Search Results (586)

Aluminum-activated malate transporter (ALMT) channels mediate root malate efflux, a key response for plant survival in acidic, aluminum-toxic soils. The acidophilic genus Vaccinium is horticulturally important, yet its ALMT family has remained uncharacterized. Leveraging two newly available chromosome-level genomes (Vaccinium darrowii and Vaccinium duclouxii), we present the first genus-wide characterization of this family. Across 11 angiosperms we identified 145 non-redundant ALMT loci, partitioned into six major subfamilies. MCScanX synteny revealed compact retention rather than expansion: diploid Vaccinium genomes encode only 8–10 ALMTs each, with at most one intra-species syntenic paralog pair, versus multiple whole-genome-duplication-derived pairs in apple (Malus domestica, 24 loci). Pairwise Ka/Ks was elevated within Vaccinium in three subfamilies, and codon-based PAML branch-site tests detected Bonferroni-significant positive selection on terminal Vaccinium branches in Subfamilies 5 and 4 (2ΔlnL = 40.65 and 12.95), yielding three Bayes Empirical Bayes candidate sites in Subfamily 5 (F249, E410, L411) and two in Subfamily 4 (E249, G399); Subfamily 2 was non-significant and is interpreted as relaxed constraint. All candidate residues map to C-terminal cytoplasmic regulatory regions rather than the transmembrane pore. These findings indicate that the compact Vaccinium ALMT repertoire retained its ancestral channel architecture while accumulating lineage-associated divergence in cytoplasmic regulatory regions; the identified residues are candidates for downstream functional validation rather than demonstrated drivers of acid-soil adaptation. Full article

Postharvest quality deterioration of Korla fragrant pear (Pyrus sinkiangensis Yu) severely constrains its market value, yet the regulatory role of preharvest soil management in shaping postharvest performance remains poorly understood. Although green manure is widely adopted to ameliorate orchard soil degradation, species-specific modulation of postharvest storage trajectories and the quantitative fidelity of soil-to-fruit nutrient transmission have rarely been resolved for climacteric pear species. This study investigated how green manure species modulate fruit quality at harvest and during postharvest storage life and their underlying soil–fruit linkages. Three preharvest treatments were imposed, as follows: control (CK), sweet clover (CM), and alfalfa (MX). Fruits were harvested and stored at 4 °C, with samplings at 1, 5, 10, 15, and 20 d. A critical quality transition was identified at 15 d, characterized by the concurrent peaking of soluble sugars, organic acids, vitamin C, and anthocyanins alongside an optimal sugar–acid ratio. Beyond this inflection point, CM and MX diverged markedly: CM enhanced soluble sugar accumulation, anthocyanin retention, and ester volatile production—most notably hexyl acetate, which increased over 14.4-fold—thereby generating a pronounced fruity aroma bouquet. Conversely, MX sustained higher amino acid and vitamin C levels and conferred superior late-storage stability, evidenced by a three-fold lower coefficient of variation in the sugar–acid ratio relative to CK. Partial-least-squares structural equation modeling (PLS–SEM) revealed soil fertility as the principal exploratory associative factor of fruit quality, but the fidelity of soil-to-fruit transmission was species-dependent. MX exhibited the highest observed associative strength (R² = 0.971), whereas CM exhibited attenuated transmission fidelity (R² = 0.777), with network analysis further indicating that CM exhibited divergent associative patterns of key soil–fruit correlations. These findings suggest that green manure identity is linked to postharvest quality through divergent soil–fruit coupling pathways: alfalfa shows nutrient transmission efficiency and stabilizes nutritional quality, whereas sweet clover promotes sugar-aroma accumulation at the cost of reduced soil–fruit conversion fidelity. Species-specific green manure selection thus offers a viable strategy for targeted modulation of postharvest traits in Korla fragrant pear. Full article

Rare genetic diseases are heterogeneous across mechanisms, trajectories, and treatment responses. To date, approved therapies remain available for only a small proportion of rare genetic diseases. Oligonucleotide-based RNA therapeutics, particularly antisense oligonucleotides (ASOs) and small interfering RNAs (siRNAs), offer a promising therapeutic avenue for rare genetic diseases with sequence-level precision. However, traditional preclinical paths may mis-predict human outcomes when disease biology diverges from animal models. New approach methodologies (NAMs), including patient-derived induced pluripotent stem cells (iPSCs), organoid models, and clinical-trials-in-a-dish (CTiD), aim to bring human biology earlier into the translational pipeline. NAMs enable variant-to-function studies, efficacy screening, and safety triage at clinically relevant speed and scale. While critics argue that NAMs are unvalidated and cannot replace preclinical animal models, proponents report that they are increasingly able to recapitulate human phenotypes and predict clinical liabilities, although their predictive validity remains context-dependent. Here, a front-loaded human filter refers to the use of human-derived systems early in development to support mechanistic interpretation, candidate prioritization, and early liability assessment before broader nonclinical evaluation. Recent studies pairing NAMs with ASOs support rapid, patient-specific preclinical screening in selected settings, while also showing the need for broader evidence on delivery, pharmacology, safety, and clinical relevance. This review places these developments within the translational realities of oligonucleotide-based therapeutics, including model fidelity, ASO chemistry and optimization, delivery challenges, pharmacology, regulatory pathways for individualized ASOs, and accessibility. We also propose a pragmatic validation framework to assess the scientific and translational credibility of NAMs across rare genetic diseases. Full article

Cyclin-dependent kinases (CDKs) and cyclins are master regulators of the cell cycle, playing critical roles in plant growth, development, and stress responses. While these gene families have been extensively studied in model plants, a comprehensive analysis in soybean remains underexplored. To address this gap, we performed a genome-wide identification and systematic analysis of these families in soybean using bioinformatic approaches. Expression profiles and protein interactions of selected GmCDK and GmCyclin candidates were tested by qRT-PCR and BiFC assays. A total of 28 GmCDK and 101 GmCyclin genes were identified, revealing a significant expansion compared to Arabidopsis, rice, and maize, primarily driven by whole-genome and segmental duplications. Phylogenetic analysis classified GmCDKs into seven conserved clades (CDKA-CDKG) and GmCyclins into ten distinct subfamilies. Expression profiling demonstrated dynamic, tissue-specific patterns, with distinct modules active during seed development and in tissues. Promoter analysis further linked these genes to hormonal and stress-responsive pathways. Crucially, BiFC assay identified specific interactions between GmCDKA2, GmCDKA3, GmCDKB1 and GmCYCA3-3, suggesting evolutionary divergence in soybean CDK-Cyclin regulatory networks. This study provides a foundational resource for the soybean cell cycle regulome, highlighting its evolutionary plasticity and implicating specific CDK-Cyclin pairs as potential targets for manipulating agronomic traits such as seed development and stress resilience. Full article

Accredited sensory certification is increasingly central to the credibility of EU Geographical Indications (GIs), particularly for wine, where typicity and regional identity compli-cate harmonization. In the absence of shared descriptors, reference standards, and decision rules, sensory control may produce inconsistent conformity outcomes across certification bodies. This study examines the main drivers and limitations of sensory harmonization in GI wine certification, with particular focus on Portugal’s mandatory batch-level sensory approval system. Using a structured narrative review and comparative analysis, it integrates sensory science literature with the EU regulatory framework requiring verifiable organoleptic characteristics in product specifications. National approaches from Portugal, Croatia, France, Germany, Greece, Italy, Romania and Spain are compared, alongside the harmonized IOC Panel Test model for virgin olive oil. Accreditation is analyzed through International Organization for Standardization standards ISO/IEC 17025 and ISO/IEC 17065, as well as EA guidance. Results show increasing convergence toward formal sensory certification but persistent divergence in how typicity is operationalized and translated into conformity decisions. The study proposes a conceptual framework in which harmonization focuses on evidential conditions rather than uniform sensory identities. Full article

Soil salinization represents one of the most severe abiotic constraints on global forest productivity. Populus, the most widely cultivated fast-growing timber tree and a premier model woody plant, exhibits striking intrageneric variation in salt tolerance—from the extremely halophytic Populus euphratica to highly salt-sensitive cultivated clones. Understanding the molecular basis of this variation has profound implications for saline–alkali land reclamation and salt-tolerant variety breeding. This review systematically synthesizes current knowledge on Populus salt-stress responses, covering three primary injury mechanisms (osmotic stress, ionic toxicity, and oxidative damage) and the corresponding physiological countermeasures. We further survey functional genes across four major categories: ion transporters, osmotic-adjustment enzymes, antioxidant-defense components, and transcription factors. Crucially, we extend beyond the herbaceous-plant paradigm by examining salt-tolerance strategies that are specific to the woody architecture of Populus: long-distance radial and axial Na⁺ transport through tall stems, salt sequestration in senescent bark and wood parenchyma, and deep-root ion exclusion strategies. Comparative insights from other woody genera are incorporated to highlight convergent and divergent mechanisms. On this basis, we propose an integrated multi-level regulatory model in which Na⁺ compartmentalization/efflux serves as the core, ROS homeostasis as the key regulatory axis, and osmotic adjustment as the auxiliary strategy. Outstanding challenges—including unresolved primary salt-signal perception, insufficient pathway integration, and limited in planta gene-function verification—are critically assessed, and future research priorities encompassing multi-omics integration, CRISPR-based gene editing, and natural-population genomics are outlined. Full article

Oocyte quality determines fertilization success during the reproductive period of fish and is affected by a key intrinsic factor: the level of lipid deposition in the yolk. In this study, lipidomics was used to characterize lipid profiles of adipose tissue, liver, serum and ovary to clarify divergent hepatic lipid allocation to fat depots versus ovary in Sichuan taimen (Hucho bleekeri). The results showed that glycerophospholipids (GPs) constituted the most diverse lipid class across all four tissues, with phosphatidylcholine (PC) being the most abundant in each tissue. Relative to adipose tissue, liver, and serum, the ovary exhibited significant enrichment of PC and phosphatidylethanolamine (PE) and higher accumulation of docosahexaenoic acid (DHA)- and arachidonic acid (ARA)-containing GPs and triacylglycerols (TGs), as well as enrichment in TGs, diacylglycerols (DGs), and cardiolipin (CL), compared to adipose tissue. The expression levels of fads6, elovl2, elovl5, dgat1b, dgat2, pparα, nfyb, and fabp7 were higher in the liver, while lpcat1 was highly expressed specifically in the ovary. The results of this study demonstrate that long-chain polyunsaturated fatty acids (LC-PUFAs) exhibited selective enrichment in the ovary, which facilitates yolk lipid deposition and provides structural and energetic support for oocyte development and early embryonic development. From a lipidomic perspective, this study reveals the regulatory characteristics underlying ovarian yolk deposition in H. bleekeri, providing important evidence for understanding the mechanisms of egg quality formation as well as a theoretical basis for broodstock management, artificial propagation, and the improvement of egg and larval quality. Full article

The transforming growth factor-beta (TGF-β) superfamily regulates diverse biological processes, including proliferation, differentiation, apoptosis, tissue remodeling, and reproductive signaling across metazoans. Here, we performed a genome-wide characterization of the TGF-β gene family in donkey (Equus asinus, ASM1607732v2) using comparative genomics and bioinformatics analyses, with horse (Equus caballus, EquCab3.0) as a reference to investigate evolutionary conservation and functional divergence. Genome assemblies and proteomes were retrieved from NCBI, and TGF-β genes were identified using BLASTp and HMMER searches (Pfam PF00019), followed by phylogenetic, conserved motif, synteny, Ka/Ks, mutation prediction, subcellular localization, and tissue-specific expression analyses. We identified 40 TGF-β genes in donkeys, exceeding the numbers reported in several mammals, suggesting possible lineage-specific expansion or differential gene retention within Equidae. Phylogenetic and motif analyses demonstrated strong evolutionary conservation across the two principal clades (TGF-β-like and BMP-like). Four segmental duplications were identified, with Ka/Ks ratios ranging from 0.28 to 0.43, indicating strong purifying selection on duplicated genes. Synteny analysis revealed extensive collinearity with the horse genome, supporting conserved equid genomic architecture. Comparative sequence analysis identified 160 amino acid variants, including 11 predicted deleterious mutations in key genes (GDF6, GDF9, GDF10, BMP15, and RGMA), suggesting potential functional divergence associated with reproductive and developmental pathways. Importantly, transcriptomic validation using publicly available donkey RNA-seq tissue expression data (NCBI BioProject: PRJNA1017964) revealed distinct tissue-specific expression patterns, with reproductive tissues (ovary and uterus) displaying enriched expression of TGF-β/BMP signaling components, particularly TGFBR1, TGFBR2, TGFB1, BMP2, BMP4, and BMP7, while canonical fecundity genes (GDF9 and BMP15) exhibited ovary-associated expression. This receptor-dominant signaling profile may have a coordinated TGF-β regulatory network underlying folliculogenesis, reproductive tissue remodeling, and fertility-related processes in donkeys. Subcellular localization predictions showed that most proteins (22/40) were extracellularly localized, consistent with conserved signaling functions. Together, this study provides the first integrated genomic and tissue-expression atlas of the donkey TGF-β superfamily, offering new insights into equid-specific evolutionary conservation, reproductive signaling, and functional divergence. Full article

Heat shock proteins of the 70 kDa family (HSP70s) are essential molecular chaperones that preserve proteostasis by assisting protein folding, transport, and degradation. Although the core HSP70 architecture is deeply conserved, the degree and functional significance of sequence divergence across vertebrates remain incompletely understood. Here, an integrative comparative analysis of HSP70 proteins from ten representative vertebrate species spanning mammals, birds, amphibians, and teleost fish was performed. Multiple sequence alignment, phylogenetic reconstruction, motif discovery, entropy-based conservation profiling, hydrophobicity analysis, and structural mapping reveal a strikingly conserved ATPase domain alongside a more variable substrate-binding domain and C-terminal region. Multiple Expectation Maximization for Motif Elicitation (MEME) motif analysis identifies both universally conserved motifs and lineage-specific elements, highlighting functional constraint as well as adaptive diversification. Structural projection onto the human HSP70 crystal structure (PDB 5AQV) demonstrates that conserved hydrophobic residues cluster in the protein core, whereas variable residues are predominantly surface-exposed. Together, these findings illuminate how evolutionary pressures shape both the conserved chaperone machinery and flexible regulatory regions of HSP70, and they establish a scalable analytical framework for comparative protein evolution studies. Full article

Single-use plastic (SUP) pollution has emerged as a major sustainability and environmental-governance challenge in developing and institutionally fragile regions characterized by weak waste-management systems, uneven enforcement capacity, and fragmented regional coordination. This paper examines the regulation of SUPs in the Intergovernmental Authority on Development (IGAD) member states as a case of regional environmental governance in contexts of institutional diversity and limited regulatory capacity. Using a structured doctrinal and comparative legal-analysis methodology, the study evaluates formally enacted national and selected subnational legal and policy instruments through the framework of international environmental law principles, particularly prevention, precaution, polluter-pays, cooperation, and life-cycle governance. The findings reveal substantial divergence in plastics governance across the region. Some countries, including Eritrea, Djibouti, and Somalia, rely primarily on direct bans and preventive restrictions, while others regulate plastics indirectly through broader environmental and waste-management frameworks. Kenya demonstrates the region’s most integrated governance model through preventive regulation, extended producer responsibility (EPR), recycling obligations, and circular-economy measures, whereas responsibility-based governance remains weak across much of the region. The study further shows that fragmented legal systems, weak enforcement capacity, limited recycling infrastructure, and insufficient regional coordination continue to undermine effective plastics governance in IGAD. From a sustainability-law perspective, the paper demonstrates how fragmented institutional environments and uneven governance capacities shape plastics governance in underexamined, fragile regional contexts. It concludes that progressive regional harmonization integrating prevention, producer responsibility, recycling systems, lifecycle governance, and transboundary cooperation offers the most viable pathway toward sustainable plastics governance in the IGAD region while contributing to discussions concerning SDGs 12, 13, 14, 16, and 17. The findings further suggest that IGAD institutions and member-state governments should strengthen extended producer responsibility frameworks, invest in recycling and waste-management infrastructure, enhance enforcement capacity, and promote coordinated regional policies to support a transition toward a circular economy and reduce transboundary plastic pollution. Full article

Strictosidine synthases (STRs) are catalytic enzymes involved in terpenoid indole alkaloid biosynthesis, whereas STR-like (STRL) genes in cereal crops remain poorly understood. Previous studies of the maize STR-like (STRL) gene family have mainly provided genome-wide identification, phylogenetic classification, structural annotation and expression profiling, but the evolutionary constraints and molecular mechanisms underlying STRL diversification remain insufficiently resolved. In this study, we investigated the maize STRL gene family from an evolutionary and structural perspective by integrating sequence divergence, codon usage bias, selection pressure, protein structural modelling, Gene Ontology (GO) enrichment and tissue-specific expression analysis. A total of 21 ZmSTRL genes were analyzed and their comparative and phylogenetic analyses revealed conserved lineages together with maize-associated expansion patterns. Codon usage and neutrality analyses indicated heterogeneous evolutionary constraints among ZmSTRL genes, suggesting that mutational pressure alone does not explain their sequence divergence. Protein conservation and three-dimensional structural modelling showed a generally conserved STR-related catalytic framework, while member-specific variation in terminal and loop regions suggested localized structural divergence. GO enrichment supported conserved catalytic and metabolic signatures, but these associations were interpreted as putative functional evidence rather than direct functional confirmation. Tissue-specific qRT-PCR analysis revealed divergent expression patterns among selected ZmSTRL genes in root, stem, leaf, and anther tissues, indicating possible regulatory specialization. Overall, this study provides an evolutionary-constraint-based framework for understanding STRL diversification in maize and identifies candidate genes and structural features for future functional validation. Full article

Laccases (EC 1.10.3.2, LAC) are copper-containing glycoproteins involved in lignin biosynthesis, and as such, they play important roles in plant development and stress responses. In this study, a genome-wide analysis of the LAC gene family was performed in Pinus massoniana (Chinese red pine), identifying 78 PmaLAC genes, all predicted to encode cell membrane-localized proteins. These genes were unevenly distributed across eight chromosomes, with notable clusters on chromosomes 7 and 8, indicating gene duplication-driven expansion in P. massoniana. Phylogenetic analysis revealed that PmaLAC genes are classified into five subfamilies, reflecting the lineage-specific expansion and evolutionary divergence of gymnosperm LAC genes. Conserved motif and gene structure analyses showed high conservation among PmaLAC proteins. Promoter analysis identified numerous cis-acting elements related to hormone signaling, stress, and light responses. RNA-seq analysis revealed distinct tissue-specific expression patterns for PmaLAC gene family members. Moreover, degradome analysis combined with dual-luciferase assays supported the interaction between miR397c-9 and PmaLAC31, suggesting that miR397c-9 negatively regulates PmaLAC31 and indicating a potentially conserved miRNA-mediated regulatory mechanism. Overall, this study provides a systematic overview of the composition, evolution, and potential regulation mechanisms of the PmaLAC gene family in P. massoniana, providing a useful resource for future functional characterization of PmaLAC genes. Full article

Background: Safflower (Carthamus tinctorius L.) is a high-value economic crop with broad applications in agriculture, industry, and traditional medicine. Seed germination and seedling establishment are critical stages in the safflower life cycle, as they directly influence subsequent seedling establishment, survival, and plant growth. However, the transcriptomic dynamics and regulatory mechanisms underlying these processes remain largely unexplored, and the functional roles of long non-coding RNAs (lncRNAs) in this context are also poorly understood. Methods: In this study, transcriptome sequencing was performed across five developmental stages from seed germination to seedling establishment in safflower, followed by a comprehensive transcriptomic analysis and lncRNA identification. Results: Transcriptome sequencing identified a total of 3027 lncRNAs, including 940 natural antisense transcript (NAT)-pair-associated lncRNAs, which were classified into the divergent, convergent, and chimeric categories. Among these, 767 lncNATs were differentially expressed. Further analysis identified 542 NAT pairs in which both the protein-coding gene and its corresponding lncNAT exhibited a differential expression across the five developmental stages. A functional enrichment analysis of the predicted target genes of these lncRNAs suggested their involvement in photosynthesis and hormone-related responses. An enrichment analysis of differentially expressed genes (DEGs) across developmental stages further revealed the significant enrichment of photosynthesis and plant hormone signal transduction-related pathways, suggesting that these pathways are closely associated with safflower seed germination and seedling establishment. A further analysis of photosynthesis-related genes, particularly the expression patterns of LHC family members, suggested that Stage 3 may represent an important developmental transition associated with the optimization of the light-harvesting capacity during early seedling establishment, whereas plant-hormone-related genes are involved in regulating seed germination and subsequent leaf growth during seedling establishment. In addition, a weighted gene co-expression network analysis (WGCNA) identified candidate transcription factors associated with photomorphogenesis and plant hormone responses in safflower. Conclusions: This study advances our understanding of the regulatory mechanisms underlying safflower seed germination and subsequent growth and provides valuable molecular resources for future safflower breeding programs. Full article

Soybean (Glycine max) is an important model crop for studying plant functional genes, such as the NAC transcription factor (TF) gene family. The NAC transcription factor (TF) family is one of the largest plant-specific TF families and plays critical roles in plant growth, development, and stress responses. In this study, we performed a pan-genome-wide analysis of NAC genes using 29 soybean genomes. A total of 5051 NAC genes were identified and clustered into 245 orthologous gene groups (OGGs), including 58 core, 88 soft-core, 32 shell, and 67 cloud groups. Based on phylogenetic relationships, the representative NAC OGGs were assigned to 18 subfamilies, 17 of which contained soybean NAC genes. Gene duplication analysis indicated that whole-genome duplication (WGD)/segmental duplication was the predominant driver of NAC family expansion, accounting for 90.88% of duplication events. Approximately 39.30% of NAC genes carried at least one intact transposable element (TE) within 2 kb upstream or downstream regions. NAC genes with copy number variation (CNV) harbored more nearby TEs than non-CNV genes (1.54 vs. 1.31 TEs per gene), and dispensable NAC genes contained more nearby TEs than core NAC genes (1.59 vs. 1.33 TEs per gene). These results indicate a significant association between local TE abundance and NAC gene CNV or dispensability. Selection pressure analysis showed that dispensable NAC genes had higher Ka, Ks, and Ka/Ks values than core genes, suggesting relatively relaxed evolutionary constraints. Expression profiling across six tissues revealed distinct transcriptional patterns among NAC subfamilies. Structurally conserved subfamilies generally showed broader expression, whereas structurally divergent subfamilies displayed greater expression variability. Regulatory network and Gene Ontology (GO) enrichment analyses suggested that conserved subfamilies were mainly associated with stress responses, while divergent subfamilies were related to cell wall regulation, signal transduction, and ion homeostasis. Further analysis of Wm82 drought RNA-seq data prioritized several putative drought-responsive NAC candidates, including Glyma.16G043200, Glyma.06G248900, Glyma.07G050600, Glyma.12G206900, and Glyma.18G261300. Overall, these findings elucidate the mechanisms of expansion and the functional divergence of the NAC gene family at the soybean pan-genome level, providing a theoretical basis for understanding NAC gene evolution and facilitating future crop improvement. Full article

Overexploitation of groundwater resources poses a critical challenge in major agricultural regions worldwide, yet how confined and unconfined aquifers respond differentially to governance interventions remains poorly understood. This study presents a comparative assessment of hydrochemical evolution and nitrate contamination dynamics in the Weishan Irrigation District, Shandong Province, China, contrasting pre-governance conditions (2011) with post-governance status (2022–2023) following comprehensive overexploitation control. By integrating hydrochemical characterization with stable isotope tracers (δ¹⁸O, δD, δ¹⁵N-NO₃⁻, δ¹⁸O-NO₃⁻) and Bayesian mixing models (MixSIAR), we reveal fundamentally contrasting aquifer responses to regulation. The unconfined aquifer exhibited continued degradation under persistent agricultural influence, characterized by elevated sodium, nitrate, and bicarbonate concentrations. In sharp contrast, the confined aquifer demonstrated substantial recovery, with major ion concentrations declining markedly, hydrochemical facies restored toward a pristine state, and overall water quality improving significantly to achieve full compliance with the highest-quality standards by 2023. These divergent trajectories indicate that regulatory interventions effectively restored aquitard barrier integrity, thereby shielding the confined aquifer from surface contamination, whereas the unconfined aquifer remained vulnerable to agricultural pollution. Isotope-constrained Bayesian modeling identified soil organic nitrogen, chemical fertilizers, manure/sewage, and industrial wastewater as dominant nitrate sources, with isotopic evidence confirming that the unconfined aquifer receives mixed recharge from Yellow River water and precipitation under contemporary contamination, while the confined aquifer maintains independent, pollution-free recharge. These findings demonstrate that overexploitation control can effectively rehabilitate confined aquifer systems by reestablishing natural hydrogeological barriers, but unconfined aquifers require targeted agricultural pollution mitigation. The contrasting responses highlight the necessity of aquifer-specific management strategies in irrigation-dependent regions, advancing theoretical understanding of how regulatory measures differentially affect multi-layered groundwater systems and providing a scientific basis for precision groundwater governance. Full article