Genome Annotation Research Articles

Characterization of three novel dimethyl disulfide degrading bacteria and their potential degradation pathways.

Dimethyl disulfide (DMDS) is an odor compound characterized by the lowest olfactory threshold and high toxicity. It is indispensable to explore the bacteria with high resistance and degradation efficiency to DMDS. Acinetobacter lwoffii, Pseudomonas mendocina, and Myroides odoratus were isolated from kitchen waste. After 6days of individual treatment, the removal rates were 34.22%, 40.95%, and 41.94% respectively. The DMDS metabolic pathways based on metagenomic assays were discovered to be incomplete due to the insufficient annotation of some key genes in the current database. Following 3days of treatment with bacterial consortia at ratios of 5:1 for A. lwoffii C2/ M. odoratus C7 and 1:1:1 for the three strains achieved 100% DMDS removal. Additionally, the consortia reduced hydrogen sulfide (H2S) and dimethyl sulfide (DMS).This discovery broadens the spectrum of bacteria exhibiting high tolerance and efficient degradation of DMDS, with significant implications for DMDS removal and odor treatment.

Annotation of transcription factors, chromatin-associated factors, and basal transcription machinery in the pea aphid, Acyrthosiphon pisum, and development of the ATFdb database, a resource for studies of transcriptional regulation

The pea aphid, Acyrthosiphon pisum, is an emerging model system in functional and comparative genomics, in part due to the availability of new genomic approaches and the different sequencing and annotation efforts that the community has dedicated to this important crop pest insect. The pea aphid is also used as a model to study fascinating biological traits of aphids, such as their extensive polyphenisms, their bacteriocyte-confined nutritional symbiosis, or their adaptation to the highly unbalanced diet represented by phloem sap. To get insights into the molecular basis of all these processes, it is important to have an appropriate annotation of transcription factors (TFs), which would enable the reconstruction/inference of gene regulatory networks in aphids. Using the latest version of the A. pisum genome assembly and annotation, which represents the first chromosome-level pea aphid genome, we annotated the complete repertoire of A. pisum TFs and complemented this information by annotating genes encoding chromatin-associated and basal transcription machinery proteins. These annotations were done combining information from the model Drosophila melanogaster, for which we also provide a revisited list of these proteins, and de novo prediction. The comparison between the two model systems allowed the identification of major losses or expansions in each genome, while a deeper analysis was made of ZNF TFs (with certain families expanded in the pea aphid), and the Hox gene cluster (showing reorganization in gene position in the pea aphid compared to D. melanogaster). All annotations are available to the community through the Aphid Transcription Factors database (ATFdb), consolidating the various annotations we generated. ATFdb serves as a valuable resource for gene regulation studies in aphids.

Whole genome sequencing characterization and comparative genome analysis of Acinetobacter baumannii JJAB01: A comprehensive insights on antimicrobial resistance and virulence genotype.

The emergence of antibiotic resistance has significantly elevated the threat posed by Acinetobacter baumannii as an opportunistic pathogen. A.baumannii, a notorious bacterium, poses a serious threat to health care, leading to severe nosocomial infections, particularly in immunocompromised individuals. Whole-Genome Sequencing studies are efficient in providing accurate genetic information, aiding in detecting outbreaks, surveillance of resistance, and controlling infection transmission. In this study, we investigated the whole genome of a clinical isolate A. baumannii JJAB01 which sourced from a urine sample of an Intensive Care Unit (ICU) patient. This strain showed resistance to 24 available antibiotics, signifying Extremely Drug Resistant (XDR) and high potential for pathogenicity. Whole Genome Sequencing was performed using Illumina, and the raw reads were evaluated using the FastQC tool. Genome assembly and annotation were performed with Unicycler and the RAST server. The JJAB01 genome is 4.07Mb with a GC content of 38.9%. A total of 51 and 31 virulence factors and antimicrobial-resistant (AMR) genes were predicted using the VFDB and CARD databases. Comparative genome studies were carried out on virulence factors, resistance genes, prophages, and Multi-Locus Sequence Typing (MLST) across twelve closely related A. baumannii genomes, including JJAB01, X4-584, X4-705, 2023CK-00423, 2023CK-00890, 2023CK-00127, 2022CK-00066, B20AB01, B20AB10, F20AB03, G20AB08, and X4-65. These computational investigations in this study emphasis the multidimensional nature of the ICU strain JJAB01 and its genetic similarity to other strains, thereby enhancing our understanding of drug resistance and the pathogenicity associated with A. baumannii infections.

Draft genome dataset of Streptomyces griseoincarnatus strain R-35 isolated from tidal pool sediments.

The marine isolate, Streptomyces griseoincarnatus strain R-35, was isolated from marine sediments collected from the Glencairn Tidal Pool, Table Mountain National Park, Cape Town, South Africa. The genomic DNA was sequenced using the Ion Torrent GeneStudio™ S5 platform, and the de novo assembly was performed using the SPAdes assembler on the Centre for High Performance Computing (CHPC) Lengau Cluster located at the CSIR, Rosebank, South Africa. The draft genome assembly consisted of 722 contigs totaling 7,625,174 base pairs and a G+C% content of 72.2 mol%. Genome completeness and genome contamination were determined as 99.12% and 0.92%, respectively. Genome annotations performed using the Rapid Annotation with Subsystem Technology (RAST) and the Bacterial and Viral Bioinformatics Resource Centre (BV-BRC) determined the presence of 7996 coding sequences (CDS), 63 transfer RNAs (tRNAs), and six ribosomal RNAs (rRNAs). A total of 2570 hypothetical proteins were assigned, and 5246 proteins were assigned to function. The phylogenomic positioning of S. griseoincarnatus strain R-35 was determined using the Type Strain Genome Server (TYGS) and was found to be related to S. griseoincarnatus JCM 4381T, with a digital DNA-DNA hybridisation (dDDH) value of 84.1%, and an OrthoANIu value of 98.22%. The CARD RGI algorithm on Proksee predicted the presence of 6,107 antimicrobial resistance (AMR) features, 27 biosynthetic gene clusters (BGCs) were predicted using antiSMASH, while 189 carbohydrate-active enzymes (CAZymes) were predicted using dbCAN3. The raw genome sequencing data has been submitted to the National Center for Biotechnology (NCBI) under the BioProject ID PRJNA1129156 (BioSample ID Accession Number: SAMN42145163; Short Read Archive (SRA) Accession: SRR29633055; https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1129156).

Full-length transcriptome sequencing of seven tissues of GuShi chickens.

Chickens are vital economic poultry and serve as exemplary models for avian research. The incomplete reference genome of chickens and the limited availability of full-length cDNA impede the identification of alternatively spliced transcripts, thereby delaying many fundamental chicken studies. We utilized PacBio Iso-seq technology on various chicken tissues, obtaining 170,162 full-length transcripts through comprehensive transcriptome sequencing and annotation. Of the identified transcripts, 38,925 lncRNAs were predicted and categorized into five models. Our data significantly increases the known number of chickens' lncRNAs and alternatively spliced transcripts are crucial in enhancing current genome annotations. Furthermore, this data will be invaluable for functional studies of other avian species.

Chromosome-Level Genome Assembly and Comparative Genomic Analysis of Planiliza haematocheilus: Insights into Environmental Adaptation and Hypoxia Tolerance Mechanisms.

Planiliza haematocheilus, a teleostan species noted for its ecological adaptability and economic significance, thrives in both freshwater and marine environments. This study presents a novel chromosome-level genome assembly through Hi-C, PacBio CCS, and Illumina sequencing methods. The assembled genome has a final size of 651.58Mb, with 24 chromosomes anchoring 91.94% of contigs. Contig N50 and scaffold N50 are respectively measured at 25.52Mb and 28.59Mb. Of the 22,476 protein-coding genes identified in the genome, 21,834 have functional annotations. BUSCO (Benchmarking Universal Single-Copy Orthologs) genome and gene annotation assessments yielded scores of 96% and 96.6%, respectively. The genome of P. haematocheilus revealed 228 expanded and 1433 contracted gene families. Comparative genomic analyses highlight adaptations and hypoxia tolerance, linked to protein synthesis, immune response, and metabolic regulation. The high-quality genome assembly supports advanced studies on gene expression patterns under different environmental stressors, contributing to genetic enhancement efforts for this economically important aquaculture species.

Integrative multi-omics analysis reveals the translational landscape of the plant-parasitic nematode Meloidogyne incognita

Root-knot nematodes (RKNs) of the genus Meloidogyne pose the most significant threats to global food security due to their destructive nature as plant-parasitic nematodes. Although significant attention has been devoted to investigating the gene transcription profiling of RKNs, our understanding of the translational landscape of RKNs remains limited. In this study, we elucidated the translational landscape of Meloidogyne incognita through the integration of translatome, transcriptome and quantitative proteome analyses. Our findings revealed numerous previously unannotated translation events and refined the genome annotation. By investigating the genome-wide translational dynamics of M. incognita during parasitism, we revealed that the genes of M. incognita undergo parasitic stage-specific regulation at the translational level. Interestingly, we identified 470 micropeptides (containing fewer than 100 amino acids) with the potential to function as effectors. Additionally, we observed that the effector-coding genes in M. incognita exhibit higher translation efficiency (TE). Further analysis suggests that M. incognita has the potential to regulate the TE of effector-coding genes without simultaneous alterations in their transcript abundance, facilitating effector synthesis. Collectively, our study provides comprehensive datasets and explores the genome-wide translational landscape of M. incognita, shedding light on the contributions of translational regulation during parasitism.

Probiotic Limosilactobacillus reuteri DSM 17938 Alleviates Acute Liver Injury by Activating the AMPK Signaling via Gut Microbiota-Derived Propionate.

Limosilactobacillus reuteri DSM 17938 (L. reuteri DSM 17938) was one of the most widely used probiotics in humans for gastrointestinal disorders, but few studies have investigated its role in drug-induced liver injury (DILI). Here, we evaluated the efficacy of L. reuteri DSM 17938 using a mouse model of DILI induced by triptolide. Pregavage of L. reuteri DSM 17938 for 1 week remarkably lowered hepatic inflammatory cytokines level and oxidative stress, with diminished serum alanine transaminase and aspartate aminotransferase levels. Metabolomics and RT-qPCR analysis confirmed its ability in ameliorating TP-disrupted hepatic fatty acid β oxidation. Genome annotation of L. reuteri showed its ability to modulate energy metabolism. Targeted metabolomics demonstrated that L. reuteri DSM 17938 modified the short fatty acid profiles in cecum, especially enhancing propionate levels. Further experiments found that L. reuteri DSM 17938 can activate AMPK signaling by upregulating gut microbiota-derived propionate level, thus restoring impaired mitochondrial biogenesis and energy supply processes to recover energy homeostasis, which leads to diminished ROS production and oxidative stress injury in hepatocytes. Besides, AMPK inhibitor dorsomorphin abolished all the effects on propionate protecting mitochondria and energy metabolism. This study established probiotic therapy of L. reuteri DSM 17938 as a preventive intervention for DILI in clinical. We also revealed that L. reuteri DSM 17938 can activate AMPK signaling by propionate, facilitating a deeper understanding of the action mechanism of L. reuteri DSM 17938 against acute liver injury and contributing to the development of its postbiotics and wider applications.

Fish Evo‐Devo: Moving Toward Species‐Specific and Knowledge‐Based Interactome

ABSTRACTA knowledge‐based interactome maps interactions among proteins and molecules within a cell using experimental data, computational predictions, and literature mining. These interactomes are vital for understanding cellular functions, pathways, and the evolutionary conservation of protein interactions. They reveal how interactions regulate growth, differentiation, and development. Transitioning to functionally validated interactomes is crucial in evolutionary developmental biology (Evo‐Devo), especially for non‐model species, to uncover unique regulatory networks, evolutionary novelties, and reliable gene interaction models. This enhances our understanding of complex trait evolution across species. The European Evo‐Devo 2024 conference in Helsinki hosted the first fish‐specific Evo‐Devo symposium, highlighting the growing interest in fish models. Advances in genome annotation, genome editing, imaging, and molecular screening are expanding fish Evo‐Devo research. High‐throughput molecular data have enabled the deduction of gene regulatory networks. The next steps involve creating species‐specific interactomes, validating them functionally, and integrating additional molecular data to deepen the understanding of complex regulatory interactions in fish Evo‐Devo. This short review aims to address the logical steps for this transition, as well as the necessities and limitations of this journey.

Description of the genome sequence of Corynebacterium species (Marseille-Q4381).

In 2020, we isolated a Corynebacterium strain, Marseille-Q4381, from healthy skin. We describe herein its genome sequence and annotation characteristics.

The genome sequence of Dolichopus griseipennis Stannius, 1831

We present a genome assembly from an individual male Dolichopus griseipennis (Arthropoda; Insecta; Diptera; Dolichopodidae). The genome sequence has a total length of 897.50 megabases. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 16.12 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,532 protein-coding genes.

A cross-tissue transcriptome-wide association study identifies new susceptibility genes for benign prostatic hyperplasia

Benign prostatic hyperplasia (BPH) is a prevalent urinary system disorder. Despite evidence of a significant genetic component from previous studies, the specific pathogenic genes and biological mechanisms are still largely unknown. The study utilized the FinnGen R10 dataset, encompassing 177,901 individuals (36,601 cases and 141,300 controls), and the GTEx v8 EQTLs files to conduct single-tissue and cross-tissue transcriptome-wide association studies (TWAS). FUSION method was utilized to validate the findings in specific tissues. Gene Analysis and Multi-marker Analysis of Genomic Annotation (MAGMA) were used to identify potential susceptibility genes. The intersection genes of the above results were analyzed by Mendelian randomization (MR), summary data-based MR (SMR) and colocalization studies. Fine-mapping of causal gene sets (FOCUS) software was employed to accurately locate risk genes. Gene Expression Omnibus (GEO) analysis explores the differential expression of genes. Finally, The GeneMANIA tool was utilized to further understand the functional roles of these susceptibility genes. The cross-tissue TWAS analysis revealed 28 genes associated with BPH susceptibility. Single-tissue TWAS and MAGMA further refined eight genes, and subsequent MR, SMR, FOCUS and colocalization analysis pinpointed INO80B as the key gene. The differential expression of this result was verified by GEO. INO80B may help prevent excessive prostatic cell proliferation by regulating cell cycle-related gene expression. Our research identified the INO80B gene, whose predicted expression is associated with BPH risk and hence provided a new insight into the genetic basis of this disease. However, further functional studies are necessary to elucidate the potential biological activity of these significant signals.

Genome Wide Identification of Structure Variations in Five Italian Turkey Populations

Structural variants (SVs) are one of the main sources of genetic variants and have a significant impact on phenotype evolution, disease susceptibility, and environmental adaptations. We used 73 whole genome sequencing (12x) to apply a mapping approach to identify SVs in five turkey populations. A notable degree of genetic isolation was observed between the Basilicata and Apulian populations, as indicated by principal component analysis and admixture results. A total of 11,733 SVs were detected, including 6712 deletions, 2671 duplications, 1430 inversions, and 920 translocations. The Variant Effect Predictor (VEP) analysis predicted various consequences of filtered SVs as follows: intron variants (35.8%), intergenic variants (9.6%), coding sequence variants (8.3%), downstream gene variants (7.5%), and transcript ablations (7.3%). Our functional annotation of genes overlapping with SVs was mainly enriched in recognized pathways governing positive regulation of nucleoplasm, protein binding, mitochondrion, negative regulation of cell population proliferation, identical protein binding, and calcium signaling. We produced a comprehensive SV catalog utilizing unique whole-genome turkey data. This SV catalog not only increases our understanding of genetic diversity in turkeys but also enhances our knowledge of the role of SVs in their phenotypic traits.

Mettannotator: a comprehensive and scalable Nextflow annotation pipeline for prokaryotic assemblies.

In recent years there has been a surge in prokaryotic genome assemblies, coming from both isolated organisms and environmental samples. These assemblies often include novel species that are poorly represented in reference databases creating a need for a tool that can annotate both well-described and novel taxa, and can run at scale. Here, we present mettannotator-a comprehensive, scalable Nextflow pipeline for prokaryotic genome annotation that identifies coding and non-coding regions, predicts protein functions, including antimicrobial resistance, and delineates gene clusters. The pipeline summarises the results of these tools in a GFF (General Feature Format) file that can be easily utilised in downstream analysis or visualised using common genome browsers. Here, we show how it works on 200 genomes from 29 prokaryotic phyla, including isolate genomes and known and novel metagenome-assembled genomes, and present metrics on its performance in comparison to other tools. The pipeline is written in Nextflow and Python and published under an open source Apache 2.0 licence. Instructions and source code can be accessed at https://github.com/EBI-Metagenomics/mettannotator. The pipeline is also available on WorkflowHub: https://workflowhub.eu/workflows/1069. Supplementary data are available at Bioinformatics online.

Genome-wide association studies with prolapsed gland of the third eyelid in dogs

Cherry eye, the common name for the prolapse of the third eyelid gland in dogs, is a widespread ophthalmic disease affecting dogs of various breeds. This condition severely affects the quality of life of affected dogs, and its underlying cause remains unresolved. In this study, 170K SNP microarray data were collected from 653 brachycephalic dogs and 788 brachycephalic and mesocephalic dogs. These two datasets were analyzed separately in genome-wide association studies (GWAS) involving 12 dog breeds affected by cherry eye. The GWAS analysis of 653 short-headed dogs revealed that four SNPs in the CFA3:15627075-15983629 bp region exceeded the genome-level significance threshold. Association analysis of this region also indicated that these four SNPs were strongly associated. Gene annotation showed that the region contained genes such as KIAA0825, FAM172A, and NR2F1, of which NR2F1 was associated with eye development. The results showed that GWAS analysis performed on 788 short- and medium-headed dogs identified five SNPs in the CFA22:15627075-15983629 bp region that exceeded the genome-level significance threshold, and association analysis was performed in this region, which showed that these five SNPs were strongly associated. In addition, 104 annotated genes were identified in both GWAS. To explore the genes involved in cherry eyes, we performed GO functional enrichment analysis. The genes involved in the high pathway were DIO3 and TTC8. In addition, an in-depth analysis revealed 33 genes associated with eye development and diseases. Our study provides new perspectives for further understanding cherry eye in dogs.

Evolutionary constrained genes associated with autism spectrum disorder across 2,054 nonhuman primate genomes

BackgroundSignificant progress has been made in elucidating the genetic underpinnings of Autism Spectrum Disorder (ASD). However, there are still significant gaps in our understanding of the link between genomics, neurobiology and clinical phenotype in scientific discovery. New models are therefore needed to address these gaps. Rhesus macaques (Macaca mulatta) have been extensively used for preclinical neurobiological research because of remarkable similarities to humans across biology and behaviour that cannot be captured by other experimental animals.MethodsWe used the macaque Genotype and Phenotype (mGAP) resource consisting of 2,054 macaque genomes to examine patterns of evolutionary constraint in known human neurodevelopmental genes. Residual variation intolerance scores (RVIS) were calculated for all annotated autosomal genes (N = 18,168) and Gene Set Enrichment Analysis (GSEA) was used to examine patterns of constraint across ASD genes and related neurodevelopmental genes.ResultsWe demonstrated that patterns of constraint across autosomal genes are correlated in humans and macaques, and that ASD-associated genes exhibit significant constraint in macaques (p = 9.4 × 10− 27). Among macaques, many key ASD-implicated genes were observed to harbour predicted damaging mutations. A small number of key ASD-implicated genes that are highly intolerant to mutation in humans, however, showed no evidence of similar intolerance in macaques (CACNA1D, MBD5, AUTS2 and NRXN1). Constraint was also observed across genes associated with intellectual disability (p = 1.1 × 10− 46), epilepsy (p = 2.1 × 10− 33) and schizophrenia (p = 4.2 × 10− 45), and for an overlapping neurodevelopmental gene set (p = 4.0 × 10− 10).LimitationsThe lack of behavioural phenotypes among the macaques whose genotypes were studied means that we are unable to further investigate whether genetic variants have similar phenotypic consequences among nonhuman primates.ConclusionThe presence of pathological mutations in ASD genes among macaques, along with evidence of similar genetic constraints to those in humans, provides a strong rationale for further investigation of genotype-phenotype relationships in macaques. This highlights the importance of developing primate models of ASD to elucidate the neurobiological underpinnings and advance approaches for precision medicine and therapeutic interventions.

Pervasive formation of double-stranded RNAs by overlapping sense/antisense transcripts in budding yeast mitosis and meiosis.

Previous RNA profiling studies revealed co-expression of overlapping sense/antisense (s/a) transcripts in pro- and eukaryotic organisms. Functional analyses in yeast have shown that certain s/a mRNA/mRNA and mRNA/lncRNA pairs form stable double-stranded RNAs (dsRNAs) that affect transcript stability. Little is known, however, about the genome-wide prevalence of dsRNA formation and its potential functional implications during growth and development in diploid budding yeast. To address this question, we monitored dsRNAs in a Saccharomyces cerevisiae strain expressing the ribonuclease DCR1 and the RNA binding protein AGO1 from Naumovozyma castellii. We identify dsRNAs at 347 s/a loci that express partially or completely overlapping transcripts during mitosis, meiosis or both stages of the diploid life cycle. We associate dsRNAs with s/a loci previously thought to be exclusively regulated by antisense interference, and others that encode antisense RNAs, which downregulate sense mRNA-encoded protein levels. To facilitate hypothesis building we developed the Sense/Antisense double-stranded RNA (SensR) expression viewer. Users are able to retrieve different graphical displays of dsRNA and RNA expression data using genome coordinates and systematic or standard names for mRNAs and different types of stable or cryptic long non-coding RNAs (lncRNAs). Our data are a useful resource for improving yeast genome annotation and for work on RNA-based regulatory mechanisms controlling transcript and protein levels. The data are also interesting from an evolutionary perspective, since natural antisense transcripts that form stable dsRNAs have been detected in many species from bacteria to humans. The SensR viewer is freely accessible at https://sensr.genouest.org.

A meta-analysis of genome-wide association studies to identify candidate genes associated with feed efficiency traits in pigs.

Pig production is an agricultural sector of great economic and social relevance to Brazil and global markets. Feed efficiency traits directly influence the sustainability of pig production due to the economic impact of feed costs on the production system and the environmental footprint of the industry. Therefore, breeding for improved feed efficiency has been a target of worldwide pig breeding programs. Genome-wide association studies (GWAS) enable the assessment of the genetic background of complex traits, which contributes to a better understanding of the biological mechanisms regulating their phenotypic expression. In this context, the primary objective of this study was to identify and validate genomic regions and candidate genes associated with feed conversion ratio (FCR) and residual feed intake (RFI) in pigs based on a comprehensive systematic review and meta-analysis of GWAS. The METAL software was used to implement the meta-analysis and the Bonferroni multiple testing correction considering a significance threshold of 0.05. The significant single nucleotide polymorphisms (SNPs) in the meta-analysis were used to identify candidate genes, followed by a functional genomic enrichment analysis. The systematic review identified 13 studies, of which 7 evaluated FCR, 3 evaluated RFI, and 3 studies investigated both traits, with 160 and 96 SNPs identified for FCR and RFI, respectively. After the meta-analysis, 145 markers were significantly associated with FCR and 90 with RFI. The gene annotation process resulted in 105 and 114 genes for FCR and RFI, respectively. The enrichment analysis for FCR resulted in 16 significant gene ontology (GO) terms, while six terms were identified for RFI. The main GO terms were actin cytoskeleton (GO_BP:0030036), membrane (GO_CC:0016020), integral components of the peroxisomal membrane (GO_CC:0005779), and carbohydrate binding (GO_MF:0030246). The main candidate genes identified were MED18, PHACTR4, ABCC2, TRHDE, FRS2, FAR2 and FIS1 for FCR, and ADGRL2, ASGR1, ASGR2, and MAN2B1 for RFI. These findings contribute to a better understanding of the genetic mechanisms associated with feed efficiency traits in pigs, providing a foundation for future improvements in pig breeding programs.

Comprehensive genome annotation of Trilocha varians, a new model species of Lepidopteran insects

Trilocha varians is a member of the bombycid moths. Since T. varians has a considerably shorter generation period than the prevailing model species, Bombyx mori, this species would be a novel model insect in Lepidoptera. To facilitate further use of T. varians, we developed genome annotation information on the chromosome-scale assembly of T. varians previously published by our group. 9 RNA-seq datasets and 2 Iso-seq datasets were submitted for transcriptome-based gene prediction. As a result, 16,266 protein-coding genes were predicted on the latest genome assembly, and 98.6% of BUSCO sequences were present in our gene models. ATAC-seq was also conducted to determine chromatin accessibility across the genome. Finally, piRNA-targeted small RNA-seq revealed T. varians genome harbours 517 piRNA clusters (piCs). This information will encourage and facilitate potential users who plan to use this species.

Genomic and biochemical investigations in the biomineralizing potential of an isolated marine ureolytic Bacillus sp. N₉.

Microbially Induced Calcium Carbonate Precipitation (MICP) plays a significant role in coastal soil stabilization and erosion prevention. In the present study, the biomineralizing potential of a newly isolated Bacillus sp. N₉ was investigated through MICP. The isolated Bacillus sp. N₉ induced calcium carbonate (CaCO3) precipitation in broth, agar, and real beach rock was evaluated by X-ray Fluorescence, Scanning Electron Microscopy, and X-ray Diffraction. The visual appearance of calcites as white crystals on the rock and the sealing of cracks in the beach sand cubes confirmed the CaCO3 precipitation by the isolated strain. The pH changes, soluble calcium (Ca2+) concentration levels and urease activity during the growth of the isolated Bacillus sp. N₉ were studied. High urease activity was observed, resulting in an increased production of carbonate (CO32-) ions, which in turn promoted a higher rate of CaCO₃ precipitation. Further, the assembly of the 3.27Mb genome of the isolated Bacillus sp. N₉ was evaluated using Nanopore sequencing technology, and various extracellular proteins, including urease, were identified. The gene annotation through PROKKA and RAST predicted 6700 and 7317 protein-coding sequences. The pathway annotation analysis through gene ontology, KEGG, and COG inferred the presence of genes in proteolytic characteristics, carbohydrate metabolism, and amino acid derivatives. The present study provides valuable insights into the isolated native Bacillus sp. N₉, demonstrating its potential to produce high urease activity and calcite precipitation through combined genomics and in vitro techniques.