Biology subjects

Wang, T.

Publications and source records attributed to Wang, T..

At least 199 records · Page 11Linked to original sources

bioRxiv2025

Common Signatures of Neutrophils in Diverse Disease Conditions

Neutrophils are traditionally recognized for their pro-inflammatory roles. However, accumulating evidence has begun to highlight the plasticity of transcriptional states of neutrophils during pathological insults. Whether such unconventional neutrophils may share commonality across diverse disease conditions is incompletely understood. Here, we systematically profile over 500,000 neutrophils in key immune compartments and disease-inflicted tissues of the mouse models of metabolic disorder, autoimmunity, tissue damage, peripheral cancers, and intracranial gliomas. Of importance, we observe two distinct neutrophil clusters with unique immune features. The first cluster, characterized by Cd274, Vegfa, and antigen presentation, is highly enriched within the diseased tissues. In contrast, the second cluster with elevated Cd244 and type 2 immune response but reduced maturation markers primarily emerges in the peripheral blood and spleens of specific disease scenarios. These results have elucidated the common signatures of neutrophils in response to various pathological conditions, providing a valuable reference for diagnostic and therapeutic applications.

immunology↗

bioRxiv2025

Microglial GSDMD-Mediated Pyroptosis Drives Neuroinflammation in Parkinson's Disease

RationaleParkinsons disease (PD), a globally prevalent neurodegenerative disorder, is characterized by substantia nigra dopaminergic neuron degeneration and striatal dopamine depletion. While microglial pyroptosis is implicated in neuroinflammation and neural injury via inflammatory cytokine release, the role of the CASPASE-1/GSDMD pathway in PD pathogenesis remains incompletely defined. Methods1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to construct PD model in vivo, GSDMD-knockout mice was employed to assess pyroptotic mechanisms. MPP-stimulated BV2 microglia were treated with a CASPASE-1 inhibitor in vitro. Microglia-specific GSDMD conditional knockout mice were generated to evaluate cell-type contributions to neuroinflammation and motor deficits. ResultsGSDMD deficiency attenuated MPTP-induced neuroinflammation, dopaminergic neuron loss, and motor dysfunction in vivo. MPP exposure triggered NLRP3 inflammasome activation and pyroptosis in BV2 microglia, which was suppressed by CASPASE-1 inhibition. Critically, microglia-specific GSDMD ablation mitigated nigrostriatal degeneration and dyskinesia in PD mice, confirming the centrality of microglial pyroptosis. ConclusionOur findings demonstrate that microglia drive neuroinflammation in PD via CASPASE-1/GSDMD-mediated pyroptosis, directly linking this pathway to dopaminergic neurodegeneration and motor impairment. Targeting GSDMD-dependent pyroptosis represents a promising therapeutic strategy.

neuroscience↗

bioRxiv2025

The human transcription factor occupancy landscape viewed using high-resolution in situ base-conversion strand-specific single-molecule chromatin accessibility mapping

Chromatin accessibility profiling is a key tool for mapping the location of cis-regulatory elements (cREs) in the genome and tracking chromatin state dynamics during development, in response to various external and internal stimuli, and in disease contexts. Single-molecule footprinting (SMF) methods that rely on the labeling of individual accessible DNA bases have emerged in recent years as a powerful chromatin accessibility mapping approach, as they provide not just an average readout of accessibility over a given genomic position but also the distribution of accessibility states within a population at the level of individual original DNA molecules. However, methylation-based SMF approaches have often been limited either in their resolution or in labeling readout accuracy. We developed a deaminase-based high-resolution strand-specific single molecule footprinting approach that uses highly active sequence context-independent endogenous methylation-insensitive double-strand DNA (dsDNA) deaminases (CseDa01 and LbDa02 C[->]U/T-ssSMF), which convert accessible cytosines into uracils (which are converted to thymine after PCR amplification). We demonstrate the application of the method to mapping single-molecule accessibility states in human cell lines in both a short and a long-read format, and quantifying the occupancy states of individual transcription factors (TFs) as well as TF co-accessibility and strand-specific accessibility patterns.

genomics↗

bioRxiv2025

Evolutionary implications of stipule occurrence in Permo-Carboniferous wetland forests dominated by Psaroniaceae

Stipules are specialized appendages borne at the leaf base with roles as diverse as sheltering of delicate growing tissues from environmental exposure, vegetative propagation and dispersal, and modification into climbing hooks or protective spines. Stipules are widely present in extant vascular plants, but their origins in geological history remain obscure and must have involved convergent evolution more than once, perhaps as early as the late Paleozoic emergence of forested terrestrial ecosystems. Based on extraordinary collections from the early Permian Wuda Tuff Flora, the earliest known stipule structure in the plant kingdom has been identified in marattialean tree ferns Psaroniaceae, the dominant element of Permo-Carboniferous wetland forests. Psaroniaceous stipules are consistent with protection of juvenile fronds and the stem apex as well as retention of a functional role in mature and withered fronds. Furthermore, the continued and perhaps indeterminate growth and the fully laminated stipules invite speculation of a potential role in vegetative propagation following detachment from the parent frond. However, no direct fossil evidence of stipules acting as vegetative propagules is currently available.

paleontology↗

bioRxiv2025

Phased epigenetic fragility and stability accelerate the genetic evolution of B-cells

Non-genetic heterogeneity can provide population robustness when responding to threats or making developmental decisions, but when the biological process rests on specific individuals (tissue-resident macrophages or genetic evolution), non-genetic heterogeneity degrades performance. Vaccine responses depend on the Darwinian evolution of B-cells to generate high-affinity, genetically-encoded antibodies, yet B-cell decision-making is non-genetically variable. In fact, B-cell epigenetic states are fragile during selection but stable during the proliferation burst. Here we report that the iterative dynamics of epigenetic fragility and stability of B-cell fate decisions do not impair but accelerate affinity maturation, by modulating the generation and removal of high-affinity outliers on a fitness landscape that is non-monotonic defined not only by survival/proliferation but plasma cell differentiation. These insights reconcile classical B-cell clonal selection theory with experimentally observed dynamics of epigenetic variability. The resulting model correctly predicts emergent vaccine response properties in alternate mouse strains and may contribute to personalized vaccination strategies. HIGHLIGHTSO_LIB-cell epigenetic states are fragile during antigen selection but stable within proliferative bursts C_LIO_LIEpigenetic state fragility allows high affinity cells to escape plasma cell differentiation C_LIO_LIEpigenetic state stability within proliferative bursts maximizes progeny of fittest cells C_LIO_LIPhased epigenetic dynamics of B-cells predicts emergent vaccine response properties C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/661408v1_ufig1.gif" ALT="Figure 1"> View larger version (50K): org.highwire.dtl.DTLVardef@1c2af9forg.highwire.dtl.DTLVardef@980003org.highwire.dtl.DTLVardef@128d8c0org.highwire.dtl.DTLVardef@ae48b8_HPS_FORMAT_FIGEXP M_FIG C_FIG

systems biology↗

bioRxiv2025

Simulation and experimental study on the strategy of soil auxiliary heating in a solar greenhouse in northeast China

When solar greenhouses in north eastern China experience extreme weather, the internal soil requires varying auxiliary heating to ensure the normal growth of crops. In this study, considering the effects of four kinds of extreme weather on solar greenhouses, four different types of heating requirements were obtained through experiences in a Liaoshen-type solar greenhouse. Four distinct heating strategies were simulated using a soil temperature prediction model. The initial temperatures for these strategies were 13{degrees}C, 11{degrees}C, 13{degrees}C, and 14{degrees}C, respectively, while the target temperature was uniformly set at 15{degrees}C. The heating durations for the respective strategies were 432,000 seconds, 432,000 seconds, 864,000 seconds, and 1,728,000 seconds. To ensure a minimum soil temperature of 15{degrees}C within the solar greenhouse, the temperature of the heating layer located 400 mm underground needed to be maintained at or above 20{degrees}C, 28{degrees}C, 18{degrees}C, and 19{degrees}C, respectively. The simulation results were validated through experimental verification, with maximum and minimum errors of 3.1% and 0.5%, respectively. Several targeted promotional suggestions were also proposed. The model exhibited high accuracy and is capable of developing optimal and precise soil heating strategies for various types of solar greenhouses in north eastern China.

plant biology↗

bioRxiv2025

A spatial atlas of chemoradiation therapy in pancreatic cancer identifies cellular and microenvironmental determinants of persister populations

The molecular pathways involved in the response to radiation therapy in pancreatic ductal adenocarcinoma (PDAC) remain poorly understood. We aimed to elucidate the adaptive mechanisms and cellular interactions within PDAC to radiation therapy (RT). We constructed a transcriptomic landscape of the cellular subtypes and spatially resolved neighborhoods from 50 patient samples, including 16 longitudinally matched single cell RNA sequencing and 34 spatial transcriptomics specimens. To resolve shortcomings of cell-type mixtures in spatial data, we developed a novel statistical method called SpaCCI (spatially aware analysis of cell-cell interactions) to profile cell-cell interactions and ligand-receptor enrichment. This revealed CXCL12/TGF{beta}-driven persister cell niches where activated fibroblasts reprogram tumor- associated macrophages and spatially exclude stress-response CD8 T cells after RT. Persister cancer cells displayed transcriptional evidence of recalcitrance to metal-induced cell death pathways of ferroptosis and cuproptosis which were recapitulated in preclinical models. Our study reveals the selective pressures experienced by PDAC following RT that may help provide insight for future multimodal therapeutic strategies.

cancer biology↗

bioRxiv2025

A scalable, all-optical method for mapping synaptic connectivity with cell-type specificity

Single-cell transcriptomics has uncovered the vast heterogeneity of cell types that compose each region of the mammalian brain, but describing how such diverse types connect to form functional circuits has remained challenging. Current methods for measuring the presence and strength of synaptic connections principally rely on low-throughput whole-cell recording approaches. However, the development of optical tools for perturbing and observing neural activity--now notably including genetically encoded voltage indicators--presents an exciting opportunity to vastly increase the throughput of physiological connectivity mapping. Here, we combine massively parallel optical measurements of synaptic strength with a novel pipeline for thick-tissue spatial transcriptomics to assay synaptic connectivity motifs with high sensitivity, high throughput, and cell-type specificity. We apply this approach in the motor cortex to describe new cell-type-specific synaptic innervation patterns for long-range thalamic and contralateral input onto more than 1000 motor cortical neurons.

neuroscience↗

bioRxiv2025

ZIPcnv: accurate and efficient inference of copy number variations from shallow whole-genome sequencing

Shallow whole-genome sequencing (sWGS), a rapid and cost-effective sequencing technology, has gradually been widely adopted for CNV analyses. However, with genome-wide coverage of only 0.1-5x, sWGS data exhibit a pronounced zero-inflation phenomenon, whereby a large fraction of loci contains zero sequencing reads. Zero-inflation causes read counts to fluctuate by several-fold between adjacent windows. As a result, random upward blips in coverage can be misinterpreted as copy-number gains (false positives), and true deletions often become indistinguishable from pervasive zero-coverage noise. In addition, existing CNV detection tools developed for sWGS data often struggle to adapt across different CNV sizes. These combined effects severely constrain the accuracy of CNV inference. To address above challenges, we propose ZIPcnv, a novel CNV detection tool specifically designed for sWGS data. First, we apply a large sliding window to smooth the raw read depth signal, which transforms the original zero-inflated statistical characteristics into approximately normal distribution characteristics. We then design a statistical process model that robustly detects persistent shifts under high background noise using a cumulative sum strategy, classifying genomic regions into candidate and non-candidate CNV regions. Finally, dynamic sliding windows are used for one-pass detection of CNVs of varying lengths, with window size adapting to the CNV region size. We evaluated the performance of ZIPcnv on simulated data and 190 real whole-genome sequencing samples. Experimental results show that ZIPcnv consistently outperforms currently popular CNV detection tools. The ZIPcnv source code is freely available at https://github.com/Nevermore233/ZIPcnv.

bioinformatics↗

bioRxiv2025

Type-I interferon priming signal balances anti-bacterial and anti-tumor trained immunity in alveolar macrophages

Tissue-resident macrophages may be trained to confer enhanced response to heterologous re-stimulation, and thus foster versatile trained immunity (TI) against both infections and tumors. However, key priming signals that contribute to such functional versatility in trained macrophages remain not fully understood. Here we show that influenza A virus (IAV) infection in mice induces lasting transcriptional imprints of acute type-I interferon (IFN-I) signaling in lung-resident alveolar macrophages (AMs) that confer balanced anti-bacterial and anti-tumor TI responses. In both IAV-infected mice and an ex vivo cytokine-mediated training system, IFN-I signaling is found to be critical to developing anti-tumor TI functions, although it limits anti-bacterial TI functions in AMs. Moreover, human AMs carrying transcriptional imprints of IFN-I signaling are associated with anti-tumor immunity and immune activation in lung cancer tissues. Our findings highlight IFN-I as a key priming signal required to developing versatile TI functions in AMs with balanced anti-bacterial and anti-tumor potential.

immunology↗

bioRxiv2025

Sustained Lysosomal Delivery of Enhanced Cy3-Labeled Acid Nanoparticles Restores Lysosomal pH in Retinal Pigment Epithelial Cells and Astrocytes

Lysosomal pH is frequently elevated in age-dependent neurodegenerations like Age-related Macular Degeneration (AMD), Alzheimers Disease (AD), and Parkinsons Disease (PD). Tools that restore lysosomal pH to an optimal acidic range could enhance enzymatic degradation and reduce waste accumulation. Acidic nanoparticles offer a promising strategy for restoring lysosomal function, but accurate tracking of organelle delivery and long-term retention is needed to optimize dosage. To improve detection and enhance delivery, nanoparticles were synthesized from Poly(D,L-lactide-co-glycolide) (PLGA) polymers covalently linked to the fluorescent Cyanine3 amine (Cy3) probe. Nanoparticle concentration and loading times were optimized to achieve >90% delivery to lysosomes in cultured induced pluripotent stem cell-derived retinal pigment epithelial (iPS-RPE) cells. Uptake was heterogeneous, varying between adjacent cells. Once loaded into lysosomes, the nanoparticles were stably retained, with no detectable changes in concentration, distribution, or size for at least 28 days. iPS-RPE cells internalized more nanoparticles than the ARPE-19 cell line or mouse optic nerve head astrocyte cultures. Functionally, PLGA nanoparticles restored an acidic pH and cathepsin D levels in compromised lysosomes. In summary, Cy3-PLGA nanoparticles enabled improved tracking and long-term delivery to lysosomes, supporting future in vivo applications to restore lysosomal pH in aging and degenerating tissues. Graphical AbstractIncreased lysosomal pH reduces degradative enzyme efficiency and contribute to age-dependent neurodegeneration. This study describes synthesis of nanoparticles to restore an acidic lumen and degradative function. Nanoparticles were optimized for lysosomal delivery to astrocytes and iPS-derived retinal pigmented epithelial (RPE) cells. The fluorescent marker Cy3 was covalently bound to polymers for improved tracking to lysosomes. Particles were stably retained inside the lysosomal lumen for at least 28 days. Nanoparticles restored pH to compromised lysosomes to baseline levels and increased active Cathepsin D. The improved design will aid in vivo tracking and repair in models where lysosomal alkalinization contributes to pathology. Created in BioRender. Mitchell, C. (2025) https://BioRender.com/8hvj96m. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=139 SRC="FIGDIR/small/659991v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@d6fcf1org.highwire.dtl.DTLVardef@7833aborg.highwire.dtl.DTLVardef@188593corg.highwire.dtl.DTLVardef@9aa531_HPS_FORMAT_FIGEXP M_FIG C_FIG New and NoteworthyTools that restore acidic pH in compromised lysosomes can enhance autophagy and waste degradation in degenerative disorders marked by excessive accumulation. Here, we describe the novel synthesis of lysosome-targeted nanoparticles composed of PLGA polymers covalently bound to Cy3 fluorescent dye. These Cy3-PLGA nanoparticles enabled improved tracking of lysosomal delivery and demonstrated sustained long-term retention within lysosomes, supporting their potential for future applications to restore lysosomal pH in aging and degenerating tissues.

cell biology↗

bioRxiv2025

Dual effect of alpha-synuclein disease variants on condensate formation

-synuclein is a pre-synaptic protein1,2 implicated in synucleinopathies like Parkinsons disease and Dementia with Lewy Bodies, where it accumulates in intracellular aggregates termed Lewy bodies and Lewy neurites3. Recent studies have reported that -synuclein undergoes phase separation to form biomolecular condensates both in vitro and in mammalian cells4-7. -synuclein condensates are thought to contribute to disease through progressive aggregation5,8,9. Here we show that specific PD-associated -synuclein variants fail to form biomolecular condensates. We demonstrate that only two -synuclein variants associated with familial disease, E46K and E83Q, enhance condensate formation in vitro and in cells. However, variants including A30G, G51D, and A53E fail to form or have reduced levels of condensate formation in cells. The same phenotypes are reflected in the budding yeast model showing differential inclusion formation. In iPSC-derived neurons, the propensity of -synuclein variants to undergo VAMP2-mediated phase separation reflects the level of synaptic enrichment. We show that the intrinsic propensity of -synuclein to form condensates and the ability to bind lipid membranes are important to mediate condensate formation in cells. Our results emphasize that -synuclein pathology follows divergent pathways, with both increased and decreased condensate formation contributing to disease. This study establishes biomolecular condensates as a key intermediate in -synuclein dysfunction, providing a novel foundation for translational research.

neuroscience↗

bioRxiv2025

KDM5A enhances MHC-I antigen presentation and potentiates antitumor immunity unleashed by immune checkpoint blockade

While some studies have implicated the histone 3 lysine 4 demethylase KDM5A in tumorigenesis and promotion of metastasis, KDM5A has also been shown to boost immunotherapy responses in melanoma. The distinct functional role of KDM5A in the context of immunotherapy and the underlying mechanisms remain largely unknown. Here, we report that higher KDM5A expression strongly correlates with higher MHC-I expression, CD8+ T cell infiltration, and prolonged survival in colorectal cancer and gastric cancer patients receiving anti- PD-1 therapy. KDM5A promotes MHC-I-mediated antigen presentation and CD8+ T cell-mediated tumor killing across multiple murine cancer models. Mechanistically, KDM5A upregulates MHC-I expression through the SOCS1/IFN-{gamma}/STAT1 signaling and improves tumor cell-intrinsic antigen presentation capacity by inhibiting lysosomal proteases in a demethylase-dependent manner. Moreover, KDM5A directly represses lysosomal cathepsins in dendritic cells and promotes the cross-priming activity. Through a high-throughput chemical compound library-based screen, we identified Clomiphene, an FDA-approved small molecule, which significantly elevates KDM5A expression, and enhances antitumor immunity during anti-PD-1 immunotherapy in mouse models of melanoma and colon cancer. Our findings that KDM5A functions in MHC-I-mediated immune activation during anti-PD-1 therapy present an opportunity for developing KDM5A-enhancing therapies to increase tumor immunogenicity, sensitize solid tumors to immunotherapy and exclude immune evasion.

immunology↗

bioRxiv2025

Advanced adaptive strategies in an ancestral body plan: insights from a 510-Ma-old leptomitid sponge

Sponges have thrived in diverse environmental conditions since the early Cambrian till today. However, little is known about how their adaptive capability and strategies have been shaped throughout evolutionary history. Here, we explore this question based on a new leptomitid sponge fossil from the Cambrian Stage 4. The family Leptomitidae was an abundant sponge group inhabiting Cambrian soft substrates but significantly declined thereafter. The new species exhibits a sophisticated set of morphological characteristics adaptive to a shallow siliciclastic environment, which are unprecedented among leptomitids. These include (1) a robust body wall woven by spirally twisted monaxonic spicules; (2) a thick stub-like root tuft for anchoring; (3) spicules radiating out from the sponge body to prevent clogging and sinking; and (4) the inferred capability to close the osculum against unfavourable stimuli. Nevertheless, the new fossil species maintains a leptomitid body plan which lacks modularity and morphological plasticity, the two common and critical attributes in extant sponges to enhance flexibility and resilience in changing environmental conditions. This juxtaposition of evolutionary innovation and structural conservatism offers a compelling case for further exploration of the evolutionary mechanisms that shaped early sponge lineages.

paleontology↗

bioRxiv2025

Selective vulnerability of cerebral vasculature to NOTCH3 variants in small vessel disease and rescue by phosphodiesterase-5 inhibitor

NOTCH3 variants cause CADASIL the most common genetic form of small vessel disease (SVD) and vascular dementia (VaD) and increase the stroke and SVD/VaD risk. CADASIL is a systemic vasculopathy but predominantly manifests in the brain. The molecular mechanisms of CADASIL remain largely unclear with no specific available treatments. NOTCH3 is primarily expressed in vascular smooth muscle cells (VSMCs). Using human induced pluripotent stem cell (iPSC) models and developmental lineage-specific VSMC differentiation, we revealed a selective vulnerability of cerebral but not peripheral VSMC mimics to NOTCH3 variants. Transcriptomic, protein and functional analyses demonstrated a switching of CADASIL iPSC-VSMCs from a contractile to a synthetic phenotype, accompanied with extensive extracellular matrix accumulation and impairment of cell adhesion leading to anoikis. Importantly, we describe an endothelial independent nitric oxide signalling in VSMCs which was dysregulated in the CADASIL iPSC-VSMCs, and posphodiesterase-5 (PDE5) inhibition successfully rescued the abnormal VSMC function, suggesting a novel therapeutic strategy. Our findings offer new mechanistic insights into brain specific phenotype in NOTCH3-associated SVD/VaD and support patient-specific iPSCs to be a valuable model for identifying targeted treatment for NOTCH3-associated SVD/VaD.

neuroscience↗

bioRxiv2025

Kre6-Dependent β-1,6-glucan Biosynthesis Only Occurs in the Conidium of Aspergillus fumigatus

The structural role of {beta}-1,6-glucan has remained under-investigated in filamentous fungi compared to other fungal cell wall polymers, and previous studies have shown that the cell wall of the mycelium of A. fumigatus did not contain {beta}-1,6-glucans. In contrast, the current solid-state NMR investigations showed that the conidial cell wall contained a low amount of {beta}-1,6-glucan. ssNMR comparisons of the A. fumigatus and C. albicans {beta}-1,6-glucans showed they are structurally similar. Deletion of the KRE6 gene which is the only KRE gene in the A. fumigatus genome resulted in a mutant depleted of {beta}-1,6-glucan which has a growth phenotype similar to the parental strain. Even though it is not an essential polymer in A. fumigatus, {beta}-1,6-glucan play a role in cell wall organization since the kre6{Delta} mutant showed a higher sensitivity to Congo-red and Calcofluor white which are known to be general cell wall inhibitors. It is also another example of the significant structural differences seen between conidium and mycelium of filamentous fungi.

biochemistry↗

bioRxiv2025

A novel frameshift mutation in Phosphoinositide 3-kinase regulatory subunit 1 (PIK3R1) causes immunodeficiency and Amyotrophic Lateral Sclerosis (ALS)

Mutations in PIK3R1, a regulatory subunit of Class I PI3K, are implicated in immune disorders and neurological conditions. We identified a novel heterozygous pathogenic frameshift mutation (c.1710dup) in PIK3R1 in a patient with common variable immunodeficiency who developed slowly progressive Amyotrophic Lateral Sclerosis. Induced pluripotent stem cells (iPSCs) and iPSC-derived motor neurons (iMNs) demonstrated that this mutation resulted in PIK3R1 haploinsufficiency, with downstream activation of AKT, disruption of neuronal electrical function and increased apoptosis in iPSC-derived motor neurons. Single-cell RNA sequencing (scRNA-seq) and pathway analysis of differentially expressed genes showed apoptosis pathways were upregulated in neuronal clusters from iMNs harboring the PIK3R1c.1710 dup mutation. Mutated iPSC-derived brain organoids were smaller than matched controls. scRNA-seq of brain organoids showed more active apoptosis in neuronal clusters of patient-derived brain organoids. These findings identify a critical and novel role for PIK3R1 haploinsufficiency in neuronal function and survival.

neuroscience↗

bioRxiv2025

Developing a versatile technology for agroinfiltration in multiple plants

Agrobacterium-mediated transient transformation methods are widely used in plant science and molecular pharming. However, plant immune response and post-transcriptional gene silencing (PTGS) can compromise gene expression, limiting the adaptability of these methods across plant species. The lack of a robust reporter system further hinders advancements in agroinfiltration technology. To address these challenges, we developed a fungal bioluminescence pathway (FBP)-based reporter system that utilizes caffeic acid to generate self-sustained luminescence. By incorporating NahG and P19, our FBP assay enhances transient transformation efficiency in over 20 plant families, including major crops, vegetables, ornamental plants, and trees. Furthermore, we demonstrate its utility for assessing protein localization, interactions, and transcriptional regulation in multiple plant species, thereby broadening our understanding of gene function throughout the plant kingdom.

plant biology↗