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Metagenomic binning is an essential technique for genome-resolved characterization of uncultured microorganisms in various ecosystems but hampered by the low efficiency of binning tools in adequately recovering metagenome-assembled genomes (MAGs). Here, we introduce BASALT ( B inning A cross a S eries of A ssemb l ies T oolkit) for binning and refinement of short- and long-read sequencing data. BASALT employs multiple binners with multiple thresholds to produce initial bins, then utilizes neural networks to identify core sequences to remove redundant bins and refine non-redundant bins. Using the same assemblies generated from Critical Assessment of Metagenome Interpretation (CAMI) datasets, BASALT produces up to twice as many MAGs as VAMB, DASTool, or metaWRAP. Processing assemblies from a lake sediment dataset, BASALT produces ~30% more MAGs than metaWRAP, including 21 unique class-level prokaryotic lineages. Functional annotations reveal that BASALT can retrieve 47.6% more non-redundant opening-reading frames than metaWRAP. These results highlight the robust handling of metagenomic sequencing data of BASALT. Binning is an essential step in genome-resolved metagenomic analysis in which assembled contigs originating from the same source population are clustered. However it is challenging, especially for low abundance microbial species. Here the authors introduce a toolkit that integrates multiple prominent binning tools and AI for efficient and high-resolution recovery of non-redundant bins from short- and long-read metagenomic sequencing datasets.

The salt lake ecosystem, characterized by extreme environmental gradients, harbors microbes that uniquely adapt to high salt stress through natural selection. However, how abiotic and biotic factors shape the microbial community assembly in Yuncheng Salt Lakes remains unclear. Here, we investigated the assembly processes and meta co-occurrence patterns of microbiota in both water and sediment sampled from 14 distinct wide range of salinity lakes in the Shanxi Yuncheng area, Midwest of China, using 16S rRNA and 18S rRNA gene sequencing technology combined with multivariate ecological and statistical methods. Habitat differentiation led to the differences in microbial diversity, co-occurrence patterns, and community assembly between sedimentary and planktonic communities. Sedimentary prokaryotes were more shaped by deterministic processes than planktonic bacterial communities. Salinity was a major abiotic factor influencing the balance between stochastic and deterministic processes in both sediment and water. Enhanced microbial interactions within sediments exhibited a more prominent impact in shaping community assembly, as indicated by the stronger association between network-inferred species and prokaryotic βNTI. Moreover, we revealed significant differences in how core species concerning βNTI responded to biotic and abiotic factors. Our findings elucidated the ecological process underlying microbial communities in Yuncheng Salt Lakes and shed light on the mechanism of microorganisms to maintain community complexity and diversity in the extreme environment. Key points • Sedimentary prokaryotes were more shaped by deterministic processes than planktonic prokaryotic communities. • Salinity was a major factor influencing the balance between stochastic and deterministic process. • Inter-domain and intra-domain symbiotic interactions within sedimentary communities represent key biotic factors influencing their community assembly. Graphical abstract

Alkaline lakes are extreme environments inhabited by diverse microbial extremophiles. However, large-scale distribution patterns, environmental adaptations, community assembly, and evolutionary dynamics of microbial communities remain largely underexplored. This study investigated the characteristics of microbial communities on rare and abundant taxa in alkaline lake sediments in west and northwest China. We observed that abundant taxa varied significantly with geographical distance, while rare taxa remained unaffected by regional differences. The assembly process of abundant taxa was influenced by dispersal limitation, whilst rare taxa were predominantly driven by heterogeneous selection. Network analysis indicated that rare taxa as core species for community interactions and community stability. Rare taxa exhibited higher speciation and transition rate than abundant taxa, serving as a genetic reservoir and potential candidates to become abundance taxa, highlighting their crucial role in maintaining microbial diversity. These insights underscore the significant influence of rare taxa on ecosystem biodiversity and stability in alkaline lakes.

Intestinal bacterial communities are highly relevant to the digestion, nutrition, growth, reproduction, and immunity of animals, but little is known about the composition and function of intestinal microbiota in deep-sea invertebrates. In this study, the intestinal microbiota of six holothurian Molpadia musculus were investigated, showing that their midguts were predominantly occupied by Izemoplasmatales bacteria. Using metagenomic sequencing, a draft genome of 1,822,181 bp was successfully recovered. After comparison with phylogenetically related bacteria, genes involved in saccharide usage and de novo nucleotide biosynthesis were reduced. However, a set of genes responsible for extracellular nucleoside utilization and 14 of 20 amino acid synthesis pathways were completely retained. Under oligotrophic condition, the gut-associated bacterium may make use of extracellular DNA for carbon and energy supplement, and may provide essential amino acids to the host. The clustered regularly interspaced short palindromic repeat (CRISPR) and restriction–modification (RM) systems presented in the genome may provide protection against invading viruses. A linear azol(in)e-containing peptide gene cluster for bacteriocin synthesize was also identified, which may inhibit the colonization and growth of harmful bacteria. Known virulence factors were not found by database searching. On the basis of its phylogenetic position and metabolic characteristics, we proposed that the bacterium represented a novel genus and a novel family within the Izemoplasmatales order and suggested it be named “Candidatus Bathyoplasma sp. NZ”. This was the first time describing host-associated Izemoplasmatales.

In this study, the complete mitochondrial DNA sequence of a hagfish Eptatretus sp. Nan-Hai from a depth of 1000 m is presented. The complete sequence was determined using next-generation sequencing and long PCRs. The mitochondrial genome of Eptatretus sp. Nan-Hai is 17,538 bps in size and composed of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one control region (D-loop). The base composition of mitochondrial genome is biased toward A + T content, at 67.21%, with GC skew of −0.35 and AT skew of −0.03. A phylogenetic tree revealed that within the genus Eptatretus, Eptatretus sp. Nan-Hai is closely related to Eptatretus atami.

Abstract Metagenomic binning enables the in-depth characterization of microorganisms. To improve the resolution and efficiency of metagenomic binning, BASALT (Binning Across a Series of AssembLies Toolkit), a novel binning toolkit was present in this study, which recovers, compares and optimizes metagenomic assembled genomes (MAGs) across a series of assemblies from short-read, long-read or hybrid strategies. BASALT incorporates self-designed algorithms which automates the separation of redundant bins, elongate and refine best bins and improve contiguity. Evaluation using mock communities revealed that BASALT auto-binning obtained up to 51% more number of MAGs with up to 10 times better MAG quality from microbial community at low (132 genomes) and medium (596 genomes) complexity, compared to other binners such as DASTool, VAMB and metaWRAP. Using BASALT, a case-study analysis of a Salt Lake sediment microbial community from northwest arid region of China was performed, resulting in 426 non-redundant MAGs, including 352 and 69 bacterial and archaeal MAGs which could not be assigned to any known species from GTDB (ANI < 95%), respectively. In addition, two Lokiarchaeotal MAGs that belong to superphylum Asgardarchaeota were observed from Salt Lake sediment samples. This is the first time that candidate species from phylum Lokiarchaeota was found in the arid and deep-inland environment, filling the current knowledge gap of earth microbiome. Overall, BASALT is proven to be a robust toolkit for metagenomic binning, and more importantly, expand the Tree of Life. Competing Interest Statement The authors have declared no competing interest. Footnotes * ↵* Contact Information: Name: Ke Yu, Address: Shenzhen Graduate School, Peking University, Lishui Road, Nanshan district, Shenzhen, Email: yuke.szatpku.edu.cn * Abstract updated to make statement clearer.

以生活在同一生境,但具有不同进化关系和不同食性的野生哺乳类动物(鼠科、猬科和鼩鼱科)为研究对象,通过16S rRNA基因扩增子测序,分析和比较其肠道菌群.共识别出5378个操作分类单位(OTUs),主要隶属 Firmicutes(40.55％),Proteobacteria(34.60％)和 Bacteroidetes(13.67％).Firmicutes 和 Bacteroidetes 是鼠科的优势菌门,Proteobacteria是猬科和鼩鼱科的优势菌门.多样性分析表明,鼠科、猬科与鼩鼱科的肠道微生物多样性及群落结构具有显著性差异;LEfSe分析表明,鼠科中存在更多与复杂碳水化合物发酵相关的细菌,猬科和鼩鼱科中含较多氨基酸发酵菌;益生菌(如Lactobacillus和Lactococcus等)共存于这3类野生小型哺乳类动物中,起调控宿主健康的作用.研究结果揭示,宿主食性与进化关系影响着野生小型哺乳动物肠道菌群结构,而肠道菌群可能在多种方面对宿主起益生作用.