmentation. Collectively, this work provides the first comprehensive overview of the LAR gene family in sainfoin, offering important insights into its roles in PA biosynthesis and establishing a theoretical foundation for the genetic improvement of forage quality.

Licorice is an important medicinal herb worldwide, including three Chinese Pharmacopoeia species (Glycyrrhiza uralensis, G. inflata, G. glabra), with bioactive compounds crucial for disease treatment and industrial applications. However, the genetic mechanisms underlying the biosynthesis, diversification, and environmental adaptation of bioactive compounds in Glycyrrhiza species have long remained unclear. Herein, we assembled a gapless telomere-to-telomere (T2T) genome of G. uralensis with resolved telomeres and centromeres and three significantly improved high-quality chromosome-level Glycyrrhiza genomes, alongside a variation map of 188 wild accessions. Population analysis revealed evolutionary divergence among species, with selection signals linked to medicinal compound pathways. We identified 4CL5 as a key gene for stress response and compound synthesis. GWAS validation highlighted the GiPHL1-Gi4CL5 module's role in licochalcone A accumulation and enhanced stress adaptation in G. inflata. This study provides the first T2T Glycyrrhiza genome and insights into medicinal compound biosynthesis and environmental adaptation.

Objective The GDSL esterase/lipase (GELP) family comprises a group of multifunctional hydrolases with broad substrate specificity and catalytic versatility, playing an essential role in plant growth and developmental processes. The GDSL lipase gene GhGELP23D from Gossypium hirsutum is highly expressed during cotton fiber elongation development. This study aims to investigate its function in plant cell elongation and provides references for the mechanism elucidation of GELP-mediated regulation of cell growth. Method The GhGELP23D was cloned from upland cotton fibers. Bioinformatic analyses were performed to characterize its physicochemical properties, structural features, and phylogenetic relationships. The promoter region was analyzed with PlantCARE to identify putative cis-elements. Expression patterns were detected based on public transcriptomic data and RT-qPCR validation. A GUS reporter construct driven by the GhGELP23D promoter was generated to examine the tissue-specific expression of GhGELP23D in stably transformed Arabidopsis. Subcellular localization of GhGELP23D was determined through transient expression in Nicotiana benthamiana leaves. Transgenic Arabidopsis lines heterologously expressing GhGELP23D (GhGELP23D-OE) were obtained using the floral-dip method, and phenotypic analyses were conducted. Additionally, virus-induced gene silencing (VIGS) was employed to suppress GhGELP23D expression in cotton. Silencing efficiency was verified by RT-qPCR, and the resulting changes in fiber length were assessed in GhGELP23D-VIGS plants. Result GhGELP23D encodes a 356-amino-acid protein that is stable, weakly basic, and hydrophilic, containing a typical signal peptide but no transmembrane domain. Transcriptomic analysis and RT-qPCR detection showed that GhGELP23D was highly expressed during cotton fiber elongation. Subcellular localization analysis revealed that GhGELP23D is predominantly localized to the extracellular space. Analysis of the GhGELP23D promoter indicated the presence of multiple cis-elements related to light responsiveness, hormone signaling, and stress response. Histochemical GUS staining showed that the GhGELP23D promoter can drive reporter gene expression in various Arabidopsis tissues, indicating broad tissue expression activity. Heterologous expression of GhGELP23D in Arabidopsis (GhGELP23D-OE) significantly enhanced plant growth and development, resulting in increased plant height and longer primary roots and root hairs. Furthermore, complementation of GhGELP23D in the mutant resulted in a pronounced recovery of primary root and root hair length similar to that of the wild type (WT). In cotton, the fiber length of GhGELP23D-VIGS lines was significantly reduced compared with that of empty-vector control plants, with an average decrease of approximately 13.14%. Conclusion GhGELP23D encodes an extracellular GDSL lipase that is highly expressed during the fiber elongation stage of upland cotton and plays an important role in plant cell elongation development.

Verticillium wilt (VW), caused by the soil-borne fungus Verticillium dahliae, is a major disease that markedly compromises both the yield and fiber quality of cotton. In this study, we explored the function and underlying mechanism of the cotton expansin gene GhEXLB2 in response to VW infection. Expression profiling revealed that members of the GhEXL family exhibit distinct patterns across tissues and under various biotic and abiotic stresses. Notably, GhEXLB2, which encodes an extracellular protein, showed the strongest induction following V. dahliae challenge. Ectopic expression of GhEXLB2 in Arabidopsis thaliana promoted root elongation and root hair formation, and was associated with improved resistance to the pathogen. In contrast, silencing GhEXLB2 in cotton via virus-induced gene silencing (VIGS) led to pronounced vascular browning, increased pathogen recovery, and a lower level of disease resistance. In addition, RNA-seq profiling of GhEXLB2-silenced (VIGS) cotton plants revealed that most differentially expressed genes were enriched in pathways related to phytohormone signaling and plant-pathogen interactions, with salicylic acid (SA) signaling and WRKY transcription factors emerging as central regulatory components. Analysis of the GhEXLB2 promoter further identified multiple cis-acting elements associated with stress and hormone responsiveness. When integrated with protein-protein interaction (PPI) prediction data, these results suggest that GhEXLB2 may be modulated by a network of transcription factors and signaling pathways. Collectively, the evidence supports a positive association between GhEXLB2 and VW resistance. This study provides a framework for understanding expansin functions in cotton defense against VW.

Endocytosis is central to cellular trafficking and signaling across eukaryotes, yet whether and how plant viruses actively reprogram this pathway remains unclear. Here, we show that the geminiviral betasatellite-encoded βC1 reprograms the host VPS9a-Rab5 endocytic module to promote viral infection. βC1 associates with the Rab5 GTPases ARA6 and ARA7 as well as their guanine nucleotide exchange factor VPS9a, stabilizing the VPS9a-Rab5 complex and enhancing nucleotide exchange. This catalytic potentiation sustains Rab5 activation and drives endosome proliferation, which, in turn, stabilizes βC1 to support efficient viral replication. Genetic disruption of Rab5 or VPS9a compromises endocytosis, reduces βC1 accumulation, and restricts infection by multiple geminiviruses. Together, these findings define the VPS9a-Rab5 module as a central proviral hub linking host membrane dynamics to effector stability and viral DNA amplification, revealing an unanticipated strategy in which a viral effector amplifies a cellular regulatory catalyst to reprogram a fundamental cellular pathway for virus infection.

Extensive use of chemical herbicides has raised serious concerns regarding agricultural sustainability and ecological safety, highlighting the need for environment friendly bioherbicides. In this study, activity-guided fractionation led to the identification of xanthoxylin and α-santonin from the ethanol extract of a dominant desert plant, Seriphidium transiliense, with the phytotoxicity of xanthoxylin being reported for the first time. Petri dish bioassay revealed that both compounds significantly suppressed seedling growth of tested plants in a dose-dependent manner; at 1000 μg/mL, α-santonin inhibited root growth of Amaranthus retroflexus, Setaria viridis, Medicago sativa, and Oxybasis glauca by 98.25%, 79.75%, 71.40%, and 62.75%, respectively, whereas the corresponding inhibition rates for xanthoxylin were 59.15%, 89.71%, 38.80%, and 62.90%. Following pot experiments revealed that both compounds significantly increased MDA content and altered the activities of SOD, CAT, and POD of A. retroflexus seedlings, indicating the induction of oxidative stress. Treated plants also displayed chlorosis and leaf whitening, suggesting possible disturbance of photosynthetic pigment-related processes; subsequent molecular docking further implied that both compounds may interact with protoporphyrinogen IX oxidase (PPO), a key enzyme associated with tetrapyrrole metabolism and chlorophyll biosynthesis. Our results suggests that α-santonin and xanthoxylin have the potential to be developed as bio-herbicides.

Licorice (Glycyrrhiza uralensis) roots are among the most widely used medicinal raw materials in traditional medicine and in numerous industries, with their polysaccharides serving as key bioactive constituents responsible for immunomodulation, tissue repair, and drug synergy. However, the low extraction yield and quality of the polysaccharides limit their extensive utilization. Presently, a highly efficient method for the extraction of polysaccharides from G. uralensis roots is lacking. Here, we developed an ultrasound-assisted deep eutectic solvent (UDE) method for extracting high-yield and premium-quality polysaccharides from G. uralensis roots. Eight extraction methods were systematically compared using deep eutectic solvents (DESs) and water as extraction media under various processing modes, including heat conduction, alkaline treatment, enzymatic hydrolysis, microwave irradiation, and ultrasound. Comparative analysis indicated that the UDE method resulted in the highest polysaccharide yield of 18.56% with a 1.71-fold increase over hot water extraction. UDE-extracted polysaccharides exhibited lower molecular weight, smaller particle size, higher solubility, improved thermal stability, and enriched galacturonic acid content, forming a weak gel-like structure with high elastic modulus. Density functional theory and independent gradient model analyses revealed that the DESs interacted with monosaccharide units through extensive hydrogen bonding and van der Waals forces. The UDE-extracted polysaccharides also displayed strong radical scavenging activity similar to vitamin C, and they alleviated IL-13-induced inflammation in BEAS-2B cells by reducing nitric oxide production, restoring antioxidant enzyme activity, and suppressing the STAT6/MYD88 axis. Thus, a green, efficient, and scalable UDE-mediated method is established for extracting highly bioactive polysaccharides for applications in numerous fields.

Polyamines, a class of low-molecular-weight nitrogen-containing bases with high biological activity, are ubiquitous in organisms and play protective roles in plants under stress. Polyamine oxidase (PAO), a typical flavoprotein characterized as a glycoprotein, is a key enzyme in polyamine catabolism that directly mediates polyamine breakdown and maintains intracellular polyamine homeostasis. However, the specific functions of PAOs in cotton fiber development remain largely unclear. In this study, we identified 23 GhPAO genes from the upland cotton (Gossypium hirsutum L.) genome via comprehensive bioinformatics approaches. We systematically analyzed their physicochemical properties, phylogenetic relationships, gene structures, chromosomal locations, conserved motifs, cis-acting elements, and expression patterns. Quantitative real-time PCR (qPCR) analysis confirmed that GhPAO10 and GhPAO21 exhibited the most pronounced transcript accumulation during both fiber development and stress response processes. Further yeast one-hybrid (Y1H) and dual-luciferase reporter assays indicated that the GhPAO21 promoter was directly regulated by the transcription factor GhTGA1. Our findings provide a foundation for elucidating the functional roles of the PAO gene family in upland cotton and underscore potential candidate genes associated with fiber development and stress responses.

Ammopiptanthus nanus (Fabaceae) is a Class II nationally protected endangered evergreen shrub in China and is endemic to the arid regions of Central Asia. To assess how contrasting ex situ management histories are associated with sequence-variant retention at an ecologically relevant gene, we analyzed a 594 bp coding fragment of the antifreeze protein gene (AnAFP) in one wild population and two ex situ collections maintained under active versus passive management contexts. Only two variable sites were detected across 75 individuals, both represented by single-base indels near the 5 ' end of the coding region. The wild population contained both rare variants, the actively managed ex situ collection retained one of them at low frequency, and the passively maintained collection was monomorphic across the analyzed fragment. Rarefaction analysis indicated that the absence of variation in the passive collection is unlikely to be explained by sample-size disparity alone at this targeted locus. Because only one locus was analyzed, these results are interpreted as locus-specific patterns rather than evidence of genome-wide diversity change. Nevertheless, the observed pattern is consistent with reduced retention of rare sequence variants in the passive ex situ collection and with the possibility that a narrow founder base, together with the absence of subsequent genetic supplementation, contributed to this outcome. These results support the view that ex situ conservation of A. nanus may benefit from maximizing founder representation, maintaining sufficiently large managed collections, and combining neutral marker approaches with targeted monitoring of ecologically relevant loci. Targeted loci such as AnAFP should, however, be regarded as complementary indicators rather than stand-alone proxies for broader genetic diversity or adaptive potential.

Histidine triad (HIT) family proteins contain a conserved histidine triad motif and play key roles in fungal metabolism and pathogenicity. This study focused on VD9136, a member of the HIT family in Verticillium dahliae, aiming to elucidate its biological function and mechanism underlying its role in cotton pathogenesis. A systematic investigation of the VD9136 gene in V. dahliae was conducted using bioinformatics analysis, gene knockout, genetic complementation, and pathogenicity assays. The results showed that VD9136 protein consists of 136 amino acids and is a stable, neutral, and weakly hydrophilic protein that lacks transmembrane domains and signal peptides; it is localized to the extracellular space via a non-classical secretion pathway. Its secondary structure is predominantly composed of α-helices and random coils. Phylogenetic analysis revealed that VD9136 is closely related to VliHIT, a homologous protein from V. longisporum, the pathogen responsible for Verticillium wilt in rapeseed. The promoter region of VD9136 contains multiple cis-acting elements, including light-responsive, hormone-responsive, and stress-responsive elements, indicating that its transcription may be regulated by multiple signaling pathways. VD9136 was significantly upregulated during the early stage of cotton infection (6-24 h post-inoculation). Pathogenicity assays demonstrated that V. dahliae knockout mutants lacking VD9136 exhibited a significant reduction in virulence, as evidenced by a lower disease index, decreased fungal biomass within plant tissues, and attenuated vascular browning in cotton plants. The pathogenic phenotype was successfully restored in genetic complementation strains. This study identified VD9136 as a key regulatory factor in the pathogenic process of V. dahliae, and its loss of function reduces the pathogenicity of V. dahliae. The findings provide a theoretical basis for elucidating the pathogenic mechanism of cotton Verticillium wilt and for developing corresponding prevention and control strategies.

IntroductionSafflower (Carthamus tinctorius L.) is a prized medicinal species whose therapeutic value hinges on the abundance of bioactive metabolites. Accumulation of these metabolites are influenced by a range of environmental and edaphic factors, including soil physicochemical parameters, extracellular enzyme activities, composition and function of rhizosphere microbiome. However, how these factors individually and synergistically orchestrate the biosynthesis, transport, and ultimate storage of pharmaceutically active compounds within Safflower tissues remains unknown.MethodsHere, high-throughput amplicon sequencing coupled with comprehensive physiological profiling was employed to investigate soil characteristics, enzyme activities, and rhizosphere microbial communities of safflower across 36 soil samples collected at two distinct altitudes and two growth stages.ResultsThe effective component content was detected in 18 samples, and our results revealed that the safflower stigmas from the high- altitude site (YM) contained significantly elevated levels of hydroxysafflor yellow A (HSYA) compared to those from the lowland site (YF). Soils at the YM site exhibited markedly higher fertility, with available phosphorus, total nitrogen, and organic matter identified as key drivers of HSYA accumulation. Both sites showed high diversity and abundance in rhizosphere microbial communities, with Actinobacteria and Proteobacteria dominating the bacterial communities, and Ascomycota being the predominant fungal phylum.DiscussionTaken together, our findings show that soil properties, microbial communities, and climatic conditions work interactively to influence the buildup of bioactive compounds in safflower. These insights suggest that precise management of soil nutrients and the rhizosphere microbiome can improve medicinal safflower quality.

Transposable elements (TEs) constitute the largest fraction of cotton genomes, serving as primary drivers for genome expansion and species diversification. However, comprehensive chromosomal physical maps visualizing the evolutionary dynamics of TEs between allotetraploid cotton (including naturally colored and white cotton) and related diploid progenitors remain scarce. Here, we reconstructed TE landscapes by integrating a de novo assembly approach with fluorescence in situ hybridization (FISH). Comparative FISH analysis of 31 distinct TEs revealed conserved chromosomal distribution patterns across naturally colored and white cotton cultivars, with the A-subgenome exhibiting significantly higher TEs abundance and diversity than the D-subgenome. Cytological analysis between diploid progenitor and allotetraploid cotton showed that most TEs (22/31) maintained conserved evolutionary patterns post-polyploidization, while only 9 TEs underwent marked abundance shifts. Notably, cytological and genomic analyses revealed eight unique TEs that were highly abundant in the A-diploid progenitor but either absent (six TEs) or present at extremely low levels (two TEs) in the D-diploid progenitor, however, following allopolyploidization, these TEs underwent massive amplification in the D-subgenome, even producing distinct FISH signals in the interstitial regions of all D-subgenome chromosomes. This leads us to hypothesize that asymmetric TE composition between A and D-subgenomes probably activates two complementary mechanisms—expansion of pre-existing D-subgenome TEs and invasion-driven amplification from the A-subgenome—collectively driving convergent evolution of the D subgenome toward an A-subgenome-like TE pattern, thereby potentially enhancing allotetraploid cotton genome stability. These findings provide further insights into the roles of TEs in cotton allopolyploidization and genome evolution from a cytological perspective.

The rapid advancement of wearable electronics has underscored the importance of flexible thermoelectric (TE) devices as efficient platforms for continuous energy harvesting from body heat or environmental temperature gradients, due to their excellent adaptability to complex and dynamic surfaces. Among various candidates, cellulose-based materials derived from renewable biomass stand out for their outstanding processability, sustainability, and environmental compatibility, making them ideal scaffolds for flexible TE systems. Recent developments reveal that combining cellulose matrices with conductive polymers, carbon nanomaterials, and inorganic TE components can yield hybrid composites that simultaneously exhibit high thermoelectric performance and mechanical flexibility. Such innovations highlight the significant potential of cellulose materials for sustainable energy conversion and self-powered wearable applications. This review critically summarizes the latest progress in the design, fabrication, and integration of cellulose-based thermoelectric materials and devices, with a particular focus on processing strategies and performance enhancement mechanisms. Furthermore, it systematically discusses the existing challenges in material selection, thermal-electrical transport optimization, environmental degradability, and scalable manufacturing. Through this comprehensive analysis, the review aims to provide deep insights and practical guidance for developing next-generation, high-performance, and eco-friendly thermoelectric materials to advance sustainable energy harvesting technologies.

Natural polysaccharides are essential macromolecules found in both plant and animal kingdoms, playing significant roles in food, medicine, biomaterials, and packaging. Deep eutectic solvents (DES), formed through hydrogen-bond interactions, offer simple synthesis, eco-friendliness, and lower melting points. This review demonstrates DES-mediated multi-modal extraction techniques (ultrasound, microwave, pressure, etc.) achieves 1.45-4.30-fold higher polysaccharide yields versus hot water extraction. Specifically, wolfberry fruits showed 2.44-fold increase, Ganoderma lucidum exhibited 4.30-fold enhancement, and Camellia oleifera fruit shells demonstrated 1.45-fold improvement. DES modifies key structural parameters including molecular weight reduction, degree of esterification decrease, and morphology changes (smaller particle sizes with enhanced porosity). Enhanced bioactivities include: DPPH center dot scavenging capacity increased 15-30 %, anti-glycation inhibition reached 87.63 % versus 71.52 % for water extraction in Polygonatum odoratum, and superior alpha-glucosidase inhibition for diabetes management in multiple sources. This article reviews DES origins, synthesis, synergy with multi-modal techniques, and supports further research and industrial implementation.

Paraquat (PQ) causes acute lung injury in part via mitochondrial respiratory dysfunction. We evaluated whether Glycyrrhiza uralensis. extract (LE) confers holistic protection and whether liquiritin (LQ) is a key mitochondria-targeting constituent. A PQ lung-injury model was established in KM mice by a single oral dose of PQ (20 mg/kg). LE was gavaged after PQ exposure for in vivo assessment; LQ was used as pretreatment in A549 and HPAEpic cells. Outcomes included lung function, pulmonary microcirculation, histopathology, and oxidative-stress markers (MDA, SOD). UPLC-Q/TOF-MS profiled LE composition. Differential genes and pathway enrichment were derived from integrated transcriptomics (GSE171625 plus our RNA-seq). Mitochondrial superoxide, membrane potential (Δψm), mtDNA leakage, and respiratory-chain proteins were measured. Rotenone and antimycin A were used as pharmacological probes, and molecular docking was employed to assess target engagement. Eighteen major LE constituents were identified; enrichment linked them to oxidative stress, inflammation, fibrosis, and metabolism pathways. Transcriptomics converged on mitochondrial pathways. LE treatment improved clinical appearance and lung function, enhanced microcirculation, reduced alveolar wall thickening, inflammation, and early fibrosis, lowered MDA levels, and restored SOD levels in PQ-exposed mice. In vitro, LQ restored mitochondrial function, increasing Δψm and respiratory activity while limiting PQ-induced damage. Perturbation with rotenone/antimycin A supported Complex I as a core target, consistent with docking. This work establishes a prediction-to-validation chain from LE's multi-component holistic efficacy to LQ's mitochondria-targeted mechanism, validating Complex I as a mechanistic node and supporting LE/LQ as candidates against PQ-induced lung injury.

Cotton, as a globally significant economic crop, is intricately regulated in its growth and development by the key genes for SA (Salicylic acid) biosynthesis. In the present study, a systematic analysis of genes related to SA biosynthesis was conducted across four cotton species, leading to the identification of 70 genes. Specifically, the tetraploid species Gossypium hirsutum and G. barbadense were found to harbor 22 and 23 genes, respectively, representing a substantial expansion compared to the 12 and 13 genes identified in the diploid progenitors G. arboreum and G. raimondii. Comprehensive characterization of chromosomal localization, phylogeny, domain architecture, and promoter cis-elements revealed a uniform distribution of key genes involved in SA biosynthesis across A/D sub-genomes of tetraploids with extensive interspecific collinearity; whole-genome and segmental duplication act as the dominant drivers for the expansion of this gene family, while partial gene loss following polyploidization results in non-doubled gene copy numbers in tetraploids relative to diploids, which reflects the evolutionary selection for genomic dosage balance. The key genes for SA biosynthesis demonstrate a high degree of conservation in protein sequences, protein structures, and conserved motifs, which constitute the structural basis for the stable maintenance of their core functions in the SA biosynthesis pathway during plant evolution. This is closely related to their core function in the salicylic acid (SA) synthesis pathway and serves as the structural basis for the stable maintenance of gene functions during evolution. Analysis of cis-elements revealed that the expression of key genes involved in SA biosynthesis is governed by a complex interplay of phytohormones, stress signals, and transcription factors. Yeast one-hybrid (Y1H) assays confirmed the interaction between the GhPAL and GhICS gene and predicted candidate transcription factors, specifically the binding of GhWRKY21 to GhICS2-1 promoter and GhMYB12 to GhPAL1-2 promoter, thus elucidating their stage-specific regulatory mechanisms in cotton fiber development and reflecting their evolution. This study provides a fundamental basis for investigating the role of the SA signaling pathway in cotton development and offers support for cotton molecular breeding.

Sainfoin (Onobrychis viciifolia Scop.) is a valuable perennial forage legume abundant in proanthocyanidins (PAs). As important secondary metabolites in plants, PAs not only contribute to plant stress tolerance and defense processes, but also significantly improve forage quality. Leucoanthocyanidin reductase (LAR) is a key enzyme in the biosynthesis and accumulation of PAs. However, systematic research on the LAR gene family in sainfoin remains limited. Therefore, we conducted a genome-wide identification and systematic characterization of the LAR gene family in the legume forage species sainfoin. A total of 83 OvLAR genes were identified in the sainfoin genome, which were unevenly distributed across 28 chromosomes. Phylogenetic analysis classified the 83 OvLAR proteins into five evolutionary clades. Furthermore, segmental duplication served as the major driving force underlying the expansion of the OvLAR gene family. Cis-acting element profiling indicated that OvLAR gene expression is potentially regulated by both endogenous cues and environmental signals, suggesting that these genes are involved in complex regulatory networks that coordinate secondary metabolism and adaptive development. Consistently, qRT-PCR results revealed tissue-specific expression patterns among OvLAR genes, as well as differential transcriptional responses to multiple abiotic stresses. Functional assays further demonstrated that overexpression of OvLAR71 in tobacco and sainfoin significantly promoted PA biosynthesis and accumulation in leaves and flowers, accompanied by distinct changes in floral pigmentation. Collectively, this work provides the first comprehensive insight into the LAR gene family in sainfoin, offers critical insights into their roles in PA biosynthesis, and establishes a theoretical foundation for the genetic improvement of forage quality.

ABA-responsive element-binding factors (ABFs), as a crucial subfamily within the basic leucine zipper (bZIP) transcription factor family, exert a significant regulatory function in the process of plant responses to stress. However, the genome-wide characteristics and low-temperature response mechanisms of ABFs in cotton remain poorly understood. Herein, through genome-wide analysis across ten Gossypium species, we systematically identified 138 ABF transcription factors characterized by conserved bZIP domains. Evolutionary analysis demonstrated that segmental duplication events, coupled with purifying selection, contributed to family expansion. Cis-element profiling of promoters suggested their involvement in hormonal signaling and abiotic stress responses. Expression analysis displayed that GhABF3 expression level was significantly up-regulated under cold stress. Genetic functional analysis confirmed that GhABF3-overexpression in A. thaliana enhanced the cold tolerance and virus-induced gene silencing (VIGS)-mediated GhABF3-silencing in cotton compromised cold resistance. Comparative transcriptomic analysis of VIGS-GhABF3 cotton plants indicated that the pathways of hormone signaling, ascorbic acid metabolism, and flavonoid biosynthesis are the significantly enriched pathways by KEGG analysis. Integrated DAP-seq and RNA-seq analyses identified GhAO1 as a direct target of GhABF3, which was further validated by Y1H and dual-LUC assays. The contents of AsA and flavonoid were significantly reduced in both VIGS-GhABF3 and VIGS-GhAO1 cotton plants. This research conducted a systematic analysis of the evolutionary traits of the cotton ABF family, clarified the function and molecular mechanism underlying GhABF3-mediated enhancement of low-temperature adaptability, and thus laid a robust basis for further exploration of multi-level regulatory networks and the breeding of cold-tolerant cotton varieties.

This study investigated the mitochondrial response mechanisms of Syntrichia caninervis Mitt., a desiccation-tolerant moss from the Chinese Gurbantunggut Desert, to dehydration-rehydration stress at the subcellular level under three dehydration intensities: rapid drying (RD), slow drying (SD), and air drying (AD). The results showed that the moss maintained the structural integrity of mitochondrial membranes across treatments, demonstrating remarkable phenotypic plasticity. Rapid dehydration caused a pronounced decline in mitochondrial membrane potential (ΔΨm) and distortion of cristae, relying primarily on emergency antioxidant responses involving superoxide dismutase (SOD) and ascorbate peroxidase (APX) to scavenge reactive oxygen species (ROS). In contrast, slow dehydration activated a "pre-adaptive" antioxidant strategy characterized by sustained peroxidase (POD) activation, accumulation of reduced glutathione (GSH), and upregulation of alternative oxidase (AOX) activity. Principal component analysis confirmed that APX, SOD, and AOX were key contributors to drought adaptation. This study is the first to reveal dual-track adaptive mechanisms in desiccation-tolerant plants mediated through mitochondrial ultrastructure, membrane potential dynamics, and redox homeostasis regulation, and provides new targets for improving drought resistance in crops.