Genome Sequence Analysis Research Articles

Genome Sequencing and Comparative Genomic Analysis of Attenuated Strain Gibellulopsis nigrescens GnVn.1 Causing Mild Wilt in Sunflower

Genome Sequencing and Comparative Genomic Analysis of Attenuated Strain Gibellulopsis nigrescens GnVn.1 Causing Mild Wilt in Sunflower

Identification and validation of lung adenocarcinoma (LUAD)-associated targets for monascin from the extracts of Monascus purpureus-fermented rice: Compound preparation, high-throughput genome sequencing and bioinformatics analysis

Despite the remarkable bioactivities exhibited by monascin against several tumors, its therapeutic targets remain unexplored. In this study, our objective is to identify therapeutic targets based on the antitumor activity of monascin against lung adenocarcinoma (LUAD) A549 cells. The compound monascin was derived from extracts of Monascus purpureus-fermented rice. Its chemical structure was determined using spectroscopic methods, while the physicochemical properties of monascin were investigated through thermal behavior analysis. Herein, antitumor studies indicated that monascin significantly inhibited the viability of A549 cells, with an IC50 value of 2.05 μM; RNA-sequencing revealed 12 up-regulated and 29 down-regulated genes as differentially expressed genes (DEGs) for A549 cells in response to monascin; RT-qPCR validation supported the plausibility of the RNA-sequencing results. Moreover, utilizing data from the Cancer Genome Atlas (TCGA) database, univariate and multivariate Cox regression analyses provided evidence supporting GPR37 and FAM83A as potential candidate genes for predicting disease progression in LUAD patients. Additionally, the LASSO Cox regression and nomogram analyses confirmed that FAM83A and GPR37 genes had the potential to predict overall survival for LUAD patients. Finally, molecular docking studies suggested that monascin could interact with both GPR37 and FAM83A proteins through the hydrogen bonding and hydrophobic interactions; Western blotting assays indicated that monascin inhibited the expression levels of both GPR37 and FAM83A proteins. Overall, the clinical prognosis of GPR37 and FAM83A highlights the rationale for targeting these specific genes with monascin. Additionally, there findings provide compelling evidence for the ploypharmacological properties of monascin against LUAD.

Molecular characterization of ST15 carbapenem-resistant Klebsiella pneumoniae isolated in a single patient

The carbapenem-resistant K. pneumoniae (CRKP) poses a serious threat to antibiotic applicability and public health. During treatment, K. pneumoniae (KP) frequently exhibits shifts in drug-resistant phenotypes, complicating clinical treatment as it transitions from sensitivity to resistance. In this study, we analyzed the clinical and molecular characteristics of drug resistance changes in KP strains isolated from a single patient, and the potential mechanisms underlying these resistance changes. Antimicrobial susceptibility test and string test were conducted to evaluate the resistant and virulent characterization of the strains. Pulsed field gel electrophoresis (PFGE) was used to investigate the homology relationship between the strains. The whole genome sequencing and phylogenetic analysis of 9 representative isolates was also performed. The transfer ability of the drug-resistant plasmid was studied by plasmid conjugation experiment. The transconjugants were verified by PCR amplification of specific genes, antimicrobial susceptibility test and PFGE. Our results revealed that 9 KP strains, isolated from the same patient, exhibited 'resistance-sensitivity-resistance-sensitivity' alternately to carbapenems. The differences in DNA fingerprint bands among the nine KP isolates were ≤ 3, which can be classified as the same PFGE type. Phylogenetic analysis showed that these 9 strains constituted a distinct branch within the phylogenetic tree. All nine KP strains belonged to the ST15-KL19 clone. Six of the strains were classified as CRKP, all of which carried eleven drug resistance genes: oqxB, oqxA, fosA6, aac(3)-lld, blaSHV-28, blaKPC-2, blaOXA-1, mph(A), tet(A), catB3 and aac(6')-lb-cr, mediating drug resistance to quinolones, fosfomycin, aminoglycosides, β-lactam, carbapenems, macrolides and chloramphenicol, belonging to multi-drug resistant bacteria. The carbapenem-resistant plasmid p2-KP3762-1 was found to transfer within species, from CRKP to hypervirulent K. pneumoniae (hvKP) RJF293HA, carbapenem-sensitive K. pneumoniae (CSKP) KP3657 and E. coli C600 at a frequency of (1.19±1.58) ×10-6, (1.09±1.38) ×10-7 and (10.9±9.53) ×10-6 respectively, resulting in the dissemination of carbapenem resistance genes. In this study, KP strains isolated from a single patient exhibited an alternating phenotype of resistant-sensitive-resistant-sensitive to carbapenems. The 9 KP isolates share a high degree of genetic similarity. The plasmid p2-KP3762-1, harboring the carbapenem resistance gene blaKPC-2, may undergo inter-strain and inter-clone transfer via conjugation in the patient during treatment. Furthermore, our findings suggest that the pathogens in this patient are likely to have a common ancestral origin.

Bacillus methylotrophicus influences on the virulence of Aeromonas hydrophila and probiotic applications in Ctenopharyngodon idella

Probiotics are alternatives to antibiotics that resist infection and benefit the host in several aspects. Here, we isolated and identified a novel strain of Bacillus methylotrophicus (XA-8) with a broad spectrum of activity against Aeromonas hydrophila AhX040 and other aquatic bacterial pathogens. The strain was non-pathogenic as it revealed no cytotoxic effects in hepatic L8824 cells or pathophysiological signs in the tissue of Ctenopharyngodon idella (grass carp). Using a co-culture approach, XA-8 (1×109 CFU mL−1) when co-cultivated with AhX040 (1×108 CFU mL−1) revealed a reduction in the cell density, cytotoxicity, and infection rate of AhX040. More interestingly, genes such as aer, act, hylA, lafA, hcp, and luxS, associated with the virulence of AhX040, were significantly down-regulated during co-cultivation (P<0.05). Furthermore, XA-8 as a feed supplement to grass carp up-regulated the immune-related genes in the liver, spleen, kidney, and intestine. The villi length and expression level of growth-related genes in the intestine were significantly higher (P<0.05) in the XA-8 supplemented grass carp. In addition to resisting infection (52.63 %), probiotic XA-8 modulated the gut microbiota of grass carp by increasing Fusobacteriota and Bacteroidota at the phylum level, while decreasing Aeromonas at the genus level. The whole genome sequence analysis showed the strain possesses multiple gene clusters for antimicrobial substances and metabolism. Our results demonstrate that B.methylotrophicus XA-8 is a novel probiotic with potential applications for aquaculture.

Identification of biallelic POLA2 variants in two families with an autosomal recessive telomere biology disorder

AbstractPOLA2 encodes the accessory subunit of DNA polymerase α (polα)/primase, which is crucial for telomere C-strand fill-in. Incomplete fill-in of the C-rich telomeric strand after DNA replication has been proposed as a mechanism for Coats plus syndrome, a phenotype within the broader spectrum of telomere biology disorders (TBD). Coats plus syndrome has so far been associated with pathogenic variants in POT1, CTC1, and STN1. Here we report the findings of biallelic deleterious rare variants in POLA2 gene detected by whole genome sequencing and segregation analysis in five young adults from two unrelated families. All five individuals displayed abnormally short telomeres and a clinical phenotype suggesting a TBD disorder with Coats plus features including retinal and gastrointestinal telangiectasias. Our results suggest POLA2 as a novel autosomal recessive gene for a TBD with Coats plus features.

Whole Genome Sequencing of Brucella melitensis Isolates from Kuwait for the Identification of Biovars, Variants and Relationship within a Biovar.

The identification of Brucella genotypes is essential for epidemiological studies. The whole genome sequencing is emerging as a novel tool for genetic characterization of infectious microbes. The aim of this study was to genotype B. melitensis isolates from Kuwait using whole genome sequencing and variant analysis of the sequence data. DNA were purified from 15 heat-inactivated B. melitensis isolates and used to prepare sequencing libraries employing Nextera XT DNA Sample Preparation Kit (Illumina San Diego, CA, USA) and sequenced on a MiSeq (Illumina). The sequence files were aligned to three biovars of B. melitensis, i.e., biovar 1 str. 16M, biovar 2 str. 63/9, and biovar 3 str. Ether. The alignment and variant calling were performed using 'bwa mem' and SAMtools/VCFtools, respectively. The genome size of all the isolates was around 3.3 mega base pairs and resembled B. melitensis biovar 2. Single nucleotide polymorphisms (SNPs), insertions and deletions, (indels) were spread all over the genome; but 138 SNPs were common among the 14 isolates, supporting the same ancestral origin. A neighbor-joining tree analysis identified isolate 2 as an outlier. In addition, SNPs (2 - 478) specific to each isolate were also identified, which divided the B. melitensis biovar 2 into two major groups/genotypes. A further analysis showed that the Kuwaiti isolates of the present study shared phylogeny mainly with strains from the Middle Eastern countries. Among the 15 isolates studied in Kuwait, biovar 2 is the most prevalent biovar of B. melitensis. Furthermore, isolate-specific genetic variations were identified, which may be useful in epidemiological investigations.

Isolation and characterization of Salmonella enteritidis bacteriophage Salmp-p7 isolated from slaughterhouse effluent and its application in food.

Salmonella enteritidis is one of the most common pathogens that cause foodborne disease outbreaks and food spoilage, which seriously threatens human health. Bacteriophages have shown broad application prospects in controlling harmful microorganisms during food processing and preservation due to their ability to specifically infect bacteria. In this study, Salmonella enteritidis bacteriophage Salmp-p7 was isolated and characterized from slaughterhouse wastewater. Transmission electron microscopy (TEM) analysis showed that Salmp-p7 belonged to the Siphoviridae family and was active against Salmonella enteritidis and Escherichia coli. Whole genome sequence analysis showed that Salmp-p7 was a lytic bacteriophage with a total length of 60,066bp of sequence. Salmp-p7 has a short incubation period and a long burst duration, with a burst volume of 55 PFU/cell and agood lysis effect. It can maintain a stable state within the temperature range of 30-60℃ and pH range of 4-12and has the potential for application in food. In vitro, antimicrobial curves and inhibition of biofilm removal experiments showed that Salmp-p7 could effectively inhibit and eliminate Salmonella enteritidis. The application of Salmp-p7 to the whole liquid of infected eggs resulted in a significant reduction of viable bacteria. And Salmp-p7 has high stability and lytic activity and has the potential to become a new biological control agent for Salmonella enteritidis in eggs.

Whole genome analysis of Stenotrophomonas geniculata MK2 and antagonism against Botrytis cinerea in strawberry.

A novel strain isolated from soil identified as Stenotrophomonas geniculata MK2 could control strawberries' postharvest disease gray mold. An in vitro investigation showed that MK2 had significant bioactivity against Botrytis cinerea, with an observed zone of inhibition of 85%. The strain MK2 was 88% effective in controlling gray mold on detached fruits. De novo whole genome sequencing analysis showed that strain MK2 has a single circular chromosome with a genome size of 736,465bp, a G + C content of 66.34%, a coding ratio of 89.80%, and a protein-coding gene of 442. The NR database identified about 4284 genes among Stenotrophomonas spp. and S. geniculata, sharing the maximum number of 1277 genes with the MK2 strain. In COG annotation, most gene percentage was linked to general functions. In KEGG annotations, the majority of genes are associated with metabolism. According to the GO analysis, the maximum number of genes involved in the molecular process was linked to catalytic and transporter activity. CAZymes (carbohydrate-active enzymes) showed that enzymes related to glycosyl transferases (48), carbohydrate esterases (54), and glycoside hydrolases (51) are involved in the non-ribosomal synthesis of secondary metabolites. The PHI database showed that in strain MK2, the reduced virulence was 68 protein counts, and similarly unaffected pathogenicity protein counts were 52. AntiSMASH analysis for biosynthesis-related gene clusters involved in the production of secondary metabolites showed ten gene clusters coded for 2,3-dihydroxybenzoylserine, griseobactin, fuscachelin, benarthin, mirubactin, myxochelin, and bacillibactin. The MK2 strain could be a potent biocontrol agent for postharvest diseases.

A comprehensive study on the mitochondrial genome of Volva habei and exploring phylogenetic relationships in Littorinimorpha

In order to enrich our taxonomic and systematic comprehension of Ovulidae within the evolutionary framework of Littorinimorpha. we present a comprehensive analysis of the mitochondrial genome (mitogenome) sequence of Volva habei using next-generation sequencing technology (GenBank accession number OR492307). The mitogenome spans a total length of 16,519 bp, encompassing a complete set of 37 genes, including 13 protein-coding genes (PCGs), 22 tRNAs and two rRNAs, demonstrating a distinct AT bias. Notably, trnS2 lacks a dihydrouracil (DHU) arm, thus preventing the formation of a typical secondary structure. In contrast, the remaining tRNAs exhibit a characteristic cloverleaf-like secondary structure. Comparative analysis with ancestral gastropods reveals substantial differences in three gene clusters (or genes), incorporating fifteen tRNAs and eight PCGs. Of particular significance are the observed inversions and translocations, representing the predominant types of rearrangements in V. habei. Phylogenetic analysis strongly supports the monophyletic grouping of all Littorinimorpha species, with V. habei forming a distinct Ovulidae clade. It is noteworthy that V. habei forms a sister group with Cypraeidae, collectively belonging to the Cypraeoidea. In summary, this study not only advances our comprehension of the entire mitochondrial dataset for Calyptraeoidea but also provides novel insights into the phylogenetic relationships within Littorinimorpha.

Genome sequencing, phylogenomics, and population analyses of Tilletia, with recognition of one common bunt species, T. caries (synonym T. laevis), distinct from dwarf bunt, T. controversa

ABSTRACT Some species of Tilletia are responsible for diseases in economically important crops, such as wheat and rice. In this study, we sequenced, assembled, and annotated 22 new genomes for Tilletia, with a focus on species causing dwarf bunt (DB; T. controversa), common bunt (CB; T. caries and T. laevis), and rice kernel smut (RKS; T. horrida). We present the first genomes for four other species (T. bromi, T. fusca, T. goloskokovii, and T. rugispora), resulting in the largest and most diverse sample of Tilletia genomes studied to date. Depending on the species and strain, the assembly size ranged from 24.3 to 30.5 Mb and gene prediction resulted in 7138 to 8261 gene models per genome. Phylogenomic analyses with hundreds to thousands of genes revealed significant support for the relationships among certain Tilletia taxa and validated findings of previous molecular studies that employed a small number of genes. Further population-level analyses showed two distinct populations of DB and CB: T. controversa (DB) as a single population and another intermixed population of T. caries and T. laevis (CB). No evidence of geographic isolation was observed within these populations. Our phylogenomic analyses also supported previous multigene hypotheses that multiple lineages of Tilletia may cause RKS. Collectively, our results suggest that taxonomic revisions are needed for the RKS-causing pathogens and provide convincing evidence for formally recognizing the CB-causing taxa as one species, named T. caries (synonym T. laevis). Overall, our study significantly enhances genomic resources for Tilletia, offers insights into phylogenetic relationships and population structure, and provides whole genome sequences for future studies.

Comparative genomics analysis of Salmonella Enteritidis isolated from clinical cases associated with chicken

Salmonella Enteritidis is a major foodborne pathogen, and the emergence of multidrug-resistant (MDR) S. Enteritidis poses a serious public health challenge. In this study, we report the genomic characterization of five S. Enteritidis isolates from clinical. These isolates exhibited resistance to seven classes of antimicrobials with four of the five characterized as MDR. Isolate 33 A exhibited resistance to colistin and polymyxin B, while no associated antimicrobial resistance genes (ARGs) were identified in its genome. Isolate 21 A and 44 A were extended-spectrum beta-lactamases-producing (ESBLs). Whole genome sequencing analysis revealed the presence of multiple mobile genetic elements (MGEs), including plasmids, prophages, and genomic islands, which may have facilitated the acquisition and dissemination of ARGs. Notably, several ARGs, including blaCTX−M−55, blaTEM−141, blaTEM−1B, aph(3’)-IIa, aph(3’’)-Ib, aph(6)-Id, tet(A), floR, fosA3, and sul2, were identified on plasmids. In addition, chromosomal point mutations in gyrA (D87G and D87Y) and acrB (F28L and L40P) were also observed in each isolate. Multiple virulence genes associated with the type III secretion system were identified on Salmonella pathogenicity islands (SPIs) SPI-1 and SPI-2. Phylogenetic analysis revealed that the five isolates, along with a clinical and chicken origin isolates in the database, clustered together, suggesting a probable common source of infection. Our findings highlight the intricate genetic mechanisms behind MDR in S. Enteritidis, emphasizing the ongoing necessity for surveillance and appropriate antimicrobial usage. This contributes to our understanding of S. Enteritidis transmission within the food chain.

Characterization of a Novel Tn7-like Transposon Carrying blaDHA-1 in Providencia stuartii MF1 Isolated from Swine Wastewater.

Providencia stuartii is an emerging pathogen that causes nosocomial infections. In this study, a multidrug-resistant strain P. stuartii MF1 was isolated from swine wastewater. Comprehensive analysis of whole genome sequencing revealed that dozens of antibiotic resistance genes were found in MF1. A novel transposon Tn6450M which has high sequence identity to Tn6450 and the plasmid-borne Tn6765 from Proteus mirabilis was identified in the genome of MF1. Tn6450M was determined to be stably inserted into a new attTn7 site in the P. stuartii MF1 genome and contains the third-generation cephalosporins resistance-associated genes blaDHA-1. Intergeneric transmission of Tn6450 variants poses risks for the spread of antibiotic resistance genes.

Genome sequencing and comparative genomic analysis of bovine mastitis-associated non-aureus staphylococci and mammaliicocci (NASM) strains from India

We describe the whole-genome sequencing and comparative genomic analysis of 22 mastitis-associated NASM strains isolated from India. The mean genome size of the strains was 2.55 Mbp, with an average GC content of 32.2%. We identified 14 different sequence types (STs) among the 22 NASM strains. Of these, ST1 and ST6 of S. chromogenes were exclusively associated with bovine mastitis. Genome-wide SNP-based minimum spanning tree revealed the intricate phylogenetic relationships among NASM strains from India, categorizing them into five major clades. Interestingly, mastitis-associated strains formed separate subclades in all the NASM species studied, indicating distinct host-specific co-evolution. The study identified 32 antimicrobial resistance (AMR) genes and 53 virulence-associated genes, providing insights into the genetic factors that could contribute to the pathogenicity of NASM species. Some virulence and AMR genes were found in the predicted genomic islands, suggesting possible horizontal transfer events.

Epidemiological description of fire blight introduction patterns to Central Asia and the Caucasus region based on CRISPR spacer typing and genome analysis

In the last two decades, fire blight has progressively spread eastward from Europe and the Mediterranean area to several pome-fruit producing regions of Asia. Its causative agent, the bacterial pathogen Erwinia amylovora, was detected in several new countries, including Georgia, Kyrgyzstan, and Kazakhstan. In the latter two states, the disease creates a threat not only to the commercial apple and pear production, but also to the wild Malus and Pyrus species that constitute the basis of the local forest ecosystems. In this study, we investigated the genetic diversity of the pathogen in Central Asia and the Caucasus region utilizing CRISPR Repeat Regions (CRRs) genotyping and genome sequencing, with the aim to understand its dissemination patterns across the continent. Genome sequence analysis revealed that all strains from these two regions exclusively derived from the archetypal CRR1 genotype A. Our analysis revealed three main E. amylovora clades in Central Asia, with distinct yet partial overlapping geographical distributions. Genomic relationships among isolates indicate that Central Asian strains are genetically closest to those from the Persian region and the Middle East, while the Georgian population is genetically more distant and can align with strains from the Volga District in southern Russia and the Eastern Mediterranean area. Notably, this study also includes strains from the first confirmed occurrences of fire blight in Uzbekistan, Tajikistan, and China. Our findings highlight the importance of phylogenetic analysis and genome sequencing in understanding the phytopathogen epidemics and protecting key agricultural species and the genetic resources of their wild counterparts in the forest.

Major depressive disorder is associated with mitochondrial ND6 T14502C mutation in two Han Chinese families

BACKGROUND Globally, the World Health Organization ranks major depressive disorder (MDD) as the leading cause of disability. However, MDD molecular etiology is still poorly understood. AIM To explore the possible association between mitochondrial ND6 T14502C mutation and MDD. METHODS Clinical data were collected from two pedigrees, and detailed mitochondrial genomes were obtained for the two proband members. The assessment of the resulting variants included an evaluation of their evolutionary conservation, allelic frequencies, as well as their structural and functional consequences. Detailed mitochondrial whole genome analysis, phylogenetic, and haplotype analysis were performed on the probands. RESULTS Herein, we reported the clinical, genetic, and molecular profiling of two Chinese families afflicted with MDD. These Chinese families exhibited not only a range of onset and severity ages in their depression but also extremely low penetrances to MDD. Sequence analyses of mitochondrial genomes from these pedigrees have resulted in the identification of a homoplasmic T14502C (I58V) mutation. The polymorphism is located at a highly conserved isoleucine at position 58 of ND6 and distinct mitochondrial DNA (mtDNA) polymorphisms originating from haplogroups M10 and H2. CONCLUSION Identifying the T14502C mutation in two individuals with no genetic relation who exhibit symptoms of depression provides compelling evidence that this mutation may be implicated in MDD development. Nonetheless, the two Chinese pedigrees that carried the T14502C mutation did not exhibit any functionally significant mutations in their mtDNA. Therefore, the phenotypic expression of the T14502C mutation related to MDD may be influenced by the nuclear modifier gene(s) or environmental factors.

Complete genome sequencing of SARS-CoV-2 strains that were circulating in Uzbekistan over the course of four pandemic waves.

Since the rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as a global COVID-19 pandemic affecting millions of people globally, it has become one of the most urgent research topics worldwide to better understand the pathogenesis of COVID-19 and the impact of the harmful variants. In the present study, we conducted whole genome sequencing (WGS) analysis of 110 SARS-CoV-2 genomes, to give more data about the circulation of SARS-CoV-2 variants during the four waves of pandemic in Uzbekistan. The whole genome sequencing of SARS-CoV-2 samples isolated from PCR-positive patients from Tashkent, Uzbekistan, in the period of 2021 and 2022 were generated using next-generation sequencing approaches and subjected to further genomic analysis. According to our previous studies and the current genome-wide annotations of clinical samples, we have identified four waves of SARS-CoV-2 in Uzbekistan between 2020 and 2022. The dominant variants observed in each wave were Wuhan, Alpha, Delta, and Omicron, respectively. A total of 347 amino acid level variants were identified and of these changes, the most frequent mutations were identified in the ORF1ab region (n = 159), followed by the S gene (n = 115). There were several mutations in all parts of the SAR-CoV-2 genomes but S: D614G, E: T9I, M: A63T, N: G204 R and R203K, NSP12: P323L, and ORF3a(NS3): T223I were the most frequent mutations in these studied viruses. In our previous study, no mutation was found in the envelope (E) protein. In contrast, in our present study, we identified 3 (T9I, T11A and V58F) mutations that made changes to the structure and function of the E protein of SARS-CoV-2. In conclusion, our findings showed that with the emergence of each new variant in our country, the COVID-19 pandemic has also progressed. This may be due to the considerable increase in the number of mutations (Alpha-46, Delta- 146, and Omicron-200 mutations were observed in our samples) in each emerged variant that shows the SARS-CoV-2 evolution.

Fertility and early embryonic development in a CD46-edited Gir heifer with reduced susceptibility to BVDV.

Bovine viral diarrhea virus (BVDV) infection during pregnancy is a significant contributor to reproductive failures in cattle. The bovine receptor for BVDV (CD46) was previously edited with a six amino acid substitution (G82QVLAL to A82LPTFS) and shown to have significantly reduced BVDV susceptibility in a Gir heifer calf. Since a role for CD46 has been proposed in mammalian fertilization, our objective was to assess the edited heifer's fertilization rates, early embryonic development, and germline transmission conformation of the edit. Cumulus oocyte complexes were collected from the edited heifer and unedited females, fertilized with semen from an unedited bull and cultured until the blastocyst stage. Ultrasound examinations and serum progesterone concentration were also monitored to confirm estrous cyclicity in the CD46-edited heifer. Estrous cyclicity was normal with visualization of a corpus luteum and elevated progesterone concentrations. Fertilization rates and blastocyst development were not different in oocytes from edited and unedited controls. Genome sequence analysis of blastocysts confirmed germline transmission of either edited allele from the heifer. Subsequently, the CD46-edited heifer was artificially inseminated with semen from an unedited Gir bull and fertility status was confirmed with a diagnosed conception at day 35 of gestation. Thus, a six amino acid substitution in CD46 did not negatively affect fertilization of edited oocytes or early embryonic development when fertilized with semen from an unedited bull. An edited bull is still needed to similarly evaluate reproductive function of sperm cells carrying this CD46 edit.

Evaluation of sequence-based tools to gather more insight into the positioning of rhizogenic agrobacteria within the Agrobacterium tumefaciens species complex.

Rhizogenic Agrobacterium, the causative agent of hairy root disease (HRD), is known for its high phenotypic and genetic diversity. The taxonomy of rhizogenic agrobacteria has undergone several changes in the past and is still somewhat controversial. While the classification of Agrobacterium strains was initially mainly based on phenotypic properties and the symptoms they induced on plants, more and more genetic information has been used along the years to infer Agrobacterium taxonomy. This has led to the definition of the so-called Agrobacterium tumefaciens species complex (Atsc), which comprises several genomospecies. Interestingly, the rhizogenic Agrobacterium strains are found in several of these genomospecies. Nevertheless, even up until today Agrobacterium strains, and in particular rhizogenic agrobacteria, are prone to misclassification and considerable confusion in literature. In this study, we evaluated different phylogenetic analysis approaches for their use to improve Agrobacterium taxonomy and tried to gain more insight in the classification of strains into this complex genus, with a particular focus on rhizogenic agrobacteria. The genome sequence analysis of 580 assemblies, comprising Agrobacterium, Allorhizobium and Rhizobium strains demonstrated that phylogenies based on single marker genes, such as the commonly used 16S rRNA and recA gene, do not provide sufficient resolution for proper delineation of the different genomospecies within the Atsc. Our results revealed that (in silico) multi-locus sequences analysis (MLSA) in combination with average nucleotide identity (ANIb) at a 94.0% threshold delineates genomospecies accurately and efficiently. Additionally, this latter approach permitted the identification of two new candidate genomospecies.

Leveraging Multi-omics to Disentangle the Complexity of Ovarian Cancer.

To better understand ovarian cancer lethality and treatment resistance, sophisticated computational approaches are required that address the complexity of the tumor microenvironment, genomic heterogeneity, and tumor evolution. The ovarian cancer tumor ecosystem consists of multiple tumors and cell types that support disease growth and progression. Over the last two decades, there has been a revolution in -omic methodologies to broadly define components and essential processes within the tumor microenvironment, including transcriptomics, metabolomics, proteomics, genome sequencing, and single-cell analyses. While most of these technologies comprehensively characterize a single biological process, there is a need to understand the biological and clinical impact of integrating multiple -omics platforms. Overall, multi-omics is an intriguing analytic framework that can better approximate biological complexity; however, data aggregation and integration pipelines are not yet sufficient to reliably glean insights that affect clinical outcomes.

Circulation of &lt;i&gt;Mycobacterium tuberculosis&lt;/i&gt; strains of the Beijing Central Asian Outbreak genotype in the Kemerovo region — Kuzbass in 2018–2022

Introduction. The Kemerovo region — Kuzbass is characterized by a high prevalence of multidrug-resistant (MDR) tuberculosis (TB), including coinfection with HIV (HIV/TB). A previously unknown in Russia relationship between MDR and the Beijing Central Asian Outbreak (CAO) subtype has been discovered, which updates studies of Mycobacterium tuberculosis taking into account this resistant variant. Objective: to study the molecular genetic structure of the M. tuberculosis population, to assess the prevalence and possible routes of emergence of Beijing CAO strains in the Kemerovo region — Kuzbass. Materials and methods. A total of 325 M. tuberculosis strains were studied in 2018–2022 using spoligotyping, MIRU-VNTR 24 and SNP typing. Whole genome sequencing and bioinformatics analysis were performed for seven Beijing CAO strains. Results. Primary MDR and pre-extensive drug resistance (pre-XDR) were detected in 39.4% and 11.5% of strains, respectively. In the total sample, MDR was 43.4%, pre-XDR — 19.7%. In the structure of the M. tuberculosis population, the Beijing genotype prevailed (78.8%), with its subtypes Central Asian Russian (40.9%) and B0/W148 (32.6%). The Euro-American lineage (27.3%) was represented by the genotypes T (6.5%), LAM (5.8%), Ural (4.9%), H (0.9%); one strain CAS1-Delhi was detected, the genotype of 2.8% of strains was not identified. The proportion of Beijing CAO was 12.6% of the total sample; this subtype was significantly more often detected among HIV/TB (20.6%) than in HIV-negative TB patients (9.1%; p = 0.005). The results of the Beijing CAO genome analysis from the Kemerovo region indicate the absence of a direct chain of transmission between these TB cases. A hypothesis has been put forward about the introduction of Beijing CAO to the Kemerovo region from Central Asia and its endemic circulation in the region. Conclusion. A high level of MDR and pre-XDR was detected in Beijing genotype strains in the M. tuberculosis population of the Kemerovo region — Kuzbass, especially the B0/W148 (97.2%) and CAO (87.5%) subtypes. Beijing CAO strains, detected mainly in newly diagnosed HIV/TB patients, require further monitoring and control of their spreading.