Gene Cluster Research Articles

Whole Genome Sequencing and Biocontrol Potential of Streptomyces luteireticuli ASG80 Against Phytophthora Diseases

Phytophthora-induced crop diseases, commonly known as “plant plagues”, pose a significant threat to global food security. In this study, strain ASG80 was isolated from sisal roots and demonstrated a broad-spectrum antagonistic activity against several Phytophthora species and fungal pathogens. Strain ASG80 was identified as Streptomyces luteireticuli via phylogenetic analysis, digital DNA–DNA hybridization (dDDH), and average nucleotide identity (ANI). Whole-genome sequencing identified 40 biosynthetic gene clusters (BGCs) related to secondary metabolite production, including antimicrobial compounds. Strain ASG80 extract exhibited broad-spectrum inhibitory activity against Phytophthora nicotianae, P. vignae, P. cinnamomi, and P. sojae. Pot experiments showed that strain ASG80 extract significantly reduced sisal zebra disease incidence, with an efficacy comparable to the fungicide metalaxyl. These findings suggest that strain ASG80 is a promising biocontrol agent with substantial potential for managing Phytophthora-related diseases in agriculture.

Conjugation mediates large-scale chromosomal transfer in Streptomyces driving diversification of antibiotic biosynthetic gene clusters.

Streptomyces are ubiquitous soil dwelling bacteria with large, linear genomes that are of special importance as a source of metabolites used in human and veterinary medicine, agronomy and industry. Conjugative elements (Actinomycetes Integrative and Conjugative Elements, AICEs) are the main drivers of Streptomyces Horizontal Gene Transfer. AICE transfer has long been known to be accompanied by mobilization of chromosomal DNA. However, the magnitude of DNA transfer, or the localization of acquired DNA across their linear chromosome, has remained undetermined. We here show that conjugative crossings in sympatric strains of Streptomyces result in the large-scale, genome-wide distributed replacement of up to one third of the recipient chromosome, a phenomenon for which we propose the name 'Streptomyces Chromosomal Transfer' (SCT). Such chromosome blending results in the acquisition, loss and hybridization of Specialized Metabolite Biosynthetic Gene Clusters, leading to a novel metabolic arsenal in exconjugant offspring. Harnessing conjugation-mediated SMBGC diversification holds great promise in the discovery of new bioactive compounds including antibiotics.

Emergence of the mobile RND-type efflux pump gene cluster tmexCD1-toprJ1 in Klebsiella pneumoniae clinical isolates in Japan.

Tigecycline is an antimicrobial agent with a broad spectrum of activity against both Gram-positive and Gram-negative bacteria. However, mobile tigecycline resistance gene clusters, such as tnfxB-tmexCD-toprJ, have spread globally. The prevalence of tigecycline-resistant Enterobacterales in clinical settings in Japan is unknown. To investigate the tnfxB-tmexCD-toprJ gene cluster in the genome sequences of Enterobacterales clinical isolates in Japan. We investigated the tnfxB-tmexCD-toprJ cluster from the genome sequences of 5143 Enterobacterales isolates collected from 175 hospitals around Japan between 2019 and 2020 as part of a national genomic surveillance program for antimicrobial-resistant bacteria. The tnfxB1-tmexCD1-toprJ1 cluster was detected in two Klebsiella pneumoniae isolates in 2019. One isolate possessed a 299.4 kb IncFIB(K) plasmid, pJBBGAAF19431, and the other possessed a 224.9 kb IncHI1B/IncFIB(K) hybrid plasmid, pJBEAACG19501, co-carrying multiple antimicrobial resistance genes, including extended-spectrum β-lactamase genes, blaOXA-1 and blaCTX-M-27, respectively, along with tnfxB1-tmexCD1-toprJ1. The genetic context of the tnfxB1-tmexCD1-toprJ1-surrounding structure on pJBBGAAF19431 was similar to that of a K. pneumoniae plasmid pHNAH8I-1 from a chicken in China in 2017, and the cluster was embedded in an apparently intact mobile DNA element: strand-biased circularizing integrative element. The tnfxB1-tmexCD1-toprJ1 on pJBEAACG19501 was embedded in a Tn3 family transposon related to TnAs1. The plasmid pJBEAACG19501 was highly similar to that of K. pneumoniae, isolated from humans in China in 2021. tmexCD-toprJ was present in Japan as of 2019. Even in Japan, where the clinical use of tigecycline is significantly rare, tmexCD-toprJ-harbouring multidrug-resistant Enterobacterales is a public health threat and requires continuous monitoring.

Phased T2T genome assemblies facilitate the mining of disease-resistance genes in Vitis davidii

Abstract Grape is an important fruit crop, and its production faces significant threat from diseases, resulting in substantial economic loss. Wild grape relatives are valuable resources for the restoration of disease-resistance loci. However, available resistance loci in wild grape genomes remain largely unexplored. In this study, we assembled two phased genomes, including a high resistant Chinese wild grape, Vitis davidii Föex, and a susceptible cultivar, V. vinifera L. cv. “Manicure Finger”. We detected a total of 36,688 structural variations (SVs), with the genes associated with heterozygous SVs showing an enrichment in allele-specific expression (ASE). Furthermore, we identified eight subgroups of R genes and found that 74.2% of R genes overlap with transposable elements (TEs). Among R genes, NBS-type genes exhibit higher expression profiles in the wild grape genome compared with those in the grape cultivar. Additionally, five specific NBS-type R gene clusters were identified in the wild grape genome that are absent in the cultivar. Through genetic mapping, we identified four quantitative trait loci (QTLs) associated with grape white rot resistance based on the Vitis davidii genome, within which six NBS-type R genes exhibit differential expression between wild and cultivated grapes. Overall, our study revealed the landscape of resistance genes in grape genomes, providing valuable genetic resources for further breeding programs.

Data-Driven Approaches in Antimicrobial Resistance: Machine Learning Solutions

Background/Objectives: The emergence of antimicrobial resistance (AMR) due to the misuse and overuse of antibiotics has become a critical threat to global public health. There is a dire need to forecast AMR to understand the underlying mechanisms of resistance for the development of effective interventions. This paper explores the capability of machine learning (ML) methods, particularly unsupervised learning methods, to enhance the understanding and prediction of AMR. It aims to determine the patterns from AMR gene data that are clinically relevant and, in public health, capable of informing strategies. Methods: We analyzed AMR gene data in the PanRes dataset by applying unsupervised learning techniques, namely K-means clustering and Principal Component Analysis (PCA). These techniques were applied to identify clusters based on gene length and distribution according to resistance class, offering insights into the resistance genes’ structural and functional properties. Data preprocessing, such as filtering and normalization, was conducted prior to applying machine learning methods to ensure consistency and accuracy. Our methodology included the preprocessing of data and reduction of dimensionality to ensure that our models were both accurate and interpretable. Results: The unsupervised learning models highlighted distinct clusters of AMR genes, with significant patterns in gene length, including their associated resistance classes. Further dimensionality reduction by PCA allows for clearer visualizations of relationships among gene groupings. These patterns provide novel insights into the potential mechanisms of resistance, particularly the role of gene length in different resistance pathways. Conclusions: This study demonstrates the potential of ML, specifically unsupervised approaches, to enhance the understanding of AMR. The identified patterns in resistance genes could support clinical decision-making and inform public health interventions. However, challenges remain, particularly in integrating genomic data and ensuring model interpretability. Further research is needed to advance ML applications in AMR prediction and management.

Genome-wide comparative analysis of clinical and environmental strains of the opportunistic pathogen Paracoccus yeei (Alphaproteobacteria)

IntroductionParacoccus yeei is the first species in the genus Paracoccus to be implicated in opportunistic infections in humans. As a result, P. yeei strains provide a valuable model for exploring how bacteria shift from a saprophytic to a pathogenic lifestyle, as well as for investigating the role of horizontally transferred DNA in this transition. In order to gain deeper insights into the unique characteristics of this bacterium and the molecular mechanisms underlying its opportunistic behavior, a comparative physiological and genomic analysis of P. yeei strains was performed.ResultsComplete genomic sequences of 7 P. yeei isolates (both clinical and environmental) were obtained and analyzed. All genomes have a multipartite structure comprising numerous extrachromosomal replicons (59 different ECRs in total), including large chromids of the DnaA-like and RepB families. Within the mobile part of the P. yeei genomes (ECRs and transposable elements, TEs), a novel non-autonomous MITE-type element was identified. Detailed genus-wide comparative genomic analysis permitted the identification of P. yeei-specific genes, including several putative virulence determinants. One of these, the URE gene cluster, determines the ureolytic activity of P. yeei strains—a unique feature among Paracoccus spp. This activity is induced by the inclusion of urea in the growth medium and is dependent on the presence of an intact nikR regulatory gene, which presumably regulates expression of nickel (urease cofactor) transporter genes.DiscussionThis in-depth comparative analysis provides a detailed insight into the structure, composition and properties of P. yeei genomes. Several predicted virulence determinants (including URE gene clusters) were identified within ECRs, indicating an important role for the flexible genome in determining the opportunistic properties of this bacterium.

Chromosome architecture as a determinant for biosynthetic diversity in Micromonospora.

Natural products - small molecules generated by organisms to facilitate ecological interactions - are of great importance to society and are used as antibacterial, antiviral, antifungal and anticancer drugs. However, the role and evolution of these molecules and the fitness benefits they provide to their hosts in their natural habitat remain an outstanding question. In bacteria, the genes that encode the biosynthetic proteins that generate these molecules are organised into discrete loci termed biosynthetic gene clusters (BGCs). In this work, we asked the following question: How are biosynthetic gene clusters organised at the chromosomal level? We sought to answer this using publicly available high-quality assemblies of Micromonospora, an actinomycete genus with members responsible for biosynthesizing notable natural products, such as gentamicin and calicheamicin. By orienting the Micromonospora chromosome around the origin of replication, we demonstrated that Micromonospora has a conserved origin-proximal region, which becomes progressively more disordered towards the antipodes of the origin. We then demonstrated through genome mining of these organisms that the conserved origin-proximal region and the origin-distal region of Micromonospora have distinct populations of BGCs and, in this regard, parallel the organization of Streptomyces, which possesses linear chromosomes. Specifically, the origin-proximal region contains highly syntenous, conserved BGCs predicted to biosynthesize terpenes and a type III polyketide synthase. In contrast, the ori-distal region contains a highly diverse population of BGCs, with many BGCs belonging to unique gene cluster families. These data highlight that genomic plasticity in Micromonospora is locus-specific, and highlight the importance of using high-quality genome assemblies for natural product discovery and guide future natural product discovery by highlighting that biosynthetic novelty may be enriched in specific chromosomal neighbourhoods.

Degradation mechanism of difructose dianhydride III in Blautia species.

Di-fructofuranose 1,2':2,3' dianhydride (DFA-III) is a cyclic fructo-disaccharide, which is produced by the condensation of two fructose molecules via the caramelization or enzymatic reaction of inulin fructotransferase. A strain of Blautia producta was known to utilize DFA-III as a carbohydrate source; however, the mechanisms remain unclear. In this study, we characterized the glycoside hydrolase (GH) family 91 DFA-III hydrolase (DFA-IIIase) from B. parvula NBRC 113351. Recombinant BpDFA-IIIase catalyzed the reversible conversion of DFA-III to inulobiose, which is further degraded to fructose by the cooperative action of DFA-IIIase and GH32 β-D-fructofuranosidase. DFA-III was utilized in several Blautia species with a gene cluster for DFA-III degradation (e.g., B. parvula NBRC 113351, B. hydrogenotrophica JCM 14656, and B. wexlerae JCM 35486), but not by B. wexlerae JCM 31267, which does not possess the gene cluster. Furthermore, B. hansenii JCM 14655, which cannot metabolize fructose, could not utilize DFA-III; however, it could degrade DFA-III to fructose in the presence of DFA-III-degrading enzymes. Fecal fermentation tests showed that Blautia species are important gut microbe for degrading DFA-III. KEY POINTS: • BpDFA-IIIase is the first characterized DFA-IIIase in intestinal non-pathogenic bacteria. • DFA-IIIase is widely conserved in Blautia species. • DFA-III is degraded to d-fructose through inulobiose by the cooperative action of DFA-IIIase and β-d-fructofuranosidase.

Antimicrobial activities and bacteriocins of lactic acid bacteria isolated from Nigeria indigenous fermented cowmilk “nunu”

Nunu is a nutritious fermented yoghurt-like milk beverage that can serve as a source of lactic acid bacteria (LAB) with antimicrobial and bacteriocin potentials. Nunu samples were screened for LAB with antimicrobial activity against test organisms. Three LAB, namely Lactobacillus parafarraginis OCLP1, L. fermentum CSL58 and L. buchneri MZE12 out of a total of 14 isolates produced antimicrobial activity against Bacillus anthracis CHF-2011-32b, B. cereus DHM5-1, Providencia vermicola KSVDL-09-71600, P. rettgeri SIRT3, P. stuartti SILT11 and other test organisms. Antimicrobial factors were assayed in LAB cell-free extracts (CFE), neutralized CFE, and ammonium sulphate precipitants of the CFE, and Lactobacillus parafarraginis OCLP1 demonstrated highest activity with zone of inhibition = 20.33±3.51 mm­, 13.33±1.09 mm and 25.33±2.52 mm respectively. The CFE of the LAB especially L. parafarraginis OCLP1 demonstrated wide activity spectra by inhibiting both the gram-positive and gram-negative bacteria tested. Result of plasmid curing of the LAB indicate that the bacteriocins of L. parafarraginis OCLP1 was plasmid-encoded while the others have their gene clusters on the chromosome. This study concluded that bacteriocin-producing LAB can be found in “nunu” and recommend the use of the isolated LAB as starter in fermented food production after ensuring their safety through toxicological studies.

Meta-analysis of spatial genetic patterns among European saproxylic beetles

AbstractThe phylogeography of many forest-dwelling species in Europe is well understood. However, our knowledge regarding the genetics of saproxylic beetles remains insufficient. This knowledge gap extends to understanding the influence of both quaternary history and contemporary forest dynamics on population genetics. To fill this gap, we conducted a systematic review and meta-analysis of recent literature concerning saproxylic beetle taxa with available genetic data. We include both threatened and common species in our study, which enabled us to generalize our findings to the whole saproxylic community. Results suggest a latitudinal decrease in diversity in most species, likely influenced by Pleistocene glaciation and subsequent population expansions from southern refugia. Additionally, we observed an east–west gradient in diversity, with threatened species exhibiting higher diversity towards the east. This may reflect historical forest dynamics and anthropogenic pressures, such as heavy wood logging in Western Europe. Similarly, we found a pattern along altitude, with populations in higher elevation forests, which are often more natural, exhibiting higher diversity. Furthermore, we identified distinct phylogenetic units or genetic clusters in southern Europe reflecting the distribution of glacial refugia. For some taxa, distinct units were also reported in eastern Europe where populations spread from Asian refugia. Central Europe showed a high number of phylogenetic units, although unique (private) clades or clusters were absent. Most likely it is an effect of the presence of beetles that originated from various refugia belonging to different phylogenetic units.This study brings insights into general phylogeographic patterns, which have previously been examined only for single representatives of saproxylic beetles. It should also help in the proper planning of conservation and management efforts for wood-dwelling beetles.

Regulation of neutrophil associated RNASET2 expression in rheumatoid arthritis

Neutrophils (PMNs) are key players of innate immune responses through the release of cytoplasmic granule content and the formation of neutrophil extracellular traps (NETs). RNASET2 is an acidic ribonuclease, recently proposed as an alarmin signal associated with inflammatory responses. Here we show that, along the neutrophil maturation cascade, RNASET2 is expressed in segmented and mature PMNs. In human PMNs, RNASET2 colocalized with primary and tertiary granules and was found to be associated with NETs following PMA or Nigericin stimulation. Similarly, activation of PMNs by soluble immune complexes, a hallmark of several autoimmune diseases, also induced RNASET2-associated NETs. Genome-wide association studies recently identified RNASET2 among a cluster of genes associated with increased susceptibility to develop autoimmune diseases, including rheumatoid arthritis (RA). RNASET2 was found expressed by PMNs and macrophages infiltrating inflamed joints in a murine model of RA (K/BxN Serum-Transfer-Induced Arthritis, STIA), by immunostaining. Similar results were found in synovial biopsies of RA patients with active disease. In addition, we demonstrate that RNASET2 circulating levels correlated with the onset and the severity of disease in two mouse models of inflammatory arthritis, STIA and CIA (Collagen-Induced Arthritis) and in serum of RA patients. These results show that PMNs are an important source of RNASET2 and that its circulating levels are associated with RA development suggesting a role for RNASET2 in the pathogenesis of immune-mediated diseases.

Xanthomonas protegens sp. nov., a novel rice seed-associated bacterium, provides in vivo protection against X. oryzae pv. oryzae, the bacterial leaf blight pathogen.

Historically, Xanthomonas species are primarily known for their pathogenicity against plants, but recently, there have been more findings of non-pathogenic xanthomonads. In the present study, we report isolates from healthy rice seeds that belong to a new species, X. protegens, a protector of the rice plants against a serious pathogenic counterpart, i.e. X. oryzae pv. oryzae upon leaf clip co-inoculation. The new member species is non-pathogenic to rice and lacks a type III secretion system. The pangenome investigation revealed a large number of unique genes, including a novel lipopolysaccharide biosynthetic gene cluster, that might be important in its adaptation. The phylo-taxonogenomic analysis revealed that X. protegens is a taxonomic outlier species of X. sontii, a core, vertically transmitted rice seed endophyte with numerous probiotic properties. Interestingly, X. sontii is also reported as a keystone species of healthy rice seed microbiome. The findings and resources will help in the development of unique gene markers and evolutionary studies of X. sontii as a successful symbiont and X. oryzae as a serious pathogen. Here, we propose X. protegens sp. nov. as a novel species of the genus Xanthomonas with PPL118=MTCC 13396=CFBP 9164=ICMP 25181 as the type strain. PPL117, PPL124, PPL125 and PPL126 are other strains of the species.

Genomic insights into the cold adaptation and secondary metabolite potential of Pseudoalteromonas sp. WY3 from Antarctic krill

In the Antarctic marine ecosystem, krill play a pivotal role, yet the intricate microbial community intertwined with these diminutive crustaceans remains largely unmapped. In this study, we successfully isolated and characterized a unique bacterial strain, Pseudoalteromonas sp. WY3, from Antarctic krill. Genomic analysis revealed that WY3 harbors a multitude of genes associated with cold shock proteins, oxidoreductases, and enzymes involved in the osmotic stress response, equipping it with a robust molecular arsenal to withstand frigid Antarctic conditions. Furthermore, the presence of two distinct biosynthesis-related gene clusters suggests that WY3 has the potential to synthesize diverse secondary metabolites, including aryl polyenes and ribosomally synthesized and post-translationally modified peptides. Notably, the identification of genes encoding enzymes crucial for biological immunity pathways, such as apeH and ubiC, hints at a complex symbiotic relationship between WY3 and its krill host. This comprehensive study highlights the robust potential of WY3 for secondary metabolite production and its remarkable ability to thrive at extremely low temperatures in the Antarctic ecosystem, shedding light on the interplay between culturable microorganisms and their hosts in harsh environments, and providing insights into the underexplored microbial communities associated with Antarctic marine organisms and their role in environmental adaptation and biotechnological applications.

Genomic perspectives on foodborne illness

Whole-genome sequencing of bacterial pathogens is used by public health agencies to link cases of food poisoning caused by the same source of contamination. The vast majority of these appear to be sporadic cases associated with small contamination episodes and do not trigger investigations. A "contamination episode" refers to one or more contamination events from a single source over a period of time. We examine clusters of sequenced clinical isolates of Salmonella, Escherichia coli, Campylobacter, and Listeria that differ by only a small number of mutations (SNPs) to identify features of the underlying contamination episodes. These analyses provide additional evidence that the youngest age groups have greater susceptibility to infection by Salmonella, E. coli, and Campylobacter than older age groups. This age bias is weaker for the common Salmonella serovar Enteritidis than Salmonella in general. A large fraction of the contamination episodes causing sickness appear to have a long duration. For example, 50% of the Salmonella cases are in clusters that persist for almost 3 y. For all four pathogen species, the majority of the cases were part of genetic clusters with illnesses in multiple states and likely to be caused by contaminated commercially distributed foods. Salmonella infections in infants under 3 mo are predominantly acquired from the same contaminated food, pet food, or environmental sources as older individuals, rather than infant formula contaminated during production.

Construction of a Wilms tumor risk model based on machine learning and identification of cuproptosis-related clusters

BackgroundCuproptosis, a recently identified type of programmed cell death triggered by copper, has mechanisms in Wilms tumor (WT) that are not yet fully understood. This research focuses on examining the link between WT and Cuproptosis-related genes (CRGs), with the goal of developing a predictive model for WT.MethodsFour gene expression datasets related to WT were sourced from the GEO database. Subsequently, expression profiles of CRGs were extracted for differential analysis and immune infiltration studies. Utilizing 105 WT samples, clusters related to Cuproptosis were identified. This involved analyzing associated immune cell infiltration and conducting functional enrichment analysis. Disease-characteristic genes were pinpointed using weighted gene co-expression network analysis. Finally, the WT risk prediction model was constructed by four machine learning methods: random forest, support vector machine (SVM), generalized linear and extreme gradient strength model. The best-performing machine learning model was chosen, and a nomogram was created. The effectiveness of this predictive model was validated using methods such as the calibration curve, decision curve analysis, and by appiying it to the TARGET-GTEx dataset.ResultsThirteen differentially expressed Cuproptosis-related genes were identified. The infiltration level of CD8 + T cells in WT children was lower than that in Normal tissue (NT) children, and the level of M0 infiltration of macrophages and T follicular helper cells was higher than that in NT children. In addition, two clusters of cuproptosis-related WT were identified. Enrichment analysis results indicated that genes in cluster 2 were primarily involved in cell division, nuclear division regulation, DNA biosynthesis process, ubiquitin-mediated proteolysis. The SVM model was judged to be the optimal model using 5 genes. Its accuracy was confirmed through a calibration curve and decision curve analysis, demonstrating satisfactory performance on the TARGET-GTEx validation dataset. Additional analysis revealed that these five genes exhibited high expression in both the TARGET-GTEx validation dataset and sequencing data.ConclusionThis research established a link between WT and Cuproptosis. It developed a predictive model for assessing the risk of WT and pinpointed five key genes associated with the disease.

COG database update 2024.

The Clusters of Orthologous Genes (COG) database, originally created in 1997, has been updated to reflect the constantly growing collection of completely sequenced prokaryotic genomes. This update increased the genome coverage from 1309 to 2296 species, including 2103 bacteria and 193 archaea, in most cases, with a single representative genome per genus. This set covers all genera of bacteria and archaea that included organisms with 'complete genomes' as per NCBI databases in November 2023. The number of COGs has been expanded from 4877 to 4981, primarily by including protein families involved in bacterial protein secretion. Accordingly, COG pathways and functional groups now include secretion systems of types II through X, as well as Flp/Tad and type IV pili. These groupings allow straightforward identification and examination of the prokaryotic lineages that encompass-or lack-a particular secretion system. Other developments include improved annotations for the rRNA and tRNA modification proteins, multi-domain signal transduction proteins, and some previously uncharacterized protein families. The new version of COGs is available at https://www.ncbi.nlm.nih.gov/research/COG, as well as on the NCBI FTP site https://ftp.ncbi.nlm.nih.gov/pub/COG/, which also provides archived data from previous COG releases.

Terpestacin and Its Derivatives: Bioactivities and Syntheses.

Terpestacin (1), fusaproliferin (2), and their derivatives are a class of sesterterpenes featured by a trans-fused 5/15-membered ring skeleton. There are 45 natural products (1, 2, 4-27, 65-83) isolated from various wild fungi (Fusarium sp., Bipolaris sorokiniana, Arthrinium sp., etc.) or from genetic mutants via biosynthetic gene clusters mining, and 37 derivatives (28-64) produced by semi-synthetic modifications. These compounds show a diverse range of important bioactivities such as antivirus, antimicrobial, cytotoxic, phytotoxic, anti-flammatory, and brine shrimp lethal activities. To date, two racemic and five enantioselective chemical total syntheses of 1 (including 2 and their isomers) have been developed. Over the past decade, a number of biosynthetic gene clusters or their mutants, along with their encoding enzymes responsible for producing sesterterpenes such as terpestacin and its derivatives, have also been identified. This review covers the literature from the year 1993, when 1 was firstly discovered, to May 2024, focusing on the bioactivities and syntheses of 1 and its derivatives or isomers.

Biosynthesis of (−)‐Vinigrol

(−)‐Vinigrol, as one of the most complex and challenging molecules in total synthesis, the parallel biosynthetic strategy employed by nature for the synthesis of this compound has not yet been identified. Here, we identified a minimal gene cluster encoding a diterpene cyclase (VniA) and a cytochrome P450 (VniB), which completes the synthesis of (−)‐vinigrol through three steps. VniA first cyclizes geranylgeranyl diphosphate to generate an unusual vinigrol‐type diterpene skeleton, and then VniB catalyzes the allylic C(sp3)‐H iterative oxidation. Further genome mining investigation provides new fungal sources for this rare and valuable vinigrol‐type diterpene skeleton.

Polyphasic Identification of Rhizomonospora bruguierae gen. nov., sp. nov., Isolated from Mangrove Rhizosphere Soil.

A novel yellowish-white actinobacterial strain, designated as TS60-4CT, was isolated from the rhizosphere soil of the mangrove Bruguiera gymnorhiza (L.) Lam. in Okinawa, Japan, and was subjected to a polyphasic assessment. The strain could grow at 0-3.0% NaCl concentrations (w/v), pH 6.0-10.0, and 20-37°C. The 16S rRNA gene sequences-based phylogeny showed that the novel isolate belongs to the family Micromonosporaceae and that it shared the highest sequence similarity (98.4%) with Micromonospora craniellae LHW63014T. The hydrolysate of cell-wall of strain TS60-4CT contained alanine, glutamic acid, glycine, and meso-A2pm. The acyl type of muramic acid was N-glycolyl. The main (> 10%) fatty acids were anteiso-C17:0 (22.1%), iso-C16:0 (17.8%), iso-C15:0 (14.7%), and anteiso-C15:0 (10.3%) and the predominant menaquinones of the isolate were MK-9(H6) and MK-9(H4). The polar lipids of the isolate were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, an unidentified aminolipid, and three unidentified lipids. The genome size of strain TS60-4CT was 6.7 Mbp with a DNA G + C content of 72.4%. The average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity value could differentiate strain TS60-4CT from its closely related taxa. The genome of the strain had 22 putative biosynthetic gene clusters for secondary metabolites, indicating a significant potential as a producer of bioactive chemicals. Consequently, the strain is considered to represent a novel genus and a new species of the family Micromonosporaceae, for which the name Rhizomonospora bruguierae gen. nov., sp. nov. is proposed. The type species is R. bruguierae, with the type strain TS60-4CT (= NBRC 107566T = TBRC 2024T).

Biosynthesis of (-)-Vinigrol.

(-)-Vinigrol, as one of the most complex and challenging molecules in total synthesis, theparallel biosynthetic strategy employed by nature for the synthesis of this compound has not yet been identified. Here, we identified a minimal gene cluster encoding a diterpene cyclase (VniA) and a cytochrome P450 (VniB), which completes the synthesis of (-)-vinigrol through three steps. VniA first cyclizes geranylgeranyl diphosphate to generate an unusual vinigrol-type diterpene skeleton, and then VniB catalyzes the allylic C(sp3)-H iterative oxidation. Further genome mining investigation provides new fungal sources for this rare and valuable vinigrol-type diterpene skeleton.