Gene Cluster Research Articles

Interpretable adenylation domain specificity prediction using protein language models.

Natural products have long been a rich source of diverse and clinically effective drug candidates. Non-ribosomal peptides (NRPs), polyketides (PKs), and NRP-PK hybrids are three classes of natural products that display a broad range of bioactivities, including antibiotic, antifungal, anticancer, and immunosuppressant activities. However, discovering these compounds through traditional bioactivity-guided techniques is costly and time-consuming, often resulting in the rediscovery of known molecules. Consequently, genome mining has emerged as a high-throughput strategy to screen hundreds of thousands of microbial genomes to identify their potential to produce novel natural products. Adenylation domains play a key role in the biosynthesis of NRPs and NRP-PKs by recruiting substrates to incrementally build the final structure. We propose MASPR, a machine learning method that leverages protein language models for accurate and interpretable predictions of A-domain substrate specificities. MASPR demonstrates superior accuracy and generalization over existing methods and is capable of predicting substrates not present in its training data, or zero-shot classification. We use MASPR to develop Seq2Hybrid, an efficient algorithm to predict the structure of hybrid NRP-PK natural products from microbial genomes. Using Seq2Hybrid, we propose putative biosynthetic gene clusters for the orphan natural products Octaminomycin A, Dityromycin, SW-163B, and JBIR-39.

Genome-Wide Identification and Expression Profile of Farnesyl Pyrophosphate Synthase (FPS) Gene Family in Euphorbia Hirta L.

The biosynthesis of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are essential for sesquiterpenes and triterpenes, respectively, is primarily governed by the mevalonate pathway, wherein farnesyl pyrophosphate synthase (FPS) plays a pivotal role. This study identified eight members of the FPS gene family in Euphorbia hirta, designated EhFPS1-EhFPS8, through bioinformatics analysis, revealing their distribution across several chromosomes and a notable tandem gene cluster. The genes exhibited strong hydrophilic properties and key functional motifs crucial for enzyme activity. An in-depth analysis of the EhFPS genes highlighted their significant involvement in isoprenoid metabolism and lipid biosynthesis, with expression patterns influenced by hormones such as jasmonic acid and salicylic acid. Tissue-specific analysis demonstrated that certain FPS genes, particularly EhFPS1, EhFPS2, and EhFPS7, showed elevated expression levels in latex, suggesting their critical roles in terpenoid biosynthesis. Furthermore, subcellular localization studies have indicated that these proteins are primarily found in the cytoplasm, reinforcing their function in metabolic processes. These findings provide a foundational understanding of the FPS genes in E. hirta, including their gene structures, conserved domains, and evolutionary relationships. This study elucidates the potential roles of these genes in response to environmental factors, hormone signaling, and stress adaptation, thereby paving the way for future functional analyses aimed at exploring the regulation of terpenoid biosynthesis and enhancing stress tolerance in this species.

Comparative metagenomics reveals the metabolic flexibility of coastal prokaryotic microbiomes contributing to lignin degradation

Coastal wetlands are rich in terrestrial organic carbon. Recent studies suggest that microbial consortia play a role in lignin degradation in coastal wetlands, where lignin turnover rates are likely underestimated. However, the metabolic potentials of these consortia remain elusive. This greatly hinders our understanding of the global carbon cycle and the “bottom-up” design of synthetic consortia to enhance lignin conversion. Here, we developed two groups of lignin degrading consortia, L6 and L18, through the 6- and 18-month in situ lignin enrichments in the coastal East China Sea, respectively. Lignin degradation by L18 was 3.6-fold higher than L6. Using read-based analysis, 16S rRNA amplicon and metagenomic sequencing suggested that these consortia possessed varied taxonomic compositions, yet similar functional traits. Further comparative metagenomic analysis, based on metagenomic assembly, revealed that L18 harbored abundant metagenome-assembled genomes (MAGs) that encoded diverse and unique lignin degradation gene clusters (LDGCs). Importantly, anaerobic MAGs were significantly enriched in L18, highlighting the role of anaerobic lignin degradation. Furthermore, the generalist taxa, which possess metabolic flexibility, increased during the extended enrichment period, indicating the advantage of generalists in adapting to heterogenous resources. This study advances our understanding of the metabolic strategies of coastal prokaryotic consortia and lays a foundation for the design of synthetic communities for sustainable lignocellulose biorefining.

Range-wide understanding of genetic diversity and population structure of endangered Kashmir musk deer in North-Western Himalaya.

The endangered Kashmir musk deer (Moschus cupreus), native to high-altitude Himalayas, is an ecological significant and endangered ungulate, threatened by habitat loss and poaching for musk pod distributed in western Himalayan ranges of India, Nepal and Afghanistan. Despite its critical conservation status and ecological importance in regulating vegetation dynamics, knowledge gaps persist regarding its population structure and genetic diversity, hindering effective management strategies. We aimed to understand the population genetics of Kashmir musk deer in north-western Himalayas using two mitochondrial DNA (mtDNA) regions and 11 microsatellite loci. Total of 63 consensus sequences were analyzed, revealing 10 haplotypes with high haplotype diversity (Hd = 0.906 ± 0.031) and moderate nucleotide diversity (π = 0.00539 ± 0.004) at mitochondrial markers. Microsatellite analysis indicated moderate genetic variability, with observed heterozygosity (Ho = 0.445 ± 0.049) and expected heterozygosity (He = 0.764 ± 0.039), comparable to the forest musk deer but relatively lower than that observed in other ungulates. Population structure analysis identified three potential genetic clusters, with evidence of genetic admixture in the Lahaul Valley, possibly due to gene flow from neighboring areas. Historical demographic analyses using Bayesian skyline plots and mismatch distribution curves indicated stable population sizes in the past, followed by recent declines in maternal effective population size. Our findings highlight the necessity of preserving the genetic diversity of the Kashmir musk deer to ensure its long-term survival. We recommend expanding the geographic range of sampling to include regions such as Chamba and Kinnaur districts in Himachal Pradesh, potential areas in Jammu and Kashmir and Uttarakhand, for more comprehensive population-level analysis. Such efforts will be crucial for developing targeted conservation strategies and mitigating the impacts of human activities on this endangered species.

Neptunizhulides, Cryptic trans-AT Polyketide Synthase-Derived Metabolites from Paraneptunicella aestuarii NBU2194.

Genome mining of Paraneptunicella aestuarii NBU2194 resulted in the identification of a family of 17-membered macrolides, neptunizhulides A-F. Their structures were elucidated by comprehensive spectroscopic data analysis. Stereochemical assignments of the neptunizhulides were determined by J-based configuration analysis, ROESY NMR, Mosher's ester derivatization, and bioinformatic predictions. Bioinformatic analysis and a 15N-serine labeling experiment resulted in the identification of a putative trans-acyltransferase polyketide synthase (trans-AT PKS) biosynthetic gene cluster, and a biosynthetic pathway is proposed.

A microbial natural product fractionation library screen with HRMS/MS dereplication identifies new lipopeptaibiotics against Candida auris.

The rise of drug-resistant fungal pathogens, including Candida auris , highlights the urgent need for novel antifungal therapies. We developed a cost-effective platform combining microbial extract prefractionation with rapid MS/MS-bioinformatics-based dereplication to efficiently prioritize new antifungal scaffolds. Screening C. auris and C. albicans revealed novel lipopeptaibiotics, coniotins, from Coniochaeta hoffmannii WAC11161, which were undetectable in crude extracts. Coniotins exhibited potent activity against critical fungal pathogens on the WHO Fungal Priority Pathogens List, including C. albicans , C. neoformans , multidrug-resistant C. auris , and Aspergillus fumigatus , with high selectivity and low resistance potential. Coniotin A targets β-glucan, compromising fungal cell wall integrity, remodelling, and sensitizing C. auris to caspofungin. Identification of a PKS-NRPS biosynthetic gene cluster further enables the discovery of related clusters encoding potential novel lipopeptaibiotics. This study demonstrates the power of natural product prefractionation in uncovering bioactive scaffolds and introduces coniotins as promising candidates for combating multidrug-resistant fungal pathogens.

Gene cluster regulators from plants to microbes: Key role in vesicular targeting, transport of intermediate compounds and secondary metabolite biosynthesis

Gene cluster regulators from plants to microbes: Key role in vesicular targeting, transport of intermediate compounds and secondary metabolite biosynthesis

Global transcriptome changes during growth of a novel Penicillium coffeae isolate on the wheat stripe rust fungus, Puccinia striiformis f. sp. tritici.

Wheat stripe rust caused by the fungus Puccinia striiformis f. sp. tritici (Pst) is currently the most destructive disease of wheat. The major control methods which include the deployment of resistant wheat cultivars and application of chemical fungicides are losing efficiency as the fungus evolves. Natural antagonists of Pst may be an avenue for alternative and environmentally sustainable control of the disease in the field. Here we describe a novel fungus found growing on Pst pustules. We identified the fungus as a novel isolate of the plant endophyte Penicillium coffeae. We present a high-quality reference genome and a comparative transcriptomic analysis used to investigate how the fungus deploys its genes during growth amongst Pst spores. The gene content of the P. coffeae ANU01 genome is suggestive of a generalist that makes use of diverse substrates. An abundance of genes related to lipid, amino acid and carbohydrate metabolism indicate that P. coffeae ANU01 has evolved the ability to exploit nutrient stores in Pst urediniospores. P. coffeae ANU01 deploys a number of biosynthetic gene clusters during growth on Pst spores, potentially to inhibit urediniospores germination and halt defence responses. A number of genes encoding carbohydrate active enzymes are also highly upregulated, suggesting targeting and degradation of Pst urediniospores structures. Alongside carbohydrates, P. coffeae ANU01 appears to target spore lipids as a nutrient source, secreting several highly upregulated lipases. Our findings broaden the understanding of growth associated with rust spores as an evolutionary strategy and provide insight into the genes potentially required for this process.

Molecular and immune landscape of tumours in geriatric patients with non-small cell lung cancer, melanoma and renal cell carcinoma

ObjectiveCancer patients aged ≥80 years present unique characteristics affecting response to immune checkpoint inhibitors (ICIs), with unidentified molecular differences. This study aimed to explore potential biomarkers of response to ICI...

Clarification of the biosynthetic gene cluster involved in the antifungal prodrug echinocandin B and its robust production in engineered Aspergillus pachycristatus.

Echinocandin antifungals exhibit high efficacy against drug-resistant strains due to their unique mechanism of action. The production of their semi-synthetic precursors relies solely on microbial metabolism, leading to elevated production costs. Anidulafungin, an excellent echinocandin drug, is derived from echinocandin B (ECB), which is industrially produced by Aspergillus pachycristatus. However, the genes involved in the actual ECB biosynthesis remain unclear, which hinders yield improvements through engineered strains. This study systematically investigated the putative ECB biosynthetic gene cluster using genomic and transcriptomic profiling combined with gene editing. Among the 18 putative genes previously reported, only a 13-gene cluster (ecdA, ecdG-J, htyA-F) was found to be actively involved in ECB biosynthesis, while the remaining 5 genes (ecdB-F) were non-essential and functioned independently. Notably, we identified that htyC and htyD were involved in L-homotyrosine biosynthesis, while HtyF catalyzed the C4 hydroxylation of 3S-hydroxyl-L-homotyrosine. Most importantly, EcdJ was identified as a crucial global transcriptional activator regulating the ECB gene cluster. Deletion of ecdJ silenced all related genes and abolished ECB production. Accordingly, overexpressing ecdJ alone or combining ecdA and htyF together significantly enhanced ECB yield. Under optimized liquid fermentation conditions, ECB production in the OEecdJ strain achieved 841 ± 23.11 mg/L. Solid-state fermentation further enhanced the ECB yield to 1.5 g/L, which is 7.7-fold higher than that of the wild-type strain under initial liquid fermentation conditions. This study has thoroughly elucidated the functions of key genes involved in the ECB biosynthesis and provided effective strategies for enhancing antifungal prodrug-ECB production, achieving the highest ECB production in an engineering A. pachycristatus strain.

A next-generation sequencing-based universal target panel and algorithm for one-stop detection of copy number alterations and single-nucleotide variations in the HBB gene cluster for rapid diagnosis of β-thalassemia.

This study aimed to develop and validate a targeted next-generation sequencing (NGS) panel along with a data analysis algorithm capable of detecting single-nucleotide variants (SNVs) and copy number variations (CNVs) within the beta-globin gene cluster. The aim was to reduce the turnaround time in conventional genotyping methods and provide a rapid and comprehensive solution for prenatal diagnosis, carrier screening, and genotyping of β-thalassemia patients. We devised a targeted NGS panel spanning an 80.4 kb region on chromosome 11, encompassing the beta-globin gene cluster and 5' locus control region. We also developed an advanced data analysis algorithm consisting of variant calling and depth plot analysis that facilitates simultaneous detection of SNVs and CNVs in a single run. The test panel and algorithm were validated with 14 in-house β-thalassemia carrier/patient samples and cross-checked against the HbVar database. We identified seven pathogenic SNVs and five CNVs within the beta-globin gene cluster in various combinations, such as heterozygous, homozygous, and compound heterozygous conditions. Additionally, the coordinates of 169 rare deletions and 11 fusion mutations reported in the HbVar database were checked to verify the theoretical ability of our developed gene panel to detect all the CNVs within the target region. The developed panel and NGS technology can detect both SNVs and CNVs in a single run and can also be utilized for prenatal diagnosis and carrier screening for hemoglobinopathies, underscoring its versatility and clinical utility.

Global dissemination of the beta-lactam resistance gene blaTEM-1 among pathogenic bacteria.

Antibiotic resistance presents a burgeoning global health crisis, with over 70% of pathogenic bacteria now exhibiting resistance to at least one antibiotic. This study leverages a vast dataset of 618,853 pathogenic bacterial genomes from the NCBI pathogen detection database, offering comprehensive insights into antibiotic resistance patterns, species-specific profiles, and transmission dynamics of resistant pathogens. We centered our investigation on the beta-lactam resistance gene blaTEM-1, found in 43,339 genomes, revealing its extensive distribution across diverse species and isolation sources. The study unveiled the prevalence of 15 prominent antibiotic resistance genes (ARGs), including those conferring resistance to beta-lactam, aminoglycoside, and tetracycline antibiotics. Distinct resistance patterns were observed between Gram-positive and Gram-negative bacteria, indicating the influence of phylogeny on resistance dissemination. Notably, the blaTEM-1 gene demonstrated substantial prevalence across a wide array of bacterial species (8) and a high number of isolation sources (11). Genetic context analysis revealed associations between blaTEM-1 and mobile genetic elements (MGEs) like transposons and insertion sequences. Additionally, we observed recent horizontal transfer events involving clusters of blaTEM-1 genes and MGEs underscore the potential of MGEs in facilitating the mobilization of ARGs. Our findings underscore the importance of adopting a One Health approach to global genomic pathogen surveillance, aiming to uncover the transmission routes of ARGs and formulate strategies to address the escalating antibiotic resistance crisis.

Genome-Guided Identification and Characterisation of Broad-Spectrum Antimicrobial Compounds of Bacillus velezensis Strain PD9 Isolated from Stingless Bee Propolis.

The emergence of multidrug-resistant pathogens presents a significant global health challenge, which is primarily fuelled by overuse and misuse of antibiotics. Bacteria-derived antimicrobial metabolites offer a promising alternative strategy for combating antimicrobial resistance issues. Bacillus velezensis PD9 (BvPD9), isolated from stingless bee propolis, has been reported to have antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA). This study aimed to characterise and identify the antimicrobial compounds (AMCs) synthesised by BvPD9 through integration of genome mining and liquid chromatography-mass spectrometry (LC-MS) analysis. The whole-genome sequence of BvPD9 contained 4,263,351 base pairs and 4101 protein-coding sequences, with 12 potential AMC biosynthetic gene clusters. Comparative genomic analysis highlighted the unique profile of BvPD9 that possesses the largest number of unknown proteins, indicating significant potential for further exploration. The combined genomics-metabolic profiling uncovered five AMCs in BvPD9 extract, including bacillibactin, bacilysin, surfactin A, fengycin A, and bacillomycin D. The extract exhibited a broad antibacterial spectrum against 25 pathogenic bacteria, including both Gram-positive and Gram-negative bacteria, with the lowest minimum inhibitory concentration (MIC, 0.032mg/ml) against S. epidermidis ATCC 12228, and the lowest minimum bactericidal concentration (MBC; 0.128mg/ml) against MRSA ATCC 700699 and Aeromonas hydrophilia. The robust stability of BvPD9 extract was demonstrated at high temperatures, over a wide range of pH conditions (6 to 12) and in the presence of various hydrolytic enzymes. Additionally, the extract showed 50% haemolytic and cytotoxicity activity at 0.158 and 0.250mg/ml, respectively. These characteristics suggest potential applications of BvPD9 metabolites for tackling antimicrobial resistance and its applicability across diverse industries.

Lampbrush chromosomes of Danio rerio.

Danio rerio, commonly known as zebrafish, is an established model organism for the developmental and cell biology studies. Although significant progress has been made in the analysis of the D. rerio genome, cytogenetic studies face challenges due to the unclear identification of chromosomes. Here, we present a novel approach to the study of the D. rerio karyotype, focusing on the analysis of lampbrush chromosomes isolated from growing oocytes. Lampbrush chromosomes, existing during diplotene, serve as a powerful tool for high-resolution mapping and transcription analysis due to their profound decondensation and remarkable lateral loops decorated by RNA polymerases and ribonucleoprotein (RNP) matrix. In D. rerio, lampbrush chromosomes are about 20 times longer than corresponding metaphase chromosomes. We found that the lampbrush chromosome stage karyotype of D. rerio is generally undifferentiated, except for several bivalents bearing distinct marker structures, including loops with complex RNP matrix and locus-associated nuclear bodies. Locus-associated nuclear bodies were enriched for coilin and snRNAs; the loci where they formed presumably correspond to the histone gene clusters. Further, we observed the accumulation of splicing factors in giant terminal RNP aggregates on one bivalent. DAPI staining of Danio rerio lampbrush chromosomes revealed large and small chromomeres non-uniformly distributed along the axis. For example, D. rerio lampbrush chromosome 4, comprising the sex-determining region, is divided into two halves-with small chromomeres bearing long lateral loops and with large dense chromomeres bearing no or very tiny lateral loops. As centromeres were not distinguishable, we identified centromeric regions in all bivalents by FISH mapping of pericentromeric RFAL1, RFAL2, and RFAM tandem repeats. Through a combination of morphological analysis, immunostaining of marker structures, and centromere mapping, we developed cytological maps of D. rerio lampbrush chromosomes. Finally, by RNA FISH we revealed transcripts of pericentromeric and telomeric tandemrepeats at the lampbrush chromosome stage.

In Vitro Cholesterol-Lowering Bioactivity, Synthetic Pathway, and Structural Characterization of Exopolysaccharide Synthesized by Schleiferilactobacillus harbinensis Z171.

A strain identified as Schleiferilactobacillus harbinensis was isolated from Chinese sauerkraut, and its exopolysaccharide (EPS) exhibited excellent in vitro cholesterol-lowering bioactivity. Besides, the whole genome of this strain and the structure characteristics of the purified EPS were investigated in this study. S. harbinensis Z171 presented a strong EPS production capacity, with five nucleotide sugar biosynthesis pathways regulated by an EPS synthesis gene cluster. Structural characterization revealed that the purified fraction F-EPS1A was a neutral polysaccharide with a molecular weight of 6.4 × 104 Da. The structure of F-EPS1A contained a backbone that comprised blocks of four 1,2-linked and three 1,3-linked alpha mannose units. Some 1,2-linked alpha mannose residues were branched at C6 with side chains formed by single alpha mannose or a disaccharide consisting of 1,6-linked alpha mannose residues. The structural characteristics endowed F-EPS1A with a high level of cholesterol-lowering bioactivity. In addition, whole genome analysis indicated that S. harbinensis Z171 possessed a strong EPS production capacity. These findings suggested that the EPS produced by S. harbinensis Z171 could be applied as a potential cholesterol-lowering prebiotic agent or supplement in functional food.

Pangenomic analysis of the bacterial cellulose-producing genera Komagataeibacter and Novacetimonas.

Bacterial cellulose (BC) holds significant commercial potential due to its unique structural and chemical properties, making it suitable for applications in electronics, medicine, and pharmaceuticals. However, large-scale BC production remains limited by challenges in bacterial performance. In this study, we compared 79 microbial genomes from three genera-Komagataeibacter, Novacetimonas, and Gluconacetobacter-to investigate their pangenomes, genetic diversity, and evolutionary relationships. Through comparative genomic and phylogenetic analyses, we identified distinct genome compositions and evolutionary patterns that differ from previous reports. The role of horizontal gene transfer (HGT) in shaping the genetic diversity and adaptability of these bacteria was also explored. Key determinants in BC production, such as variations in the bacterial cellulose biosynthesis (bcs) operon, carbohydrate uptake genes, and carbohydrate-active enzymes, were examined. Additionally, several biosynthetic gene clusters (BGCs), including Linocin M18 and sactipeptides, which encode for antimicrobial peptides known as bacteriocins, were identified. These findings reveal new aspects of the genetic diversity in cellulose-producing bacteria and present a comprehensive genomic toolkit that will support future efforts to optimize BC production and improve microbial performance for commercial applications.

Mining Silent Biosynthetic Gene Clusters for Natural Products in Filamentous Fungi.

Filamentous fungi are of great interest due to their powerful metabolic capabilities and potentials to produce abundant various secondary metabolites as natural products (NPs), some of which have been developed into pharmaceuticals. Furthermore, high-throughput genome sequencing has revealed tremendous cryptic NPs underexplored. Based on the development of in silico genome mining, various techniques have been introduced to rationally modify filamentous fungi,awakening the silent biosynthetic gene clusters (BGCs) and visualizing the NPs originally cryptic. This review summarizes the key strategies and research advances in the activation of cryptic BGCs in filamentous fungi, including endogenous non-targeted activation, single-target activation, and heterologous expression. It also provides a critical evaluation of these strategies and offers perspectives on the current state of research.

Identifying a Biocontrol Bacterium with Disease-Prevention Potential and Employing It as a Powerful Biocontrol Agent Against Fusarium oxysporum.

Biocontrol microbes are environment friendly and safe for humans and animals. To seek biocontrol microbes effective in suppressing Fusarium oxysporum is important for tomato production. F. oxysporum is a soil-borne pathogen capable of causing wilt in numerous plant species. Therefore, we found a biocontrol bacterium with an excellent control effect from the rhizosphere soil of plant roots. In this work, we focus on two parts of work. The first part is the identification and genomic analysis of the biocontrol bacterium Y-4; the second part is the control efficiency of strain Y-4 on F. oxysporum. For this study, we identified strain Y-4 as Bacillus velezensis. It is an aerobic Gram-positive bacterium that can secrete a variety of extracellular enzymes and siderophores. Strain Y-4 also contains a large number of disease-resistant genes and a gene cluster that forms antibacterial substances. In addition, we found that it significantly inhibited the reproduction of F. oxysporum in a culture dish. In the indoor control effect test, after treatment with strain Y-4 suspension, the disease index of tomato plants decreased significantly. Furthermore, the control efficiency of the plants was 71.88%. At the same time, Y-4 bacterial suspension induced an increase in POD and SOD enzyme activities in tomato leaves, resulting in increased plant resistance. Taken together, strain Y-4 proves to be an effective means of controlling F. oxysporum in tomatoes.

Comparing microsatellites and single nucleotide polymorphisms to evaluate genetic structure and diversity in wolverines (Gulo gulo) across Alaska and western Canada

Abstract The Wolverine (Gulo gulo) is a cold-adapted species of conservation interest because it is sensitive to human development, disturbance, exploitation, and climate warming. Wolverine populations have been studied across much of their distributional range to evaluate patterns of genetic diversity, genetic structure, and gene flow. Little population structure has been detected in northwestern North America with microsatellite loci, but low genomic diversity in wolverines may limit detection of genetic differences in this highly vagile species. Here, we genotyped a relatively large sample of wolverines from across Alaska (US) and adjacent Yukon (Canada) with 12 microsatellite loci (n = 501) and 4,222 single nucleotide polymorphisms (SNPs; n = 201) identified using restriction-site associated DNA sequencing. We compared the relative ability of our microsatellite and SNP datasets to evaluate population genetic structure, genetic diversity, differentiation, and isolation by distance (IBD). We predicted that the SNP dataset would detect a higher degree of genetic structure and provide more significant support for IBD. We found evidence for multiple genetic clusters, including genetic distinctiveness of wolverines in southeast Alaska and on the Kenai Peninsula. The SNP dataset detected additional genetic clusters that align largely with ecoregions, and the SNP dataset showed stronger evidence of IBD, while the 2 datasets were generally consistent in estimates of genetic diversity and differentiation among regional groups. Our results highlight the importance of genomic methods to assess gene flow in wolverines. Identifying population genetic structure allows an assessment of the potential impacts of conservation threats and is an important precursor for designing population monitoring programs.

Genetic structure and diversity of Mycoplasma hyopneumoniae based on a MLVA typing scheme.

Several epidemiological studies have been carried out using Multiple-Locus Variable-number tandem repeat Analysis (MLVA) for M. hyopneumoniae typing. However, a global perspective on the implications of the genetic diversity of this pathogen is lacking. This study aimed to determine and to analyze the genetic structure of M. hyopneumoniae based on the p97R1-p146R3 MLVA typing scheme and to characterize, analyze and compare MLVA types among countries where the information was publicly available. A set of 797 publicly available data of M. hyopneumoniae p97R1-p146R3 MLVA genetic types from six different countries were analyzed using Genalex 6.41 software to characterize loci polymorphism and using Structure 2.3.4 software in order to identify the genetic structure. A total of 185 MLVA types were identified among the analyzed data. For the p97R1 and p146R3 loci, most of the molecular variation in M. hyopneumoniae populations was identified within countries. Three genetic clusters and their recombinations were identified globally. M. hyopneumoniae is a genetically diverse pathogen with limited clonality and three well-defined clusters and their combinations were identified in this investigation. The greatest genetic variation of M. hyopneumoniae was observed within countries.