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Theionarchaea is a recently described archaeal class within the Euryarchaeota. While it is widely distributed in sediment ecosystems, little is known about its metabolic potential and ecological features. Here, we used metagenomics and metatranscriptomics to characterize 12 theionarchaeal metagenome-assembled genomes, which were further divided into two subgroups, from coastal mangrove sediments of China and seawater columns of the Yap Trench. Genomic analysis revealed that apart from the canonical sulfhydrogenase, Theionarchaea harbor genes encoding heliorhodopsin, group 4 [NiFe]-hydrogenase, and flagellin, in which genes for heliorhodopsin and group 4 [NiFe]-hydrogenase were transcribed in mangrove sediment. Further, the theionarchaeal substrate spectrum may be broader than previously reported as revealed by metagenomics and metatranscriptomics, and the potential carbon substrates include detrital proteins, hemicellulose, ethanol, and CO2. The genes for organic substrate metabolism (mainly detrital protein and amino acid metabolism genes) have relatively higher transcripts in the top sediment layers in mangrove wetlands. In addition, co-occurrence analysis suggested that the degradation of these organic compounds by Theionarchaea might be processed in syntrophy with fermenters (e.g., Chloroflexi) and methanogens. Collectively, these observations expand the current knowledge of the metabolic potential of Theionarchaea, and shed light on the metabolic strategies and roles of these archaea in the marine ecosystems.

This study explores the comparative effect of conventional and organic treatments on the rhizosphere microbiome of Mangifera indica cv. Dashehari. The long-term exposures (about 20 years) were monitored under a subtropical ecosystem. Based on plant growth properties and acetylene reduction assay, 12 bacterial isolates (7 from G1-organic and 5 from G2-conventional systems) were identified as Pseudomonas and Bacillus spp. In the conventional system, dehydrogenase activity significantly decreased (0.053 µg TPF formed g−1 of soil h−1) and adversely affected the bacterial diversity composition. In comparison, organic treatments had a good impact on dehydrogenase activity (0.784 µg TPF formed g−1 of soil h−1), alkaline phosphatase (139.25 µg PNP g−1 soil h−1), and bacterial community composition. The Metagenomics approach targeted the V3 and V4 regions to see the impact in the phylum, order, family, genus, and species for both the treatments. Results showed that phylum Acidobacteria (13.6%), Firmicutes (4.84%), and Chloroflexi (2.56%) were dominating in the G2 system whereas phylum Bacteroides (14.55%), Actinobacteria (7.45%), and Proteobacteria (10.82%) were abundantly dominated in the G1 system. Metagenome sequences are at the NCBI-GenBank sequence read archive with SRX8289747 (G1) and SRX8289748 (G2) in the study PRJNA631113. Results indicated that conventional and organic conditions affect rhizosphere microbiome and their environment.

Recently discovered fields of seeps in western Siberia are an important, previously unknown source of methane. Seeps are located in the floodplains of small rivers and vary in shape, size and localization. Methane fluxes from seeps may be high, making them the sources of high regional importance. However, the data on the seep bacterial community composition are scarce, concentrating mainly on methanotrophic bacteria. In the present work, the overall bacterial diversity in the sediments of shallow-water seeps at the floodplain of the Bolshaya Rechka River was studied using high-throughput 16S rRNA gene sequencing. Molecular analysis revealed that Gammaproteobacteria and Actinobacteria were the most abundant bacterial groups (28.5–33.8 and 11–13.2% of total 16S rRNA genes, respectively). A significant part of the sequences belonged to Chloroflexi, Desulfobacterota, and Bacteroidota. Acidobacteriota and Verrucomicrobiota were responsible for 1.5–2.7 and 1.0–1.9%, respectively. The methanotrophic community was dominated by bacteria of the genus Methylobacter. The most numerous species-level OTU (8% of all 16S rRNA gene sequences) belonged to Methylobacter tundripaludum (97% identity). Methanotrophic Alphaproteobacteria were not detected in the seeps. Our results confirm the presence of an active bacterial community in the sediments of shallow-water seeps, with predominance of methanotrophic Gammaproteobacteria.

In the present study, we have identified an ω-transaminase (ω-TA) from Chloroflexi bacterium from the genome database by using two ω-TA sequences (ATA117 Arrmut11 from Arthrobacter sp. KNK168 and amine transaminase from Aspergillus terreus NIH2624) as templates in a BLASTP search and motif sequence alignment. The protein sequence of the ω-TA from C. bacterium (CbTA) shows 38% sequence identity to that of ATA117 Arrmut11. The gene sequence of CbTA was inserted into pRSF-Duet1 and functionally expressed in Escherichia coli BL21(DE3). The results showed that the recombinant CbTA has a specific activity of 1.19 U/mg for (R)-α-methylbenzylamine [(R)-MBA] at pH 8.5 and 45 °C. The substrate acceptability test showed that CbTA has significant reactivity to aromatic amino donors and amino receptors. More importantly, CbTA also exhibited good affinity toward some cyclic substrates. The homology model of CbTA was built by Discovery Studio, and docking was performed to describe the relative activity toward some substrates. CbTA evolved by site-specific mutagenesis and found that the Q192G mutant increased the activity to (R)-MBA by around 9.8-fold. The Q192G mutant was then used to convert two cyclic ketones, N-Boc-3-pyrrolidinone and N-Boc-3-piperidone, and both the conversions were obviously improved compared to that of the parental CbTA.

Traditionally, filamentous fungi and actinomycetes are well-known cellulolytic microorganisms that have been utilized in the commercial production of cellulase enzyme cocktails for industrial-scale degradation of plant biomass. Noticeably, the Ktedonobacteria lineage (phylum Chloroflexi) with actinomycetes-like morphology was identified and exhibited diverse carbohydrate utilization or degradation abilities. In this study, we performed genome-wide profiling of carbohydrate-active enzymes (CAZymes) in the filamentous Ktedonobacteria lineage. Numerous CAZymes (153–290 CAZymes, representing 63–131 glycoside hydrolases (GHs) per genome), including complex mixtures of endo- and exo-cellulases, were predicted in 15 available Ktedonobacteria genomes. Of note, 4–28 CAZymes were predicted to be extracellular enzymes, whereas 3–29 CAZymes were appended with carbohydrate-binding modules (CBMs) that may promote their binding to insoluble carbohydrate substrates. This number far exceeded other Chloroflexi lineages and were comparable to the cellulolytic actinomycetes. Six multi-modular extracellular GHs were cloned from the thermophilic Thermosporothrix hazakensis SK20-1T strain and heterologously expressed. The putative endo-glucanases of ThazG5-1, ThazG9, and ThazG12 exhibited strong cellulolytic activity, whereas the putative exo-glucanases ThazG6 and ThazG48 formed weak but observable halos on carboxymethyl cellulose plates, indicating their potential biotechnological application. The purified recombinant ThazG12 had near-neutral pH (optimal 6.0), high thermostability (60°C), and broad specificity against soluble and insoluble polysaccharide substrates. It also represented described a novel thermostable bacterial β-1,4-glucanase in the GH12 family. Together, this research revealed the underestimated cellulolytic potential of the Ktedonobacteria lineage and highlighted its potential biotechnological utility as a promising microbial resource for the discovery of industrially useful cellulases.

Bacteria play an important role in pollutant transformation in activated sludge-based wastewater treatment plants (WWTPs). Exploring the microbial community structure and diversity is essential to improving the performance of wastewater treatment processes. This study employed Illumina MiSeq high-throughput sequencing to investigate the microbial community composition and diversity in a cattle farm wastewater treatment plant (Cf-WWTP). The results showed that the dominant phyla in the whole process were Proteobacteria, Bacteroidetes, and Firmicutes. The principal coordinate analysis (PCoA) indicated that the different stages had a significant impact on the microbial community structure; Bacteroidetes was the dominant phylum in the anearobic stage and Proteobacteria was the dominant phylum in the anoxic-oxic stage. Redundancy analysis (RDA) revealed that total phosphorus (TP) was the most significant factor that regulated the microbial community composition, followed by chemical oxygen demand (COD), total nitrogen (TN), and pH. Proteobacteria, Patescibacteria, and Chloroflexi were simultaneously negatively correlated with TN, COD, and TP. Nitrogen metabolic pathway and transformation mechanism was elucidated by a complete denitrification function predicted with phylogenetic investigation of communities with reconstruction of unobserved states (PICRUSt), as well as detection of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). These results provide new insights into our understanding of microbial community and metabolic functions of Cf-WWTP.

To improve our understanding of early microbial colonization of pristine minerals and their group-specific C utilization, we exposed minerals (illite/goethite/quartz) amended with artificial root exudates (ARE, glucose, and citric acid) in grassland soils for a period of 24 weeks. FTIR spectra indicated that mineral-associated ARE were used within the first 2 weeks of exposure and were replaced by other carbohydrates derived from living or dead cells as well as soil-borne C sources transported into the mineralosphere after heavy rain events. Fungi and Gram-positive bacteria incorporated ARE-derived C more rapidly than Gram-negative bacteria. Gram-negative bacteria presumably profited indirectly from the ARE by cross-feeding on mineral-associated necromass of fungi and Gram-positive bacteria. The Gram-negative bacterial phyla Verrucomicrobia, Planctomycetes, Gemmatimonadetes, Armatimonadetes, and Chloroflexi showed a positive correlation with Gram-negative PLFA abundances. After 24 weeks of exposure in the grassland soils, abundances of soil microorganisms in the mineralosphere reached only 3.1% of the population density in soil. In conclusion, both bacteria and fungi slowly colonize new surfaces such as pristine minerals, but quickly assimilate artificial root exudates, creating an active microbial community in the mineralosphere.

In this study, we explored the diversity and community composition of bacteria along a vertical gradient in Lake Issyk Kul, Kyrgyzstan, one of the largest and deepest brackish lakes in in the world. We identified 4904 bacterial ASVs based on analysis of 16S rRNA gene sequences and determined significant changes in the composition responding mainly to the variables depth and salinity. A higher abundance of Proteobacteria and Bacteroidetes was observed in the surface waters and the lake tributaries. Cyanobacteria were more abundant in the deep chlorophyll maximum (28.5 to 128 m), while Planctomycetes and Chloroflexi were dominant at depths between 128 to 600 m. According to our machine learning analysis used for identifying the most critical environmental factors, depth and temperature revealed the strongest effect on members of Proteobacteria, Planctomycetes, and Chloroflexi, while oxygen is associated with the variations in Cyanobacteria. Also, a notable increase in alpha diversity estimations was observed with increasing water depth. This work evidences significant differences in the structure of bacterial communities along the depth gradient in deep, transparent lake ecosystems. Notably, there is a dominance of Planctomycetes and Chloroflexi in the deepest layers, which can only be seen in a few other lakes with similar characteristics as Lake Issyk Kul and raises questions about their ecological role.

The stages of formation and development of activated sludge biofilm carrying out the anammox process in a laboratory sequencing batch reactor (SBR) with complete biomass retention on the carrier were investigated using light, epifluorescence, and electron microscopy. Light microscopy revealed biofilm formation on the carrier to occur within one week. Rod-shaped and filamentous microorganisms were the first to attach on the carrier and acted as a skeleton for biofilm formation, thus playing an important role in colonization of the carrier. Epifluorescence microscopy revealed physiologically active anammox bacteria of the genera Candidatus “Brocadia” and Ca. “Jettenia” in the biofilms. Selective autofluorescence of the colonies of anammox bacteria Ca. “Jettenia” was observed. This autofluorescence was probably caused by specific proteins in the extracellular polymeric substances (EPS), their synthesis and/or amount depending on the colony age and the physiological state of the cells. Structural organization of the colonies of anammox bacteria was investigated by electron microscopy. High-throughput sequencing of the 16S rRNA gene fragments revealed the presence of sequences affiliated with members of the phyla Chloroflexi, Bacteroidetes, Planctomycetes, and Proteobacteria in the biofilms. Apart from anammox bacteria, anaerobic and facultatively anaerobic organotrophs, stage I nitrifiers, denitrifiers, and sulfate reducers were detected. Members of the genus Ca. “Brocadia” were predominant among the anammox bacteria, probably due to better adhesion of their cells to the carrier or to their competitive advantage over Ca. “Jettenia” in the presence of organic acids (acetate and formate) in the medium.

Caves are considered to be extreme and challenging environments. It is believed that the ability of microorganisms to produce secondary metabolites enhances their survivability and adaptiveness in the energy-starved cave environment. Unfortunately, information on the genetic potential for the production of secondary metabolites, such as polyketides and nonribosomal peptides, is limited. In the present study, we aimed to identify and characterize genes responsible for the production of secondary metabolites in the microbial community of one of the deepest caves in the world, Krubera-Voronja Cave (43.4184 N 40.3083 E, Western Caucasus). The analysed sample materials included sediments, drinkable water from underground camps, soil and clay from the cave walls, speleothems and coloured spots from the cave walls. The type II polyketide synthases (PKSs) ketosynthases α and β and the adenylation domains of nonribosomal peptide synthetases (NRPSs) were investigated using a metagenomic approach. Taxonomic diversity analysis showed that most PKS sequences could be attributed to Actinobacteria followed by unclassified bacteria and Acidobacteria, while the NRPS sequences were more taxonomically diverse and could be assigned to Proteobacteria, Actinobacteria, Cyanobacteria, Firmicutes, Chloroflexi, etc. Only three putative metabolites could be predicted: an angucycline group polyketide, a massetolide A-like cyclic lipopeptide and a surfactin-like lipopeptide. The absolute majority of PKS and NRPS sequences showed low similarity with the sequences of the reference biosynthetic pathways, suggesting that these sequences could be involved in the production of novel secondary metabolites.

Uncultured members of the Chloroflexi phylum are highly enriched in numerous subseafloor environments. Their metabolic potential was evaluated by reconstructing 31 Chloroflexi genomes from six different subseafloor habitats. The near ubiquitous presence of enzymes of the Wood–Ljungdahl pathway, electron bifurcation, and ferredoxin‐dependent transport‐coupled phosphorylation indicated anaerobic acetogenesis was central to their catabolism. Most of the genomes simultaneously contained multiple degradation pathways for complex carbohydrates, detrital protein, aromatic compounds, and hydrogen, indicating the coupling of oxidation of chemically diverse organic substrates to ubiquitous CO₂ reduction. Such pathway combinations may confer a fitness advantage in subseafloor environments by enabling these Chloroflexi to act as primary fermenters and acetogens in one microorganism without the need for syntrophic H₂ consumption. While evidence for catabolic oxygen respiration was limited to two phylogenetic clusters, the presence of genes encoding putative reductive dehalogenases throughout the phylum expanded the phylogenetic boundary for potential organohalide respiration past the Dehalococcoidia class.

A previous study indicated that gut microbiota changed notably in Graves’ orbitopathy (GO) patients as compared to controls. However, the characteristics of intestinal bacteria in Graves’ disease (GD) and GO are unclear. The present study aimed to identify specific intestinal bacteria of GD and GO, respectively. The gut microbial communities of the fecal samples of 30 GD patients without GO, 33 GO subjects, and 32 healthy subjects were analyzed and compared by 16S rRNA gene sequencing. At the phylum level, the proportion of Deinococcus-Thermus and Chloroflexi was decreased significantly in GO patients as compared to GD. At the genus level, the proportion of Subdoligranulum and Bilophila was increased while that of Blautia, Anaerostipes, Dorea, Butyricicoccus, Romboutsia, Fusicatenibacter, unidentified_ Lachnospiraceae, unidentified_Clostridiales, Collineslla, Intestinibacter, and Phascolarctobacterium was decreased in the GO group as compared to the GD group. Random forest analysis was used for the identification of specific intestinal microbiota, and Deinococcus-Thermus, Cyanobacteria and Chloroflexi were ranked in the top ten according to their contributions to sample classification. Moreover, compared to the control, there were multiple gut bacterial enrichment metabolic pathways in GO and GD patients, including nucleotide metabolism, enzyme family, and energy metabolism. Compared to GO, the only enrichment metabolic pathway found in GD was the viral protein family. This study highlighted the significant differences in the intestinal microbiota and predictive functions of GD with GO, thereby providing new insights into the role of the gut bacteria that might contribute to the development of GO in GD patients.

All purple photosynthetic bacteria contain RC–LH1 ‘Core’ complexes. The structure of this complex from Rhodobacter sphaeroides, Rhodopseudomonas palustris and Thermochromatium tepidum has been solved using X-ray crystallography. Recently, the application of single particle cryo-EM has revolutionised structural biology and the structure of the RC–LH1 ‘Core’ complex from Blastochloris viridis has been solved using this technique, as well as the complex from the non-purple Chloroflexi species, Roseiflexus castenholzii. It is apparent that these structures are variations on a theme, although with a greater degree of structural diversity within them than previously thought. Furthermore, it has recently been discovered that the only phototrophic representative from the phylum Gemmatimonadetes, Gemmatimonas phototrophica, also contains a RC–LH1 ‘Core’ complex. At present only a low-resolution EM-projection map exists but this shows that the Gemmatimonas phototrophica complex contains a double LH1 ring. This short review compares these different structures and looks at the functional significance of these variations from two main standpoints: energy transfer and quinone exchange.

BACKGROUND: Microbes that govern a unique biochemical process of oxidizing ammonia into dinitrogen gas, such as anaerobic ammonium oxidation (anammox) have been reported to play a pivotal role in agricultural soils and in oceanic environments. However, limited information for anammox bacterial abundance and distribution in the terrestrial habitats has been known. METHODS AND RESULTS: Phylogenetic and next-generation sequencing analyses of bacterial 16S rRNA gene were performed to examine potential anammox bacteria in paddy soils. Through clone libraries constructed by using the anammox bacteria-specific primers, some clones showed sequence similarities with Planctomycetes (87% to 99%) and anammox bacteria (94% to 95%). Microbial community analysis for the paddy soils by using Illumina Miseq sequencing of 16S rRNA gene at phylum level was dominated by unclassified Bacteria at 33.2 ± 7.6%, followed by Chloroflexi at 20.4 ± 2.0% and Acidobacteria at 17.0 ± 6.5%. Planctomycetes that anammox bacteria are belonged to was 1.5% (± 0.3) on average from the two paddy soils. CONCLUSION: We suggest evidence of anammox bacteria in the paddy soil. In addition to the relatively well-known microbial processes for nitrogen-cycle, anammox can be a potential contributor on the cycle in terrestrial environments such as paddy soils.

Phytoplankton growth and degradation are associated with bacterial communities in phycosphere. We compared the bacterial communities in two algal blooms (Microcystis aeruginosa and Euglena sanguinea). In particular, large particle-associated, small particle-associated, and free-living bacteria were collected in succession using membranes with three different pore sizes (10 µm, 1 µm, and 0.22 µm) in this study. The results showed that Proteobacteria and Bacteroidetes were the dominant phyla in both blooms. Significant differences in the structure, function, and assembly were observed between large particle-associated and free-living bacterial community in both blooms. Specifically, relative abundance of Roseomonas was higher in the large-size (> 10 µm) aggregates, while Candidate_division_TM7 and Candidate_division_SR1 were more abundant in the free-living bacterial community (0.22–1 µm). The large particle-associated bacterial community was mainly driven by heterogeneous selection and dispersal limitation, whereas the small particle-associated and free-living bacterial communities were mostly determined by dispersal limitation. Besides, some specific bacterial subgroups were found between M. aeruginosa and E. sanguinea blooms. Chlorobi, Chloroflexi, and Fusobacteria were abundant in the M. aeruginosa blooms, whereas Deinococcus–Thermus was abundant in the E. sanguinea blooms. Those specific bacteria provide competitive advantages for the dominated algae. Altogether, these findings indicate that the phycosphere of bloom-forming algae provides different ecological niches where different functional bacterial groups can adapt.

In this study, a yeast strain with an outstanding NH3-N degradation ability was isolated from the sediment of a black-odor water channel in Guangdong Province, China. Based on phenotypic and phylogenetic analysis, this strain was identified as Pichia kudriavzevii GW1. The optimum conditions for NH3-N degradation by the GW1 strain were as follows: 0.3% inoculum concentration, 1.5 L/min aeration, pH 7, and a temperature of 35°C. Under optimized conditions, the GW1 strain degraded 95.5% of the NH3-N. The strain was then added to simulated black-odor water under optimal degradation conditions to investigate changes to the bacterial community over time. 16S rRNA sequencing of samples collected on days 0, 7, 14, and 21 showed that, in the presence of the GW1 strain, the relative abundances of the phyla Proteobacteria, Bacteroidetes, Chloroflexi, and Firmicutes increased in the black-odor water. In addition, the relative abundance of Propionivibrio, a known NH3-N degrading genus, increased. This study will facilitate the use of microbiological methods to repair black-odor water.

The trace amounts (0.53 ppmv) of atmospheric hydrogen gas (H2) can be utilized by microorganisms to persist during dormancy. This process is catalyzed by certain Actinobacteria, Acidobacteria, and Chloroflexi, and is estimated to convert 75 × 1012 g H2 annually, which is half of the total atmospheric H2. This rapid atmospheric H2 turnover is hypothesized to be catalyzed by high-affinity [NiFe] hydrogenases. However, apparent high-affinity H2 oxidation has only been shown in whole cells, rather than for the purified enzyme. Here, we show that the membrane-associated hydrogenase from the thermoacidophilic methanotroph Methylacidiphilum fumariolicum SolV possesses a high apparent affinity (Km(app) = 140 nM) for H2 and that methanotrophs can oxidize subatmospheric H2. Our findings add to the evidence that the group 1h [NiFe] hydrogenase is accountable for atmospheric H2 oxidation and that it therefore could be a strong controlling factor in the global H2 cycle. We show that the isolated enzyme possesses a lower affinity (Km = 300 nM) for H2 than the membrane-associated enzyme. Hence, the membrane association seems essential for a high affinity for H2. The enzyme is extremely thermostable and remains folded up to 95 °C. Strain SolV is the only known organism in which the group 1h [NiFe] hydrogenase is responsible for rapid growth on H2 as sole energy source as well as oxidation of subatmospheric H2. The ability to conserve energy from H2 could increase fitness of verrucomicrobial methanotrophs in geothermal ecosystems with varying CH4 fluxes. We propose that H2 oxidation can enhance growth of methanotrophs in aerated methane-driven ecosystems. Group 1h [NiFe] hydrogenases could therefore contribute to mitigation of global warming, since CH4 is an important and extremely potent greenhouse gas.

The Ikogosi Warm Spring is a unique ecological niche in Western Nigeria with an average temperature and pH of 38°C and 5.8 respectively. It mixes with an adjacent cold spring (28°C & pH 7.6), about 100 meters from source, yielding a confluence body of water of 32°C and pH 7.7. To explore the bacterial community structure of this uncommon environment and to scan for potentially useful bacteria, metagenomes extracted directly from five samples (source and mid-point of warm spring; source and midpoint of cold spring, and the confluence) were analyzed. Using the MiSeq Illumina next generation sequencing protocols, the V3-V4 region of the 16S rRNA gene pool was sequenced and analyzed by QIIME (Quantitative Insights into Microbial Ecology) and R software. At least 11% (47,446) of all the sequences were unknown to any of the databases employed. Bacterial diversity and abundance at the source of both springs were extremely low, accounting for less than 0.07% of the total sequence reads at the confluence, 100 m downstream. In contrast to the highly diversified mesophilic confluence community where 21 different phyla were identified, only 4 and 5 phyla were recovered from the source-point of the warm spring and cold spring respectively. The most prevalent phyla in all samples were members of the versatile Proteobacteria (35% - 50% relative abundance), and the hardy Firmicutes (33% - 40%). Operational taxonomic units (OTUs) obtained from all the samples averaged at 1414. Temperature and pH were equally significant predictors of genomic diversity and richness, with the warm and cold spring sources having less than 5 bacteria phyla. Exiguobacterium sp. (a potential plastic degrader) and other deep rooted bacteria were found in the warm spring while the cold spring outflow contained among others such as Rubrobacter sp. and Chloroflexi sp. (which is close to the phylogenetic root of the domain Bacteria). Many taxonomically unresolved sequences could indicate the presence of potentially novel bacteria in this unique body of water and underscores the need to systematically mine these rare genetic reservoirs for biotechnological applications. Moreover, such tropical hydrothermal ecosystems could contain some unknown primitive bacteria at the origins of life.

The present study investigated the disregarded impact of microbial community composition, diversity and abundance of the starter biomass on lab-scale bioreactor performance, steady-state long time operation, biodegradation outcome and microbial kinetics. Standard lab-scale experiments are usually initiated with inoculum taken from wastewater treatment plants (WWTPs) without considering microbial community composition. Two identical sequencing batch bioreactors (SBR-1, SBR-2) were operated with inocula obtained from two domestic WWTPs to assess the role of inoculum composition on reactor performance and treatment efficacy. Filamentous microorganisms were dominated in SBR-1 (SVI: 265 mL/g), whereas their supremacy was diminished in SBR-2 (SVI: 160 mL/g). Next generation sequencing analyses verified high abundance of Candidatus Microthrix (14%) and Kouleothrix spp. (3%) in SBR-1. Although Chloroflexi C10-SB1A metagenome accounted for 16% of WWTP-2 community, SBR-2 was operated at steady-state for longer period without deterioration. Statistical indicators based on molecular studies showed that WWTP-2 has higher diversity than WWTP-1. Higher biodiversity in WWTP-2 community supported the stable process performance. Modelling studies denoted an increase in half saturation coefficients (Ks) in filamentous microorganisms dominated reactor. The results of this study provide insight on how the filamentous organisms typically found in WWTPs become a fundamental constituent during the bioreactor operation.

Nonribosomal peptides (NRPs) and polyketides (PKs) have widely been widely applied in clinical drugs in use today, and one of the main sources for novel NRPs and PKs is soil microbes. However, the diversity of soil bacteria and distribution of their NRPs and PKs biosynthetic genes in different landscapes in extreme environments have not been reported. In this study, 16S rRNA gene fragments from five landscape belts in Changbaishan were analyzed. The results demonstrated that Acidobacteria, Proteobacteria and Chloroflexi accounted for 65% of all bacterial species. Furthermore, the diversities of the PKs and NRPs in the different landscape belts of Changbaishan were compared based on the diversity of gene fragments of the polyketide synthase (PKs) ketosynthase (KS) domain and nonribosomal peptide synthetase (NRPs) adenylation domain (AD). The results showed that NRPs and PKs had highest richness in alpine tundra and birch forest belts, which were negatively correlated with the contents of the available phosphorus, nitrogen, calcium, potassium, and soil organic matter. NRPs and PKs mainly existed in unidentified bacteria, Proteobacteria and Actinobacteria. In addition, a 16S rRNA KEGG pathway analysis showed that the soil microorganisms of Changbaishan have the potential to promote secondary metabolite biosynthesis, resist diseases and regulate immune systems. These combined results suggest that soils of alpine tundra and birch forest belts in Changbaishan may be the main sources for searching the novel antibacterial drugs.

A novel bacterial α 1-2-fucosyltransferase (α 2FT) from Thermosynechococcus sp. NK55a (Ts2FT) has been discovered and characterized. It shares 28-62% protein sequence homology to α 2FTs reported previously. The Ts2FT was cloned as an N-terminal His6-tagged recombinant protein (His6-Ts2FT) and expressed in E. coli BL21 (DE3). It was expressed at a level of 6.2mg/L culture after induction with 0.05mM of isopropylβ-d-1-thiogalactoside (IPTG) at 16°C for 20h. It showed the optimal activity at a reaction temperature of 40°C and pH of 7.0. The presence of a Mg2+ improved its catalytic efficiency. Ts2FT displayed a strict acceptor specificity and could recognize only β1-3-galatoside acceptors. It was used efficiently for one-pot multienzyme synthesis of fucosylated oligosaccharides. One of the products, lacto-N-fucopentaose I was shown to promote the growth of intestinal probiotics including those belonging to Acidobacteria, Actinobacteria, Proteobacteria, Planctomycetes, and Chloroflexi.

In the present study, we explored four different geothermal spots of the Deulajhari spring cluster at a proximity of 10–20 meters with temperatures of 43 to 65°C to unravel their genesis, bacterial diversity and CAZyme potential. However, minor variations in physicochemical properties; TOC, sodium, chloride, zinc and nitrate were observed, including the pH of the spring openings. Illumina based amplicon sequencing revealed Firmicutes, Proteobacteria and Chloroflexi as the major bacterial phylum with higher abundance in the DJ04 sample. The alpha diversity of all the springs was almost same, whereas beta diversity revealed variations in the degree of uniqueness of OTUs at different temperatures. Statistical analysis established a positive correlation between sulfur content with Heliobacterium, Thermodesulfovibrio, Thermodesulfobacterium and Herpetosipho as well as TOC and HCO3 with Thermoanaerobacter, Desulfovibrio, Candidatus solibacter and Dehalogenimona. The major hydrocarbon family genes and Carbohydrate Active Enzyme pathways were predicted to be highest in DJ04 with elevated concentrations of HCO3 and TOC. Higher homogeneity in geo-physicochemical and microbial features direct the possibility of the common origin of these springs through plumbing systems. However, the minor variations in diversity and functionality were due to variations in temperature in spring openings through the mixing of subsurface water contaminated with carbohydrates from leaf biomass litter. Functional characterization of the thermophilic bacteria of this spring provides essential scope for further industrial applications. The biogeochemical reasons hypothesized for the genesis of unique multiple openings in the cluster are also of interest to conservation scientists for taking measures toward necessary laws and regulations to protect and preserve these springs.

Different rice farming systems affect the soil microbial communities. Here, we report the results of 16S rRNA gene amplicon sequencing of soils collected from intensive rice cultivation and rice-shrimp farming systems in Soc Trang, Vietnam. The dominant phyla in these systems were Firmicutes , Actinobacteriota , Chloroflexi , Myxococcota , and Acidobacteriota .

Microorganisms in clay barriers could affect the long-term performance of waste containers in future deep geological repositories (DGR) for used nuclear fuel through production of corrosive metabolites (e.g., sulfide), which is why clay materials are highly compacted: to reduce both physical space and access to water for microorganisms to grow. However, the highly compacted nature of clays and the resulting low activity or dormancy of microorganisms complicate the extraction of biomarkers (i.e., PLFA, DNA etc.) from such barriers for predictive analysis of microbial risks. In order to overcome these challenges, we have combined culture- and 16S rRNA gene amplicon sequencing-based approaches to describe the functional diversity of microorganisms in several commercial clay products, including two different samples of Wyoming type MX-80 bentonite (Batch 1 and Batch 2), the reference clay for a future Canadian DGR, and Avonlea type Canaprill, a clay sample for comparison. Microorganisms from as-received bentonites were enriched in anoxic 10% w/v clay microcosms for three months at ambient temperature with addition of 10% hydrogen along with presumable indigenous organics and sulfate in the clay. High-throughput sequencing of 16S rRNA gene fragments indicated a high abundance of Gram-positive bacteria of the phylum Firmicutes (82%) in MX-80 Batch 1 incubations. Bacterial libraries from microcosms with MX-80 Batch 2 were enriched with Firmicutes (53%) and Chloroflexi (43%). Firmicutes also significantly contributed (70%). Sequence analysis revealed presence of the bacterial families Peptostreptococcaceae, Clostridiaceae, Peptococcaceae, Bacillaceae, Enterobacteriaceae, Veillonellaceae, Tissierellaceae and Planococcaceae in MX-80 Batch 1 incubations; Bacillaceae, along with unidentified bacteria of the phylum Chloroflexi, in MX-80 Batch 2 clay microcosms, and Pseudomonadaceae, Hydrogenophilaceae, Bacillaceae, Desulfobacteraceae, Desulfobulbaceae, Peptococcaceae, Pelobacteraceae, Alcaligenaceae, Rhodospirillaceae in Canaprill microcosms. Exploration of potential metabolic pathways in the bacterial communities from the clay microcosms suggested variable patterns of sulfur cycling in the different clays with the possible prevalence of bacterial sulfate-reduction in MX-80 bentonite, and probably successive sulfate-reduction/sulfur-oxidation reactions in Canaprill microcosms. Furthermore, analysis of potential metabolic pathways in the bentonite enrichments suggested that bacteria with acid-producing capabilities (i.e., fermenters and acetogens) together with sulfide-producing prokaryotes might perhaps contribute to corrosion risks in clay systems. However, the low activity or dormancy of microorganisms in highly compacted bentonites as a result of severe environmental constraints (e.g., low water activity and high swelling pressure in the confined bentonite) in situ would be expected to largely inhibit bacterial activity in highly compacted clay-based barriers in a future DGR., Bentonite microcosms; Bacterial diversity; High-throughput sequencing; QIIME (quantitative insights into microbial ecology); PICRUST (phylogenetic investigation of communities by reconstruction of unobserved states); Potential bacterial activity.

International audience; Microbes are important component in terrestrial ecosystem, which are believed to play vital roles in biogeochemical cycles of metalloids in mining and smelting surroundings. Many studies on microbial diversity and structures have been investigated around mining and smelting sites, whereas the ecological processes and co-occurrence patterns that influence the biogeographic distributions of microbial communities is yet poorly understood. Herein, microbial biogeography, assembly mechanism and co-occurrence pattern around mining and smelting zone were systematically unraveled using 16S rRNA gene sequencing. The 66 microbial phyla co-occurring across all the samples were dominated by Proteobacteria, Chloroflexi, Acidobacteria and Crenarchaeota. Obvious distance-decay (r = 0.3448, p < 0.001) of microbial community was observed across geographic distances. Differences in microbial communities were driven by the joint impacts of soil factors, spatial and metalloids levels. Dispersal limitation dominated the microbial assemblies in whole, SC and GX sites while homogeneous selection governed that in YN site. The changes in pH and Sb level significantly influenced the deterministic and stochastic processes of microbial communities. Network analysis suggested a typical module distribution, which had apparent ecological links among taxa in modules. This study provides first insight of the mechanism to maintain microbial diversity in metalloids-laden biospheres.

The roles of microorganisms in enhancing crop production have been demonstrated for a range of cropping systems. Most studies to date, however, have been confined to a limited number of locations, making it difficult to identify general soil biotic and abiotic characteristics underpinning the yield-promotion across various locations. This knowledge gap limits our capacity to harness soil microbiome to improve crop production. Here we used high-throughput amplicon sequencing to investigate the common features of bacterial community composition, ecological networks and physicochemical properties in six yield-invigorating and adjacent yield-debilitating orchards. We found that yield-invigorating soils exhibited higher contents of organic matter than yield-debilitating soils and harboured unique bacterial communities. Greater alpha diversity and higher relative abundances of Planctomycetes and Chloroflexi were observed in yield-debilitating soils. Co-occurrence network analysis revealed that yield-invigorating soils displayed a greater number of meta-modules and a higher proportion of negative links to positive links. Chloroflexi was recognized as a keystone taxon in manipulating the interaction of bacterial communities in yield-invigorating soils. Structural equation modelling showed that soil organic matter, beta diversity of bacterial community, and network connector (Chloroflexi) were key factors supporting high-yield pear production. Altogether, we provide evidence that yield-invigorating soils across a range of locations appear to share common features, including accumulation of soil organic matter, higher microbial diversity, enrichment of key taxa like Chloroflexi, and maintaining a competitive network. These findings have implications for science-based guidance for sustainable food production.

The production of bioactive metabolites is increasingly recognized as an important function of host-associated bacteria. An example is defensive symbiosis that might account for much of the chemical richness of marine invertebrates including sponges (Porifera), one of the oldest metazoans. However, as most complex microbiomes remain largely uncultivated and lack reference genomes, unequivocally linking metabolic functions to a cellular source is a challenge. Here we report an analysis pipeline of microfluidic encapsulation, Raman microscopy, and integrated digital genomics (MERMAID) for an efficient identification of uncultivated producers. We applied this method to the chemically rich bacteriosponge Theonella swinhoei, previously shown to contain ‘Entotheonella’ symbionts providing most of its bioactive substances except for the antifungal aurantosides that lacked biosynthetic gene candidates in the metagenome. Raman-guided single-bacterial analysis and sequencing revealed a cryptic, distinct multiproducer, ‘Candidatus Poriflexus aureus’ from a new Chloroflexi lineage. Its exceptionally large genome contains numerous biosynthetic loci and suggested an even higher chemical richness of this sponge than previously appreciated. This study highlights the importance of complementary technologies to uncover microbiome functions, reveals remarkable parallels between distantly related symbionts of the same host, and adds functional support for diverse chemically prolific lineages being present in microbial dark matter.Significance StatementThe production of bioactive metabolites is increasingly recognized as an important function of host-associated bacteria. However, the acquisition of integrated genomic and metabolic data from uncultivated environmental bacteria is still challenging. In this work, we explored the combination of Raman microscopy and single-cell sequencing to localize chemical features to a specific bacterium in an uncultivated microbiome, and we specified the bacteria in the uncultured lineage as a producer of aurantoside, an antifungal natural product, from a chemically and microbially complex sponge. This study offers a new methodology as well as insights into chemical functions of uncultivated life.

The Chloroflexi superphylum have been investigated primarily from the perspective of reductive dehalogenation of toxic compounds, anaerobic photosynthesis and wastewater treatment, but remain relatively little studied compared to their close relatives within the larger Terrabacteria group, including Cyanobacteria, Actinobacteria, and Firmicutes. Here, we conducted a detailed phylogenetic analysis of the phylum Chloroflexota, the phylogenetically proximal candidate phylum Dormibacteraeota, and a newly defined sibling phylum proposed in the current study, Eulabeiota. These groups routinely root together in phylogenomic analyses, and constitute the Chloroflexi supergroup. Chemoautotrophy is widespread in Chloroflexi. Two Form I Rubisco ancestral subtypes that both lack the small subunit are prevalent in ca. Eulabeiota and Chloroflexota, suggesting that the predominant modern pathway for CO2 fixation evolved in these groups. The single subunit Form I Rubiscos are inferred to have evolved prior to oxygenation of the Earth’s atmosphere and now predominantly occur in anaerobes. Prevalent in both Chloroflexota and ca. Eulabeiota are capacities related to aerobic oxidation of gases, especially CO and H2. In fact, aerobic and anaerobic CO dehydrogenases are widespread throughout every class-level lineage, whereas traits such as denitrification and reductive dehalogenation are heterogeneously distributed across the supergroup. Interestingly, some Chloroflexota have a novel clade of group 3 NiFe hydrogenases that is phylogenetically distinct from previously reported groups. Overall, the analyses underline the very high level of metabolic diversity in the Chloroflexi supergroup, suggesting the ancestral metabolic platform for this group enabled highly varied adaptation to ecosystems that appeared in the aerobic world.

Background The native crop bacterial microbiota of the rhizosphere is envisioned to be engineered for sustainable agriculture. This requires the identification of keystone rhizosphere Bacteria and an understanding on how these govern crop-specific microbiome assembly from soils. We identified the metabolically active bacterial microbiota (SSU RNA) inhabiting two compartments of the rhizosphere of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), rye (Secale cereale), and oilseed rape (Brassica napus L.) at different growth stages. Results Based on metabarcoding analysis the bacterial microbiota was shaped by the two rhizosphere compartments, i.e. close and distant. Thereby implying a different spatial extent of bacterial microbiota acquirement by the cereals species versus oilseed rape. We derived core microbiota of each crop species. Massilia (barley and wheat) and unclassified Chloroflexi of group ‘KD4-96’ (oilseed rape) were identified as keystone Bacteria by combining LEfSe biomarker and network analyses. Subsequently, differential associations between networks of each crop species’ core microbiota revealed host plant-specific interconnections for specific genera, such as the unclassified Tepidisphaeraceae ‘WD2101 soil group’. Conclusions Our results provide keystone rhizosphere Bacteria derived from for crop hosts and revealed that cohort subnetworks and differential associations elucidated host species effect that was not evident from differential abundance of single bacterial genera enriched or unique to a specific plant host. Thus, we underline the importance of co-occurrence patterns within the rhizosphere microbiota that emerge in crop-specific microbiomes, which will be essential to modify native crop microbiomes for future agriculture and to develop effective bio-fertilizers.

A mesophilic filamentous anoxygenic phototrophic bacterium, designated M50-1, was isolated from a microbial mat of the Chukhyn Nur soda lake (northeastern Mongolia) with salinity of 5−14 g/L and pH 8.0−9.3. The organism is a strictly anaerobic phototrophic bacterium, which required sulfide for phototrophic growth. The cells formed short undulate trichomes surrounded by a thin sheath and containing gas vesicles. Motility of the trichomes was not observed. The cells contained chlorosomes. The antenna pigments were bacteriochlorophyll d and β- and γ-carotenes. Analysis of the genome assembled from the metagenome of the enrichment culture revealed all the enzymes of the 3-hydroxypropionate bi-cycle for autotrophic CO2 assimilation. The genome also contained the genes encoding a type IV sulfide:quinone oxidoreductase (sqrX). The organism had no nifHDBK genes, encoding the proteins of the nitrogenase complex responsible for dinitrogen fixation. The DNA G + C content was 58.6%. The values for in silico DNA‒DNA hybridization and average nucleotide identity between M50-1 and a closely related bacterium ‘Ca. Chloroploca asiatica’ B7-9 containing bacteriochlorophyll c were 53.4% and 94.0%, respectively, which corresponds to interspecies differences. Classification of the filamentous anoxygenic phototrophic bacterium M50-1 as a new ‘Ca. Chloroploca’ species was proposed, with the species name ‘Candidatus Chloroploca mongolica’ sp. nov.

The Kilianstollen Marsberg (Rhenish Massif, Germany) has been extensively mined for copper ores, dating from early medieval period until 1945. The exposed organic-rich alum shale rocks influenced by the diverse mine drainages at an ambient temperature of 10 ∘C could naturally enrich biogeochemically distinct heavy metal resistant microbiota. This amplicon-sequence-based study evaluates the microbially colonized subterranean rocks of the abandoned copper mine Kilianstollen to characterize the colonization patterns and biogeochemical pathways of individual microbial groups. Under the selective pressure of the heavy metal contaminated environment at illuminated sites, Chloroflexi (Ktedonobacteria) and Cyanobacteria (Oxyphotobacteria) build up whitish–greenish biofilms. In contrast, Proteobacteria, Firmicutes and Actinobacteria dominate rocks around the uncontaminated spring water streams. The additional metagenomic analysis revealed that the heavy metal resistant microbiome was evidently involved in redox cycling of transition metals (Cu, Zn, Co, Ni, Mn, Fe, Cd, Hg). No deposition of metals or minerals, though, was observed by transmission electron microscopy in Ktedonobacteria biofilms which may be indicative for the presence of different detoxification pathways. The underlying heavy metal resistance mechanisms, as revealed by analysis of metagenome-assembled genomes, were mainly attributed to transition metal efflux pumps, redox enzymes, volatilization of Hg, methylated intermediates of As3+, and reactive oxygen species detoxification pathways.

International audience; Two 915-MHz microwave treatments (2 kW-8 min and 4 kW-4 min) were applied to soil and their effects were monitored just after treatments (T0) and 26 days (T26) in soil microcosms. Densities of culturable bacteria, fluorescent pseudomonads and nematodes, hydrolysis activity and soil DNA content declined by over 50% immediately after both microwave treatments (T0), excluding the total fungal 18S rRNA (-13 or -17%) and bacterial 16S rRNA copies (non-significant). A rapid shift in bacterial community composition occurred from T0 towards a large increase in the relative abundance of Firmicutes (+1650%) and a concomitant decrease in various phyla (e.g. Acidobacteria, Actinobacteria, Bacteroidetes, Proteobacteria) from -85 to -61%. At T26 and for both treatments, fluorescein diacetate hydrolysis, density of culturable bacteria, 18S rRNA gene numbers, Simpson diversity, relative abundances of Bacteroidetes and Proteobacteria regained levels similar to controls. Alpha, Delta and Gamma classes Proteobacteria also end up reaching a similar level. In contrast, fluorescent pseudomonad density, nematode diversity and abundance, soil DNA content and relative abundances of some phylum (Acidobacteria, Actinobacteria, Chloroflexi) did not increase in such proportions. Most of the soil biological properties have not been permanently impacted. Nevertheless, the recovery kinetics highly differed according properties, with resilience indices at T26 varying from -87 to +99%.

This study assessed the effects of changes in organic carbon content on soil bacterial community composition and diversity in the Antarctic Fildes Peninsula. 16S rRNA gene sequencing was performed to investigate bacterial community composition. Firstly, we found that organic carbon (OrC) and nutrients showed an increasing trend in the lake area. Secondly, soil geochemistry changes affected microbial composition in the soil. Specifically, we found 3416 operational taxonomical units (OTUs) in 300 genera in five main phyla: Proteobacteria, Actinobacteria, Acidobacteria, Chloroflexi, and Bacteroidetes. Although the diversity was similar among the four sites, the composition was different. Among them, Hungateii content changed very significantly, from 16.67% to 33.33%. Canonical correspondence analysis showed that most measured geochemical factors were relevant in structuring microbiomes, and organic carbon concentration showed the highest correlation, followed by NO3−-N. Hungateii was significantly correlated with the content of organic carbon. Our finding suggested organic carbon played an important role in soil bacterial communities of the Antarctic coastal lake region.

Soil ecosystems surrounding chromium slag undergo continuous harsh physicochemical conditions due to multiple heavy metals contamination. Previous studies of soil microbial communities mainly focused on surface soil layer, while little was known about the depth-related distributions of the microbial communities in chromium (Cr)-contaminated soil. In this study, a comprehensive analysis of depth-related distributions of microbial communities in Cr-contaminated soil was carried out by Illumina sequencing of 16s rRNA genes. The results revealed that bacterial diversities at 0 cm depth layer were significantly higher than those below 20 cm depths. And there was a remarkable difference in bacterial compositions along with the sampling depths especially for the dominant phyla of Proteobacteria, Actinobacteria, Chloroflexi and Fimicutes (p 0.05). While the archaea accounted for a relatively low proportion of the microbes and showed stability in the compositions with the predominant phyla of Thaumarchaeota and Euryarchaeota. The linear discriminate analysis (LDA) and effect size (LEfSe) analysis showed that there were thirty-seven kinds of biomarker microbes existing in the five soil layers with LDA threshold of 4.0, and each layer showed distinct microbial divisions, indicating that microbes with different biological functions might survive along with the sampling depths. The environmental variables including total chromium (Cr), Cr(Ⅵ), Mn, Ni, and Zn had considerable influences on microbial community composition in the contaminated soil. A total of 25 Cr(Ⅵ)-reducing strains were further isolated and identified, which were phylogenetically affiliated to Proteobacteria, Actinobacteria and Firmicutes. Among the isolated Cr(Ⅵ)-reducing strains, Bacillus stratosphericus was the first time to be reported with Cr(Ⅵ) reducing capacity.

Recently, both sand and fly ash have been used for nutrient removal in bioretention systems. However, the improvement in nutrient removal was hampered by a lack of data about of microbial diversity and metabolism effects in the mentioned materials based bioretention systems. Therefore, a mixture with sand, soil and fly ash (1:1:1) was selected as the base in bioretention systems. The investigation of microbial diversity implied that 11 dominant microflora were found, which changed weakly at phylum level but significantly at genus level. The analysis for both urease and extracellular polymer (EPS) showed that urease levels increased with the increase of submerged zone height, which was in line with nitrogen removal, while EPS had the opposite situation. Overall evaluation of microbial role suggested that the enhancement of dominant microflora in the used bioretention systems, like Chloroflexi and Nitrospirae, could strengthen nitrogen removal.

The thermal waters of Gellert Hill discharge area of the Buda Thermal Karst System (Hungary) are characterized by high (up to 1000 Bq/L) 222Rn-activity due to the radium-accumulating biogeochemical layers. Samples were taken from these ferruginous and calcareous layers developed on spring cave walls and water surface. Accumulation of potentially toxic metals (e.g. As, Hg, Pb, Sn, Sr, Zn) in the dense extracellular polymeric substance containing bacterial cells and remains was detected by inductively coupled plasma mass spectrometry. The comparison of bacterial phylogenetic diversity of the biofilm samples was performed by high throughput next generation sequencing (NGS). The analysis showed similar sets of mainly unidentified taxa of phyla Chloroflexi, Nitrospirae, Proteobacteria, Planctomycetes; however, large differences were found in their abundance. Cultivation-based method complemented with irradiation assay was performed using 5, 10 and 15 kGy doses of gamma-rays from a 60Co-source to reveal the extreme radiation-resistant bacteria. The phyla Actinobacteria, Firmicutes, Proteobacteria (classes Alpha- Beta- and Gammaproteobacteria), Bacteriodetes and Deinococcus-Thermus were represented among the 452 bacterial strains. The applied irradiation treatments promoted the isolation of 100 different species, involving candidate novel species, as well. The vast majority of the isolates belonged to bacterial taxa previously unknown as radiation-resistant microorganisms. Members of the genera Paracoccus, Marmoricola, Dermacoccus and Kytococcus were identified from the 15 kGy dose irradiated samples. The close relatives of several known radiation-tolerant bacteria were also detected from the biofilm samples, alongside with bacteria capable of detoxification by metal accumulation, adsorption and precipitation in the form of calcium-carbonate which possibly maintain the viability of the habitat. The results suggest the establishment of a unique, extremophilic microbiota in the studied hydrothermal spring caves.

Rusty root is a widespread problem in ginseng production, which can reduce ginseng quality and root price. The formation of rusty root is a complex process caused by multiple factors, primarily soil physicochemical factors and microorganisms. However, the precise roles played by these factors in the development of this disease remain unclear, especially the role of microorganisms. Deciphering the structure of the rhizosphere microbial communities will provide a more comprehensive and deeper understanding of rusty root disease. Rhizosphere microbial communities from five ginseng fields with different rusty root grades and indices were characterized by MiSeq sequencing. The pattern of beta diversity indicated that rusty root disease might be positively correlated with bacterial community structure, but not with fungal community structure. Rusty root may be caused by excessive Fe (II) oxidation and Fe (III) deposition accompanied by aluminum (Al) and manganese (Mn), in which nitrate-dependent Fe (II)-oxidizing bacteria, including Acidobacteria, Chloroflexi, and Nitrospirae, play a driving role. Moreover, the accumulation of iron (Fe), Al, and Mn could enhance growth and weak invasion of some fungi, thus exacerbating the rust symptoms. However, no correlation was found between the relative abundance of the genus Ilyonectria and rusty root grade or index (P > 0.05); suggesting that Ilyonectria species may not be essential for rusty root disease. Thus, the oxidation and deposition of rhizosphere Fe, Al, and Mn facilitated by nitrate-dependent Fe (II)-oxidizing bacteria might be key factors contributing to rusty root disease. Further studies on the inhibition of Fe, Al, and Mn transformation and accumulation, and application of iron/nitrogen-cycling related bacteria in ginseng soil might elucidate effective methods for preventing rusty root.

This study aimed to determine the methane oxidation rates (MOR), pmoA gene abundance and diversity, and microbial community composition using Illumina high-throughput sequencing. Mudflats located within Yancheng City, divided into different plots with 0-, 11-, and 20-year successive rice planting histories, were selected and sampled. The study found that the relative MOR (normalized with the 16S rRNA gene) increased dramatically after 11-year cultivation and remained stable in 20-year treatment, indicating that long-term rice cultivation in mudflats promoted MOR. The sequencing data analysis revealed that high MOR was related to the synergistic growth of methane-producing archaea (MPA) and aerobic and facultative methane-consuming bacteria (MCB) mainly belonging to Proteobacteria. Redundancy and correlation analyses showed that Methylophilaceae and Methylococcaceae affiliated within β- and γ-Proteobacterial methanotrophs were closely related to the relative MOR. Methane-oxidizing archaea (MOA) coupled to sulfate and nitrite reductions contributed more to the high and stable MOR compared with Proteobacterial MCB. Chloroflexi and Geobacter were the potential hydrogen donors for hydrogenotrophic MPA. The results showed that long-term rice cultivation in mudflats promoted the relative MOR. The unknown MOA coupled to sulfate and nitrite reductions, besides the necessary hydrogenotrophic MPA and their hydrogen donors (Chloroflexi and Geobacter) collectively contributed to methane cycling.

This study applied high-throughput sequencing technology and PICRUSt (phylogenetic investigation of communities by reconstruction of unobserved state) to examine the microbial population dynamics during the composting of dairy manure and rice straw in a static (without turning) composting system. The results showed that the composition of the bacterial community varied significantly during the composting process. The dominant phyla included Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Chloroflexi. Biodiversity indices showed that bacterial community diversity had the peak value during the mesophilic phase. Redundancy analysis indicated that nitrogen was the most important factor in the distribution of genera during the composting process. Finally, the Pearson correlation results suggested that Thermomonospora and Thermopolyspora could be the biomarkers of the composting maturation phase. The metabolic characteristics of the bacterial communities were predicted by PICRUSt. The result showed that metabolism of amino acids, lipids, and most of the carbohydrates increased during the whole composting treatment. However, methane metabolism, carbon fixation pathways in prokaryotes, and nucleotide metabolism decreased after the thermophilic phase. The present study provides a better understanding for bacterial community composition and function succession in dairy manure composting.

A novel slow-growing, facultatively anaerobic, filamentous bacterium, strain MO-CFX2T, was isolated from a methanogenic microbial community in a continuous-flow bioreactor that was established from subseafloor sediment collected off the Shimokita Peninsula of Japan. Cells were multicellular filamentous, non-motile and Gram-stain-negative. The filaments were generally more than 20 µm (up to approximately 200 µm) long and 0.5–0.6 µm wide. Cells possessed pili-like structures on the cell surface and a multilayer structure in the cytoplasm. Growth of the strain was observed at 20–37 °C (optimum, 30 °C), pH 5.5–8.0 (pH 6.5–7.0), and 0–30 g l−1 NaCl (5 g l−1 NaCl). Under optimum growth conditions, doubling time and maximum cell density were estimated to be approximately 19 days and ~105 cells ml−1, respectively. Strain MO-CFX2T grew chemoorganotrophically on a limited range of organic substrates in anaerobic conditions. The major cellular fatty acids were saturated C16 : 0 (47.9 %) and C18 : 0 (36.9 %), and unsaturated C18 : 1ω9c (6.0 %) and C16 : 1ω7 (5.1 %). The G+C content of genomic DNA was 63.2 mol%. 16S rRNA gene-based phylogenetic analysis showed that strain MO-CFX2T shares a notably low sequence identity with its closest relatives, which were Thermanaerothrix daxensis GNS-1T and Thermomarinilinea lacunifontana SW7T (both 85.8 % sequence identity). Based on these phenotypic and genomic properties, we propose the name Aggregatilinea lenta gen. nov., sp. nov. for strain MO-CFX2T (=KCTC 15625T, =JCM 32065T). In addition, we also propose the associated family and order as Aggregatilineaceae fam. nov. and Aggregatilineales ord. nov., respectively.

Most aerobic bacteria exist in dormant states within natural environments. In these states, they endure adverse environmental conditions such as nutrient starvation by decreasing metabolic expenditure and using alternative energy sources. In this study, we investigated the energy sources that support persistence of two aerobic thermophilic strains of the environmentally widespread but understudied phylum Chloroflexi. A transcriptome study revealed that Thermomicrobium roseum (class Chloroflexia) extensively remodels its respiratory chain upon entry into stationary phase due to nutrient limitation. Whereas primary dehydrogenases associated with heterotrophic respiration were downregulated, putative operons encoding enzymes involved in molecular hydrogen (H2), carbon monoxide (CO), and sulfur compound oxidation were significantly upregulated. Gas chromatography and microsensor experiments showed that T. roseum aerobically respires H2 and CO at a range of environmentally relevant concentrations to sub-atmospheric levels. Phylogenetic analysis suggests that the hydrogenases and carbon monoxide dehydrogenases mediating these processes are widely distributed in Chloroflexi genomes and have probably been horizontally acquired on more than one occasion. Consistently, we confirmed that the sporulating isolate Thermogemmatispora sp. T81 (class Ktedonobacteria) also oxidises atmospheric H2 and CO during persistence, though further studies are required to determine if these findings extend to mesophilic strains. This study provides axenic culture evidence that atmospheric CO supports bacterial persistence and reports the third phylum, following Actinobacteria and Acidobacteria, to be experimentally shown to mediate the biogeochemically and ecologically important process of atmospheric H2 oxidation. This adds to the growing body of evidence that atmospheric trace gases are dependable energy sources for bacterial persistence.

Phototrophic communities forming in the bottom of the Berikei highly mineralized mesothermal sulfide springs (Kayakent region, Dagestan, Russia) were investigated. The Berikei springs are an interesting example of combined effect of such factors as temperature, salinity, pH, and sulfide of occurrence and structure of phototrophic microbial communities. The water was of the sodium chloride type with salinity of 48‒97 g/L, near-neutral brine pH, and sulfide concentration of ~1 mM. The temperature at the stream bottom was as high as 60°C. Elevated temperature and high salinity limited the development of phototrophic communities. Formation of cyanobacterial mats occurred at temperatures below 54°C. Phormidium-like cyanobacteria and unicellular Synechocystis sp. predominated in the mats. The number of cyanobacterial species increased at desalination to 48‒57 g/L with emergence of the species morphologically resembling Spirulina sp., Leptolyngbya sp., and Oscillatoria sp. Among anoxygenic phototrophic bacteria, halophilic purple bacteria Ectothiorhodospira sp., Marichromatium sp., and Rhodovulum sp., green sulfur bacteria Prosthecochloris sp., and unidentified Chloroflexi were present. Oxygenic photosynthesis in the mats was not inhibited by sulfide. Production of cyanobacterial mats was up to 4.7‒53.8 µg/(cm2 h). The contribution of anoxygenic phototrophic bacteria to photosynthetic production varied from 0 to 100%. The composition of this microbial community was compared to those of the phototrophic microbial communities of the mesothermal springs of the Dead Sea coast (Israel), the Washington warm lake (United States), and the Paoha Island hot springs (Mono Lake, United States.

Extracellular proteins are of paramount importance in the cell-cell interactions of anammox biofilms. However, the inherent aggregation mechanisms of anammox have largely remained elusive. Herein, using a quartz sand extraction protocol and follow-up iTRAQ-based quantitative proteomics, we identified 367 extracellular proteins from initial colonizers, mature biofilm, and detached biofilm. The extracellular proteins were mainly membrane-associated. Most of the recovered proteins (226, 72.5%) originated from the phylum Planctomycetes. In summary, 215 and 190 of the 367 proteins recovered were up- and/or downregulated at least 1.2-fold during the biofilm formation and detachment periods, respectively. These differentially expressed proteins were dominantly involved in metal ion binding, which was regarded as strong evidence for their abilities to enhance ionic bridges in extracellular polymeric substances (EPS). Scanning electron microscopy-energy-dispersive X-ray spectroscopy (SEM-EDX) analysis of the biofilms further showed substantial levels of calcium and iron minerals. Critically, representative Sec-dependent secretory proteins affiliated with coccoid Planctomycetes, rod-shaped Proteobacteria, and filamentous Chloroflexi (11, 4, and 2 with differential expression, respectively) were found to have typical and abundant inner β-sheet structures, wherein hydrophobic moieties can promote anammox aggregation. Overall, these findings highlight links between differentially expressed protein functions and morphologic traits of anammox consortia during biofilm development.

The bacterial community of an anaerobic granular sludge associated with uranium depletion was investigated following its exposure to uranium under different initial pH conditions (pH 4.5, 5.5, and 6.5). The highest uranium removal efficiency (98.1%) was obtained for the sample with an initial pH of 6.5, which also supported the highest bacterial community richness and diversity. Venn diagrams visualized the decrease in the number of genera present in both the inoculum and the uranium-exposed biomass as the initial pH decreased from 6.5 to 4.5. Compared with the inoculum, a significant increase in the abundances of the phyla Chloroflexi and Proteobacteria was observed following uranium exposure. At initial pH conditions of 6.5 to 4.5, the proportions of the taxa Anaerolineaceae, Chryseobacterium, Acinetobacter, Pseudomonas, and Sulfurovum increased significantly, likely contributing to the observed uranium removal. Uranium exposure induced a greater level of dynamic diversification of bacterial abundances than did the initial pH difference.

Nitrogen removal and microbial interactions in two combined nitritation and Anammox systems with or without the addition of organics were examined. Two systems were successfully started up by adopting intermittent aeration. Organics addition deteriorated nitrogen removal, and total inorganic nitrogen and ammonium removal percentages decreased by 16.4% and 26.3%, respectively. Organics addition promoted the growth of Chloroflexi and Proteobacteria while suppressed the growth of Candidatus Kuenenia. Organics addition decreased activities of fatty acid biosynthesis and metabolism, amino acid metabolism and biofilm formation, while increased activities of steroid metabolism and glycosaminoglycan biosynthesis. Heterotrophs and Candidatus Kuenenia might interact with other organisms by using diverse quorum sensing systems. Chloroflexi and Proteobacteria interacted with Candidatus Kuenenia in nitrogen metabolism and biofilm formation. Proteobacteria played a key role in building a nitrite loop with Candidatus Kuenenia and nitrifiers. These results clarified microbial interactions in the autotrophic nitrogen removal process and advance its application.

Settleability of particles in activated sludge systems can be impaired by an overgrowth of filamentous bacteria, a problem known as bulking. These filaments are often members of the phylum Chloroflexi, sometimes reaching abundances in excess of 30% of the biovolume. The uncultured Chloroflexi phylotype, Candidatus Amarolinea, has been observed in high abundances in Danish full-scale activated sludge systems by 16S rRNA gene amplicon surveys, where it has been associated with bulking. In this study, fluorescence in situ hybridization was applied to confirm their high abundance, filamentous morphology, and contribution to the interfloc bridging that characterizes filamentous bulking. Furthermore, genome-centric metagenomics using both Illumina and Oxford Nanopore sequencing was used to obtain a near complete population genome (5.7 Mbp) of the Ca. Amarolinea phylotype, which belongs to the proposed novel family Amarolineaceae within the order Caldilineales of Chloroflexi. Annotation of the genome indicated that the phylotype is capable of aerobic respiration, fermentation, and dissimilatory nitrate reduction to ammonia. The genome sequence also gives a better insight into the phylogenetic and evolutionary relationships of the organism. The name Candidatus Amarolinea aalborgensis is proposed for the species.

The anaerobic ammonium oxidation (anammox) process has gained much popularity in recent years following its success in nitrogen removal. However, not much has been reported on techniques to promote anammox bacteria immobilization and associated microbial community evolution. In this study, a novel upflow porous-plate anaerobic reactor (UPPAR) was developed and explored to promote biomass (anammox) retention and growth. To comprehend the performance of the UPPAR, its nitrogen removal efficiencies, as well as the microbial community dynamics involved in the nitrogen removal process, was evaluated and reported. When NLR ranging 0.98–1.08 kg m−3 d−1 was introduced at various stages of the UPPAR operation, a rapid start-up was achieved in 63 d, and the overall nitrogen removal rate could reach 90–95%. By the end of the start-up period, it was revealed that Proteobacteria abundance had reduced by 43.92% as opposed Planctomycetes which increased from 2.95% to 43.52%. Conversely, after the UPPAR had been operated for 124 d, thus at steady-state, the most pronounced phylum observed was Planctomycetes (43.52%) followed by Proteobacteria (26.63%), Chloroflexi (5.87%), Ignavibacteriae (5.55%), and Bacteroidetes (4.9%). Predominant genera observed included Candidatus Kuenenia – (25.46%) and Candidatus Brocadia – (3.15%), an indication that nitrogen removal mechanism within the UPPAR was mainly conducted via autotrophic anammox process. Scanning electron microscopy (SEM) revealed that sludge samples obtained at steady-state were predominantly in granular form with sizes ranging between 2 mm to 5 mm. Granules surfaces were dominated with normal to coccoid-shaped cells as revealed by the SEM.

Organic matter degradation and bacterial communities associated to the thermophilic phase of composting were compared using two different types of anaerobic digestates, one from a sewage sludge digester (SD), and the other from an agricultural digester (AD). The composting process exhibited similar variations in temperature, pH, moisture content and bacterial profiles, despite the inherent feedstock differences along with distinctive initial bacterial composition. According to the data obtained from 16S rRNA gene amplicon sequencing, SD constituted more than 20 bacterial phyla with Proteobacteria (21%) and Chloroflexi (21%) being predominant, meanwhile AD was represented by only 7 phyla in which Firmicutes was the most abundant phylum (73%). Nevertheless, bacterial community profiles of the two composting systems became more similarly represented at the phylum level, both dominated by Proteobacteria (65% in AD and 61% in SD), whereas Chromatiaceae and Sphingomonadaceae were the most abundant families in AD and SD, respectively. Highly diverse but similar bacterial communities were detected during the composting of different anaerobic digestates at the thermophilic phase.

Aims The phylum Chloroflexi is frequently found in high abundance in methanogenic reactors, but their role is still unclear as most of them remain uncultured and understudied. Hence, a detailed analysis was performed in samples from five up-flow anaerobic sludge blanket (UASB) full-scale reactors fed different industrial wastewaters. Methods and results Quantitative PCR show that the phylum Chloroflexi was abundant in all UASB methanogenic reactors, with higher abundance in the reactors operated for a long period of time, which presented granular biomass. Both terminal restriction fragment length polymorphism and 16S rRNA gene amplicon sequencing revealed diverse Chloroflexi populations apparently determined by the different inocula. According to the phylogenetic analysis, the sequences from the dominant Chloroflexi were positioned in branches where no sequences of the cultured representative strains were placed. Fluorescent in situ hybridization analysis performed in two of the reactors showed filamentous morphology of the hybridizing cells. Conclusions While members of the Anaerolineae class within phylum Chloroflexi were predominant, their diversity is still poorly described in anaerobic reactors. Due to their filamentous morphology, Chloroflexi may have a key role in the granulation in methanogenic UASB reactors. Significance and impact of the study Our results bring new insights about the diversity, stability, dynamics and abundance of this phylum in full-scale UASB reactors which aid in understanding their function within the reactor biomass. However, new methodological approaches and analysis of bulking biomass are needed to completely unravel their role in these reactors. Combining all this knowledge with reactor operational parameters will allow to understand their participation in granulation and bulking episodes and design strategies to prevent Chloroflexi overgrowth.

The application of straw and biochar can effectively improve soil quality, but whether such application impacts paddy soil bacterial community development remains to be clarified. Herein, the impacts of three different field amendment strategies were assessed including control (CK) treatment, rice straw (RS) application (9000 kg ha−1), and biochar (BC) application (3150 kg ha−1). Soil samples were collected at five different stages of rice growth, and the bacterial communities therein were characterized via high-throughput 16S rDNA sequencing. The results of these analyses revealed that soil bacterial communities were dominated by three microbial groups (Chloroflexi, Proteobacteria and Acidobacteria). Compared with the CK samples, Chloroflexi, Actinobacteria, Nitrospirae and Gemmatimonadetes levels were dominated phyla in the RS treatment, and Acidobacteria, Actinobacteria, Nitrospirae and Patescibacteria were dominated phyla in the BC treatment. Compared with the RS samples, Chloroflexi, Acidobacteria, Actinobacteria, and Verrucomicrobia levels were increased, however, Proteobacteria, Gemmatimonadetes, Nitrospirae, and Firmicute levels were decreased in the BC samples. Rhizosphere soil bacterial diversity rose significantly following RS and BC amendment, and principal component analyses confirmed that there were significant differences in soil bacterial community composition among treatment groups when comparing all stages of rice growth other than the ripening stage. Relative to the CK treatment, Gemmatimonadaceae, Sphingomonadaceae, Thiovulaceae, Burkholderiaceae, and Clostridiaceae-1 families were dominant following the RS application, while Thiovulaceae and uncultured-bacterium-o-C0119 were dominant following the BC application. These findings suggest that RS and BC application can improve microbial diversity and richness in paddy rice soil in Northeast China.