Biology subjects

Hu, Z.

Publications and source records attributed to Hu, Z..

At least 199 records · Page 11Linked to original sources

bioRxiv2023

Establishment of a novel non-integrated human pluripotent stem cell-based gastruloid model

Embryo loss and pregnancy disorders are prevalent worldwide, with both conditions critically associated with dysfunctioning gastrulation processes. Gastrulation and post-gastrulation organogenesis are crucial stages of embryonic development that establish the blueprint for body part formation. These processes involve the sequential generation of three germ layer cells and primordial germ cells, as well as the assembly of the precursor tissues for body parts. However, due to ethical limitations associated with studying human embryogenesis, a more detailed understanding of gastrulation and post-gastrulation organogenesis remains elusive. To ensure that the knowledge obtained from gastruloids is biologically meaningful and clinically relevant, it is critical to create high-fidelity human embryo models that closely mimic embryogenesis in vivo. Here, we developed a two-stage derivation gastruloids in vitro based on human pluripotent stem cells. Morphological tracking mimicks the developmental processes of models from Carnegie Stage 4 (CS4) to early CS7. Our gastruloids exhibit key structures characteristic of human embryos, including amniotic cavity, embryonic disc, primitive streak, primary yolk sac, secondary yolk sac, and blood islets. Comparison of our cell lineage development maps showed that gastruloids closely resembled human natural CS7 gastrula. Our gastruloids exhibited transcriptional characteristics that mimicked the molecular pathways observed in natural embryos development. Importantly, we found that in our model, extraembryonic mesoderm originates from the yolk sac and primordial germ cells originate from the posterior epiblast of the embryonic disc. Moreover, we found that thalidomide affects the differentiation of three germ layer cells, resulting in the arrest of human gastruloid development. In conclusion, by establishing a human gastruloid, we were able to gain valuable insights into the mechanisms responsible for human gastrulation and shed light on the causes of early embryo loss and pregnancy disorders.

developmental biology↗

bioRxiv2023

Predicting the impact of rare variants on RNA splicing in CAGI6

BackgroundVariants which disrupt splicing are a frequent cause of rare disease that have been under-ascertained clinically. Accurate and efficient methods to predict a variants impact on splicing are needed to interpret the growing number of variants of unknown significance (VUS) identified by exome and genome sequencing. Here we present the results of the CAGI6 Splicing VUS challenge, which invited predictions of the splicing impact of 56 variants ascertained clinically and functionally validated to determine splicing impact. ResultsThe performance of 12 prediction methods, along with SpliceAI and CADD, was compared on the 56 functionally validated variants. The maximum overall accuracy achieved was 82% from two different approaches, one weighting SpliceAI scores by minor allele frequency, and one applying the recently published Splicing Prediction Pipeline (SPiP). SPiP performed optimally in terms of sensitivity, while an ensemble method combining multiple prediction tools and information from databases exceeded all others for specificity. ConclusionsSeveral challenge methods equalled or exceeded the performance of SpliceAI, with ultimate choice of prediction method likely to depend on experimental or clinical aims. One quarter of the variants were incorrectly predicted by at least 50% of the methods, highlighting the need for further improvements to splicing prediction methods for successful clinical application.

bioinformatics↗

bioRxiv2023

C1q Enables Influenza HA Stem Binding Antibodies to Block Viral Attachment and Broadens the Antibody Escape Repertoire

Broadly neutralizing, anti-hemagglutinin stem antibodies (Abs) are a promising universal influenza vaccine target. While anti-stem Abs are not believed to block viral attachment, we show that C1q confers attachment inhibition and boosts fusion and neuraminidase inhibition, greatly enhancing virus neutralization activity in vitro and in mice challenged with influenza virus via the respiratory route. These effects reflect increased steric interference and not increased Ab avidity. Remarkably, C1q greatly expands the anti-stem Ab viral escape repertoire to include residues throughout the hemagglutinin. Some substitutions cause antigenic alterations in the globular region or modulate HA receptor avidity. We also show that C1q enhances the neutralization activity of non-RBD anti-SARS-CoV-2 Spike Abs, an effect dependent on Spike density on the virion surface. Together, our findings show that first, Ab function must be considered in a physiological context and second, inferring the exact selection pressure for Ab-driven viral evolution is risky business, at best.

microbiology↗

bioRxiv2023

Whole genome sequencing reveals autooctoploidy in the Chinese sturgeon and its evolutionary trajectories

The Order Acipenseriformes, which include sturgeons and paddlefishes, represent "living fossils" with complex genomes that are good models for understanding whole genome duplication (WGD) and ploidy evolution in fishes. Here we sequenced and assembled the first high-quality chromosome-level genome for the complex octoploid Acipenser sinensis (Chinese sturgeon), a critically endangered species that also represents a poorly understood ploidy group in Acipenseriformes. Our results show that A. sinensis is a complex autooctoploid species containing four kinds of octovalents (8 n), a hexavalent (6 n), two tetravalents (4 n), and a divalent (2 n). We propose based on an analysis taking into account delayed rediploidization that its octoploid genome composition results from two rounds of homologous whole genome duplications (WGDs), and further provide insight into the timing of its ploidy evolution. This study provides the first octoploid genome resource of Acipenseriformes for understanding ploidy composition and evolutionary trajectories of polyploidy fishes.

genomics↗

bioRxiv2023

IL-1R signaling drives enteric glia-macrophage interaction in colorectal cancer progression

Enteric glial cells (EGCs) have been implicated in colorectal cancer (CRC) pathogenesis. However, their precise mechanisms of interaction with the CRC immune cell compartment and pro-tumorigenic role remain unclear. This study aimed to investigate the immunomodulatory effects of EGCs on tumor-associated macrophages (TAMs) and their involvement in CRC progression. Using EGC depletion and supplementation models, we assessed the impact of EGCs on the immunomodulation of orthotopic murine CRC. Furthermore, by making use of Bulk RNA-sequencing of CRC EGCs and single-cell sequencing of the tumor microenvironment, we identified the factors involved in the EGC-TAM crosstalk. Findings demonstrate that EGCs acquire a reactive and immunomodulatory phenotype in both murine CRC models and patients, influencing TAM differentiation. Mechanistically, secretion of IL-1 by tumor-infiltrating monocytes and macrophages triggers the phenotypic and functional switch of CRC EGCs via IL-1R. Consequently, tumor EGCs secrete IL-6, promoting the differentiation of monocytes into pro-tumorigenic SPP1+ TAMs. Importantly, the reactive tumor EGCs phenotype correlates with worse disease outcomes in preclinical models and CRC patients. Here we uncover a previously unexplored neuroimmune interaction between EGCs and TAMs within the colorectal tumor microenvironment, informing potential therapeutic strategies and enhancing our understanding of CRC progression. eTOC SummaryOur study unveils a novel neuroimmune interaction between enteric glia and TAMs in colon carcinoma. Monocyte/Macrophage-derived IL-1 activates enteric glia, leading to the differentiation of pro-tumorigenic SPP1+ TAMs via glial-derived IL-6. Blocking glial IL-1R-signaling reduces colonic tumor lesions, highlighting IL-1R as a potential therapeutic target.

cancer biology↗

bioRxiv2023

scATAnno: Automated Cell Type Annotation for single-cell ATAC-seq Data

Recent advances in single-cell epigenomic techniques have created a growing demand for scATAC-seq analysis. One key analysis task is to determine cell type identity based on the epigenetic data. We introduce scATAnno, a python package designed to automatically annotate scATAC-seq data using large-scale scATAC-seq reference atlases. This workflow generates the reference atlases from publicly available datasets enabling accurate cell type annotation by integrating query data with reference atlases, without the use of scRNA-seq data. To enhance annotation accuracy, we have incorporated KNN-based and weighted distance-based uncertainty scores to effectively detect cell populations within the query data that are distinct from all cell types in the reference data. We compare and benchmark scATAnno against 7 other published approaches for cell annotation and show superior performance in multiple data sets and metrics. We showcase the utility of scATAnno across multiple datasets, including peripheral blood mononuclear cell (PBMC), Triple Negative Breast Cancer (TNBC), and basal cell carcinoma (BCC), and demonstrate that scATAnno accurately annotates cell types across conditions. Overall, scATAnno is a useful tool for scATAC-seq reference building and cell type annotation in scATAC-seq data and can aid in the interpretation of new scATAC-seq datasets in complex biological systems.

bioinformatics↗

bioRxiv2023

Developmental Alterations in Brain Network Asymmetry in 3- to 9-Month Infants with Congenital Sensorineural Hearing Loss

Auditory exposure plays crucial roles in shaping healthy brain development and generating lateralization of functional network organization. However, little is known about whether and how an initial lack of auditory exposure in early infancy may disrupt development of functional network lateralization. We addressed this issue by recruiting 55 infants with congenital sensorineural hearing loss (SNHL) and 60 typically developing (TD) controls. Resting-state fNIRS imaging data were acquired to construct hemispheric cerebral networks, and graph theory was applied to quantify the topological characteristics of hemispheric networks. The infants with SNHL exhibited efficient small-world characteristic within each hemispheric network, however, the lateralization of functional network efficiency was substantially disrupted. Compared with TD infants with significantly increased network efficiency lateralized toward left hemisphere with age, the SNHL infants did not exhibit the emergence and development of such cerebral lateralization. Furthermore, the increased leftward asymmetry in nodal efficiency with age was found in TD but not in SNHL infants. Interestingly, the degree of hearing loss had no correlation with lateralization strength in the SNHL group. These results suggest that SNHL infants exhibited disrupted development of cortical lateralization in functional network organization, and highlight the importance of auditory stimulation-promoted multisensory functional integration in early infancy.

neuroscience↗

bioRxiv2023

Molecular Dynamics simulation of NRAS-Q61 oncogenes and new strategies for in silico drug design

The NRAS-mutant subset of melanoma represents the most aggressive and the deadliest types associated with poorer overall survival. Unfortunately, for more than 40 years, no therapeutic agent directly targeting NRAS mutations have been clinically approved yet. Herein, based on microsecond scale molecular dynamics simulations, the concept of NRAS-Q61 mutation classification was firstly proposed. NRAS Q61 positively charged mutations (Q61R and Q61K) was classified together, with a specific targetable pocket, while NRAS-Q61L classified into another category. Moreover, the isomer-sourced structure iteration (ISSI) method was developed for the in silico design of potential inhibitors (HM-516) targeting NRAS mutations. Overall, through this article, we hope to provide the academic and clinical community with new perspective and understanding of NRAS oncoproteins, and propose possible solution to this challenge.

cancer biology↗

bioRxiv2023

Deep flanking sequence engineering for efficient promoter design

Human experts are good at summarizing explicit strong patterns from small samples, while deep learning models can learn implicit weak patterns from big data. Biologists have traditionally described the sequence patterns of promoters via transcription factor binding sites (TFBSs), while the flanking sequences among TFBSs, which can also significantly influence promoter activity and function, remain largely uncharacterized. Thus, current synthetic promoters are mainly designed by the manipulation of TFBSs, while the flanking sequence is often chosen arbitrarily or by previous experience due to a lack of well-summarized optimization criteria. Here, we introduced an AI-aided promoter design framework, DeepSEED, that employs expert knowledge and deep learning methods to efficiently design synthetic promoters that have various desirable functions. DeepSEED incorporates the user-defined cis-regulatory sequences as seeds and generates flanking sequences that match the seeds. We showed that DeepSEED can automatically capture k-mer frequencies and DNA shape features from active promoters in the training set and efficiently optimize the flanking sequences to better match desired properties in synthetic promoters. We validated the effectiveness of this framework for diverse synthetic promoter design tasks in both prokaryotic and eukaryotic cells. DeepSEED successfully designed E. coli constitutive, isopropyl-beta-D-1-thiogalactopyranoside (IPTG)-inducible, and mammalian cell doxycycline (Dox)-inducible promoters with significant performance improvements, suggesting that DeepSEED has potential as an efficient AI-aided flanking sequence optimization approach for promoter design that may be of great benefit in synthetic biology applications. Availabilityhttps://github.com/WangLabTHU/deepseed

bioengineering↗

bioRxiv2023

Discovery of a new drug using lipid-based formulation targeting G12D mutated KRAS4B through non-covalent bonds

One of the most common drivers in human cancer is KRAS4B. In recent years, the promising KRAS targeted drug development has attracted significant new research interest and reignited the field of RAS therapeutics. To signal, oncogenic KRAS4B not only requires a sufficient nucleotide exchange, but also needs to recruit effectors by exposing its effector-binding sites while anchoring to plasma membrane where KRAS4B-mediated signaling events occur. Phosphodiesterase-{delta} plays an important role in sequestering KRAS4B from the cytoplasm and targeting it to cellular membranes. In this work, we have designed a drug LIG1 using lipid-based formulation to directly target both the switch-II pocket of KRAS4B-G12D and phosphodiesterase-{delta}. LIG1 was found to lock KRAS4B in its GDP-bound state while the effector-binding domain is blocked by the interface of the plasma membrane which hinders the nucleotide exchange while simultaneously it can affect the GTP-bound KRAS4B to shift from an active state to its inactive state. LIG1 is also observed to stably accommodate itself in the prenyl-binding pocket of phosphodiesterase-{delta} which impairs KRAS4B enrichment at the membrane and suppress the proliferation of KRAS4B-dependent cancer cells. In this work we report a drug based on lipid-based formulation that can foster drug discovery efforts for the targeting of oncogenes of the RAS family and beyond.

molecular biology↗

bioRxiv2023

Vangl2 suppresses NF-κB signaling and ameliorates sepsis by targeting p65 for NDP52-mediated autophagic degradation

Van Gogh-like 2 (Vangl2), a core planar cell polarity (PCP) component, plays an important role in polarized cellular and tissue morphology induction, growth development and cancer. However, its role in regulating inflammatory responses remains elusive. Here, we report that Vangl2 is upregulated in patients with sepsis and identify Vangl2 as a negative regulator of NF-{kappa}B signaling by regulating the protein stability and activation of the core transcription component p65. Mice with myeloid-specific deletion of Vangl2 (Vangl2{Delta}M) are hypersusceptible to lipopolysaccharide (LPS)-induced septic shock. Vangl2 deficient myeloid cells exhibit enhanced phosphorylation and expression of p65, therefore, promoting the secretion of pro-inflammatory cytokines after LPS stimulation. Mechanistically, NF-{kappa}B signaling-induced-Vangl2 recruits E3 ubiquitin ligase PDLIM2 to catalyze K63-linked ubiquitination on p65, which serves as a recognition signal for cargo receptor NDP52-mediated selective autophagic degradation. Taken together, these findings demonstrate Vangl2 as a suppressor of NF-{kappa}B mediated inflammation and provide insights into the crosstalk between autophagy and inflammatory diseases.

immunology↗

bioRxiv2023

The Potential of Low Press and Hypoxia Environment in Assisting Pan-cancer Treatment

ObjectiveA low incidence and mortality rate of cancer has been observed in high-altitude regions, suggesting a potential positive effect of low press and hypoxia (LPH) environment on cancer. Based on this finding, our study aimed to construct a pan-cancer prognosis risk model using a series of ADME genes intervened by low oxygen, to explore the impact of LPH environment on the overall survival (OS) of various kinds of cancers, and to provide new ideas and approaches for cancer prevention and treatment. Datasets and MeasuresThe study used multiple sources of data to construct the pan-cancer prognosis risk model, including gene expression and survival data of 8,628 samples from the cancer genome atlas, and three gene expression omnibus databases were employed to validate the prediction efficiency of the prognostic model. The AltitudeOmics dataset was specifically used to validate the significant changes in model gene expression in LPH. To further identify the biomarkers and refine the model, various analytical approaches were employed such as single-gene prognostic analysis, weighted gene co-expression network analysis, and stepwise cox regression. And LINCS L1000, AutoDockTools, and STITCH were utilized to explore effective interacting drugs for model genes. Main Outcomes and ConclusionsThe study identified eight ADME genes with significant changes in the LPH environment to describe the prognostic features of pan-cancer. Lower risk scores calculated by the model were associated with better prognosis in 25 types of tumors, with a p-value of less than 0.05. The LPH environment was found to reduce the overall expression value of model genes, which could decrease the death risk of tumor prognosis. Additionally, it is found that the low-risk group had a higher degree of T cell infiltration based on immune infiltration analysis. Finally, drug exploration led to the identification of three potential model-regulating drugs. Overall, the study provided a new approach to construct a pan-cancer survival prognosis model based on ADME genes from the perspective of LPH and offered new ideas for future tumor prognosis research.

cancer biology↗

bioRxiv2023

Redefinition of archetypal phytoplankton-associated bacteria taxa based on globally distributed dinoflagellates and diatoms

Bacteria colonizing in the phycosphere formed by phytoplankton exudates play important roles in marine ecosystems, yet their taxonomy is poorly defined. Here, we customized the analytical approaches for the microalga-attached microbiotas from 110 diatom and 86 dinoflagellate samples to reveal key bacterial players and their ecological significance in the phycosphere. The results demonstrated a co-occurrence of host-specificity and conservation of phytoplankton-associated bacterial communities, defined 8 diatom- and 23 dinoflagellate-affiliated characteristic genera, as well as identifying 14 core genera prevalent with phytoplankton populations. Further classification of these 14 core genera into three tiers showed their distinct ecological features regarding occupancy, connectivity and community-stabilizing, whilst also matching their inherent metabolic properties. Our study redefines the archetypal phytoplankton-associated bacteria taxa more specifically up to the genus level, highlighting the significance of rarely noticed bacteria in the phycosphere, which is invaluable when selecting target bacteria for studying phytoplankton-bacteria interactions.

microbiology↗

bioRxiv2023

The remarkable complexity of the brain microbiome in health and disease

Microbes in human brain and their potential contribution to neurodegenerative conditions such as Alzheimers disease (AD) have long been debated. We recently developed a new method (the electronic tree of life, eToL) based on small subunit ribosomal RNA (rRNA) probes, further confirmed by large subunit rRNA analysis, to comprehensively address the spectrum of microorganisms in control and AD brain. We report a remarkable diversity of brain microbes in control brain. The most abundant are fungi, bacteria, and chloroplastida, and we report detailed identification of representative microbial species. The pattern is substantially conserved across different bilateran species from Drosophila to human. In terms of diversity, the human brain microbiome appears to be a subset (~20%) of the gut microbiome. Adenovirus type C was the major virus found in human brain; other viruses were not well represented. However, the spectrum of brain microbes differed between individuals as well as between brain regions examined from single individuals (amygdala, cingulate cortex, hippocampus, hypothalamus); of these four regions, the highest microbial burden was in cingulate cortex. There was evidence of spreading of pathogens between brain regions in single individuals. Some microbes are over-represented in AD brain according to two measures: (i) absolute number of microbes normalized to endogenous human transcripts, and (ii) the number of brain specimens showing overabundance versus control. Species over-represented in AD brain according to both measures notably include bacteria (Streptococcus, Staphylococcus/Bacillus, Sphingomonas/Ralstonia) and fungi (Acrocalymma/Altenaria/Aureobasidium of the Aspergillus group; Komagataella of the Candida group, Cortinarius of the Schizophyllum group, and Tausonia of the Cryptococcus group), that are all related to known human pathogens. In addition, an uncharacterized chloroplastida (algae-related) species was more abundant in AD brain samples. Although these findings point to diverse microbial species, indicative of multiple causation, similar absolute levels of bacteria and fungi in AD brain samples could suggest synergy between pathogens. However, it is important to stress that not all AD samples were positive for these microbes, but this could be because the affected brain region(s) was not examined. These findings support the contention that infection, perhaps associated with declining immunity with age, may contribute to AD development.

neuroscience↗

bioRxiv2023

Neutralization of SARS-CoV-2 BQ.1.1 and XBB.1.5 by Breakthrough Infection Sera from Previous and Current Waves in China

SARS-CoV-2 is continuing to evolve and diversify, with an array of various Omicron sub-lineages, including BA.5, BA.2.75, BN.1, BF.7, BQ.1, BQ.1.1, XBB and XBB.1.5, now circulating globally at recent time. In this study, we evaluated the neutralization sensitivity of a comprehensive panel of Omicron subvariants to sera from different clinical cohorts, including individuals who received homologous or heterologous booster vaccinations, vaccinated people who had Delta or BA.2 breakthrough infection in previous waves, and patients who had BA.5 or BF.7 breakthrough infection in the current wave in China. All the Omicron subvariants exhibited substantial neutralization evasion, with BQ.1, BQ.1.1, XBB.1, and XBB.1.5 being the strongest escaped subvariants. Sera from Omicron breakthrough infection, especially the recent BA.5 or BF.7 breakthrough infection, exhibited higher neutralizing activity against all Omicron sub-lineages, indicating the chance of BA.5 and BF.7 being entirely replaced by BQ or XBB subvariants in China in a short-term might be low. We also demonstrated that the BQ and XBB subvariants were the most resistant viruses to monoclonal antibodies. Continuing to monitor the immune escape of SARS-CoV-2 emerging variants and developing novel broad-spectrum vaccines and antibodies are still crucial.

immunology↗

bioRxiv2023

Robust biomarker discovery through multiplatform multiplex image analysis of breast cancer clinical cohorts

AO_SCPLOWBSTRACTC_SCPLOWSpatial profiling of tissues promises to elucidate tumor-microenvironment interactions and enable development of spatial biomarkers to predict patient response to immunotherapy and other therapeutics. However, spatial biomarker discovery is often carried out on a single patient cohort or imaging technology, limiting statistical power and increasing the likelihood of technical artifacts. In order to analyze multiple patient cohorts profiled on different platforms, we developed methods for comparative data analysis from three disparate multiplex imaging technologies: 1) cyclic immunofluorescence data we generated from 102 breast cancer patients with clinical follow-up, in addition to publicly available 2) imaging mass cytometry and 3) multiplex ion-beam imaging data. We demonstrate similar single-cell phenotyping results across breast cancer patient cohorts imaged with these three technologies and identify cellular abundance and proximity-based biomarkers with prognostic value across platforms. In multiple platforms, we identified lymphocyte infiltration as independently associated with longer survival in triple negative and high-proliferation breast tumors. Then, a comparison of nine spatial analysis methods revealed robust spatial biomarkers. In estrogen receptor-positive disease, quiescent stromal cells close to tumor were more abundant in good prognosis tumors while tumor neighborhoods of mixed fibroblast phenotypes were enriched in poor prognosis tumors. In triple-negative breast cancer (TNBC), macrophage proximity to tumor and B cell proximity to T cells were greater in good prognosis tumors, while tumor neighborhoods of vimentin-positive fibroblasts were enriched in poor prognosis tumors. We also tested previously published spatial biomarkers in our ensemble cohort, reproducing the positive prognostic value of isolated lymphocytes and lymphocyte occupancy and failing to reproduce the prognostic value of tumor-immune mixing score in TNBC. In conclusion, we demonstrate assembly of larger clinical cohorts from diverse platforms to aid in prognostic spatial biomarker identification and validation. SO_SCPLOWTATEMENTC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWOFC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWSIGNIFICANCEC_SCPLOWOur single-cell spatial analysis of multiple clinical cohorts uncovered novel biomarkers of patient outcome in breast cancer. Additionally, our data, software, and methods will help advance spatial characterization of the tumor microenvironment.

cancer biology↗

bioRxiv2023

SunTag-PE: a modular prime editing system enables versatile and efficient genome editing

Prime editing (PE) holds tremendous potential in the treatment of genetic diseases because it can install any desired base substitution or local insertion/deletion. However, the full-length PE effector size (6.3-kb) was beyond the packaging capacity of adeno-associated virus (AAV), hindering its clinical transformation. Various splitting strategies have been used to improve its delivery, but always accompanied by compromised PE efficiency. Here, we developed a modular and efficient SunTag-PE system that splits PE effectors into GCN4-nCas9 and single-chain variable fragment (scFv) tethered reverse transcriptase (RT). We observed that SunTag-PEs with 1xGCN4 in the N terminus of nCas9 was the most efficient configuration rather than multiple copies of GCN4. This SunTag-PE strategy achieved editing levels comparable to canonical fused-PE and higher than other split-PE strategies (including sPE and MS2-PE) in both PE2 and PE3 forms with no increase in insertion-deletion (indel) byproducts. Moreover, we successfully validated the modularity of SunTag-PE system in the Cas9 orthologs of SauCas9 and FrCas9. Finally, we employed dual AAVs to deliver SunTag-ePE3 and efficiently corrected the pathogenic mutation in HBB mutant cell line. Collectively, our SunTag-PE system provides an efficient modular splitting strategy for prime editing and further facilitate its transformation in clinics.

bioengineering↗

bioRxiv2023

Leafhopper transmits soybean stay-green associated virus to legume plants

A novel geminivirus, soybean stay-green associated virus (SoSGV), was previously shown to cause soybean delayed senescence and is associated with the incidences of soybean stay-green syndrome. The transmission methods of SoSGV were not yet understood. We captured insects belonging to 24 distinct species in a soybean field with the SoSGV outbreak and detected the presence of SoSGV only in leafhoppers and bean bugs (Riptortus pedestris). Caged feeding experiments using captured leafhoppers and bean bugs from soybean fields showed that leafhoppers, but not bean bugs, are vectors transmitting SoSGV. The common brown leafhopper (Orosius orientalis) is identified as the dominant leafhopper species and can establish colonies feeding on soybean plants in experimental conditions. An investigation of SoSGV defective DNA revealed that soybean genomic DNA could be inserted into the SoSGV genome, while sequences from wild soybean, red bean, and cowpea were also identified. We further showed that the common brown leafhopper could transmit SoSGV to wild soybean and red bean plants, emphasizing the vector role of the leafhopper in the transmission of SoSGV in the field.

microbiology↗