Biology subjects

Wang, L.

Publications and source records attributed to Wang, L..

At least 217 records · Page 12Linked to original sources

bioRxiv2026

Sex-determination cascade orchestrates male-male pheromone synthesis in the bean bug

Volatile pheromones are vital for insect intraspecific communication, yet the genetic basis of homosexual recognition remains elusive. In the bean bug Riptortus pedestris, we show that two previously identified aggregation pheromone components, (E)-2-hexenyl-(Z)-3-hexenoate (E2HZ3H) and (E)-2-hexenyl-(E)-2-hexenoate (E2HE2H), serve as key chemosensory cues for male-male recognition during mate selection. Their biosynthesis is governed by the sex-determination cascade Rpfmd-Rpdsx. The male-specific isoform Rpdsx_M promotes pheromone production in males and induces ectopic synthesis in females upon knockdown of the feminizing switch gene Rpfmd. Knockdown of Rpdsx_M in males abolishes both compounds, prompting wild-type males to court them as if they were females. Metathoracic gland cells act as the production hub. Behaviourally, E2HZ3H or E2HE2H disrupt mating when applied to females: males avoid such females. E2HZ3H reduces female mobility in the presence of the male-derived primary aggregation pheromone tetradecyl isobutyrate (14:iBu), whereas E2HE2H shows no obvious such effect. These differential effects ensure mating accuracy. The discovery of volatile pheromones functioning in male-male recognition and of their synthesis being governed by the sex-determination cascade updates our understanding of mating accuracy in insect chemical communication.

animal behavior and cognition↗

bioRxiv2026

CESAR: High-Sensitivity Detection of Copy Number Variations in ctDNA Using Segmentation and Anchor Recalibration

BackgroundDetecting copy number variations (CNVs) in circulating tumor DNA (ctDNA) is crucial for the companion diagnosis and resistance monitoring of various solid tumors (e.g., NSCLC, Glioblastoma). However, when tumor-derived DNA fractions are extremely low (often <1%), traditional depth-based methods frequently fail due to non-linear sequencing depth fluctuations and probe-specific capture biases inherent to targeted Next-Generation Sequencing (NGS). MethodsWe developed CESAR (CNV Estimation with Segmentation and Anchor Recalibration), a computational tool for tumor-only CNV detection in targeted NGS panels. CESAR uses Circular Binary Segmentation (CBS) to re-partition target regions according to relative capture efficiency, then applies a data-driven "anchor" selection procedure that, for each target segment, identifies a personalized set of co-varying genomic segments. By selecting the anchor set that minimizes the coefficient of variation (CV) of the anchor-recalibrated depth ratio across a panel of normals, CESAR recalibrates the per-segment baseline and suppresses probe-specific technical noise. Copy-number status is then called from the deviation of the observed ratio against this trained baseline. ResultsUsing standard DNA reference materials, CESAR identified amplifications of MET, ERBB2, and EGFR at low tumor fractions. CESAR resolved focal alterations as subtle as 2.18 copies (a 1.09-fold change relative to the diploid baseline) while reporting no false-positive amplifications in diploid control regions. Applied to a clinical cohort of nine NSCLC ctDNA samples profiled on a 94-gene panel, CESAR successfully separated three MET-amplified samples from six matched negatives, where the standard CNVkit-based pipeline failed to distinguish them. In head-to-head benchmarking on identical data, CESAR reduced technical variance relative to the widely used CNVkit and more reproducibly resolved low-level copy-number gains, particularly in the depth-heterogeneous MET amplicon. ConclusionsCESAR provides a stable and sensitive framework for tumor-only CNV calling in liquid biopsies. On reference standards it outperforms CNVkit in bias and reproducibility, and on a clinical NSCLC cohort it recovered MET CNV abnormalities that a standard CNVkit pipeline missed. Validation on larger and more diverse cohorts is warranted.

bioinformatics↗

bioRxiv2026

Sex-Specific Vulnerability to Radiofrequency Electromagnetic Radiation-Induced Reproductive and Neurological Impairment in Mice

Electromagnetic radiation (EMR) constitutes a pervasive environmental stressor, yet the sex-specific vulnerabilities to its biological effects remain inadequately characterized. This study investigated the differential impacts of 3.2 GHz pulsed EMR on reproductive and neurological functions in male and female mice following a four-week exposure regimen. We identified a pronounced sexual dimorphism in systemic and organ-specific responses. Male mice exhibited a primary vulnerability in the reproductive axis, manifesting as severe spermatogenic impairment, including depletion of the spermatogonial stem cell pool, disruption of sperm chromatin remodeling, and consequent declines in sperm quality and quantity. In contrast, exposed females showed preserved ovarian function but displayed marked neurobehavioral deficits, including increased anxiety- and depressive-like behaviors, and cognitive impairment, which were correlated with neurodegeneration in the hippocampus and prefrontal cortex. Proteomic analyses revealed distinct sex-specific biomarker profiles, with serum levels of Mdk, histone H1 variants (H1-1, H1-4, H1-5), Hp1bp3, Ncf2, and Rhoc strongly associated with male testicular dysfunction, while decreased levels of the synaptic motor protein KIF13A in serum and brain tissue were linked to female neurological impairment. Our findings delineate clear sex-divergent pathological pathways that target male spermatogenesis and female neurocognitive function and propose novel circulating protein biomarkers. This work provides a critical foundation for sex-aware risk assessment and the development of targeted preventive strategies against EMR-associated health risks.

pathology↗

bioRxiv2026

CEACAM5/6+ Tumor Cells and IL-1β+ Macrophages Drive Resistance to Chemo-immunotherapy in Gastric Cancer

Chemo-immunotherapy is a first-line treatment for advanced gastric cancer, yet response rates remain limited and resistance mechanisms are poorly defined. Here we generate a single-cell atlas of 542,121 cells from 35 patients treated with anti-PD-1 plus chemotherapy, profiling pre- and post-treatment tumors linked to clinical response. Integrating spatial transcriptomics, immunohistochemistry, and bulk RNA sequencing, we identify two temporally distinct resistance programs. Intrinsic resistance in pre-treatment non-responders is marked by enrichment of CEACAM5/6 tumor cells that form immune-excluded spatial niches characterized by macrophage recruitment and CD8 T-cell exhaustion. Acquired resistance in post-treatment non-responders is driven by expansion of IL-1{beta} macrophages, which induces coordinated NF-{kappa}B activation across tumor and stromal compartments, promoting PD-L1 upregulation, epithelial-mesenchymal transition, and chronic inflammation. These findings delineate an evolutionary trajectory of resistance and nominate CEACAM5/6 and IL-1{beta} as predictive biomarkers and therapeutic targets to improve anti-PD-1-based combination strategies.

cancer biology↗

bioRxiv2026

Presenilin 1 (PS1) located at mitochondrial inner membrane regulates mitochondrial cristae junction proteins arrangement and cristae formation in HEK293 cells

Presenilin 1 (PS1), a key pathogenic factor in familial Alzheimers disease, is implicated in regulation of mitochondrial functions, yet its precise sub-mitochondrial localization and underlying mechanisms remain poorly understood. In this study, we generated PS1 knockout (PS1 KO) cell lines to investigate the role of PS1 in mitochondrial structure and function. Our results demonstrated that PS1 is directly localized to the mitochondrial inner membrane. PS1 deficiency led to reduced ATP production, impaired mitochondrial respiration capacity, decreased mitochondrial membrane potential, disrupted Ca2+ homeostasis, and elevated reactive oxygen species (ROS) accumulation. Moreover, loss of PS1 caused abnormal mitochondrial cristae structure. Further analysis revealed that PS1 interacts with mitochondrial inner membrane proteins. Its absence promotes ATAD3A oligomerization and disrupts its arrangement at mitochondrial cristae junctions, leading to expansion of the mitochondria-associated membrane (MAM) and instability of mitochondrial DNA (mtDNA). Our findings demonstrate that PS1 acts as a central regulator of mitochondrial cristae morphogenesis by modulating protein interaction networks at cristae junctions, thereby illuminating fundamental molecular mechanisms contributing to mitochondrial dysfunctions in Alzheimers disease.

cell biology↗

bioRxiv2026

Enteric sensory neurons for nutrient detection and gut motility

The enteric nervous system (ENS) orchestrates gastrointestinal reflexes and brain-gut communication via molecularly diverse neurons. Among these, intrinsic primary afferent neurons (IPANs) are essential for detecting luminal nutrients and irritants, yet their molecular identities, sensory properties, and functions remain poorly resolved. Here, we establish a segment-resolved single-cell atlas of the murine ENS, including a comprehensive characterization of the gastric ENS. This resource defines a refined taxonomy of enteric neurons and glia and enabled the development of a genetic toolkit for molecularly defined IPANs. Using chemogenetics and calcium imaging, we discovered that myenteric neurons detect a wide range of nutrients, irritants, and cytokines. Nutrient detection depends on a functional connection between chemosensory epithelial cells and enteric neurons mediated by 5-HT--HTR3 axis. Through optogenetic analysis, we demonstrated segment-specific regulation of gut motility by different IPANs. Our work establishes a genetic and physiological framework for enteric-specific sensory mechanisms.

neuroscience↗

bioRxiv2026

Toward Computationally Complete Spatial Omics

Multimodal spatial omics has transformed biology by mapping molecular complexity within intact tissues, yet current technologies remain limited in the number of modalities measured simultaneously and often produce lower-quality data than single-modality assays. We present COSIE, a computational framework that generates high-resolution, multilayered molecular landscapes across tissue sections, individuals, and platforms. COSIE integrates histology, epigenome, transcriptome, proteome, and metabolome into a unified representation. Applied to 12 datasets spanning 10 spatial technologies, eight modalities, and nine tissue types, ranging from thousands of spots to millions of cells, COSIE outperforms existing methods. It resolves tissue structures, enhances noisy measurements, predicts unmeasured modalities, and captures dynamic processes. In human tumors, COSIE identifies invasive subregions linked to clinical outcomes and predicts spatial gene expression in TCGA samples using only histology images. By transforming fragmented data into comprehensive spatial maps, COSIE advances computationally complete spatial omics and the creation of digital tissue twins for biomedicine.

genomics↗

bioRxiv2026

Primary cilia promote resistance to EGFR tyrosine kinase inhibitor, osimertinib, in non-small cell lung cancer

Patients with advanced non-small cell lung cancer (NSCLC) and mutations in epidermal growth factor receptor (EGFR) benefit from EGFR tyrosine kinase inhibitors (TKIs). Osimertinib, a third-generation EGFR TKI, is standard first-line therapy for EGFR-mutated NSCLC, but most patients develop resistance to it. Here, we demonstrate that increased formation of primary cilia, microtubule-based sensory organelles, is associated with osimertinib-refractory NSCLC progression. EGFR-mutated, osimertinib-resistant human NSCLC cells had increased cilia formation and acetylation of -tubulin and reduced histone deacetylase 6 (HDAC6) activity compared to their osimertinib-sensitive counterparts. HDAC6 inhibition increases cilia formation in osimertinib-sensitive NSCLC cells, and overexpression of exogenous HDAC6 sensitized osimertinib-resistant NSCLC cells to osimertinibs anti-proliferative effects. Because intraflagellar transport (IFT) proteins are essential for primary cilium formation and function, we knocked down IFT88 in osimertinib-resistant NSCLC cells, which reversed osimertinib resistance in orthotopic and subcutaneous mouse models of lung cancer. Mechanistically, increased sodium influx during osimertinib-induced inhibition of EGFR signalling promotes cilia formation through sustained HDAC6 inactivity and greater -tubulin acetylation. Inhibition of sodium influx with dibutyryl-cAMP decreased cilium formation, increased sensitivity to osimertinib, and reduced tumor progression in mice bearing osimertinib-resistant lung tumors. Collectively, our findings suggest that enhanced primary cilium formation mediates EGFR TKI resistance and that targeted inhibition of ciliogenesis may prevent or overcome osimertinib resistance.

cancer biology↗

bioRxiv2026

Wiz regulates clustered protocadherin genes by restricting CTCF/cohesin loop extrusion in a genomic-distance biased manner

Zinc finger (ZF) proteins (ZFPs) constitute the largest family of transcription factors in mammals; however, their regulatory mechanism remains largely elusive. Here we propose COP (C2H2-ZFP occupancy predictor), a deep learning framework that integrates DNA sequence information with protein primary and secondary features to systematically predict ZFP genomic enrichments. Applying COP to the mouse clustered protocadherin (cPcdh) gene locus, we identified dozens of C2H2-ZFPs potentially involved in CTCF-mediated gene regulation with Wiz having the highest number of 12 ZFs. We confirmed Wiz enrichments at all of the CBS (CTCF-binding site) elements across the Pcdh clusters by Myc-tagging the endogenous Wiz gene. Genetic experiments revealed significant increases of expression levels of the cPcdh genes upon Wiz deletion in both neuronal cells in vitro and in mouse brain in vivo. Finally, integrated ChIP-seq, RNA-seq, and 4C-seq analyses demonstrated that Wiz regulates CTCF/cohesin occupancy and long-range enhancer-promoter contacts in a genomic-distance biased manner. Together, these findings have interesting implications regarding molecular mechanism of C2H2-ZFPs in cPcdh gene regulation.

cell biology↗

bioRxiv2026

Exploration of the screening and regulatory mechanisms of biomarkers related to ac4C modification in laryngeal squamous cell carcinoma patients based on single-cell analysis and machine learning

BackgroundN4-acetylcytidine (ac4C) modification plays a critical role in cancer development. Exploring ac4C modification in laryngeal squamous cell carcinoma (LSCC) may help elucidate its pathogenesis. MethodsLSCC-related datasets were obtained from GEO. After preprocessing and annotating single-cell data, malignant cells were identified by CNV scoring and further divided into subpopulations. Malignant epithelial cells (MECs) were identified and subclustered based on ac4C-related gene activity. Prognostic genes were screened using Cox regression and machine-learning approaches, followed by validation in clinical samples using qPCR. The biological and immunological relevance of these genes was further explored through immune infiltration, immunotherapy response, and mutation analyses. ResultsThe 14,465 identified MECs were classified into five subgroups (MEC1-5), among which MEC3 showed the strongest association with the ac4C gene set. Machine-learning analysis of MEC3-derived genes yielded seven prognostic markers, including BARX1, FHL2, NXPH4, PKMYT1, TNFAIP8L1, CRLF1, and CENPP. qPCR confirmed their differential expression between tumor and adjacent normal tissues. These genes were significantly associated with alterations in the tumor immune microenvironment, with high-risk patients showing increased immune infiltration and immune activity. ConclusionSeven ac4C-related prognostic genes were identified that may contribute to LSCC progression by modulating the tumor immune microenvironment, providing potential therapeutic insights.

developmental biology↗

bioRxiv2026

Early electrical stimulation promotes functional recovery after volumetric muscle loss

Volumetric muscle loss (VML) injuries overwhelm the inherent regenerative capacity of skeletal muscle, causing persistent functional deficits with no routinely effective therapies. Electrical stimulation (ES) has been shown to preserve muscle structure in other injury models, but technical barriers have prevented daily delivery during the acute post-injury window when critical regenerative programs are established. Here, we developed a fully implantable bioelectronic system with nanoporous platinum-modified electrodes enabling daily therapeutic stimulation and electromyographic recording without repeated anesthesia in a rat tibialis anterior VML model. Animals receiving ES during the acute post-injury period (10 sessions over days 0-14) showed sustained functional improvement, reaching 90% of baseline torque at 8 weeks compared to 71% in unstimulated controls. This recovery reflected enhanced remodeling of injured muscle rather than synergistic muscle compensation. Histological analysis revealed coordinated early increases in vascularization, pro-regenerative macrophages, and satellite cells. These findings establish early ES as a promising intervention for promoting muscle regeneration after catastrophic injury.

bioengineering↗

bioRxiv2026

Multiscale confidence quantification for virtual spatial transcriptomics with UTOPIA

Virtual spatial transcriptomics (ST) methods predict gene expression or cell types from histology images, extending molecular readouts beyond the limited regions or samples directly measured by ST platforms. However, the statistical reliability of these predictions remains unclear. Here, we present UTOPIA, a model-agnostic framework for multiscale confidence quantification in virtual ST. UTOPIA assigns statistically calibrated confidence scores to predictions across spatial resolutions and biological granularities, ranging from single genes to metagenes and from specific cell types to broader cell classes. UTOPIA controls false discovery rates for detecting genes, metagenes, or cell types while accounting for local tissue context. We show that prediction confidence depends critically on both spatial resolution and biological granularity, with reliable inference often emerging only at coarser, biologically meaningful scales. Across multiple ST platforms and in both in-sample and out-of-sample settings, UTOPIA enhances interpretability, prevents false biological conclusions, and enables more trustworthy downstream analyses of virtual ST.

genomics↗

bioRxiv2026

Targeting CPSF73, the mRNA 3' End Processing Endonuclease, Moves Cancer Cells Away from the Mesenchymal State

BackgroundMetastasis significantly contributes to cancer-related mortality and therapeutic failure. Cancer cells acquire metastatic potential by losing epithelial characteristics and gaining mesenchymal properties through the epithelial-mesenchymal transition (EMT). Differential poly(A) site (PAS) usage, known as alternative polyadenylation (APA), generates mRNA isoforms differing in coding sequence, subcellular localization, stability, or translation efficiency. In cancer, 3'UTR shortening increases expression of proto-oncogenes by escaping miRNA-mediated repression. High expression of CPSF73, which cleaves mRNA precursors at PASs, is associated with unfavorable prognoses in cancer patients. However, the role of APA in regulating EMT remains poorly understood. MethodsIn this study, to investigate the role of APA in EMT, we employed JTE-607, a small-molecule inhibitor of CPSF73 activity, to examine the impact of catalytic inhibition of CPSF73 on proliferation and EMT in MDA-MB-231, MCF7, A549, and HepG2 cancer cells. To identify differential usage of PASs, global profiling of APA changes, and differential gene expression analysis were performed in MDA-MB-231 cells. Additionally, antisense oligonucleotides were used to block the use of a specific PAS whose APA change may be a driver of EMT reversal. ResultsOur findings showed that catalytic inhibition of CPSF73 not only attenuates cancer cell proliferation but also moves the cells away from the mesenchymal state across all four cell lines tested. Global profiling of APA changes following CPSF73 inhibition revealed widespread 3'UTR lengthening and suppression of intronic PASs in MDA-MB-231 cells. APA shifts were observed in key EMT-related genes, accompanied by decreased expression of corresponding proteins across all four cell lines. We used antisense morpholino oligonucleotides to block the proximal PAS of AKT2, shifting the balance of AKT2 mRNA isoforms toward the long isoform. This shift caused EMT reversal, marked by reduced AKT2 protein expression, changes in EMT-related markers, and impaired invasion by MDA-MB-231 cells. ConclusionTogether, these findings identify APA-mediated 3UTR lengthening, with functional consequences in EMT-related genes, as a coordinated mechanism leading to an attenuated EMT phenotype, highlighting a significant connection between APA and the EMT process. Interfering with these APA changes may offer a promising therapeutic strategy to suppress metastasis, with potential efficacy across multiple pathways. Statement of SignificanceOur findings highlight APA-mediated 3 UTR lengthening as a coordinated mechanism that promotes EMT reversal and support CPSF73 inhibition or APA-targeting strategies as potential therapeutic approaches to suppress metastasis across multiple pathways.

cancer biology↗

bioRxiv2026

Refinement of Nucleus Accumbens Neuronal Dynamics During Cocaine Self-Administration Training

Drug addiction is an acquired motivational-behavioral state that begins with drug taking, which is comprised of a series of phases, including initial acquisition, stabilization, habituation, and maintenance. In rodent models of cocaine self-administration, the forebrain region nucleus accumbens (NAc) has been critically implicated in the acquisition-maintenance process of drug taking and seeking behaviors. However, it remains unknown how NAc neurons shift their activity patterns in response to these phasic transitions during cocaine taking. To examine this, we used GCaMP6m-based in vivo Ca2+ imaging to monitor activities of principal medium spiny neurons (MSNs) in the NAc across eleven days of cocaine self-administration. Behaviorally, mice exhibited progressive stabilization of operant responding and locomotion across 11 days of cocaine self-administration. During the early training days, we detected a portion of NAc neurons--a potential neuronal ensemble--that exhibited increased activities temporally contingent to the lever-press for cocaine. The number of NAc neurons exhibiting contingent activity increased progressively over the first three training days and then decreased gradually during the later training days, exhibiting expansion-refinement dynamics that may correspond to the acquisition and subsequent stabilization/maintenance of cocaine self-administration. Using a neuron-tracking technique, we found that the lever-press-contingent NAc ensemble exhibited substantial compositional dynamics, with neurons dropping into and out across training days. These activity features of lever-press-contingent neurons may represent key circuit dynamics of the NAc that transition the acquisition toward the maintenance of cocaine-taking behavior.

neuroscience↗

bioRxiv2026

CsFDL1-CsFTL3 complex represses CsFTL3 via negative feedback to fine-tune flowering in Chrysanthemum seticuspe

In many flowering plants, the transition from vegetative growth to reproductive development is regulated by seasonal changes in photoperiod. Under inductive photoperiods, leaves produce the florigen FT (FLOWERING LOCUS T), which is transported to the shoot apex to promote flowering. The photoperiod is known to have a major effect on the flowering of chrysanthemum. In the perennial short-day (SD) plant Chrysanthemum seticuspe, the expression of CsFTL3 (FT-like gene) does not increase immediately after shifting from long-day (LD) to SD conditions but gradually accumulates under continuous SD conditions, peaking during inflorescence development. However, the underlying mechanism remains elusive. We show that CsFDL1 (an ortholog of FD) and CsFTL3 exhibit a significant inverse expression pattern in leaves during the initial stage of short-day inductions. Furthermore, the expression of CsFTL3 is upregulated in the leaves of CsFDL1-knockdown transgenic lines. CsFDL1 is expressed in leaves and forms a complex with CsFTL3 to recognize several TCGA- and ACGT-containing motifs in the CsFTL3 promoter. The CsFTL3-CsFDL1 complex downregulates CsFTL3 expression, thereby preventing its excessive induction by SD signals and inhibiting precocious floral transition. This study reveals that CsFDL1 acts as a key early repressor in the photoperiodic flowering pathway of chrysanthemum leaf, mediating negative feedback regulation by forming a complex with CsFTL3 to achieve precise temporal control of short-day-dependent flowering responses.

plant biology↗

bioRxiv2026

Kindlin-2-Moesin interaction orchestrates sprouting angiogenesis via modulating endothelial membrane mechanics and VEGF signaling

Membrane mechanics play a crucial role in cellular signaling and fate determination, yet their impact on angiogenesis remains poorly understood. Here, we identify Kindlin-2 as a key regulator of sprouting angiogenesis via regulating endothelial membrane tension through its interaction with Moesin, a crucial linker protein between the cell membrane and the actin cortex. Mechanistically, Kindlin-2 binds to the N62 residue of Moesin, limiting its overactivation and maintaining proper membrane tension to facilitate VEGFR2 endocytosis and downstream signaling. Using both developmental and pathological models, we demonstrate that the interaction of Kindlin-2 and Moesin is enhanced in high angiogenic conditions, and endothelial Kindlin-2 deletion reduces angiogenesis. Furthermore, mutation of Moesin at N62 phenocopies the effects of Kindlin-2 loss. Together, these findings uncover a previously unrecognized mechanism linking membrane tension regulation to angiogenesis and provide new insights into targeting the Kindlin-2-Moesin axis for therapeutic intervention in neovascular diseases.

developmental biology↗

bioRxiv2026

CellSwarm: LLM-Driven Cell Agents Recapitulate Tumor Microenvironment Dynamics and Sense Indirect Genetic Perturbations

Agent-based models of the tumor microenvironment (TME) traditionally rely on hand-coded rules that cannot generalize beyond their programmed logic. Here we present CELLSWARM, a framework that replaces rule-based cell decision-making with large language model (LLM)-driven autonomous agents. Each simulated cell maintains persistent state, 14 signal pathways, and a memory stream, with an LLM serving as its cognitive core. Using structured knowledge bases for cancer-specific context, CELLSWARM recapitulates TNBC microenvironment composition with fidelity comparable to hand-coded rules (Jensen-Shannon divergence 0.144 vs. 0.146; P=0.012 vs. random, Mann-Whitney U test). Beyond matching rule-based performance, LLM-driven agents demonstrate three capabilities absent from rule-based models: cross-cancer generalization by swapping knowledge base entries, treatment response prediction concordant with clinical data (anti-PD-1: 17.6% simulated vs. 21% clinical), and sensing of indirect genetic perturbations that propagate through intermediate signaling cascades (IFN-{gamma} KO: Agent +15.7% vs. Rules +0.3%; P=0.005). CELLSWARM demonstrates that LLM-driven cell agents can recapitulate and extend TME simulation beyond the reach of hand-coded rules.

systems biology↗

bioRxiv2026

Heterozygote Advantage of a Single-Copy SNAP18 Truncation Allele Enables Dominant SCN Resistance and Yield Preservation in Soybean

Soybean cyst nematode (SCN, Heterodera glycines) is the most devastating pathogen threatening global soybean production. The long-term over-reliance on rhg1-a/b resistance has led to the rapid emergence of virulent SCN populations, creating an urgent demand for novel genetic resources to safeguard soybean yield. Here, we identified a rare gain-of-function allele, SNAP18lmm3, via map-based cloning of the lesion-mimic mutant (lmm3) in the SCN-susceptible Williams 82 (rhg1-c) background. This allele encodes a 24-amino-acid C-terminal truncation of SNAP18 and exhibits an unusual mixed dominance genetic profile: the autoimmune-related leaf lesion phenotype is recessive, whereas SCN resistance is dominant. Homozygous lmm3 plants suffer significant yield losses due to spontaneous cell death, but heterozygous (LMM3+/-) plants maintain normal growth, agronomic performance, and seed yield comparable to wild-type Williams 82 in SCN-free field environments. Critically, under artificial SCN inoculation in both field trials and controlled environments, LMM3+/- heterozygotes confer robust resistance, achieving a [~]4-fold yield advantage over SCN-infested Williams 82 and a [~]44% reduction in cyst formation relative to Williams 82. This resistance profile aligns with the broad-spectrum activity of the lmm3 homozygous mutant, supported by the conserved resistance mechanism of SNAP18lmm3. Transgenic expression of SNAP18lmm3 driven by the nematode-responsive HIP promoter (HIPpro) potently arrested nematode development at the J2 stage in transgenic hairy roots, as evidenced by the drastically reduced proportion of advanced-stage nematodes. Our findings establish SNAP18lmm3 as a potent, "plug-and-play" genetic resource that circumvents the limitations of traditional dosage-dependent resistance loci. This allele enables the development of high-yielding, SCN-resistant soybean cultivars through marker-assisted selection for the heterozygous state or precision genome editing, providing a practical solution for sustainable SCN management.

plant biology↗