Comparative genomics reveals the adaptation of ammonia-oxidising Thaumarchaeota to arid soils.

Thaumarchaeota are predominant in oligotrophic habitats such as deserts and arid soils, but their adaptations to these arid conditions are not well understood. In this study, we assembled 23 Thaumarchaeota genomes from arid and semi-arid soils collected from the Inner Mongolia Steppe and the Qinghai-Tibet Plateau. Using a comparative genomics approach, integrated with 614 Thaumarchaeota genomes from public databases, we identified the traits and evolutionary forces that contribute to their adaptations to aridity. Our results showed that the newly assembled genomes represent an early diverging group within the lineage of ammonia-oxidising Thaumarchaeota. While the genomic functions previously identified in arid soil lineages were conserved across terrestrial, shallow-ocean and deep-ocean lineages, several traits likely contribute to Thaumarchaeota's adaptation to aridity. These include chlorite dismutase, arsenate reductase, V-type ATPase and genes dealing with oxidative stresses. The acquisition and loss of traits at the last common ancestor of arid soil lineages may have facilitated the specialisation of Thaumarchaeota in arid soils. Additionally, the acquisition of unique adaptive traits, such as a urea transporter, Ca2+ :H+ antiporter, mannosyl-3-phosphoglycerate synthase and phosphatase, DNA end-binding protein Ku and phage shock protein A, further distinguishes arid soil Thaumarchaeota. This study provides evidence for the adaptations of Thaumarchaeota to arid soil, enhancing our understanding of the nitrogen and carbon cycling driven by Thaumarchaeota in drylands.

Consistent community assembly but contingent species pool effects drive ß-diversity patterns of multiple microbial groups in desert biocrust systems.

One of the key goals of ecology is to understand how communities are assembled. The species co-existence theory suggests that community ß-diversity is influenced by species pool and community assembly processes, such as environmental filtering, dispersal events, ecological drift and biotic interactions. However, it remains unclear whether there are similar ß-diversity patterns among different soil microbial groups and whether all these mechanisms play significant roles in mediating ß-diversity patterns. By conducting a broad survey across Chinese deserts, we aimed to address these questions by investing biological soil crusts (biocrusts). Through amplicon-sequencing, we acquired ß-diversity data for multiple microbial groups, that is, soil total bacteria, diazotrophs, phoD-harbouring taxa, and fungi. Our results have shown varying distance decay rates of ß-diversity across microbial groups, with soil total bacteria showing a weaker distance-decay relationship than other groups. The impact of the species pool on community ß-diversity varied across microbial groups, with soil total bacteria and diazotrophs being significantly influenced. While the contributions of specific assembly processes to community ß-diversity patterns varied among different microbial groups, significant effects of local community assembly processes on ß-diversity patterns were consistently observed across all groups. Homogenous selection and dispersal limitation emerged as crucial processes for all groups. Precipitation and soil C:P were the key factors mediating ß-diversity for all groups. This study has substantially advanced our understanding of how the communities of multiple microbial groups are structured in desert biocrust systems.

Core root-associated prokaryotic community and its relationship to host traits across wheat varieties.

The root-associated microbiomes play important roles in plant growth. However, it is largely unknown how wheat variety evolutionary relatedness shapes each subcommunity in the root microbiome and, in turn, how these microbes affect wheat yield and quality. Here we studied the prokaryotic communities associated with the rhizosphere and root endosphere in 95 wheat varieties at regreening and heading stages. The results indicated that the less diverse but abundant core prokaryotic taxa occurred among all varieties. Among these core taxa, we identified 49 and 108 heritable amplicon sequence variants, whose variations in relative abundances across the root endosphere and rhizosphere samples were significantly affected by wheat variety. The significant correlations between phylogenetic distance of wheat varieties and prokaryotic community dissimilarity were only observed in non-core and abundant subcommunities in the endosphere samples. Again, wheat yield was only significantly associated with root endosphere microbiota at the heading stage. Additionally, wheat yield could be predicted using the total abundance of 94 prokaryotic taxa as an indicator. Our results demonstrated that the prokaryotic communities in the root endosphere had higher correlations with wheat yield and quality than those in the rhizosphere; thus, managing root endosphere microbiota, especially core taxa, through agronomic practices and crop breeding, is important for promoting wheat yield and quality.

The Responses of Ammonia-Oxidizing Microorganisms to Different Environmental Factors Determine Their Elevational Distribution and Assembly Patterns.

The assembly mechanisms shaping the elevational patterns of diversity and community structure in ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) are not well understood. We investigated the diversities, co-occurrence network patterns, key drivers, and potential activities of AOA and AOB communities along a large altitudinal gradient. The α-diversity of the AOA communities exhibited a monotonically decreasing pattern with increasing elevation, whereas a sinusoidal pattern was observed for the AOB communities. The mean annual temperature was the single factor that most strongly influenced the α-diversity of the AOA communities; however, the interactions of plant richness, soil conductivity, and total nitrogen made comparable contributions to the α-diversity of the AOB communities. Moreover, the ß-diversities of the AOA and AOB communities were divided into two distinct clusters by elevation, i.e., low- (1800-2600 m) and high-altitude (2800-4100 m) sections. These patterns were attributed mainly to the soil pH, followed by variations in plant richness along the altitudinal gradient. In addition, the AOB communities were more important to the soil nitrification potential in the low-altitude section, whereas the AOA communities contributed more to the soil nitrification potential in the high-altitude section. Overall, this study revealed the key factors shaping the elevational patterns of ammonia-oxidizing communities and might predict the consequences of changes in ammonia-oxidizing communities.

Interconnected versus unconnected microorganisms: Does it matter in anaerobic digestion functioning.

Microorganisms in anaerobic digestion (AD) are essential for wastes/pollutants treatment and energy recovery. Due to microbial enormous diversity, developing effective perspectives to understand microbial roles therein is urgent. This study conducted AD of swine manure, used an ensemble-based network analysis to distinguish interconnected, unconnected, copresence (positively interconnected) and mutual-exclusion (negatively interconnected) microorganisms within microbial communities, and explored their importance towards AD performances, using amplicon sequencing of 16S rRNA and 16S rRNA gene. Our analyses revealed greater importance of interconnected than unconnected microorganisms towards CH4 production and AD multifunctionality, which was attributed to higher niche breadth, deterministic community assembly, community stability and phylogenetic conservatism. The diversity was higher in unconnected than interconnected microorganisms, but was not linked to AD performances. Compared to copresence microorganisms, mutual-exclusion microorganisms showed greater and equal importance towards CH4 production and AD multifunctionality, which was attributed to their roles in stabilizing microbial communities. The increased feedstock biodegradability, by replacing part of manure with fructose or apple waste, hardly affected the relative importance of interconnected versus unconnected microorganisms towards CH4 production or AD multifunctionality. Our findings develop a new framework to understand microbial roles, and have important implications in targeted manipulation of critical microorganisms in waste-treatment systems.

Low C/N promotes stable partial nitrification by enhancing the cooperation of functional microorganisms in treating high-strength ammonium landfill leachate.

Partial nitrification is an effective process for treating high-strength ammonium landfill leachate with low C/N ratio, for the cooperation with denitrification can save almost 40% carbon addition in biological nitrogen removal. However, high ammonia loading often causes the instability of partial nitrification process. Less carbon addition can promote the stability of partial nitrification and increase the nitrite accumulation ratio (NAR). Nevertheless, the microbial mechanisms within remain further elusive. In this study, two laboratory-scale sequencing batch reactors were constructed and operated for 125 days, which were fed with ammonia synthetic wastewater with C/N of 0.6 (CN system) and C/N of 0.0 as the control (N system). CN system performed more stably and had the highest NAR of 100%. Extracellular polymeric substances (EPS) generated from carbon source provided spatial and nutrient niches to tighten the cooperation of functional microorganisms, thus, enhanced the stability and efficiency of partial nitrification. Thauera was the dominant denitrifier in CN system. Nitrosomonas was one of the most important autotrophic ammonia oxidizing bacteria, while Paracoccus and Flavobacterium were the main heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria in CN system. The enrichment of HN-AD bacteria outcompeted nitrite oxidizing bacteria (NOB), therefore leaded to higher nitrite accumulation in CN system. The findings of this study may be conducive to increasing the understanding of the microbial collaboration mechanisms of partial nitrification, thereby provides theoretical support for the improvement of biological nitrogen removal technology.

A simulated ecological restoration of bare cut slope reveals the dosage and temporal effects of cement on ecosystem multifunctionality in a mountain ecosystem.

Cement is a critical building material used in the restorations of bare cut slopes. Yet, how cement affects ecosystem's functions and their undertakers remains elusive. Here, we revealed the dosage and temporal effects of cement on plant and soil traits, extracellular enzymes, greenhouse gas fluxes and microbiome using simulation experiments. The results showed that soil pH increased with the cement content at 1st day but relatively constant values around 7 to 7.5 were detected in the flowing days. The ß-1,4-glucosidase, phenol oxidase, leucine aminopeptidase and acid phosphatase showed high activities under high cement content, and they generally increased with the cultivations except for acid phosphatase. CH4 fluxes at 16th day were less than zero, and they increased to peak around at 37th to 44th days followed by decreasing until reaching to relatively stable fluctuations around 0. Despite of decrease patterns, N2O fluxes stayed around zero across the temporal gradient except for the maximum around at 30th day in 2%, 5% and 8% cement treatment. Microbial diversity decreased with the cement content, in which there were a recovery trend for bacteria. By integrating above- and belowground ecosystem traits into a multifunctionality index, we identified a potential optimum cement content (11%). PLSPM showed that multifunctionality was affected by the shifts in soil bacterial community, enzyme activity and greenhouse gases while these components were effected by other environmental changes resulted from cement. Our results demonstrate that cement determines multifunctionality through mediating microbial community and activity, providing new insights for designing in situ experiments and ecological restoration strategies for bare cut slopes.

Biocrust diazotrophs and bacteria rather than fungi are sensitive to chronic low N deposition.

Anthropogenic long-term nitrogen (N) deposition may dramatically impact biocrusts due to the overarching N limitation of soil biota in deserts. Even low levels of N may reach a critical loading threshold altering biocrust constituents and function. To identify the impact of chronic and continuous low levels of N deposition on biocrusts, we created a realistic gradient mirroring anthropogenic N addition rate (2:1 NH4 + : NO3 - rates: 0.3, 0.5, 1.0, 1.5, 3 g N m-2  yr-1 ) and measured the response of bacteria and fungi within cyanobacterial-dominated biocrusts over 8 years in a temperate desert, the Gurbantunggut Desert, China. We found that once N deposition reached 1.5 g N m-2  yr-1 biocrust bacterial communities, including diazotrophs, were altered while no such tipping point existed for fungi. Above the threshold, bacterial richness was enhanced, the relative abundance of Chloroflexi, FBP and Gemmatimonadetes was elevated, and diazotrophs shifted from being dominated by Nostocaceae and Scytonemataceae (Cyanobacteria) to free-living Bradyrhizobiaceae (Alphaproteobacteria). Alternatively, the relative recovery of a few fungal species within the Lecanorales, Pleosporales and Verrucariales became either enriched or diminished due to N deposition. The chronic addition of N resulted in a dense and interconnected bacterial co-occurrence network that accentuated a functional shift from networks dominated by phototrophic species within the Nostocaceae, Xenococcaceae, Phormidiaceae and Scytonemataceae (Cyanobacteria) to ammonia-oxidizing species within the Nitrosomonadaceae (Betaproteobacteria) and nitrifying bacteria [i.e. Nitrospiraceae (Nitrospirae)]. Based on structural equation models, the effects of N additions on biocrust constituents were imposed through indirect effects on pH, soil electrical conductivity and ammonium concentrations. In summary, biocrust constituents are generally insensitive to chronic low levels of N depositions until rates reach above 1.5 g N m-2  yr-1 with diazotrophs being the most sensitive biocrust constituents followed by bacteria and finally fungi. Ultimately once the threshold is reached N deposition favours biocrust constituents utilizing inorganic N and other C sources over relying on phototrophic and/or N-fixing cyanobacteria for C and N.

The adjustment of life history strategies drives the ecological adaptations of soil microbiota to aridity.

Soil microbiota increase their fitness to local habitats by adjusting their life history strategies. Yet, how such adjustments drive their ecological adaptations in xeric grasslands remains elusive. In this study, shifts in the traits that potentially represent microbial life history strategies were studied along two aridity gradients with different climates using metagenomic and trait-based approaches. The results indicated that resource acquisition (e.g., higher activities of ß-d-glucosidase and N-acetyl-ß-d-glucosidase, higher degradation rates of cellulose and chitin, as well as genes involved in cell motility, biodegradation, transportation and competition) and growth yield (e.g., higher biomass and respiration) strategies were depleted at higher aridity. However, maintenance of cellular and high growth potential (e.g., higher metabolic quotients and genes related to DNA replication, transcription, translation, central carbon metabolism and biosynthesis) and stress tolerance (e.g., genes involved in DNA damage repair, cation transportation, sporulation and osmolyte biosynthesis) strategies were enriched at higher aridity. This implied that microbiota have lower growth yields but are probably well primed for rapid responses to pulses of rainfall in more arid soils, whereas those in less arid soils may have stronger resource acquisition and growth yield abilities. By integrating a large amount of evidence from taxonomic, metagenomic, genomic and biochemical investigations, this study demonstrates that the ecological adaptations of soil microbiota to aridity made by adjusting and optimizing their life history strategies are universal in xeric grasslands and provides an underlying mechanistic understanding of soil microbial responses to climate changes.

Corpse decomposition of freshwater economic fish leads to similar resistomes and the enrichment of high-risk antibiotic resistance genes in different water types.

Animal carcass decay produces many poisonous metabolites and chemical pollutants, which pose potential ecological risks to the aquatic environment and human health. However, the effects of animal cadaver decomposition on high-risk antibiotic resistance genes (ARGs) and potential pathogens in different water types are still unknown. In this study, fifteen freshwater economic fish (Carassius auratus) corpses were put into three types of water (i.e., pond water, tap water, and domestic sewage) for a 100-day decomposition. Next generation sequencing and HT-qPCR were used to illustrate how corpse decomposition affected microbial communities and ARG profiles. Our results revealed that fish corpse degradation caused similar resistomes and microbiome in different water types. MLSB (Macrolide-Lincosamide-Streptogramin B), ß-lactamase, sulfonamide, tetracycline resistance genes and transposase genes in the experimental groups were increased. Among them, tetracycline resistance genes were enriched by 224 to 136,218-fold during the process of corpse degradation. Furthermore, high-risk ARGs (ermB, floR and dfrA1), which resist to MLSB, multidrug and sulfonamide respectively, were significantly enriched in the cadaver groups and had co-occurrence patterns with opportunistic pathogens, such as Bacteroidetes, which was more than 37 times in carcass groups than that in control groups. The study is able to draw a general conclusion that cadaver decomposition of freshwater economic fish deteriorates the aquatic environment by affecting high-risk ARGs and pathogenic microorganisms regardless of water types, which poses potential threats to human health. Therefore, timely management and treatment of animal carcasses is of great significance to the protection of water environment.

The assembly of wheat-associated fungal community differs across growth stages.

Plant-associated fungal communities play a vital role in plant adaptations, physiological functions, and productivity. Therefore, it is important to reveal the mechanisms driving the assembly of these communities. Yet it is still not fully understood how community assembly and structure differentiate in plant compartments, growth seasons, and varieties at large geographic distances. In this study, we analyzed bulk soil and plant-associated fungal communities of five wheat varieties across two growth stages in three biogeographic sites with distances of about 324, 534, or 800 km apart between any two locations. Our results indicated that the fungal community varied primarily across the sample types (leaf endosphere, root endosphere, rhizosphere, and bulk soil), followed by growth stage. Compared with the regreening stage, lower α-diversity and more dominance by abundant species in the fungal community were observed in wheat-associated compartments (four sample types except for bulk soil) at the heading stage. Additionally, within each wheat-associated compartment across every growth stage, location had stronger effects on fungal community assembly than the wheat variety. The effects of variety on fungal community assembly were location specific as were the growth-stage patterns of varietal effects on leaf endosphere and rhizosphere fungal communities. We further detected a less diverse but abundant core fungal taxa that could be grouped into three clusters associated mainly with location. This study characterized the interplay effects between plant selection (compartment, growth stage, variety) and environment (location) on wheat-associated mycobiomes by determining drivers of fungal community assembly and core fungal taxa in field conditions. KEY POINTS: • Fungal community assembly was mainly shaped by sample type and growth stage • A lower diversity and more abundant core fungal taxa were shown at heading stage • Location had stronger effects on fungal community assembly than variety.

Elevation rather than season determines the assembly and co-occurrence patterns of soil bacterial communities in forest ecosystems of Mount Gongga.

Seasonal dynamics of soil microbial communities may influence ecosystem functions and services. However, few observations have been conducted on the dynamics of a bacterial community assembly across seasons in different elevations in mountain forest ecosystems. In this study, the diversity, compositions, community assembly processes, and co-occurrence interactions of soil bacterial communities were investigated using Illumina sequencing of 16S rRNA genes across different seasons during two consecutive years (2016 and 2017) at two elevational sites in Mount Gongga, China. These two sites included an evergreen broad-leaved forest (EBF, 2100 m a.s.l.) and a dark coniferous forest (DCF, 3000 m a.s.l.). The results showed that bacterial diversity and structure varied considerably between the two elevational sites with only limited seasonal variations. Interannuality had a significant effect on the diversity and structure of soil bacterial communities. The bacterial alpha diversity was significantly higher at site EBF(e.g., OTUs richness, 2207 ± 276) than at site DCF(e.g., OTUs richness, 1826 ± 315). Soil pH, temperature, elevation, and water content were identified as important factors shaping soil bacterial communities in the mountain forests. Bacterial community assembly was primarily governed by deterministic processes regardless of elevation and season. Deterministic processes were stronger at site DCF than at EBF. The soil bacterial community at site EBF harbored a more complex and connected network with less resistance to environmental changes. Overall, this study showed that seasonal dynamics of bacterial communities were much weaker than those along elevations, implying that a single-season survey on a bacterial community along an elevational gradient can represent overall changes in the bacterial community. KEY POINTS: • Seasonal dynamics of soil bacterial communities were studied in Mount Gongga. • The bacterial community was mainly affected by elevation rather than season. • Deterministic processes dominated bacterial community assembly. • The bacterial network was more complex but less stable at EBF than at DCF.

Studies on the cold tolerance of ratoon 'Chaling' common wild rice.

BACKGROUND: Rice is the staple food of many people around the world. However, most rice varieties, especially widely grown indica varieties and hybrids, are sensitive to cold stress. In order to provide a basis for the utilization of a common wild rice (CWR, Oryza rufipogon Griff.) named 'Chaling' CWR in cold-tolerant rice breeding and deepen the understanding of rice cold tolerance, the cold tolerance of ratoon 'Chaling' CWR was studied under the stress of the natural low temperature in winter in Changsha, Hunan province, China, especially under the stress of abnormal natural low temperature in Changsha in 2008, taking other ratoon CWR accessions and ratoon cultivated rice phenotypes as control. RESULTS: The results showed that ratoon 'Chaling' CWR can safely overwinter under the natural conditions in Changsha (28° 22' N), Hunan province, China, which is a further and colder northern place than its habitat, even if it suffers a long-term low temperature stress with ice and snow. In 2008, an extremely cold winter appeared in Changsha, i.e., the average daily mean temperature of 22 consecutive days from January 13 to February 3 was - 1.0 °C, and the extreme low temperature was - 4.7 °C. After subjected to this long-term cold stress, the overwinter survival rate of ratoon 'Chaling' CWR was 100%, equals to that of ratoon 'Dongxiang' CWR which is northernmost distribution in the word among wild rice populations, higher than those of ratoon 'Fusui' CWR, ratoon 'Jiangyong' CWR, and ratoon 'Liujiang' CWR (63.55-83.5%) as well as those of ratoon 'Hainan' CWR, ratoon 'Hepu' CWR, and all the ratoon cultivated rice phenotypes including 3 japonica ones, 3 javanica ones, and 5 indica ones (0.0%). CONCLUSIONS: The results indicate that ratoon 'Chaling' CWR possesses strong cold tolerance and certain freezing tolerance.

Dynamics and potential roles of abundant and rare subcommunities in the bioremediation of cadmium-contaminated paddy soil by Pseudomonas chenduensis.

Microbial bioremediation of heavy metal-contaminated soil is a potential technique to reduce heavy metals in crop plants. However, the dynamics and roles of the local microbiota in bioremediation of heavy metal-contaminated soil following microbial application are rarely reported. In this study, we used Pseudomonas chenduensis strain MBR for bioremediation of Cd-contaminated paddy soil and investigated its effects on the dynamics of the local soil bacterial community and Cd accumulation in rice. Cd accumulation in rice grains and roots were significantly reduced by the addition of the strain MBR. The addition of the strain MBR caused greater changes in bacterial communities in rhizosphere soil than in bulk soil. MBR enhanced the roles of microbial communities in transformation of Cd fractions, especially in rhizosphere soil. The strain MBR likely regulated abundant subcommunities more than rare subcommunities to improve Cd bioremediation, especially in rhizosphere soil. Consequently, the dynamics and functional roles of the local microbial communities differed significantly during bioremediation between abundant and rare subcommunities and between rhizosphere soil and bulk soil. This study provides new insight into the microbiota-related mechanisms underlying bioremediation.

Forest gaps influence fungal community assembly in a weeping cypress forest.

The forest gap crucially influences forest environments, but its effects on local fungal community assembly are not fully understood. In this study, the fungal community in a weeping cypress forest was investigated as a function of forest gap locations based on forest clearing, using amplicon sequencing of the ITS2 region. The results showed that the fungal community significantly varied with the variations in soil properties related to gap location. Deterministic processes played pivotal roles in fungal community assembly, which was mainly driven by the temperature, moisture, available nitrogen, and microbial carbon in soil. Beta diversity of the fungal community increased from the gap center to the closed canopy. The relative abundances of dominant orders such as Microascales, Sordariales, and Chaetothyriales regularly varied as a function of gap location, and they were potential indicators for different gap locations. Based on network analysis, gap locations caused distinct co-occurrence patterns of fungal communities. This study shed light on the roles of forest gaps in the assembly of local fungal communities and provided additional strategies to manage forest ecosystems.

Dynamics of the rice rhizosphere microbial community under continuous and intermittent flooding treatment.

Flooding regime is an important agronomic strategy, and widely applied in heavy metal-contaminated soil for controlling heavy metal uptake through biochemical processes. However, the dynamics of the microbial community in the rhizosphere of rice under different flooding systems are not well understood. In the present study, we investigated the dynamics between the microbial community and heavy metal ions under continuous flooding (CF) and intermittent flooding (IF) conditions to decipher the relationship between microbial community and environmental factors. The results showed that, for the complex Cd, Pb, and Zn contaminated soil under CF treatment did not significantly suppress Cd uptake, but promoted Pb and Ni accumulation into the rice root. Soil pH and bio-available phosphate content appeared to be the key factors impacting heavy metal mobility. When observing the microbial community in the rhizosphere, long term flooding resulted in an abundance of Anaeromyxobactersp., Geobacter and Desulfovibrio, and the abundance of these taxa displayed a significant relationship to Pb and Zn content of rice roots. From the study, we observed that the flooding regime could have a significant impact on concentrations of Cd, Pb and Ni in rice roots, and the different richness of SRB and FeRB may contribute to uptake of these heavy metals. In future work, the impact of Fe cycling on heavy metal bioavailability in the plant rhizosphere should be investigated further.

New Insights into the Function and Global Distribution of Polyethylene Terephthalate (PET)-Degrading Bacteria and Enzymes in Marine and Terrestrial Metagenomes.

Polyethylene terephthalate (PET) is one of the most important synthetic polymers used today. Unfortunately, the polymers accumulate in nature and to date no highly active enzymes are known that can degrade it at high velocity. Enzymes involved in PET degradation are mainly α- and ß-hydrolases, like cutinases and related enzymes (EC 3.1.1). Currently, only a small number of such enzymes are well characterized. In this work, a search algorithm was developed that identified 504 possible PET hydrolase candidate genes from various databases. A further global search that comprised more than 16 Gb of sequence information within 108 marine and 25 terrestrial metagenomes obtained from the Integrated Microbial Genome (IMG) database detected 349 putative PET hydrolases. Heterologous expression of four such candidate enzymes verified the function of these enzymes and confirmed the usefulness of the developed search algorithm. In this way, two novel and thermostable enzymes with high potential for downstream application were partially characterized. Clustering of 504 novel enzyme candidates based on amino acid similarities indicated that PET hydrolases mainly occur in the phyla of Actinobacteria, Proteobacteria, and Bacteroidetes Within the Proteobacteria, the Betaproteobacteria, Deltaproteobacteria, and Gammaproteobacteria were the main hosts. Remarkably enough, in the marine environment, bacteria affiliated with the phylum Bacteroidetes appear to be the main hosts of PET hydrolase genes, rather than Actinobacteria or Proteobacteria, as observed for the terrestrial metagenomes. Our data further imply that PET hydrolases are truly rare enzymes. The highest occurrence of 1.5 hits/Mb was observed in sequences from a sample site containing crude oil.IMPORTANCE Polyethylene terephthalate (PET) accumulates in our environment without significant microbial conversion. Although a few PET hydrolases are already known, it is still unknown how frequently they appear and with which main bacterial phyla they are affiliated. In this study, deep sequence mining of protein databases and metagenomes demonstrated that PET hydrolases indeed occur at very low frequencies in the environment. Furthermore, it was possible to link them to phyla that were previously not known to harbor such enzymes. This work contributes novel knowledge on the phylogenetic relationships, the recent evolution, and the global distribution of PET hydrolases. Finally, we describe the biochemical traits of four novel PET hydrolases.

Improvement of n-caproic acid production with Ruminococcaceae bacterium CPB6: selection of electron acceptors and carbon sources and optimization of the culture medium.

BACKGROUND: Global energy and resource shortages make it necessary to quest for renewable resources. n-Caproic acid (CA) production based on carboxylate platform by anaerobic fermentation is booming. Recently, a novel Ruminococcaceae bacterium CPB6 is shown to be a potential biotransformation factory for CA production from lactate-containing wastewater. However, little is known about the effects of different electron acceptors (EAs) on the fermentative products of strain CPB6, as well as the optimum medium for CA production. RESULTS: In this study, batch experiments were performed to investigate the fermentative products of strain CPB6 in a lactate medium supplemented with different EAs and sugars. Supplementation of acetate, butyrate and sucrose dramatically increased cell growth and CA production. The addition of propionate or pentanoate resulted in the production of C5 or C7 carboxylic acid, respectively. Further, a Box-Behnken experiment was conducted to optimize the culture medium for CA production. The result indicated that a medium containing 13.30 g/L sucrose, 22.35 g/L lactate and 16.48 g/L butyrate supported high-titer CA production (16.73 g/L) with a maximum productivity of 6.50 g/L/day. CONCLUSIONS: This study demonstrated that strain CPB6 could produce C6-C7 carboxylic acids from lactate (as electron donor) with C2-C5 short-chain carboxylic acids (as EAs), but CA (C6 carboxylic acid) was the most major and potential product. Butyrate and sucrose were the most significant EA and carbon source respectively for CA production from lactate by strain CPB6. High titer of CA can be produced from a synthetic substrate containing sucrose, lactate and butyrate. The work provided significant implications for improving CA production in industry-scale.

Diet simplification selects for high gut microbial diversity and strong fermenting ability in high-altitude pikas.

The gut microbiota in mammals plays a key role in host metabolism and adaptation. However, relatively little is known regarding to how the animals adapts to extreme environments through regulating gut microbial diversity and function. Here, we investigated the diet, gut microbiota, short-chain fatty acid (SCFA) profiles, and cellulolytic activity from two common pika (Ochotona spp.) species in China, including Plateau pika (Ochotona curzoniae) from the Qinghai-Tibet Plateau and Daurian pika (Ochotona daurica) from the Inner Mongolia Grassland. Despite a partial diet overlap, Plateau pikas harbored lower diet diversity than Daurian pikas. Some bacteria (e.g., Prevotella and Ruminococcus) associated with fiber degradation were enriched in Plateau pikas. They harbored higher gut microbial diversity, total SCFA concentration, and cellulolytic activity than Daurian pikas. Interestingly, cellulolytic activity was positively correlated with the gut microbial diversity and SCFAs. Gut microbial communities and SCFA profiles were segregated structurally between host species. PICRUSt metagenome predictions demonstrated that microbial genes involved in carbohydrate metabolism and energy metabolism were overrepresented in the gut microbiota of Plateau pikas. Our results demonstrate that Plateau pikas harbor a stronger fermenting ability for the plant-based diet than Daurian pikas via gut microbial fermentation. The enhanced ability for utilization of plant-based diets in Plateau pikas may be partly a kind of microbiota adaptation for more energy requirements in cold and hypoxic high-altitude environments.

RpoN (σ54) Is Required for Floc Formation but Not for Extracellular Polysaccharide Biosynthesis in a Floc-Forming Aquincola tertiaricarbonis Strain.

Some bacteria are capable of forming flocs, in which bacterial cells become self-flocculated by secreted extracellular polysaccharides and other biopolymers. The floc-forming bacteria play a central role in activated sludge, which has been widely utilized for the treatment of municipal sewage and industrial wastewater. Here, we use a floc-forming bacterium, Aquincolatertiaricarbonis RN12, as a model to explore the biosynthesis of extracellular polysaccharides and the regulation of floc formation. A large gene cluster for exopolysaccharide biosynthesis and a gene encoding the alternative sigma factor RpoN1, one of the four paralogues, have been identified in floc formation-deficient mutants generated by transposon mutagenesis, and the gene functions have been further confirmed by genetic complementation analyses. Interestingly, the biosynthesis of exopolysaccharides remained in the rpoN1-disrupted flocculation-defective mutants, but most of the exopolysaccharides were secreted and released rather than bound to the cells. Furthermore, the expression of exopolysaccharide biosynthesis genes seemed not to be regulated by RpoN1. Taken together, our results indicate that RpoN1 may play a role in regulating the expression of a certain gene(s) involved in the self-flocculation of bacterial cells but not in the biosynthesis and secretion of exopolysaccharides required for floc formation.IMPORTANCE Floc formation confers bacterial resistance to predation of protozoa and plays a central role in the widely used activated sludge process. In this study, we not only identified a large gene cluster for biosynthesis of extracellular polysaccharides but also identified four rpoN paralogues, one of which (rpoN1) is required for floc formation in A. tertiaricarbonis RN12. In addition, this RpoN sigma factor regulates the transcription of genes involved in biofilm formation and swarming motility, as previously shown in other bacteria. However, this RpoN paralogue is not required for the biosynthesis of exopolysaccharides, which are released and dissolved into culture broth by the rpoN1 mutant rather than remaining tightly bound to cells, as observed during the flocculation of the wild-type strain. These results indicate that floc formation is a regulated complex process, and other yet-to-be identified RpoN1-dependent factors are involved in self-flocculation of bacterial cells via exopolysaccharides and/or other biopolymers.