Biology subjects

Shin, G. Y.

Publications and source records attributed to Shin, G. Y..

8 recordsLinked to original sources

bioRxiv2026

Phylogenomic Taxonomic Analysis of Ralstonia solanacearum Strains causing Bacterial Wilt Disease in Northeastern Argentina.

Ralstonia solanacearum species complex (RSSC) is a genetically diverse group of plant pathogens, yet genomic data from South America remain limited. Here, we characterize 13 RSSC strains isolated from tomato, pepper, and eggplant in northeastern Argentina. Phylogenetic analysis of the egl marker gene assigned these strains to phylotype IIA and suggested two closely related lineages. Complete genomes (5.63-5.76 Mb) were generated for four representative strains, yielding high-quality (99.94% completeness with f_Burkholderiaceae CheckM markers), closed assemblies with canonical bipartite architecture. Phylogenetic analysis of the egl marker, 49 conserved bacterial genes, and average nucleotide identity (ANI) analyses, consistently assigned one lineage to sequevar IIA-50, forming a coherent and monophyletic group. In contrast, although egl analysis suggested the second lineage was related to one sequevar IIA-38 reference strain, genomic analysis did not support this assignment. Further, the genomic analysis revealed significant genomic distance between the genomes for two sequevar 38 representative strains, supporting a conclusion that sequevar 38 itself was not monophyletic and instead appears paraphyletic. These findings highlight limitations of single-locus classification and support genome-informed refinement of RSSC sub-phylotype taxonomy. Outcome statementReports of bacterial wilt disease in Argentina had not yet been published in the international literature although the disease has been long-standing. This study provides complete genome sequences for four Ralstonia solanacearum strains from Northern Argentina and places them within a global phylogenomic framework. The Argentine strains cluster into two closely related phylotype IIA lineages, indicating that bacterial wilt in this regional dataset is associated with genetically similar populations. For clear communication of which strains are present in Northern Argentina, we attempted to classify the lineages to the long-standing sequence variant (sequevar) system for naming R. solanacearum species complex (RSSC) strains. One lineage was confidently assigned to IIA-50 with genomic support that confirmed phylogenetic analysis of the classical genetic marker egl. However, newly available genomes for sequevar reference strains revealed an issue where two distantly related strains are currently recognized as references for sequevars. Overall, these results provide evidence supporting the need for genome-informed refinement of sub-phylotype classification and expand genomic representation of South American RSSC populations. Data summaryComplete genome assemblies and raw reads for INTABV18, INTABV29, INTABV624 and INTABV2657 are deposited to NCBI under the project number PRJNA1407867. The curated dataset of public RSSC genomes is available to users who register a free account on KBase via a KBase narrative (https://narrative.kbase.us/narrative/189849). The narrative described in a living BioRxiv pre-print [1]. Supplemental files such as Figure S1, rectangular versions of all trees (Figure 2 and 3 and S1) and supplementary table S1, S2, S3 and S4 are available on Zenodo at doi.org/10.5281/zenodo.19502890 O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=172 SRC="FIGDIR/small/721750v1_figS1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1ac3168org.highwire.dtl.DTLVardef@1dfd0d6org.highwire.dtl.DTLVardef@107ae42org.highwire.dtl.DTLVardef@141937c_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure S1.C_FLOATNO Maximum-likelihood phylogenetic tree inferred from 471 bp of endoglucanase (egl) gene sequences assigned Argentine strains as phylotype II sequevar 38 and sequevar 50. The tree was constructed using PhyML v3.0 under the GTR nucleotide substitution model with gamma-distributed rate heterogeneity ( = 0.33), as selected by the SMART model selection procedure implemented in PhyML (Lefort et al., 2017). The egl sequences from Argentine strains are highlighted in blue, and their corresponding GenBank accession numbers for both the egl nucleotide sequence and the whole-genome assembly are shown in parentheses. Reference egl sequences representing sequevars IIA-38 (CFBP6801 and CIP120) and IIA-50 (T1-UY and ACH1076) are also shown in bold and marked with yellow circles. A searchable PDF of this tree in rectangular format is available on Zenodo (doi.org/10.5281/zenodo.19502890). C_FIG O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=196 SRC="FIGDIR/small/721750v1_fig2.gif" ALT="Figure 2"> View larger version (53K): org.highwire.dtl.DTLVardef@39d776org.highwire.dtl.DTLVardef@170bd89org.highwire.dtl.DTLVardef@aba166org.highwire.dtl.DTLVardef@1f156dd_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 2.C_FLOATNO Maximum-likelihood phylogenetic tree inferred from 710 bp of endoglucanase (egl) gene sequences assigned Argentine strains as phylotype II sequevar 38 and sequevar 50. The phylogenetic tree was constructed using PhyML v3.0 under the GTR+R nucleotide substitution model, as selected by the SMART model selection procedure (Lefort et al., 2017). egl sequences from four Argentine strains (INTABV18, INTABV29, INTABV624, and INTABV2657) are shown in bold and highlighted in blue. Reference egl sequences representing sequevars IIA-38 (CFBP6801 and CIP120) and IIA-50 (T1-UY and ACH1076) are also shown in bold and marked with yellow circles. Two USA strains identified as IIA-38 (UCD576 and RS124) are shown in bold. A searchable PDF of this tree in rectangular format is available on Zenodo (doi.org/10.5281/zenodo.19502890). C_FIG O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/721750v1_fig3.gif" ALT="Figure 3"> View larger version (37K): org.highwire.dtl.DTLVardef@17dd372org.highwire.dtl.DTLVardef@1c5156corg.highwire.dtl.DTLVardef@179d9org.highwire.dtl.DTLVardef@e6d529_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 3.C_FLOATNO Approximate maximum-likelihood phylogeny based on a concatenated alignment of 49 conserved genes places four Argentine genomes (INTABV18, INTABV29, INTABV624 and INTABV2657) within the phylotype IIA clade. The tree was constructed using the SpeciesTreeBuilder v0.1.4 application on the KBase platform, incorporating the four Argentine genomes into a reference dataset of 825 genomes representing the known global diversity of the RSSC. The tree was visualized and annotated using iTOL v7.4.2. Argentine genomes are shown in bold and highlighted in blue, and egl reference strains for the sequevar IIA-38 (CIP120 and CFBP6801) and IIA-50 (T1-UY) are shown in bold and marked with yellow circles. Branches with approximate likelihood-ratio support values higher than >70% are colored in blue. A searchable PDF of this tree in rectangular format is available on Zenodo (doi.org/10.5281/zenodo.19502890). C_FIG

microbiology↗

bioRxiv2025

Lifestyle-associated variation in type IV secretion systems between phytopathogenic and environmental Ralstonia

Type IV secretion systems (T4SSs) are versatile machines with variable functions including DNA uptake and release, protein translocation, and DNA conjugation. However, the diversity, distribution, and functional roles of the T4SS in the Ralstonia genus remain poorly understood. The Ralstonia solanacearum species complex (RSSC) comprises three species of plant-pathogenic bacteria that cause bacterial wilt disease. The Ralstonia genus also includes non-RSSC species that are primarily environmental bacteria and rare opportunistic human pathogens. This study compared the diversity and phylogenetic distribution of T4SSs in the RSSC phytopathogens vs. non-RSSC environmentals. Phylogenetic analysis of VirB4 sequences and synteny analysis revealed 16 distinct T4SS clusters in Ralstonia, with ten clusters found in RSSC phytopathogen genomes, twelve in non-RSSC environmental genomes, and six clusters in both groups. Collectively, these gene clusters were more prevalent in non-RSSC environmental genomes. The presence of type IV coupling protein and relaxase genes suggests that at least 14 of these T4SS gene clusters could be putative DNA conjugation systems. The clusters were encoded on accessory plasmids of various sizes or as integrative and conjugative elements (ICEs) on the chromosome or megaplasmid. The putative regions of transfer for T4SS gene clusters in the RSSC phytopathogen genomes often contained type III effectors, type VI secretion toxin/antitoxin clusters, and hemagglutinin gene clusters. In contrast, the non-RSSC environmentals were enriched in heavy metal metabolism and resistance genes. One of the 16 T4SS clusters, cluster i, exhibited evidence of specialization for the RSSC phytopathogens. These findings shed light on the eco-evolutionary differences in the Ralstonia genus. Impact statementThe Ralstonia genus contains the Ralstonia solanacearum species complex (RSSC), a group of globally important plant pathogens that impact food security. These pathogens are known to have an expansive, open pangenome with high levels of gene flow. To shed light on the eco-evolutionary differences between RSSC phytopathogens and closely related environmental species, we explored the potential role of type IV secretion systems (T4SSs) in horizontal gene flow within the Ralstonia genus. Surprisingly, these mobile genetic elements were less common in the phytopathogens than the environmentals. Nevertheless, we identified a particular T4SS gene cluster encoded on an accessory plasmid that appears to be specialized to the pathogenic lifestyle of the RSSC. This specialized cluster harbors genes that drive RSSC host range (type III effectors) and others that likely allow these pathogens to antagonize the plant microbiota (type VI secretion toxins and hemagglutinin two partner secretion systems). Moreover, this study sheds light on the cryptic lifestyles of the poorly studied environmental Ralstonia species, revealing that their conjugative T4SSs frequently carry heavy metal metabolism and resistance genes. Overall, this study provides a foundation to investigate the functional roles of T4SS clusters and their cargo genes in the fitness of phytopathogenic and environmental Ralstonia. Data summarySupplemental Table S1 lists details of the RSSC phytopathogen, non-RSSC environmental, and Burkholderiaceae family genomes used in this study. Supplemental Table S2A is the final list of 501 VirB4 sequences collectively identified in 636 RSSC phytopathogen genomes and 143 non-RSSC environmental genomes. Supplemental Table S2B includes the annotations and NCBI accessions for every gene and protein sequence in reference clusters a-p. Supplemental Table S2C is the list of putative T4SS cargo genes identified in complete Ralstonia genomes. Supplemental Table S2D is the list of putative T4SS cargo genes identified in cluster i regions from draft and complete Ralstonia genomes. Supplemental File S1 is the multiple sequence alignment (MSA) input for HMMER, made from the 753 VirB4 protein sequences from the Burkholderiaceae family genomes. Supplemental Files S2 and S3 are PDF-format versions of the species trees displaying the presence of T4SS gene clusters in RSSC phytopathogens and non-RSSC environmentals, respectively. Supplemental File S4 is the species tree displaying the presence of RSp0179 and RSp1521 in the Ralstonia genus. Supplemental File S5 is the MSA input for FastTree 2, made from the VirB4 protein sequences from Ralstonia genomes. Supplemental Files S6-S9 are various formats of the VirB4 protein tree in Figure 1. Supplemental Files S10-S25 are the GBK files for reference clusters a-p. Supplemental File S26 is the clinker output comparing the reference clusters. Supplemental Files S27-S36 are the clinker outputs comparing the putative regions of transfer of T4SSs in complete Ralstonia genomes. Supplemental File S37 is an MSA analysis comparing RSp1521 to AcvB homologs. Supplemental File S38 is the clustered average nucleotide identity (ANI) matrix for the putative regions of transfer of cluster i T4SSs in draft and complete Ralstonia genomes. Supplemental File S39 is the clinker output comparing the cluster i regions in draft and complete Ralstonia genomes. All Supplemental Tables and Files are available through Zenodo (DOI: 10.5281/zenodo.14873518). O_FIG O_LINKSMALLFIG WIDTH=199 HEIGHT=200 SRC="FIGDIR/small/675681v2_fig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@1640850org.highwire.dtl.DTLVardef@5de6f5org.highwire.dtl.DTLVardef@e605feorg.highwire.dtl.DTLVardef@1937bac_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 1.C_FLOATNO Sequence diversity of the T4SS marker VirB4 in the Ralstonia genus. VirB4 protein sequences (n=496) were identified by a custom hidden Markov model (HMM) search, and an approximately-maximum-likelihood tree was built with FastTree 2 from a MUSCLE multiple sequence alignment (MSA) of the VirB4 sequences. Bootstrap values (0 to 1) from 1,000 resamples are shown for key nodes. The unrooted tree was visualized in iTOL and labeled using Affinity Designer. The 16 distinct T4SS gene clusters (a-p) are outlined and labeled. Circles denote clusters that are present in the RSSC phytopathogens and triangles denote clusters that are present in the non-RSSC environmentals. Colored clusters were found in RSSC phytopathogen genomes, and clusters with dashed outlines were not found in the RSSC phytopathogens. Supplemental files contain the virB4 gene list (Supplemental Table S2A), the VirB4 protein sequence MSA (Supplemental File S5), the Newick file for the tree (File S6), a PDF of the unrooted tree with branch labels (File S7), and PDFs of the rectangular tree with branch labels, without and with colors (Files S8 and S9). C_FIG This studys newly sequenced genomes, Gazipur 4 and Gazipur 5, are available on NCBI with the following accessions: GCF_049860735.1 and GCF_049860725.1, respectively. The two contigs containing the cluster j T4SS were submitted to NCBI through the Third Party Annotation (TPA) section of the DDBJ/ENA/GenBank databases with the following accessions: BK072068-BK072069. These contigs were assembled from SRA reads under the accession: SRR18649448.

microbiology↗

bioRxiv2025

Ecologically Significant Genetic Loci of P. allii: Contributions to Pathogenesis and Competition

Pantoea allii, one of four Pantoea species known to cause onion center rot, is infrequently isolated from onion compared to its closely related onion-pathogenic sister taxa. To better understand the genomic diversity and genetic determinants of pathogenicity in this species, we analyzed a collection of 38 P. allii strains isolated from two primary ecological niches, plants and water, across three continents using comparative genomics and phylogenetic approaches. Core-genome phylogeny, average nucleotide identity (ANI), and gene presence-absence analyses revealed three genetically distinct lineages. All strains harbored conserved biosynthetic gene clusters (BGCs) for quorum sensing, carotenoid production, siderophores, and thiopeptides. In contrast, two phosphonate BGCs, key determinants of onion pathogenicity, exhibited lineage-specific distributions. Onion-associated strains from Lineages 1 and 2 carried the Halophos BGC associated with onion tissue necrosis, and onion isolates encoded the alt gene cluster conferring thiosulfinate tolerance. Lineage 3 strains, isolated from both onion and rainwater, either lacked a phosphonate BGC loci or carried the HiVir phosphonate BGC. In addition, Lineage 3 strains lacked the alt cluster altogether. The localization of these virulence genes in the genome varied, with Halophos integrated in the chromosome, HiVir encoded on the conserved Large Pantoea Plasmid, and alt located on a small, variable plasmids (plasmid B). The Type IV and Type VI secretion systems showed variable genomic architectures, with plasmid-borne T4SSs and two chromosomal T6SS loci differing in conservation and gene content. Additionally, conserved Pantailocin phage islands were detected in most genomes. Overall, this study reveals that while core metabolic and competitive traits are conserved across P. allii, virulence-associated loci display lineage-specific partitioning, reflecting ecological differentiation and evolutionary plasticity within the species. Impact StatementThis study presents a comprehensive comparative genomic analysis of available Pantoea allii genomes, a known onion pathogen. By analyzing 38 strains isolated from plant and water sources across three continents, we uncovered lineage-specific distributions of key virulence genes, alongside conserved genetic traits associated with competition and environmental resilience. These findings clarify the genetic basis of P. allii pathogenesis and highlight its potential as a biocontrol agent, offering broader insights into how ecologically significant loci contribute to the dual roles of plant-associated bacteria as both pathogens and beneficial microbes. Data SummaryPENDING

genomics↗

bioRxiv2024

Plasmids encode and can mobilize onion pathogenicity in Pantoea agglomerans

Pantoea agglomerans is one of four Pantoea species for which strains have been reported in the United States to cause bacterial rot of onion bulbs. However, not all P. agglomerans strains are pathogenic to onion. We characterized onion-associated strains of P. agglomerans to elucidate the genetic and genomic signatures of onion-pathogenic P. agglomerans. We collected >300 P. agglomerans strains associated with symptomatic onion plants and bulbs from public culture collections, research laboratories, and a multi-year survey in 11 states in the USA. Genome assemblies were generated for 87 P. agglomerans strains that showed a range in onion virulence phenotypes. Combining the 87 genome assemblies with 100 high-quality, public P. agglomerans genome assemblies identified two well-represented and well-supported P. agglomerans phylogroups. Strains causing severe symptoms on onion leaves and bulbs were only identified in Phylogroup II and encoded the HiVir biosynthetic cluster for the phytotoxin pantaphos, supporting the role of HiVir as a crucial pathogenicity factor. Using a MASH-based plasmid classification system, the P. agglomerans HiVir cluster was determined to be encoded in two distinct plasmid contexts: 1) as an accessory gene cluster on a conserved P. agglomerans plasmid (pAggl), or 2) on a mosaic cluster of plasmids common among onion strains (pOnion). Analysis of closed genomes of P. agglomerans revealed that the pOnion plasmids harbored alt genes responsible for encoding tolerance to the thiosulfinate defensive chemistry in Allium spp. Additionally, many of these pOnion plasmids harbored cop gene clusters, which confer resistance to copper. However, the pOnion plasmids encoded the HiVir cluster less frequently. We demonstrated that the pOnion plasmid pCB1C, encoding HiVir and alt clusters as well as an intact conjugative type IV secretion system (T4SS), can act as a natively mobilizable pathogenicity plasmid that transforms P. agglomerans Phylogroup I strains, including environmental strains, into virulent pathogens of onion. This work indicates a central role for plasmids and plasmid ecology in mediating P. agglomerans interactions with onion plants, with potential implications for onion bacterial disease management.

microbiology↗

bioRxiv2023

Pantailocins: Phage-derived Bacteriocins from Pantoea ananatis and Pantoea stewartii subsp. indologenes

Phage-derived bacteriocins are highly specific and effective antimicrobial molecules which have successfully been used as prophylactic treatments to prevent phytopathogen infections. Given the specificity of tailocins, a necessary step for broadening the tailocin catalog and for extending applicability across systems and diseases is the screening of new clades of phytopathogens for production of molecules with tailocin-like killing activity. Here, we describe production by and sensitivity of strains to tailocins produced by Pantoea ananatis and Pantoea stewartii subsp. indologenes. Phylogenetic evidence suggests that these tailocins are derived from Myoviridae family phage like many previously described R type syringacins and R type pyocins, but also suggests that cooption from phage occurred independently of previously described tailocins. Since these tailocin encoding loci are present in the same genomic locations across multiple strains of both species and display a level of divergence that is consistent with other shared regions between the genomes and with vertical inheritance of the locus, we refer to them broadly as pantailocins.

microbiology↗

bioRxiv2022

The genetic requirements for HiVir-mediated onion necrosis by Pantoea ananatis, a necrotrophic plant pathogen

Pantoea ananatis is an unusual bacterial pathogen that lacks typical virulence determinants yet causes extensive necrosis in onion foliage and bulb tissues. The onion necrosis phenotype is dependent on the expression of a phosphonate toxin, pantaphos that is catalyzed by putative enzymes encoded by the HiVir gene cluster. The genetic contributions of individual hvr genes in HiVir-mediated onion necrosis remain largely unknown except for the first gene hvrA (phosphoenolpyruvate mutase, pepM) whose deletion resulted in the loss of onion pathogenicity. In this study, using gene deletion mutation and complementation, we report that of the ten remaining genes, hvrB-hvrF are also strictly required for the HiVir-mediated onion necrosis and in planta bacterial growth whereas hvrG-hvrJ partially contributed to these phenotypes. As the HiVir gene cluster is a common genetic feature shared among the onion-pathogenic P. ananatis strains, and as it could serve as a useful diagnostic marker of onion pathogenicity, we sought to understand the genetic basis of HiVir positive yet phenotypically deviant (non-pathogenic) strains. We identified and genetically characterized inactivating single nucleotide polymorphisms (SNPs) in essential hvr genes of six phenotypically deviant P. ananatis strains. Finally, inoculation of the cell-free spent medium of Ptac-driven HiVir strain caused P. ananatis-characteristic red onion scale necrosis (RSN) as well as cell death symptoms in tobacco. The co-inoculation of the spent medium with essential hvr mutant strains restored strains in planta populations to the wild-type level, suggesting that necrosis is important for proliferation of P. ananatis in onion tissue.

microbiology↗

bioRxiv2020

Pre-existing and de novo humoral immunity to SARS-CoV-2 in humans

Several related human coronaviruses (HCoVs) are endemic in the human population, causing mild respiratory infections1. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the etiologic agent of Coronavirus disease 2019 (COVID-19), is a recent zoonotic infection that has quickly reached pandemic proportions2,3. Zoonotic introduction of novel coronaviruses is thought to occur in the absence of pre-existing immunity in the target human population. Using diverse assays for detection of antibodies reactive with the SARS-CoV-2 spike (S) glycoprotein, we demonstrate the presence of pre-existing humoral immunity in uninfected and unexposed humans to the new coronavirus. SARS-CoV-2 S-reactive antibodies were readily detectable by a sensitive flow cytometry-based method in SARS-CoV-2-uninfected individuals and were particularly prevalent in children and adolescents. These were predominantly of the IgG class and targeted the S2 subunit. In contrast, SARS-CoV-2 infection induced higher titres of SARS-CoV-2 S-reactive IgG antibodies, targeting both the S1 and S2 subunits, as well as concomitant IgM and IgA antibodies, lasting throughout the observation period of 6 weeks since symptoms onset. SARS-CoV-2-uninfected donor sera also variably reacted with SARS-CoV-2 S and nucleoprotein (N), but not with the S1 subunit or the receptor binding domain (RBD) of S on standard enzyme immunoassays. Notably, SARS-CoV-2-uninfected donor sera exhibited specific neutralising activity against SARS-CoV-2 and SARS-CoV-2 S pseudotypes, according to levels of SARS-CoV-2 S-binding IgG and with efficiencies comparable to those of COVID-19 patient sera. Distinguishing pre-existing and de novo antibody responses to SARS-CoV-2 will be critical for our understanding of susceptibility to and the natural course of SARS-CoV-2 infection.

immunology↗

bioRxiv2020

Extraction-free COVID-19 (SARS-CoV-2) diagnosis by RT-PCR to increase capacity for national testing programmes during a pandemic

Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) causes Coronavirus disease 2019 (COVID-19), a respiratory tract infection. The standard molecular diagnostic test is a multistep process involving viral RNA extraction and real-time quantitative reverse transcriptase PCR (qRT-PCR). Laboratories across the globe face constraints on equipment and reagents during the COVID-19 pandemic. We have developed a simplified qRT-PCR assay that removes the need for an RNA extraction process and can be run on a real-time thermal cycler. The assay uses custom primers and probes, and maintains diagnostic sensitivity within 98.0% compared to the assay run on a high-throughput, random-access automated platform, the Panther Fusion (Hologic). This assay can be used to increase capacity for COVID-19 testing for national programmes worldwide.

molecular biology↗