Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play...Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.展开更多
This study was conducted to investigate the phylogenetic diversity of archaea in the rumen of adult and elderly yaks. Six domesticated female yaks, 3 adult yaks((5.3±0.6) years old), and 3 elderly yaks((10.7±...This study was conducted to investigate the phylogenetic diversity of archaea in the rumen of adult and elderly yaks. Six domesticated female yaks, 3 adult yaks((5.3±0.6) years old), and 3 elderly yaks((10.7±0.6) years old), were used for the rumen contents collection. Illumina Mi Seq high-throughput sequencing technology was applied to examine the archaeal composition of rumen contents. A total of 92 901 high-quality archaeal sequences were analyzed, and these were assigned to 2 033 operational taxonomic units(OTUs). Among these, 974 OTUs were unique to adult yaks while 846 OTUs were unique to elderly yaks; 213 OTUs were shared by both groups. At the phylum level, more than 99% of the obtained OTUs belonged to the Euryarchaeota phylum. At the genus level, the archaea could be divided into 7 archaeal genera. The 7 genera(i.e., Methanobrevibacter, Methanobacterium, Methanosphaera, Thermogymnomonas, Methanomicrobiu, Methanimicrococcus and the unclassified genus) were shared by all yaks, and their total abundance accounted for 99% of the rumen archaea. The most abundant archaea in elderly and adult yaks were Methanobrevibacter and Thermogymnomonas, respectively. The abundance of Methanobacteria(class), Methanobacteriales(order), Methanobacteriaceae(family), and Methanobrevibacter(genus) in elderly yaks was significantly higher than in adult yaks. In contrast, the abundance of Thermogymnomonas in elderly yaks was 34% lower than in adult yaks, though the difference was not statistically significant. The difference in abundance of other archaea was not significant between the two groups. These results suggested that the structure of archaea in the rumen of yaks changed with age. This is the first study to compare the phylogenetic differences of rumen archaeal structure and composition using the yak model.展开更多
The objective of the present study was to examine the effect of different weaning methods on the ruminal methanogenic archaea composition and diversity in Holstein calves.Thirty-six newborn Holstein bull calves were a...The objective of the present study was to examine the effect of different weaning methods on the ruminal methanogenic archaea composition and diversity in Holstein calves.Thirty-six newborn Holstein bull calves were assigned to 1 of 3 treatments:(1)conventional weaning(d 56)and fed a high proportion of solid feed(CWS);(2)conventional weaning(d 56)and fed a high proportion of liquid feed(CWL);(3)early weaning(d 42)and fed with a high proportion of solid feed(EWS).High-throughput sequencing of the methyl coenzyme M reductase(mcr A)gene,which encodes theα-subunit of methyl coenzyme M reductase-the enzyme that catalyzes the final step in methanogenesis was used to determine the composition and diversity of rumen methanogens.No significant difference(P>0.05)was observed for operational taxonomic units(OTUs)or richness indices,but diversity indices increased(P<0.05)for calves fed high dietary solids.Predominant families across the three treatments were Methanobacteriaceae,Thermoplasmataceae and Methanomassiliicoccaceae.Calves in the EWS treatment had a higher(P<0.05)relative abundance of Methanobrevibacter sp.strain AbM4 and Methanosphaera stadtmanae,while calves in the CWL treatment had a higher(P<0.05)abundance of Methanosphaera sp.strain SM9.A positive(P<0.05)relationship was identified between butyrate and Methanobrevibacter sp.strain AbM4.In conclusion,the composition and diversity of methanogens in the rumen of Holstein calves varied under the different weaning methods.This study identified a positive relationship between butyrate and Methanobrevibacter sp.strain AbM4,potentially reflecting correlations between ruminal fermentation variables and methanogenesis function.These in-depth analyses provide further understanding of weaning methods for intensified production systems.展开更多
Metagenomic studies have demonstrated the existence of ammonia-oxidizing archaea(AOA) and revealed they are responsible for ammoxidation in some extreme environments. However, the changes in compositional structure an...Metagenomic studies have demonstrated the existence of ammonia-oxidizing archaea(AOA) and revealed they are responsible for ammoxidation in some extreme environments. However, the changes in compositional structure and ammonia-oxidation capacity of AOA communities in biological soil crusts(BSCs) of desert ecosystems remain poorly understood.Here, we utilized Illumina MiSeq sequencing and microbial functional gene array(GeoChip 5.0) to assess the above changes along a 51-year revegetation chronosequence in the Tengger Desert, China. The results showed a significant difference in AOA-community richness between 5-year-old BSCs and older ones. The most dominant phylum during BSC development was Crenarchaeota, and the corresponding species were ammonia-oxidizing_Crenarchaeote and environmental_samples_Crenarchaeota. Network analysis revealed that the positive correlations among dominant taxa increased, and their cooperation was reinforced in AOA communities during BSC succession. Redundancy analysis showed that the dominant factor influencing the change in AOA-community structure was soil texture. GeoChip 5.0 indicated that the amoA gene abundances of AOA and ammonia-oxidizing bacteria(AOB) were basically the same, demonstrating that AOA and AOB played an equally important role during BSCs development. Our study of the long-term succession of BSC demonstrated a persistent response of AOA communities to revegetation development in desert ecosystems.展开更多
Depth-dependent distribution patterns of bacterial and archaeal communities in deep sea water column around the Ninetyeast Ridge in the Indian Ocean were investigated using 16S rRNA gene profiling.Sampling was conduct...Depth-dependent distribution patterns of bacterial and archaeal communities in deep sea water column around the Ninetyeast Ridge in the Indian Ocean were investigated using 16S rRNA gene profiling.Sampling was conducted at the northern Ninetyeast Ridge(1°59.89′N–9°59.70′S,87°58.90′E–88°00.03′E)from September to November 2016 where samples were collected from the bathyal(1000 m)to bathypelagic depths(>4000 m)in four different stations.A total of 1565405 clean data falling into 6712 bacterial OTUs and 1452727 clean data falling into 806 archaeal OTUs based on 97%similarity level were analyzed.Most of the bacterial 16S rRNA gene sequences were affiliated with Gammaproteobacteria,followed by Alphaproteobacteria and Bacteroidia.The archaeal 16S rRNA gene sequences mostly affiliated to Nitrososphaeria(Thaumarchaeota)dominated with relative abundances ranging from 52.68%to 97.2%,followed by Thermoplasmata(Euryarchaeota).Vertical partitioning of bacterial and archaeal communities among different water layers was observed.Canonical correspondence analysis(CCA)and Spearman’s correlations revealed that depth(P=0.003),dissolved oxygen(P=0.019),and nitrite(P=0.033)were the main environmental factors affecting bacterial community structure at genus level in the Ninetyeast Ridge.On the other hand,the first two CCA axes accounted for 74.4%of the explained total variance,it seems that the archaeal communities at genus level were heavily influenced by the environmental variables including depth,dissolved oxygen(DO),nitrite,salinity,phosphate,ammonia,nitrate,and silicate,but none of them exhibited any significant correlation on the structuring(P>0.1).展开更多
The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high orga...The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high organic matter content, fertility, and poor rocky soil. Using domain-specific primers, archaeal 16S rRNA gene sequences were PCR amplified for clone library construction, and a total of 185 archaeal clones were examined. The archaeal clones were affiliated with the phyla Thaumarchaeota (96.2%) and Euryarchaeota (3.8%). The most abundant thaumarchaeal group (90.3% of the clones) was the group I.1b clade, which includes soil ammonia-oxidizing archaea. The unique characteristics of Gotjawal soil, including basalt morphology, vegetation, and groundwater aquifer penetration, may be reflected in the archaeal community composition. Further study is necessary to understand the unique factors of Gotjawal soils that influence archaeal abundance, composition, and diversity.展开更多
There are five hot springs named separately SHANGT,ZHONGT,WENT,XIAT and SHENT in Lushan County,Pingdingshan City,Henan Province.The archaeal diversity of the five hot springs was investigated and analyzed.16 S rD NA v...There are five hot springs named separately SHANGT,ZHONGT,WENT,XIAT and SHENT in Lushan County,Pingdingshan City,Henan Province.The archaeal diversity of the five hot springs was investigated and analyzed.16 S rD NA v4-5 genes of archaea were sequenced by Illumina Hisiq2500 PE250 technology.The numbers of sample sequences and OTUs were sorted by using Uparse,Mothur,SILVA,MUSCLE and Qiime software.Finally,the archaeal abundance and diversity in the five hot springs were analyzed.87993,67326,44400,48611 and 48495 effective reads have been obtained from SHANGT,ZHONGT,WENT,XIAT and SHENT respectively,and 520,280,678,451 and 437 OTUs have been acquired.The analysis of archaeal classification showed that the archaea from the five hot springs could be classified into 11 phyla,in which Thaumarchaeota accounted for 95.25%and was the dominant archaea.In the genera level,the dominant archaea of the SHANGT,ZHONGT and WENT was Candidatus-Nitrosocaldus,and unidentified-Thaumarchaeota was the dominant archaea in XIAT and SHENT.The results showed that the five hot springs in Lushan are rich in microbial resources.The research provided basic data for further research and utilization of the archaea resources in the five hot springs of Lushan.展开更多
Erhualian and Landrace breeds are typical genetically obese and lean pigs, respectively. To compare the fecal methanogenic Archaeal community between these two pig breeds, fecal samples from different growth phase pig...Erhualian and Landrace breeds are typical genetically obese and lean pigs, respectively. To compare the fecal methanogenic Archaeal community between these two pig breeds, fecal samples from different growth phase pigs were collected and used for PCR-denaturing gradient gel electrophoresis(DGGE) with two primer pairs(344fGC/519r and 519f/915rGC) and real-time PCR analysis. Results showed that a better separation and higher quality of bands pattern were obtained in DGGE profiles using primers 344fGC/519r as compared with primers 519f/915rGC. Sequencing of DGGE bands showed that the predominant methanogens in the feces of Erhualian and Landrace pigs belonged to Methanobrevibacter spp. and Methanosphaera spp. Real-time PCR analysis revealed that there was no significant difference in the numbers of fecal total methanogens between Erhualian and Landrace pigs; however, pig growth phase affected the numbers of 16S rRNA genes of total methanogens and Methanobrevibacter smithii. Dissociation curves of methyl coenzyme-M reductase subunit A(mcrA) gene fragments amplified with real-time PCR showed all samples possessed a single peak at 82?C, which might be associated with M. smithii. Samples from the same growth phase of each breed showed good replicative dissociation curves. The results suggest that the growth phase(including diet factor) other than genotype of pig may affect the fecal methanogenic Archaeal community of pigs.展开更多
The number of completely sequenced archaeal genomes has been sufficient for a large-scale bioinformatic study.We have conducted analyses for each coding region from 36 archaeal genomes using the original CGS algorithm...The number of completely sequenced archaeal genomes has been sufficient for a large-scale bioinformatic study.We have conducted analyses for each coding region from 36 archaeal genomes using the original CGS algorithm by calculating the total GC content(G+C),GC content in first,second and third codon positions as well as in fourfold and twofold degenerated sites from third codon positions,levels of arginine codon usage(Arg2:AGA/G;Arg4:CGX),levels of amino acid usage and the entropy of amino acid content distribution.In archaeal genomes with strong GC pressure,arginine is coded preferably by GC-rich Arg4 codons,whereas in most of archaeal genomes with G+C<0.6,arginine is coded preferably by AT-rich Arg2 codons.In the genome of Haloquadratum walsbyi,which is closely related to GC-rich archaea,GC content has decreased mostly in third codon positions,while Arg4>>Arg2 bias still persists.Proteomes of archaeal species carry characteristic amino acid biases:levels of isoleucine and lysine are elevated,while levels of alanine,histidine,glutamine and cytosine are relatively decreased.Numerous genomic and proteomic biases observed can be explained by the hypothesis of previously existed strong mutational AT pressure in the common predecessor of all archaea.展开更多
A total of 26 isolates were obtained from solar salt ponds of different salinities(100, 150, 200, and 250) in Hangu Saltworks Co. Ltd., Tianjin, China. Phylogenetic analysis of 16 SrRNA gene sequences indicated that f...A total of 26 isolates were obtained from solar salt ponds of different salinities(100, 150, 200, and 250) in Hangu Saltworks Co. Ltd., Tianjin, China. Phylogenetic analysis of 16 SrRNA gene sequences indicated that five bacteria genera H alomonas, Salinicoccus, Oceanobacillus, Gracibacillus, and Salimicrobium and one archaea genera H alorubrum were present. The genus H alomonas was predominant with eight strains distributed in a salinity range of 100–200, followed by H alorubrum with six strains in salinity 250. Based on the genus and original sampling salinity, eight bacterial and two archaeal isolates were selected for further morphological, physiological, and biochemical characterization. All of the bacterial strains were moderately halophilic with the optimal salinity for growth being either 50 or 100, while two archaeal strains were extremely halophilic with an optimal growth salinity of 200. Additionally, we put forth strain SM.200-5 as a new candidate S alimicrobium species based on the phylogenic analysis of the 16 SrRNA gene sequence and its biochemical characteristics when compared with known related species.展开更多
Halophilic archaea comprise the maj ority of microorganisms found in hypersaline environments.C50 carotenoids accumulated in archaea cells are considered potential biotechnological products and possess a number of bio...Halophilic archaea comprise the maj ority of microorganisms found in hypersaline environments.C50 carotenoids accumulated in archaea cells are considered potential biotechnological products and possess a number of biological functions.Ten red colonies were isolated from brine water in a saltern crystallizer pond of the Hangu Saltworks,China.16 S rRNA gene sequence analysis showed that the colonies belonged to the extremely halophilic archaea genera Halobacterium and Halorubrum.Two representative strains,Halobacterium strain SP-2 and Halorubrum strain SP-4,were selected for further study on the phenotypic characteristics and effects of salinity and pH on accumulation and composition of pigments in their cells.The archaeal strains were isolated and grown in a culture medium prepared by dissolving yeast extract(10 g/L) and acid-hydrolyzed casein(7.5 g/L) into brine water obtained from a local salt pond.Their optimum salinity and pH for growth were 250 and 7,respectively,although pigment accumulation(OD490/mL broth) was highest at pH 8.In addition,at 150-300 salinity,increasing salinity resulted in decreasing pigment accumulation.Analysis of the UV-Vis spectrum,TLC and HLPC chromatograms showed that C50 carotenoid bacterioruberin is the major pigment in both strains.展开更多
The genetic codon UGA has a dual function: serving as a terminator and encoding selenocysteine. However, most popular gene annotation programs only take it as a stop signal, resulting in misannotation or completely mi...The genetic codon UGA has a dual function: serving as a terminator and encoding selenocysteine. However, most popular gene annotation programs only take it as a stop signal, resulting in misannotation or completely missing selenoprotein genes. We developed a computational method named Asec-Prediction that is specific for the prediction of archaeal selenoprotein genes. To evaluate its effectiveness, we first applied it to 14 archaeal genomes with previously known selenoprotein genes, and Asec-Prediction identified all reported selenoprotein genes without redundant results. When we applied it to 12 archaeal genomes that had not been researched for selenoprotein genes, Asec-Prediction detected a novel selenoprotein gene in Methanosarcina acetivorans. Further evidence was also collected to support that the predicted gene should be a real selenoprotein gene. The result shows that Asec-Prediction is effective for the prediction of archaeal selenoprotein genes.展开更多
The expression of mitogen-activated protein kinase(mapk) double-stranded RNA in cucumber is effective in controlling infestations of the root-knot nematode Meloidogyne incognita. However,little is known about the ecol...The expression of mitogen-activated protein kinase(mapk) double-stranded RNA in cucumber is effective in controlling infestations of the root-knot nematode Meloidogyne incognita. However,little is known about the ecological effects of transgenic plants. Here,we analyzed the diversity of 16 S r DNA genes derived from the rhizosphere archaea of transgenic cucumber plants as an indicator of ecological change. A total of 17 and 18 operational taxonomic units were detected in the rhizospheres of non-transgenic cucumber and mapk ds RNA-expressing plants,respectively. No significant difference was observed between the two groups according to Shannon and Simpson indices. In soil samples of the two rhizospheres,the dominant group was Crenarchaeota at the phylum level,with Staphylothermus,Methanococcus,Pyrodictium and Sulfolobus the abundant taxa at the genus level. These results suggest that expressing mapk double-stranded(ds) RNA in cucumber has no apparent effect on the diversity of rhizosphere archaea,and provide powerful evidence for the ecological safety of transgenic cucumber expressing mapk ds RNA.展开更多
The gastrointestinal tract,as the major digestive and absorbing organ,plays an important role in chicken growth.Numerous studies have been undertaken to investigate the gut bacterial community in broilers,but little i...The gastrointestinal tract,as the major digestive and absorbing organ,plays an important role in chicken growth.Numerous studies have been undertaken to investigate the gut bacterial community in broilers,but little is known about the archaeal community composition.To improve our understanding of the composition of intestinal archaea and the relationship between archaea and chicken growth,a feeding trial was conducted with 10 000 Ross 308 chicks in a commercial production facility under standard management.At 38 d of age,15 birds with the highest body weight( H group),15 birds with average body weight( M group),and15 birds with the lowest body weight( L group) were selected among 600 healthy male individuals.Cecal archaeal community was detected based on 16 S rRNA gene sequencing.As a result,3 321 229 archaeal sequences were clustered into 1 281 operational taxonomic units( OTUs) and classified into three phyla( Crenarchaeota,Euryarchaeota and Thaumarchaeota) and six known genera: Natronomonas,Halolamina,Salinarchaeum,Cenarchaeum,Candidatus,Nitrosoarchaeum and Nitrososphaera.The alpha indices showed that,the richness of archaea in L group was the highest,and followed by that in M group and H group,while the archaea in H group were more diverse,followed by those in M group and L group.However,the archaeal composition in all chicken samples was similar,with the vast majority of the sequences belonging to Cenarchaeum,and only a few belonging to the other five genera.展开更多
Fluorescence in-situ hybridization(FISH) and 16S rRNA gene clone library analyses were used to determine the abundance and diversity of archaea in Prydz Bay, Antarctica. Correlation analysis was also performed to asse...Fluorescence in-situ hybridization(FISH) and 16S rRNA gene clone library analyses were used to determine the abundance and diversity of archaea in Prydz Bay, Antarctica. Correlation analysis was also performed to assess links between physicochemical parameters and archaeal abundance and diversity within the sea-ice. Samples of sea-ice and seawater were collected during the 26th Chinese National Antarctic Research Expedition. The results of FISH showed that archaea were relatively abundant within the top layer of the sea-ice, and correlation analysis suggested that the concentration of 4NH+ might be one of the main factors underlying this distribution pattern. However, using 16S rRNA gene libraries, archaea were not detected in the top and middle layers of the sea-ice. All archaeal clones obtained from the bottom layer of the sea-ice were grouped into the Marine Group I Crenarchaeota while the archaeal clones from seawater were assigned to Marine Group I Crenarchaeota, Marine Group II Euryarchaeota, and Marine Group III Euryarchaeota. Overall, the findings of this study showed that the diversity of archaea in the sea-ice in Prydz Bay was low.展开更多
Box C/D RNAs guide the site-specific formation of 2′-O-methylated nucleotides(Nm)of RNAs in eukaryotes and archaea.Although C/D RNAs have been profiled in several archaea,their targets have not been experimentally de...Box C/D RNAs guide the site-specific formation of 2′-O-methylated nucleotides(Nm)of RNAs in eukaryotes and archaea.Although C/D RNAs have been profiled in several archaea,their targets have not been experimentally determined.Here,we mapped Nm in r RNAs,t RNAs,and abundant small RNAs(s RNAs)and profiled C/D RNAs in the crenarchaeon Sulfolobus islandicus.The targets of C/D RNAs were assigned by analysis of base-pairing interactions,in vitro modification assays,and gene deletion experiments,revealing a complicated landscape of C/D RNA-target interactions.C/D RNAs widely use dual antisense elements to target adjacent sites in r RNAs,enhancing modification at weakly bound sites.Two consecutive sites can be guided with the same antisense element upstream of box D or D′,a phenomenon known as doublespecificity that is exclusive to internal box D′in eukaryotic C/D RNAs.Several C/D RNAs guide modification at a single non-canonical site.This study reveals the global landscape of RNA-guided 2′-O-methylation in an archaeon and unexpected targeting rules employed by C/D RNA.展开更多
With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including ...With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.展开更多
Kinase,putative Endopeptidase,and Other Proteins of Small size(KEOPS)is a multisubunit protein complex conserved in eukaryotes and archaea.It is composed of Pcc1,Kae1,Bud32,Cgi121,and Gon7 in eukaryotes and is primari...Kinase,putative Endopeptidase,and Other Proteins of Small size(KEOPS)is a multisubunit protein complex conserved in eukaryotes and archaea.It is composed of Pcc1,Kae1,Bud32,Cgi121,and Gon7 in eukaryotes and is primarily involved in N^(6)-threonylcarbamoyl adenosine(t^(6)A)modification of transfer RNAs(tRNAs).Recently,it was reported that KEOPS participates in homologous recombination(HR)repair in yeast.To characterize the KEOPS in archaea(aKEOPS),we conducted genetic and biochemical analyses of its encoding genes in the hyperthermophilic archaeon Saccharolobus islandicus.We show that aKEOPS also possesses five subunits,Pcc1,Kae1,Bud32,Cgi121,and Pcc1-like(or Gon7-like),just like eukaryotic KEOPS.Pcc1-like has physical interactions with Kae1 and Pcc1 and can mediate the monomerization of the dimeric subcomplex(Kae1-Pcc1-Pcc1-Kae1),suggesting that Pcc1-like is a functional homolog of the eukaryotic Gon7 subunit.Strikingly,none of the genes encoding aKEOPS subunits,including Pcc1 and Pcc1-like,can be deleted in the wild type and in a t^(6)A modification complementary strain named TsaKI,implying that the aKEOPS complex is essential for an additional cellular process in this archaeon.Knock-down of the Cgi121 subunit leads to severe growth retardance in the wild type that is partially rescued in TsaKI.These results suggest that aKEOPS plays an essential role independent of the cellular t^(6)A modification level.In addition,archaeal Cgi121 possesses dsDNA-binding activity that relies on its tRNA 3ʹCCA tail binding module.Our study clarifies the subunit organization of archaeal KEOPS and suggests an origin of eukaryotic Gon7.The study also reveals a possible link between the function in t^(6)A modification and the additional function,presumably HR.展开更多
Archaea,one of the three domains of life along with Bacteria and Eukarya,contains ancient life forms such as methanogen that are observed today on Earth,and one lineage Asgard archaea is also considered the closest an...Archaea,one of the three domains of life along with Bacteria and Eukarya,contains ancient life forms such as methanogen that are observed today on Earth,and one lineage Asgard archaea is also considered the closest ancestor of Eukarya.Recently,with the development of interdisciplinary studies from Earth and Life sciences,archaeal organisms are considered to play pivotal roles in geochemical cycling in nature.However,our understanding of the attributes,origin and evolution,geochemical and ecological functions of Archaea is hampered by the scarcity of archaeal isolates,which has represented a challenge to researchers for the last 40 years.Cultivation-independent sequencing and phylogenomic analyses have demonstrated a considerable diversity of Archaea with more than 20 novel phyla.However,only four archaeal phyla have cultured representatives,leaving large gaps in our knowledge of the metabolic capabilities and ecological functions of the majority of archaeal strains identified exclusively by DNA sequencing.In this review,we summarize the discovery and development of archaeal research,highlight the knowledge gap between uncultured and cultured archaeal microbes,and call on the importance of devoting greater research efforts to archaeal cultivation.Finally,we outlined new ideas and strategic approaches,namely,(1)genome-based methods,(2)microbial network information-based methods,(3)genome-scale model-guided methods,and(4)machine learning methods,to enable the cultivation of uncultivated archaeal microbes using accumulated high-throughput sequencing data.展开更多
基金The National Key R&D Program of China under contract No.2022YFE0209300the National Natural Science Foundation of China under contract No.42006152+1 种基金the Zhejiang Provincial Natural Science Foundation of China under contract No.LQ21D060005the 111 Project under contract No.BP0820020.
文摘Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.
基金the International Cooperation Project of the Ministry of Sciences and Technology of China(2014DFA32860)the National Natural Science Foundation of China(31402104)for their financial support
文摘This study was conducted to investigate the phylogenetic diversity of archaea in the rumen of adult and elderly yaks. Six domesticated female yaks, 3 adult yaks((5.3±0.6) years old), and 3 elderly yaks((10.7±0.6) years old), were used for the rumen contents collection. Illumina Mi Seq high-throughput sequencing technology was applied to examine the archaeal composition of rumen contents. A total of 92 901 high-quality archaeal sequences were analyzed, and these were assigned to 2 033 operational taxonomic units(OTUs). Among these, 974 OTUs were unique to adult yaks while 846 OTUs were unique to elderly yaks; 213 OTUs were shared by both groups. At the phylum level, more than 99% of the obtained OTUs belonged to the Euryarchaeota phylum. At the genus level, the archaea could be divided into 7 archaeal genera. The 7 genera(i.e., Methanobrevibacter, Methanobacterium, Methanosphaera, Thermogymnomonas, Methanomicrobiu, Methanimicrococcus and the unclassified genus) were shared by all yaks, and their total abundance accounted for 99% of the rumen archaea. The most abundant archaea in elderly and adult yaks were Methanobrevibacter and Thermogymnomonas, respectively. The abundance of Methanobacteria(class), Methanobacteriales(order), Methanobacteriaceae(family), and Methanobrevibacter(genus) in elderly yaks was significantly higher than in adult yaks. In contrast, the abundance of Thermogymnomonas in elderly yaks was 34% lower than in adult yaks, though the difference was not statistically significant. The difference in abundance of other archaea was not significant between the two groups. These results suggested that the structure of archaea in the rumen of yaks changed with age. This is the first study to compare the phylogenetic differences of rumen archaeal structure and composition using the yak model.
基金supported by the Key Program for International S&T Cooperation Projects of China(2016YFE0109000)the National Key R&D Program of China(2017YFF0211702)+1 种基金the National Natural Science Foundation of China(41475126 and 31802085)the Young Scientist Lifting Project,China(2017–2019)
文摘The objective of the present study was to examine the effect of different weaning methods on the ruminal methanogenic archaea composition and diversity in Holstein calves.Thirty-six newborn Holstein bull calves were assigned to 1 of 3 treatments:(1)conventional weaning(d 56)and fed a high proportion of solid feed(CWS);(2)conventional weaning(d 56)and fed a high proportion of liquid feed(CWL);(3)early weaning(d 42)and fed with a high proportion of solid feed(EWS).High-throughput sequencing of the methyl coenzyme M reductase(mcr A)gene,which encodes theα-subunit of methyl coenzyme M reductase-the enzyme that catalyzes the final step in methanogenesis was used to determine the composition and diversity of rumen methanogens.No significant difference(P>0.05)was observed for operational taxonomic units(OTUs)or richness indices,but diversity indices increased(P<0.05)for calves fed high dietary solids.Predominant families across the three treatments were Methanobacteriaceae,Thermoplasmataceae and Methanomassiliicoccaceae.Calves in the EWS treatment had a higher(P<0.05)relative abundance of Methanobrevibacter sp.strain AbM4 and Methanosphaera stadtmanae,while calves in the CWL treatment had a higher(P<0.05)abundance of Methanosphaera sp.strain SM9.A positive(P<0.05)relationship was identified between butyrate and Methanobrevibacter sp.strain AbM4.In conclusion,the composition and diversity of methanogens in the rumen of Holstein calves varied under the different weaning methods.This study identified a positive relationship between butyrate and Methanobrevibacter sp.strain AbM4,potentially reflecting correlations between ruminal fermentation variables and methanogenesis function.These in-depth analyses provide further understanding of weaning methods for intensified production systems.
基金supported financially by the Strategic Priority Research Program of Chinese Academy of Sciences (Grant No. XDA2003010301)the National Natural Science Foundation of China (Grant No. 41621001)
文摘Metagenomic studies have demonstrated the existence of ammonia-oxidizing archaea(AOA) and revealed they are responsible for ammoxidation in some extreme environments. However, the changes in compositional structure and ammonia-oxidation capacity of AOA communities in biological soil crusts(BSCs) of desert ecosystems remain poorly understood.Here, we utilized Illumina MiSeq sequencing and microbial functional gene array(GeoChip 5.0) to assess the above changes along a 51-year revegetation chronosequence in the Tengger Desert, China. The results showed a significant difference in AOA-community richness between 5-year-old BSCs and older ones. The most dominant phylum during BSC development was Crenarchaeota, and the corresponding species were ammonia-oxidizing_Crenarchaeote and environmental_samples_Crenarchaeota. Network analysis revealed that the positive correlations among dominant taxa increased, and their cooperation was reinforced in AOA communities during BSC succession. Redundancy analysis showed that the dominant factor influencing the change in AOA-community structure was soil texture. GeoChip 5.0 indicated that the amoA gene abundances of AOA and ammonia-oxidizing bacteria(AOB) were basically the same, demonstrating that AOA and AOB played an equally important role during BSCs development. Our study of the long-term succession of BSC demonstrated a persistent response of AOA communities to revegetation development in desert ecosystems.
基金Supported by the China Ocean Mineral Resources Research and Development Association Program(Nos.DY135-E2-1-01,DY135-E2-4-00)the China Global Sea-Atmosphere Interaction Research Program(No.GASI-02-IND-STSsum)the S&T Innovation Project of Qingdao National Laboratory for Marine Science and Technology(No.2016ASKJ14)。
文摘Depth-dependent distribution patterns of bacterial and archaeal communities in deep sea water column around the Ninetyeast Ridge in the Indian Ocean were investigated using 16S rRNA gene profiling.Sampling was conducted at the northern Ninetyeast Ridge(1°59.89′N–9°59.70′S,87°58.90′E–88°00.03′E)from September to November 2016 where samples were collected from the bathyal(1000 m)to bathypelagic depths(>4000 m)in four different stations.A total of 1565405 clean data falling into 6712 bacterial OTUs and 1452727 clean data falling into 806 archaeal OTUs based on 97%similarity level were analyzed.Most of the bacterial 16S rRNA gene sequences were affiliated with Gammaproteobacteria,followed by Alphaproteobacteria and Bacteroidia.The archaeal 16S rRNA gene sequences mostly affiliated to Nitrososphaeria(Thaumarchaeota)dominated with relative abundances ranging from 52.68%to 97.2%,followed by Thermoplasmata(Euryarchaeota).Vertical partitioning of bacterial and archaeal communities among different water layers was observed.Canonical correspondence analysis(CCA)and Spearman’s correlations revealed that depth(P=0.003),dissolved oxygen(P=0.019),and nitrite(P=0.033)were the main environmental factors affecting bacterial community structure at genus level in the Ninetyeast Ridge.On the other hand,the first two CCA axes accounted for 74.4%of the explained total variance,it seems that the archaeal communities at genus level were heavily influenced by the environmental variables including depth,dissolved oxygen(DO),nitrite,salinity,phosphate,ammonia,nitrate,and silicate,but none of them exhibited any significant correlation on the structuring(P>0.1).
文摘The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high organic matter content, fertility, and poor rocky soil. Using domain-specific primers, archaeal 16S rRNA gene sequences were PCR amplified for clone library construction, and a total of 185 archaeal clones were examined. The archaeal clones were affiliated with the phyla Thaumarchaeota (96.2%) and Euryarchaeota (3.8%). The most abundant thaumarchaeal group (90.3% of the clones) was the group I.1b clade, which includes soil ammonia-oxidizing archaea. The unique characteristics of Gotjawal soil, including basalt morphology, vegetation, and groundwater aquifer penetration, may be reflected in the archaeal community composition. Further study is necessary to understand the unique factors of Gotjawal soils that influence archaeal abundance, composition, and diversity.
基金Supported by the Key Scientific Research Project of Colleges and Universities in Henan Province(16B180004)Science and Technology Public Relations Project from Department of Science and Technology in Henan Province(172102310211,172102310529)+1 种基金Scientific Research Start-up Foundation of High-level Talents in Pingdingshan University(PXY-BSQD-2015006)National Natural Fund Cultivation Project of Pingdingshan University(PXYPYJJ2016004)
文摘There are five hot springs named separately SHANGT,ZHONGT,WENT,XIAT and SHENT in Lushan County,Pingdingshan City,Henan Province.The archaeal diversity of the five hot springs was investigated and analyzed.16 S rD NA v4-5 genes of archaea were sequenced by Illumina Hisiq2500 PE250 technology.The numbers of sample sequences and OTUs were sorted by using Uparse,Mothur,SILVA,MUSCLE and Qiime software.Finally,the archaeal abundance and diversity in the five hot springs were analyzed.87993,67326,44400,48611 and 48495 effective reads have been obtained from SHANGT,ZHONGT,WENT,XIAT and SHENT respectively,and 520,280,678,451 and 437 OTUs have been acquired.The analysis of archaeal classification showed that the archaea from the five hot springs could be classified into 11 phyla,in which Thaumarchaeota accounted for 95.25%and was the dominant archaea.In the genera level,the dominant archaea of the SHANGT,ZHONGT and WENT was Candidatus-Nitrosocaldus,and unidentified-Thaumarchaeota was the dominant archaea in XIAT and SHENT.The results showed that the five hot springs in Lushan are rich in microbial resources.The research provided basic data for further research and utilization of the archaea resources in the five hot springs of Lushan.
基金funding from the National Basic Research Program of China (2012CB124705,2013CB127603)the European Community’s Seventh Framework Programme (FP7/2007-2013) under the grant agreement n° 227549+1 种基金the Royal Dutch Academy of Sciences (KNAW) through Project 09CDP006the Fundamental Research Funds for the Central Universities,China (KYZ201153)
文摘Erhualian and Landrace breeds are typical genetically obese and lean pigs, respectively. To compare the fecal methanogenic Archaeal community between these two pig breeds, fecal samples from different growth phase pigs were collected and used for PCR-denaturing gradient gel electrophoresis(DGGE) with two primer pairs(344fGC/519r and 519f/915rGC) and real-time PCR analysis. Results showed that a better separation and higher quality of bands pattern were obtained in DGGE profiles using primers 344fGC/519r as compared with primers 519f/915rGC. Sequencing of DGGE bands showed that the predominant methanogens in the feces of Erhualian and Landrace pigs belonged to Methanobrevibacter spp. and Methanosphaera spp. Real-time PCR analysis revealed that there was no significant difference in the numbers of fecal total methanogens between Erhualian and Landrace pigs; however, pig growth phase affected the numbers of 16S rRNA genes of total methanogens and Methanobrevibacter smithii. Dissociation curves of methyl coenzyme-M reductase subunit A(mcrA) gene fragments amplified with real-time PCR showed all samples possessed a single peak at 82?C, which might be associated with M. smithii. Samples from the same growth phase of each breed showed good replicative dissociation curves. The results suggest that the growth phase(including diet factor) other than genotype of pig may affect the fecal methanogenic Archaeal community of pigs.
文摘The number of completely sequenced archaeal genomes has been sufficient for a large-scale bioinformatic study.We have conducted analyses for each coding region from 36 archaeal genomes using the original CGS algorithm by calculating the total GC content(G+C),GC content in first,second and third codon positions as well as in fourfold and twofold degenerated sites from third codon positions,levels of arginine codon usage(Arg2:AGA/G;Arg4:CGX),levels of amino acid usage and the entropy of amino acid content distribution.In archaeal genomes with strong GC pressure,arginine is coded preferably by GC-rich Arg4 codons,whereas in most of archaeal genomes with G+C<0.6,arginine is coded preferably by AT-rich Arg2 codons.In the genome of Haloquadratum walsbyi,which is closely related to GC-rich archaea,GC content has decreased mostly in third codon positions,while Arg4>>Arg2 bias still persists.Proteomes of archaeal species carry characteristic amino acid biases:levels of isoleucine and lysine are elevated,while levels of alanine,histidine,glutamine and cytosine are relatively decreased.Numerous genomic and proteomic biases observed can be explained by the hypothesis of previously existed strong mutational AT pressure in the common predecessor of all archaea.
基金Supported by the International Cooperation Research Program of the Ministry of Science & Technology of China(No.2010DFA32300)the Natural Science Foundation of Tianjin(No.13JCZDJC28700)the Foundation of Tianjin Key Laboratory of Marine Resources and Chemistry(No.201105)
文摘A total of 26 isolates were obtained from solar salt ponds of different salinities(100, 150, 200, and 250) in Hangu Saltworks Co. Ltd., Tianjin, China. Phylogenetic analysis of 16 SrRNA gene sequences indicated that five bacteria genera H alomonas, Salinicoccus, Oceanobacillus, Gracibacillus, and Salimicrobium and one archaea genera H alorubrum were present. The genus H alomonas was predominant with eight strains distributed in a salinity range of 100–200, followed by H alorubrum with six strains in salinity 250. Based on the genus and original sampling salinity, eight bacterial and two archaeal isolates were selected for further morphological, physiological, and biochemical characterization. All of the bacterial strains were moderately halophilic with the optimal salinity for growth being either 50 or 100, while two archaeal strains were extremely halophilic with an optimal growth salinity of 200. Additionally, we put forth strain SM.200-5 as a new candidate S alimicrobium species based on the phylogenic analysis of the 16 SrRNA gene sequence and its biochemical characteristics when compared with known related species.
基金Supported by the International Cooperation Research Program of the Ministry of Science&Technology of China(No.2010DFA32300)the Natural Science Foundation of Tianjin(No.13JCZDJC28700)
文摘Halophilic archaea comprise the maj ority of microorganisms found in hypersaline environments.C50 carotenoids accumulated in archaea cells are considered potential biotechnological products and possess a number of biological functions.Ten red colonies were isolated from brine water in a saltern crystallizer pond of the Hangu Saltworks,China.16 S rRNA gene sequence analysis showed that the colonies belonged to the extremely halophilic archaea genera Halobacterium and Halorubrum.Two representative strains,Halobacterium strain SP-2 and Halorubrum strain SP-4,were selected for further study on the phenotypic characteristics and effects of salinity and pH on accumulation and composition of pigments in their cells.The archaeal strains were isolated and grown in a culture medium prepared by dissolving yeast extract(10 g/L) and acid-hydrolyzed casein(7.5 g/L) into brine water obtained from a local salt pond.Their optimum salinity and pH for growth were 250 and 7,respectively,although pigment accumulation(OD490/mL broth) was highest at pH 8.In addition,at 150-300 salinity,increasing salinity resulted in decreasing pigment accumulation.Analysis of the UV-Vis spectrum,TLC and HLPC chromatograms showed that C50 carotenoid bacterioruberin is the major pigment in both strains.
基金supported by the National Natu-ral Science Foundation of China under Grant No.30525037the National 863 Program under Grant No. 2004AA231060
文摘The genetic codon UGA has a dual function: serving as a terminator and encoding selenocysteine. However, most popular gene annotation programs only take it as a stop signal, resulting in misannotation or completely missing selenoprotein genes. We developed a computational method named Asec-Prediction that is specific for the prediction of archaeal selenoprotein genes. To evaluate its effectiveness, we first applied it to 14 archaeal genomes with previously known selenoprotein genes, and Asec-Prediction identified all reported selenoprotein genes without redundant results. When we applied it to 12 archaeal genomes that had not been researched for selenoprotein genes, Asec-Prediction detected a novel selenoprotein gene in Methanosarcina acetivorans. Further evidence was also collected to support that the predicted gene should be a real selenoprotein gene. The result shows that Asec-Prediction is effective for the prediction of archaeal selenoprotein genes.
基金supported by the National Natural Science Foundation of China (30900926)funded by the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-IVFCAAS)the China Agriculture Research System (CARS-23-D01)
文摘The expression of mitogen-activated protein kinase(mapk) double-stranded RNA in cucumber is effective in controlling infestations of the root-knot nematode Meloidogyne incognita. However,little is known about the ecological effects of transgenic plants. Here,we analyzed the diversity of 16 S r DNA genes derived from the rhizosphere archaea of transgenic cucumber plants as an indicator of ecological change. A total of 17 and 18 operational taxonomic units were detected in the rhizospheres of non-transgenic cucumber and mapk ds RNA-expressing plants,respectively. No significant difference was observed between the two groups according to Shannon and Simpson indices. In soil samples of the two rhizospheres,the dominant group was Crenarchaeota at the phylum level,with Staphylothermus,Methanococcus,Pyrodictium and Sulfolobus the abundant taxa at the genus level. These results suggest that expressing mapk double-stranded(ds) RNA in cucumber has no apparent effect on the diversity of rhizosphere archaea,and provide powerful evidence for the ecological safety of transgenic cucumber expressing mapk ds RNA.
基金Supported by the Special Fund for Public Projects of Zhejiang Province(2016C32073)Agriculture Project of Guizhou Province(20103047)International Cooperation Program of Zhejiang Academy of Agricultural Sciences
文摘The gastrointestinal tract,as the major digestive and absorbing organ,plays an important role in chicken growth.Numerous studies have been undertaken to investigate the gut bacterial community in broilers,but little is known about the archaeal community composition.To improve our understanding of the composition of intestinal archaea and the relationship between archaea and chicken growth,a feeding trial was conducted with 10 000 Ross 308 chicks in a commercial production facility under standard management.At 38 d of age,15 birds with the highest body weight( H group),15 birds with average body weight( M group),and15 birds with the lowest body weight( L group) were selected among 600 healthy male individuals.Cecal archaeal community was detected based on 16 S rRNA gene sequencing.As a result,3 321 229 archaeal sequences were clustered into 1 281 operational taxonomic units( OTUs) and classified into three phyla( Crenarchaeota,Euryarchaeota and Thaumarchaeota) and six known genera: Natronomonas,Halolamina,Salinarchaeum,Cenarchaeum,Candidatus,Nitrosoarchaeum and Nitrososphaera.The alpha indices showed that,the richness of archaea in L group was the highest,and followed by that in M group and H group,while the archaea in H group were more diverse,followed by those in M group and L group.However,the archaeal composition in all chicken samples was similar,with the vast majority of the sequences belonging to Cenarchaeum,and only a few belonging to the other five genera.
基金supported by the Natural Science Foundation of China(Grant nos.40876097,41076131 and 41376191)the Chinese Polar Environment Comprehensive Investigation & Assessment Programs(Grant no.CHINARE 2014-02-01)the State High-Tech Research and Development Project(863) of the Ministry of Science and Technology of China(Grant no.2012AA021706)
文摘Fluorescence in-situ hybridization(FISH) and 16S rRNA gene clone library analyses were used to determine the abundance and diversity of archaea in Prydz Bay, Antarctica. Correlation analysis was also performed to assess links between physicochemical parameters and archaeal abundance and diversity within the sea-ice. Samples of sea-ice and seawater were collected during the 26th Chinese National Antarctic Research Expedition. The results of FISH showed that archaea were relatively abundant within the top layer of the sea-ice, and correlation analysis suggested that the concentration of 4NH+ might be one of the main factors underlying this distribution pattern. However, using 16S rRNA gene libraries, archaea were not detected in the top and middle layers of the sea-ice. All archaeal clones obtained from the bottom layer of the sea-ice were grouped into the Marine Group I Crenarchaeota while the archaeal clones from seawater were assigned to Marine Group I Crenarchaeota, Marine Group II Euryarchaeota, and Marine Group III Euryarchaeota. Overall, the findings of this study showed that the diversity of archaea in the sea-ice in Prydz Bay was low.
基金supported by the Strategic Priority Research Program of Chinese Academy of Sciences(XDB0570000,XDB37010201)the Basic Research Program Based on Major Scientific Infrastructures of Chinese Academy of Sciences(JZHKYPT-2021-05)+1 种基金the National Natural Science Foundation of China(91940302,91540201,31430024,31325007)the National Key Research and Development Program of China(2017YFA0504600)。
文摘Box C/D RNAs guide the site-specific formation of 2′-O-methylated nucleotides(Nm)of RNAs in eukaryotes and archaea.Although C/D RNAs have been profiled in several archaea,their targets have not been experimentally determined.Here,we mapped Nm in r RNAs,t RNAs,and abundant small RNAs(s RNAs)and profiled C/D RNAs in the crenarchaeon Sulfolobus islandicus.The targets of C/D RNAs were assigned by analysis of base-pairing interactions,in vitro modification assays,and gene deletion experiments,revealing a complicated landscape of C/D RNA-target interactions.C/D RNAs widely use dual antisense elements to target adjacent sites in r RNAs,enhancing modification at weakly bound sites.Two consecutive sites can be guided with the same antisense element upstream of box D or D′,a phenomenon known as doublespecificity that is exclusive to internal box D′in eukaryotic C/D RNAs.Several C/D RNAs guide modification at a single non-canonical site.This study reveals the global landscape of RNA-guided 2′-O-methylation in an archaeon and unexpected targeting rules employed by C/D RNA.
文摘With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.
基金supported by the National Key Research and Development Program of China(No.2020YFA0906800)the National Natural Science Foundation of China(Nos.31970546 and 31670061 to Y.S.,31900055 to Q.H.,31970119 to J.N.,and 31771380 to Q.S.)the State Key Laboratory of Microbial Technology.
文摘Kinase,putative Endopeptidase,and Other Proteins of Small size(KEOPS)is a multisubunit protein complex conserved in eukaryotes and archaea.It is composed of Pcc1,Kae1,Bud32,Cgi121,and Gon7 in eukaryotes and is primarily involved in N^(6)-threonylcarbamoyl adenosine(t^(6)A)modification of transfer RNAs(tRNAs).Recently,it was reported that KEOPS participates in homologous recombination(HR)repair in yeast.To characterize the KEOPS in archaea(aKEOPS),we conducted genetic and biochemical analyses of its encoding genes in the hyperthermophilic archaeon Saccharolobus islandicus.We show that aKEOPS also possesses five subunits,Pcc1,Kae1,Bud32,Cgi121,and Pcc1-like(or Gon7-like),just like eukaryotic KEOPS.Pcc1-like has physical interactions with Kae1 and Pcc1 and can mediate the monomerization of the dimeric subcomplex(Kae1-Pcc1-Pcc1-Kae1),suggesting that Pcc1-like is a functional homolog of the eukaryotic Gon7 subunit.Strikingly,none of the genes encoding aKEOPS subunits,including Pcc1 and Pcc1-like,can be deleted in the wild type and in a t^(6)A modification complementary strain named TsaKI,implying that the aKEOPS complex is essential for an additional cellular process in this archaeon.Knock-down of the Cgi121 subunit leads to severe growth retardance in the wild type that is partially rescued in TsaKI.These results suggest that aKEOPS plays an essential role independent of the cellular t^(6)A modification level.In addition,archaeal Cgi121 possesses dsDNA-binding activity that relies on its tRNA 3ʹCCA tail binding module.Our study clarifies the subunit organization of archaeal KEOPS and suggests an origin of eukaryotic Gon7.The study also reveals a possible link between the function in t^(6)A modification and the additional function,presumably HR.
基金supported by the State Key R&D Project of China(Grant No.2016YFA0601102)the China Ocean Mineral Resources R&D Association(Grant No.DY135-B2-12)+6 种基金COMRA Project DY135-B2-12the National Nature Science Foundation of China(Grant Nos.41525011,41921006,41902313,91751205&92051116)the Natural Science Foundation of Shanghai(Grant No.20ZR1428000)the Shanghai Jiao Tong University Interdisciplinary Grant(Grant No.20CX-01)Japan Society for the Promotion of Science(JSPS)(KAKENHI Grant 18H05295)This is also a contribution to the Center for Ocean Mega-Science,Chinese Academy of Sciences,the Senior User Project of RV KEXUE(Grant No.KEXUE2019GZ06)the International Center for Deep-life Investigation(IC-DLI).
文摘Archaea,one of the three domains of life along with Bacteria and Eukarya,contains ancient life forms such as methanogen that are observed today on Earth,and one lineage Asgard archaea is also considered the closest ancestor of Eukarya.Recently,with the development of interdisciplinary studies from Earth and Life sciences,archaeal organisms are considered to play pivotal roles in geochemical cycling in nature.However,our understanding of the attributes,origin and evolution,geochemical and ecological functions of Archaea is hampered by the scarcity of archaeal isolates,which has represented a challenge to researchers for the last 40 years.Cultivation-independent sequencing and phylogenomic analyses have demonstrated a considerable diversity of Archaea with more than 20 novel phyla.However,only four archaeal phyla have cultured representatives,leaving large gaps in our knowledge of the metabolic capabilities and ecological functions of the majority of archaeal strains identified exclusively by DNA sequencing.In this review,we summarize the discovery and development of archaeal research,highlight the knowledge gap between uncultured and cultured archaeal microbes,and call on the importance of devoting greater research efforts to archaeal cultivation.Finally,we outlined new ideas and strategic approaches,namely,(1)genome-based methods,(2)microbial network information-based methods,(3)genome-scale model-guided methods,and(4)machine learning methods,to enable the cultivation of uncultivated archaeal microbes using accumulated high-throughput sequencing data.
