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The simultaneous enhancement of electron transfer and Li+ migration is of particular importance to achieve high electrochemical performance in lithium-ion batteries.

Pigeon breeding is associated with exposure to airborne microorganisms and endotoxin and with symptoms of the airways. Antibiotic resistance is a threat to human health. Some pigeons participate in national and international indoor exhibitions. This study aims to obtain knowledge about the potential human exposure to dust, endotoxin, fungi, and bacteria including the methicillin-resistant Staphylococcus aureus (MRSA) in a pigeon exhibition in Denmark. In walking areas for visitors, airborne microorganisms in different size fractions able to enter the airways were sampled and following identified. The average concentrations were: 5000 cfu fungi/m3, 1.8 × 104 cfu bacteria/m3, 37 endotoxin units/m3, and 0.18 mg dust/m3 air with the highest concentrations in-between rows with pigeon cages. The fungal species Wallemia sp. and Aspergillus versicolor and the bacterial species S. equorum and S. aureus were found in high concentrations. MRSA spa type t034 described to be associated with livestock was found in the air. Most of the S. aureus was present in the size fraction of 1.1–2.1 μm, which are particles able to enter the human terminal bronchi. In conclusion, fungi, bacteria, and endotoxin, respectively, were found in concentrations 10, 2000, and 200 times higher than outdoor references. The airborne bacteria in the exhibition were mainly species found previously in pigeon coops showing that the pigeons are the sources of exposure. The presence of airborne MRSA in the pigeon exhibition highlights the importance of also considering this environment as a potential place of exchange of resistant bacteria between animals and between animals and humans.

Ion-specific effects of cations (Li+, Na+, K+, Mg2+, Ca2+) and anions (F-, Cl-) on the hydrogen bond structure and dynamics of the coordination waters in the hydration shells have been studied using molecular dynamics simulations. Our simulations indicate that the hydrogen bonds between the first and second hydration shell waters show binary structural and dynamic properties. The hydrogen bond with a first shell water as the donor (HD) is strengthened, while those with a first shell water as the acceptor (HA) are weakened. For a hydrated anion, this binary effect reverses, but is less significant. This ion-specific binary effect correlates with the size and the valence of the ion, and is more significant for the strong kosmotropic ions of high charge density.

Phototrophic Gemmatimonadetes evolved the ability to use solar energy following horizontal transfer of photosynthesis-related genes from an ancient phototrophic proteobacterium. The electron cryo-microscopy structure of the Gemmatimonas phototrophica photosystem at 2.4 Å reveals a unique, double-ring complex. Two unique membrane-extrinsic polypeptides, RC-S and RC-U, hold the central type 2 reaction center (RC) within an inner 16-subunit light-harvesting 1 (LH1) ring, which is encircled by an outer 24-subunit antenna ring (LHh) that adds light-gathering capacity. Femtosecond kinetics reveal the flow of energy within the RC-dLH complex, from the outer LHh ring to LH1 and then to the RC. This structural and functional study shows that G. phototrophica has independently evolved its own compact, robust, and highly effective architecture for harvesting and trapping solar energy.

Phyllosphere is a habitat to a variety of viruses, bacteria, fungi, and other microorganisms, which play a fundamental role in maintaining the health of plants and mediating the interaction between plants and ambient environments. A recent addition to this catalogue of microbial diversity was the aerobic anoxygenic phototrophs (AAPs), a group of widespread bacteria that absorb light through bacteriochlorophyll α (BChl a) to produce energy without fixing carbon or producing molecular oxygen. However, culture representatives of AAPs from phyllosphere and their genome information are lacking, limiting our capability to assess their potential ecological roles in this unique niche. In this study, we investigated the presence of AAPs in the phyllosphere of a winter wheat (Triticum aestivum L.) in Denmark by employing bacterial colony based infrared imaging and MALDI-TOF mass spectrometry (MS) techniques. A total of ∼4,480 colonies were screened for the presence of cellular BChl a, resulting in 129 AAP isolates that were further clustered into 21 groups based on MALDI-TOF MS profiling, representatives of which were sequenced using the Illumina NextSeq and Oxford Nanopore MinION platforms. Seventeen draft and four complete genomes of AAPs were assembled belonging in Methylobacterium, Rhizobium, Roseomonas, and a novel Alsobacter. We observed a diverging pattern in the evolutionary rates of photosynthesis genes among the highly homogenous AAP strains of Methylobacterium (Alphaproteobacteria), highlighting an ongoing genomic innovation at the gene cluster level.

Aerobic anoxygenic phototrophic bacteria (AAnPB) are photoheterotrophs, which use light as auxiliary energy and require organic carbon (OC) for growth. Herein, we report the unusual strain B3, which is a true AAnPB because it requires oxygen for growth, harbours genes for cbb3- and bd-type cytochromes and acsF, and produces bacteriochlorophyll. The B3 genome encodes the complete metabolic pathways for AAnPB light utilization, CO2 fixation via Calvin-Benson-Bassham (CBB) cycle and oxidation of sulfite and H2, and the transcriptome indicated that all components of these pathways were fully transcribed. Expression of the marker genes related to photosynthesis, including pufM for light harnessing and rbcL for CO2 fixation, and the activity of RubisCO, the key enzyme in the Calvin-Benson-Bassham (CBB) cycle, increased in response to decreased OC supply. Large amounts of cell biomass were obtained in liquid BG11 medium under illumination. The strain thus likely photoautotrophically grows using sulfite or H2 as an electron donor. Similar GC contents between photosynthesis, the CBB cycle and 16S rRNA genes and the consistency of their phylogenetic topologies implied that light harnessing and carbon fixation genes evolved vertically from an anaerobic phototrophic ancestor of Rhodospirillaceae in Alphaproteobacteria. In conclusion, strain B3 represents a novel AAnPB characterized by photoautotrophy using the CBB cycle. This kind of AAnPB may be ecologically significant in the global carbon cycle.

Over the course of evolution for billions of years, bacteria that are capable of light-driven energy production have occupied every corner of surface Earth where sunlight can reach. Only two general biological systems have evolved in bacteria to be capable of net energy conservation via light harvesting: one is based on the pigment of (bacterio-)chlorophyll and the other is based on proton-pumping rhodopsin. There is emerging genomic evidence that these two rather different systems can coexist in a single bacterium to take advantage of their contrasting characteristics in the number of genes involved, biosynthesis cost, ease of expression control, and efficiency of energy production and thus enhance the capability of exploiting solar energy. Our data provide the first clear-cut evidence that such dual phototrophy potentially exists in glacial bacteria. Further public genome mining suggests this understudied dual phototrophic mechanism is possibly more common than our data alone suggested., Conserving additional energy from sunlight through bacteriochlorophyll (BChl)-based reaction center or proton-pumping rhodopsin is a highly successful life strategy in environmental bacteria. BChl and rhodopsin-based systems display contrasting characteristics in the size of coding operon, cost of biosynthesis, ease of expression control, and efficiency of energy production. This raises an intriguing question of whether a single bacterium has evolved the ability to perform these two types of phototrophy complementarily according to energy needs and environmental conditions. Here, we report four Tardiphaga sp. strains (Alphaproteobacteria) of monophyletic origin isolated from a high Arctic glacier in northeast Greenland (81.566° N, 16.363° W) that are at different evolutionary stages concerning phototrophy. Their >99.8% identical genomes contain footprints of horizontal operon transfer (HOT) of the complete gene clusters encoding BChl- and xanthorhodopsin (XR)-based dual phototrophy. Two strains possess only a complete XR operon, while the other two strains have both a photosynthesis gene cluster and an XR operon in their genomes. All XR operons are heavily surrounded by mobile genetic elements and are located close to a tRNA gene, strongly signaling that a HOT event of the XR operon has occurred recently. Mining public genome databases and our high Arctic glacial and soil metagenomes revealed that phylogenetically diverse bacteria have the metabolic potential of performing BChl- and rhodopsin-based dual phototrophy. Our data provide new insights on how bacteria cope with the harsh and energy-deficient environment in surface glacier, possibly by maximizing the capability of exploiting solar energy.

Phototrophic organisms are key components of many natural environments. There exist two main phototrophic groups: species that collect light energy using various kinds of (bacterio)chlorophylls and species that utilize rhodopsins., Photoheterotrophic bacteria represent an important part of aquatic microbial communities. There exist two fundamentally different light-harvesting systems: bacteriochlorophyll-containing reaction centers or rhodopsins. Here, we report a photoheterotrophic Sphingomonas strain isolated from an oligotrophic lake, which contains complete sets of genes for both rhodopsin-based and bacteriochlorophyll-based phototrophy. Interestingly, the identified genes were not expressed when cultured in liquid organic media. Using reverse transcription quantitative PCR (RT-qPCR), RNA sequencing, and bacteriochlorophyll a quantification, we document that bacteriochlorophyll synthesis was repressed by high concentrations of glucose or galactose in the medium. Coactivation of photosynthesis genes together with genes for TonB-dependent transporters suggests the utilization of light energy for nutrient import. The photosynthetic units were formed by ring-shaped light-harvesting complex 1 and reaction centers with bacteriochlorophyll a and spirilloxanthin as the main light-harvesting pigments. The identified rhodopsin gene belonged to the xanthorhodopsin family, but it lacks salinixanthin antenna. In contrast to bacteriochlorophyll, the expression of xanthorhodopsin remained minimal under all experimental conditions tested. Since the gene was found in the same operon as a histidine kinase, we propose that it might serve as a light sensor. Our results document that photoheterotrophic Sphingomonas bacteria use the energy of light under carbon-limited conditions, while under carbon-replete conditions, they cover all their metabolic needs through oxidative phosphorylation. IMPORTANCE Phototrophic organisms are key components of many natural environments. There exist two main phototrophic groups: species that collect light energy using various kinds of (bacterio)chlorophylls and species that utilize rhodopsins. Here, we present a freshwater bacterium Sphingomonas sp. strain AAP5 which contains genes for both light-harvesting systems. We show that bacteriochlorophyll-based reaction centers are repressed by light and/or glucose. On the other hand, the rhodopsin gene was not expressed significantly under any of the experimental conditions. This may indicate that rhodopsin in Sphingomonas may have other functions not linked to bioenergetics.

Bacterial populations rely on quorum sensing (QS) to coordinate their behaviors and are often challenged by the fluctuation in oxygen concentrations in their habitats. Oxygen is a crucial factor that affects bacterial metabolism in multiple ways. However, little is known about whether and how oxygen availability affects QS activities. To fill this gap, we used the facultative anaerobe Novosphingobium pentaromativorans US6-1 as a model system, and observed that the QS signal acyl homoserine-lactones (AHLs) were produced only in anoxic environments, such as biofilm, or liquid medium that initially contained less than 2 mg/L dissolved oxygen, but not in highly oxic environments. Comparative transcriptome analysis revealed that oxygen availability significantly affected the physiological activities in US6-1, including fatty acid metabolism, oxidative phosphorylation, citrate cycle, QS activities, and flagellar assembly. The absence of AHLs in the oxic culture was not due to degradation, but to the very low expression of the AHL synthase gene novI. High concentration of NADH during the middle log phase under static cultivation may be a trigger for AHL synthesis. This is the first report that production of AHLs is coupled with anoxic metabolism in a facultative anaerobe, which extends our knowledge on factors affecting bacterial QS occurrence. Key points: • AHL production is anoxic cultivation related. • Oxygen availability affects AHL synthesis by influencing novI expression. • Oxygen availability changes many metabolism activities including NADH production.

Molecular dynamics (MD) simulations based on the multistate empirical valence bond model have been performed to study the proton transfer (PT) process in aqueous solution. This study focuses on the details of the hydrogen bond (HB) dynamics in the solvation shells of an excess proton accompanied by PT events. The HB dynamics analyses show that the three water molecules in the first solvation shell of hydronium (H

Glacier is the dominant cold habitat in terrestrial environments, providing a model ecosystem to explore extremophilic strategies and study early lives on Earth. The dominant form of life in glaciers is bacteria. However, little is known about past evolutionary processes that bacteria underwent during adaptation to the cryosphere and the connection of their genomic traits to environmental stressors. Aiming to test the hypothesis that bacterial genomic content and dynamics are driven by glacial environmental stressors, we compared genomes of 21 psychrophilic Cryobacterium strains, including 14 that we isolated from three Tibetan ice cores, to their mesophilic counterparts from the same family Microbacteriaceae of Actinobacteria. The results show that psychrophilic Cryobacterium underwent more dynamic changes in genome content, and their genomes have a significantly higher number of genes involved in stress response, motility, and chemotaxis than their mesophilic counterparts (P < 0.05). The phylogenetic birth-and-death model imposed on the phylogenomic tree indicates a vast surge in recent common ancestor of psychrophilic Cryobacterium (gained the greatest number of genes by 1,168) after the division of the mesophilic strain Cryobacterium mesophilum. The expansion in genome content brought in key genes primarily of the categories “cofactors, vitamins, prosthetic groups, pigments,” “monosaccharides metabolism,” and “membrane transport.” The amino acid substitution rates of psychrophilic Cryobacterium strains are two orders of magnitude lower than those in mesophilic strains. However, no significantly higher number of cold shock genes was found in psychrophilic Cryobacterium strains, indicating that multi-copy is not a key factor for cold adaptation in the family Microbacteriaceae, although cold shock genes are indispensable for psychrophiles. Extensive gene acquisition and low amino acid substitution rate might be the strategies of psychrophilic Cryobacterium to resist low temperature, oligotrophy, and high UV radiation on glaciers. The exploration of genome evolution and survival strategies of psychrophilic Cryobacterium deepens our understanding of bacterial cold adaptation.

We reported the draft genome of Elstera cyanobacteriorum, containing 3.95 Mb. The G + C content of the genome is 62.4%. This strain possesses motility and chemotaxis genes, multiple carbon substrate utilization gene clusters and denitrifying reductase gene clusters. The sequence will provide the genetic basis for better understanding of adaptation to cyanobacterial aggregates and nitrogen removal during cyanobacterial blooming.

An aerobic, yellow-pigmented, bacteriochlorophyll a-producing strain, designated AAP5 (=DSM 111157=CCUG 74776), was isolated from the alpine lake Gossenköllesee located in the Tyrolean Alps, Austria. Here, we report its description and polyphasic characterization. Phylogenetic analysis of the 16S rRNA gene showed that strain AAP5 belongs to the bacterial genus Sphingomonas and has the highest pairwise 16S rRNA gene sequence similarity with Sphingomonas glacialis (98.3%), Sphingomonas psychrolutea (96.8%), and Sphingomonas melonis (96.5%). Its genomic DNA G + C content is 65.9%. Further, in silico DNA-DNA hybridization and calculation of the average nucleotide identity speaks for the close phylogenetic relationship of AAP5 and Sphingomonas glacialis. The high percentage (76.2%) of shared orthologous gene clusters between strain AAP5 and Sphingomonas paucimobilis NCTC 11030T, the type species of the genus, supports the classification of the two strains into the same genus. Strain AAP5 was found to contain C18:1ω7c (64.6%) as a predominant fatty acid (&gt, 10%) and the polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, six unidentified glycolipids, one unidentified phospholipid, and two unidentified lipids. The main respiratory quinone was ubiquinone-10. Strain AAP5 is a facultative photoheterotroph containing type-2 photosynthetic reaction centers and, in addition, contains a xathorhodopsin gene. No CO2-fixation pathways were found.

The Tibetan Plateau, also termed ‘the Third Pole’ harbors the largest number of high-altitude lakes in the world. Due to the presence of extreme conditions such as low temperature and oligotrophy, the lakes of the Tibetan Plateau share environmental features in common with lakes in the polar regions. However, the extent to which these environments are analogous, or indeed whether they harbor similar microbial communities or a high level of endemic species is poorly understood. Here we compared high-throughput 16S rRNA gene sequencing data from the lakes of the three different regions in order to characterize their taxonomic diversity, the community composition and biogeography. Our results showed despite the similarity in environmental conditions, the spatial distribution of the bacterial communities was distinct with only 3.1% of all operational taxonomic units (OTUs) being present in all three regions (although these OTUs did account for a considerable proportion of the total sequences, 36.4%). Sequences belonging to Burkholderiales and Actinomycetales dominated the shared OTUs across all three regions. Scale dependent distance decay patterns provided evidence of dispersal limitation. Climatic variables and dispersal limitation were apparently both important in controlling the spatial distribution of bacterial communities across regions. This work expands our understanding of the diversity and biogeography of lake bacterial communities across the Tibetan Plateau and provides insights into how they compare to those of the Antarctic and Arctic.

Anoxygenic phototrophs represent an environmentally important and phylogenetically diverse group of organisms. They harvest light using bacteriochlorophyll-containing reaction centers. Recently, a novel phototrophic bacterium, Gemmatimonas phototrophica , belonging to a rarely studied phylum, Gemmatimonadetes , was isolated from a freshwater lake in the Gobi Desert. To obtain more information about the environmental distribution of phototrophic Gemmatimonadetes , we collected microbial samples from the water column, upper sediment, and deeper anoxic sediment of Lake Taihu, China. MiSeq sequencing of the 16S rRNA, pufM , and bchY genes was carried out to assess the diversity of local phototrophic communities. In addition, we designed new degenerate primers of aerobic cyclase gene acsF , which serves as a convenient marker for both phototrophic Gemmatimonadetes and phototrophic Proteobacteria . Our results showed that most of the phototrophic species in Lake Taihu belong to Alpha - and Betaproteobacteria . Sequences of green sulfur and green nonsulfur bacteria (phototrophic Chlorobi and Chloroflexi , respectively) were found in the sediment. Using the newly designed primers, we identified a diverse community of phototrophic Gemmatimonadetes forming 30 operational taxonomic units. These species represented 10.5 and 17.3% of the acsF reads in the upper semiaerobic sediment and anoxic sediment, whereas their abundance in the water column was IMPORTANCE Photosynthesis is one of the most fundamental biological processes on Earth. Recently, the presence of photosynthetic reaction centers has been reported from a rarely studied bacterial phylum, Gemmatimonadetes , but almost nothing is known about the diversity and environmental distribution of these organisms. The newly designed acsF primers were used to identify phototrophic Gemmatimonadetes from planktonic and sediment samples collected in Lake Taihu, China. The Gemmatimonadetes sequences were found mostly in the upper sediments, documenting the preference of Gemmatimonadetes for semiaerobic conditions. Our results also show that the phototrophic Gemmatimonadetes present in Lake Taihu were relatively diverse, encompassing 30 operational taxonomic units.

Gemmatimonadetes represents a poorly understood bacterial phylum with only a handful of cultured species. Recently, one of its few representatives, Gemmatimonas phototrophica, was found to contain purple bacterial photosynthetic reaction centres. However, almost nothing is known about the environmental distribution of phototrophic Gemmatimonadetes bacteria. To fill this gap, we took advantage of fast-growing public metagenomic databases and performed an extensive survey of metagenomes deposited into the NCBI's WGS database, the JGI's IMG webserver and the MG-RAST webserver. By employing Mg protoporphyrin IX monomethyl ester oxidative cyclase (AcsF) as a marker gene, we identified 291 AcsF fragments (24-361 amino acids long) that are closely related to G. phototrophica from 161 metagenomes originating from various habitats, including air, river waters/sediment, estuarine waters, lake waters, biofilms, plant surfaces, intertidal sediment, soils, springs and wastewater treatment plants, but none from marine waters or sediment. Based on AcsF hit counts, phototrophic Gemmatimonadetes bacteria make up 0.4-11.9% of whole phototrophic microbial communities in these habitats. Unexpectedly, an almost complete 37.9 kb long photosynthesis gene cluster with identical gene composition and arrangement to those in G. phototrophica was reconstructed from the Odense wastewater metagenome, only differing in a 7.2 kb long non-photosynthesis-gene insert. These data suggest that phototrophic Gemmatimonadetes bacteria are much more widely distributed in the environment and exhibit a higher genetic diversity than previously thought.

Aquincola tertiaricarbonis strain MIMtkpLc11 was isolated from biological soil crusts in Inner Mongolia, China. The strain contains photosynthesis gene clusters., Aquincola tertiaricarbonis strain MIMtkpLc11 was isolated from biological soil crusts in Inner Mongolia, China. The strain contains photosynthesis gene clusters. Here, we report the draft genome sequence of strain MIMtkpLc11, which comprises 98 contigs (N50, 233,472 bp) and 5,573 protein-coding sequences.

A Gram-reaction-negative, aerobic, non-flagellated, non-gliding, rod-shaped and yellow-pigmented bacterium, designated strain TH021T, was isolated from cyanobacterial aggregates in a eutrophic lake, Taihu Lake, China. Optimal growth occurred at pH 7.0 (range: 5.0–10.0), 28 °C (range, 4–32 °C) and 0 % (w/v) NaCl (range, 0–1.0 %) in Reasoner’s 2A broth. No growth was observed at 37 °C. The cells were found to be positive for oxidase and catalase activities. The major respiratory quinone was menaquinone-6. The major fatty acids (>10 %) were identified as iso-C15 : 0 and C16 : 1ω7c/C16 : 1ω6c. The major polar lipid was phosphatidylethanolamine. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate was affiliated with the genus Flavobacterium , with the highest sequence similarity found to Flavobacterium hauense BX12T (94.92 %), followed by Flavobacterium suzhouense XIN-1T (94.85 %), Flavobacterium arcticum SM1502T (94.79 %) and Flavobacterium beibuense F44-8T (94.30 %). The genomic G+C content of strain TH021T was 41.9 mol% based on total genome calculations. Average nucleotide identities and digital DNA–DNA hybridizations values for complete genomes ranged from 69.4 to 72.8 and 18.0 to 23.8 % between strain TH021T and strains within the genus Flavobacterium . The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis suggested that strain TH021T represents a novel species within the genus Flavobacterium , for which the name Flavobacterium cyanobacteriorum sp. nov. is proposed. The type strain is TH021T (=LMG 29720T=CGMCC 1.16325T).

A bacterial strain, designated TH057T, was isolated from cyanobacterial aggregates in a eutrophic lake in China. Cells were observed to be slightly curved, rod-shaped, capsule-forming and stained Gram-negative. Optimal growth was obtained at pH 7.0 (range: pH 5–9) and 30 °C (range: 20–37 °C) in R2A broth. According to the absorption spectrum, carotenoids (455 and 490 nm) and light-harvesting complex LHI (857 nm) were present in the cells. The cells were found to be positive for oxidase and catalase activities. The major respiratory quinone was ubiquinone Q-10. The major fatty acids were identified as C17 : 1ω6c, C16 : 1ω7c/C16 : 1ω6c, C18 : 1ω6c/C18 : 1ω7c and C16 : 0. The major polar lipids were found to consist of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, unidentified glycolipid and two sphingoglycolipids. Strain TH057T shared highest 16S rRNA gene sequence similarity to Sandarakinorhabdus limnophila so42T (96.8 %), followed by Polymorphobacter fuscus D40PT (95.8 %). The genomic G+C content of strain TH057T was 66.1 mol% based on total genome calculations. The average nucleotide identity and the digital DNA–DNA hybridization value for the complete genomes were 81.0 and 23.0 % between strain TH057T and Sandarakinorhabdus limnophila so42T. The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis suggested that strain TH057T represents a novel species within the genus Sandarakinorhabdus , for which the name Sandarakinorhabdus cyanobacteriorum sp. nov. is proposed. The type strain is TH057T (=CGMCC 1.15803T=LMG 30294T).

The phyllosphere is a habitat to a variety of viruses, bacteria, fungi and other microorganisms, which play a fundamental role in maintaining the health of plants and mediating the interaction between plants and ambient environments. A recent addition to this catalogue of microbial diversity was the aerobic anoxygenic phototrophs (AAPs), a group of widespread bacteria that absorb light through bacteriochlorophyll α (BChl a) to produce energy without fixing carbon or producing molecular oxygen. However, culture representatives of AAPs from phyllosphere and their genome information are lacking, limiting our capability to assess their potential ecological roles in this unique niche. In this study, we investigated the presence of AAPs in the phyllosphere of a winter wheat (Triticum aestivum L.) in Denmark by employing bacterial colony based infrared imaging and MALDI-TOF mass spectrometry (MS) techniques. A total of ~4480 colonies were screened for the presence of cellular BChl a, resulting in 129 AAP isolates that were further clustered into 25 groups based on MALDI-TOF MS profiling, representatives of which were sequenced using the Illumina NextSeq and Oxford Nanopore MinION platforms. Twenty draft and five complete genomes of AAPs were assembled belonging in Methylobacterium, Rhizobium, Roseomonas and a novel strain in Methylocystaceae. We observed a diverging pattern in the evolutionary rates of photosynthesis genes among the highly homogenous AAP strains of Methylobacterium (Alphaproteobacteria), highlighting an ongoing genomic innovation at the gene cluster level.

The genus Limnohabitans ( Comamonadaceae , Betaproteobacteria ) is a common and a highly active component of freshwater bacterioplanktonic communities. To date, the genus has been considered to contain only heterotrophic species. In this study, we detected the photosynthesis genes pufLM and bchY in 28 of 46 strains from three Limnohabitans lineages. The pufM sequences obtained are very closely related to environmental pufM sequences detected in various freshwater habitats, indicating the ubiquity and potential importance of photoheterotrophic Limnohabitans in nature. Additionally, we sequenced and analyzed the genomes of 5 potentially photoheterotrophic Limnohabitans strains, to gain further insights into their phototrophic capacity. The structure of the photosynthesis gene cluster turned out to be highly conserved within the genus Limnohabitans and also among all potentially photosynthetic Betaproteobacteria strains. The expression of photosynthetic complexes was detected in a culture of Limnohabitans planktonicus II-D5 T using spectroscopic and pigment analyses. This was further verified by a novel combination of infrared microscopy and fluorescent in situ hybridization. IMPORTANCE The data presented document that the capacity to perform anoxygenic photosynthesis is common among the members of the genus Limnohabitans , indicating that they may have a novel role in freshwater habitats.

A bacterial strain, 1-14 T, was isolated from cyanobacterial aggregates in a eutrophic lake, Taihu Lake, China. Cells were observed to be slightly curved, rod-shaped, aerobic and Gram-stain-negative. Optimal growth occurred at pH 7.0 (range: 5.0-9.0), 28 °C (range: 20-32 °C) and 0% (w/v) NaCl (range: 0-1.0%) in R2A broth. No growth is observed at 37 °C. The cells were found to be positive for oxidase and catalase activities. The major respiratory quinone was ubiquinone Q-10. The major fatty acids (>10%) were identified as C 18:1ω6c/C 18:1ω7c, C1 6:0 3-OH and C 18:1 2-OH. The major polar lipids were found to consist of phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol and phosphatidylcholine. Within the genus Niveispirillum, strain 1-14T was most closely related to Niveispirillum cyanobacteriorum TH16 T (98.3% 16S rRNA gene sequence similarity), followed by Niveispirillum irakense DSM 11586 T (97.8%) and Niveispirillum fermenti CC-LY736 T (97.0%). The genomic G+C content of strain 1-14 T was 62.2 mol% based on total genome calculations. Genes coding for light-harvesting complexes LHI and LHII, and a photosynthetic reaction centre were detected in the genome. Average nucleotide identities and digital DNA-DNA hybridizations for complete genomes ranged from 76.4 to 83.5 and from 21.5 to 27.4% between strain 1-14 T and strains within the genus Niveispirillum. The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis suggested that strain 1-14 T represents a novel species within the genus Niveispirillum, for which the name Niveispirillum lacus sp. nov. is proposed. The type strain is 1-14 T (=CGMCC 1.12980 T=LMG 28363 T).

A bacterial strain, TH019(T), was isolated from cyanobacterial aggregates in a eutrophic lake, Taihu Lake, China. Cells were observed to be slightly curved rod-shaped and stained Gram-negative. Optimal growth was obtained at pH 7.0 (range, 5.0-9.0) and 28 °C (range, 20-37 °C) in R2A broth. The cells were found to be positive for oxidase and catalase activities, nitrate reduction and denitrification. The major respiratory quinone was ubiquinone Q-10. The major fatty acids were identified as C18 : 1 ω6c/C18 : 1 ω7c and C18 : 1 2OH. The 16S rRNA gene sequence of strain TH019(T) was phylogenetically related to Elstera litoralis Dia-1(T) (97.9 % similarity). The genomic G+C content of strain TH019(T) was 62.4 mol% based on total genome calculations. Average nucleotide identity and digital DNA-DNA hybridization values for complete genomes were 78.8 and 19.8 %, respectively, between strain TH019(T) and Elstera litoralis Dia-1(T). The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis suggested that strain TH019(T) represented a novel species within the genus Elstera, for which the name Elstera cyanobacteriorum sp. nov. is proposed. The type strain is TH019(T) (=CGMCC 1.15802(T)=LMG 29721(T)).

A characteristic feature of the order Rhodobacterales is the presence of a large number of photoautotrophic and photoheterotrophic species containing bacteriochlorophyll. Interestingly, these phototrophic species are phylogenetically mixed with chemotrophs. To better understand the origin of such variability, we sequenced the genomes of three closely related haloalkaliphilic species, differing in their phototrophic capacity and oxygen preference: the photoheterotrophic and facultatively anaerobic bacterium Rhodobaca barguzinensis, aerobic photoheterotroph Roseinatronobacter thiooxidans, and aerobic heterotrophic bacterium Natronohydrobacter thiooxidans. These three haloalcaliphilic species are phylogenetically related and share many common characteristics with the Rhodobacter species, forming together the Rhodobacter-Rhodobaca (RR) group. A comparative genomic analysis showed close homology of photosynthetic proteins and similarity in photosynthesis gene organization among the investigated phototrophic RR species. On the other hand, Rhodobaca barguzinensis and Roseinatronobacter thiooxidans lack an inorganic carbon fixation pathway and outer light-harvesting genes. This documents the reduction of their photosynthetic machinery towards a mostly photoheterotrophic lifestyle. Moreover, both phototrophic species contain 5-aminolevulinate synthase (encoded by the hemA gene) incorporated into their photosynthesis gene clusters, which seems to be a common feature of all aerobic anoxygenic phototrophic Alphaproteobacteria. Interestingly, the chrR-rpoE (sigma24) operon, which is part of singlet oxygen defense in phototrophic species, was found in the heterotrophic strain Natronohydrobacter thiooxidans. This suggests that this organism evolved from a photoheterotrophic ancestor through the loss of its photosynthesis genes. The overall evolution of phototrophy among the haloalkaliphilic members of the RR group is discussed.

Photosynthesis first emerged in prokaryotes over three billion years ago and represents one of the most fundamental biological processes on Earth. So far, species capable of performing (bacterio)chlorophyll-based phototrophy have been reported in seven bacterial phyla, i.e., Cyanobacteria, Proteobacteria, Chlorobi, Chloroflexi, Firmicutes, Acidobacteria, and Gemmatimonadetes. Here we review the discovery, physiology, genomic characteristics, environmental distribution, and possible evolutionary origin of the bacterium Gemmatimonas phototrophica strain AP64, so far the only phototrophic member of the phylum Gemmatimonadetes. This organism was isolated from a freshwater lake in the Gobi Desert, North China in 2011. It contains fully functional type-2 photosynthetic reaction centers, but they seem to only serve as an auxiliary energy source. Its photosynthesis genes are located in a 42.3 kb long photosynthesis gene cluster which appear to originate from an ancient horizontal gene transfer from a purple phototrophic bacterium. A survey of biomarker genes of phototrophic Gemmatimonadetes bacteria (PGB) in public environmental genomics databases suggests that PGB are widely distributed in diverse environments, including air, river waters/sediment, estuarine waters, lake waters, biofilms, plant surfaces, intertidal sediments, soils, springs, and wastewater treatment plants, but none from marine waters or sediment. PGB make up roughly 0.4-11.9 % of whole phototrophic microbial communities in these habitats. The discovery of PGB presents a strong evidence that genes for anoxygenic phototrophy can be transferred between distant bacterial phyla, providing new insights into the evolution of bacterial photosynthesis.

comparison of proportions of COG categories between A. paucihalophilus TH1–2 T, E. coli K12, P. putida KT2440, and S. oneidensis MR-1. (DOCX 13 kb)

To probe the genomic properties of microbes thriving in desert lakes, we sequenced the full genome of a betaproteobacterial strain (SL110) belonging to the understudied genus Caenimonas of the family Comamonadaceae. This strain was isolated from a freshwater lake in the western Gobi Desert, Northern China. Its genome contains genes encoding carbon monoxide dehydrogenase, nitrate reductase, nitrite reductase, nitric oxide reductase, and sulfur oxidation enzymes, highlighting the potentially important contribution of this group of bacteria to the cycling of inorganic elements in nature. Unexpectedly, a coenzyme F420 biosynthesis gene cluster was identified. A further search for F420 biosynthesis gene homologs in genomic databases suggests the possible widespread presence of F420 biosynthesis gene clusters in proteobacterial genomes.

Aims To perform a systematic evaluation of the applicability, validity and reliability of the long PCR-RFLP of 16S-ITS-23S rRNA genes for bacterial genotyping using both sequences retrieved from public genome databases and the experimental data obtained on bacterial cultures. Methods and Results 3301 Full-length sequences of 16S-ITS-23S rRNA genes were retrieved from 885 published bacterial genomes. Copy numbers of the whole set of 16S-ITS-23S rRNA genes per genome ranged from 1 (n = 161) to 14 (n = 4) with an average of 3·71. Their length varied greatly, from 4319 to 6568 bp with an average of 4952 bp. Computer-simulated RFLP analyses of the 16S-ITS-23S fragments flanked by the conserved primers 27F and 2241R suggested MspI, RsaI, HhaI and TaqI as the most appropriate enzymes for long PCR-RFLP analysis of the 16S-ITS-23S sequence. MspI was used to screen over 900 bacterial cultures isolated from the Huguangyan Maar Lake in southern China. An experimental sequencing of 16S rRNA genes of the isolates possessing a unique RFLP band pattern proved the broad applicability and high resolution of this approach. Conclusions These results indicate that long PCR-RFLP of 16S-ITS-23S rRNA genes is a potentially universal and reliable bacterial genotyping tool with a high resolution. Significance and Impact of the Study The methodology of long PCR-RFLP of 16S-ITS-23S rRNA genes will facilitate the exploration and tracing of cultivable microbial diversity in natural environments.

Bacterial abundances and diversity in the surface water of Lake Namco, the largest oligosaline lake on the Tibetan Plateau, were examined using flow cytometry approach and constructing 16S rRNA gene clone libraries. Bacterial abundances were from 0.08 × 106 to 1.6 × 106 cells mL−1, and were in the reported range of other lakes of the Tibetan Plateau and high mountain regions. Bacterial abundances were significantly correlated with the concentrations of chlorophyll a (chl a), but showed no significant relationship with the dissolved organic carbon (DOC), which suggested that the amount of DOC released by algae was the key factor determining the bacterial abundance rather than the total DOC. The total trace elements concentrations also obviously connected with bacterial abundances, and 9 of 20 elements showed significant relationship. Bacterial 16S rRNA gene clone sequences were affiliated to the α-, β-, γ-, δ-, and ϵ-Proteobacteria, Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Acidobacteria, P...

The study aims to reveal phylogenetic and evolutionary relationship between aerobic anoxygenic phototrophic bacteria (AAnPB) and their relatives, anaerobic anoxygenic phototrophic bacteria (AnAnPB) and nonphototrophic bacteria (NPB, which had high homology of 16S rDNA gene with AAnPB and fell into the same genus), and validate reliability and usefulness of farnesyl pyrophosphate synthase (FPPS) gene for the phylogenetic determination. FPPS genes with our modified primers and 16S rDNA genes with general primers, were amplified and sequenced or retrieved from GenBank database. In contrast to 16S rDNA gene phylogenetic tree, AAnPB were grouped into two clusters and one branch alone with no intermingling with NPB and AnAnPB in the tree constructed on FPPS. One branch of AAnPB, in both trees, was located closer to outgroup species than AnAnPB, which implicated that some AAnPB would be diverged earlier in FPPS evolutionary history than AnAnPB and NPB. Some AAnPB and NPB were closer located in both trees and this suggested that they were the closer relatives than AnAnPB. Combination codon usage in FPPS with phylogenetic analysis, the results indicates that FPPS gene and 16S rRNA gene have similar evolutionary pattern but the former seems to be more reliable and useful in determining the phylogenic and evolutionary relationship between AAnPB and their relatives. This is the first attempt to use a molecular marker beside 16S rRNA gene for studying the phylogeny of AAnPB, and the study may also be helpful in understanding the evolutionary relationship among phototrophic microbes and the trends of photosynthetic genes transfer.

NSFC [40632013, 40521003, 30900045]; MOST [200805068]; State Key Laboratory of Marine Environmental Science (Xiamen University) [MEL0701]; Natural Science Foundation of Guangdong Province, China [B09292]

A Gram-negative, non-motile, rod-shaped bacterial strain, JLT354-WT, that accumulates poly-β-hydroxybutyrate granules was isolated from the South China Sea. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was related to members of the genera Antarctobacter, Sagittula, Oceanicola and Loktanella; levels of similarity between strain JLT354-WT and members of the above genera were less than 92.0 %. The predominant fatty acid of strain JLT354-WT was C18 : 1 ω7c (83.1 %); significant amounts of C18 : 0 (7.9 %) and C12 : 1 3-OH (3.7 %) were also present. The predominant respiratory ubiquinone was Q-10. The DNA G+C content of strain JLT354-WT was 63.7 mol%. The isolate was distinguishable from members of the order Rhodobacterales based on phenotypic and biochemical characteristics. On the basis of the taxonomic data presented, strain JLT354-WT is considered to represent a novel species of a new genus, for which the name Mameliella alba gen. nov., sp. nov. is proposed. The type strain of Mameliella alba is JLT354-WT (=LMG 24665T=CGMCC 1.7290T).

The vertical community structure of bacteria along a depth profile in the Eastern Tropical North Pacific Ocean (13 degrees N, 104 degrees W) was studied by flow cytometry measurement and 16S rRNA gene clone libraries analysis. Picoeukaryotes and Synechococcus peaked at 30 m and decreased sharply below 50 m, while Prochlorococcus peaked at both 30 and 100 m layers and disappeared below 200 m. Heterotrophic bacteria peaked above shallow thermocline and decreased along the depth profile. Sequences of total 322 clones from four clone libraries (10, 100, 1000, and 3000 m) clustered into nine major lineages. gamma-Proteobacteria dominated all the depths and occupied almost the whole bacterial community at the 3000 m. alpha-Proteobacteria was abundant throughout the water column except near the sea bottom, and delta-Proteobacteria peaked at the 1000 m depth. Cyanobacteria were primarily limited to the photic zone, and the genetic diversity of Prochlorococcus showed a good correlation with niche adaptation. The appearance of the Cytophaga-Flexibacter-Bacteroides (CFB) group did not show a clear relationship with depth. Actinobacteria were found both in the photic zone and in deep water. Planctomyetes, Acidobacteria, and Verrucomicrobia were present as minor groups and more dominant in the deeper layers of water.

In order to explore the responses of the bacterioplankton community to different types of aquaculture environments, three mariculture ponds comprised of groupers (Epinephelus diacanthus, ED), prawns (Penaeus vannamei, PV), and abalone (Haliotis diversicolor supertexta, HDS) in southeast, coastal China were investigated. The free-living bacterial diversity was analyzed through the construction of 16S rDNA clone library. A total of 203 16S rDNA sequences from three clone libraries were classified into 118 operational taxonomic units (OTUs), of which 51, 31, and 42 OTUs were distributed in the ED, PV, and HDS pond, respectively, with Bacteroidetes (30.6%), Actinobacteria (55.2%), and Cyanobacteria (32.8%) as the dominant division in the respective ponds. Meanwhile, each pond occupied some unique OTUs that were affiliated with uncommon (sub-)phyla, such as candidate OP11 division, Acidobacteria, Deltaproteobacteria, Planctomycetes, and Verrucomicrobia. Bacterial diversity in the ED pond was the richest, followed by the HDS and the PV pond. OTUs of 61.9% and 94.9% have less than 90% and 97% similarity to their nearest neighbors in public databases, respectively. All OTUs were grouped into 67 clusters, covering 11 (sub-)phyla. The OTUs only from single pond distributed in 53 clusters (79.1%), the OTUs shared by two ponds were affiliated with 14 clusters (20.9%), and none of clusters was formed by the OTUs which commonly originated from the three pond libraries, suggesting that the composition of bacterial populations in these ponds were significantly different. These results indicate that the aquatic environment created by different mariculture animals may foster very special and complex bacterial communities.

Ministry of Science and Technology of the People's Republic of China [2005CB422004]; National Natural Science Foundation of China [40671044]

Aerobic anoxygenic phototrophic bacteria (AAPB) represent a widespread mixotrophic bacterial group in marine ecosystems. Here we investigated AAPB genetic diversity in the surface waters and upper twilight zones of the central Pacific, Atlantic, and Indian oceans by amplifying an AAPB marker gene (pufM, encoding photosynthetic reaction center small subunit) directly from bacterioplankton community DNA. Phylogenetic and statistical analysis of 267 pufM partial sequences in six clone libraries revealed a high diversity pattern in open ocean AAPB communities. Various AAPB subgroups belonging to Alpha- and Gamma-proteobacteria were found in both surface and upper twilight zone waters. In most samples, subgroups in which no pure culture was isolated as yet were predominant. By sampling a wide size range of bacterioplankton (0.22–200 μm) and introducing nested PCR to amplification, we retrieved abundant pufM fragments (136 sequences in 37 OTUs) directly from upper twilight zone samples. AAPB populations in upper twilight zones covered major subgroups found in surface waters and had a slightly lower diversity, higher dominance, and lower GC and GC3 contents in pufM genes than those in surface AAPB populations. These diversity data combined with previous BChl.a data in upper twilight zones support the hypothesis that AAPB may be present below euphotic zones based on the speculation that AAPB can utilize the dim light in twilight zones as a supplement to energy supply in their heterotrophic lives.