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The host recognition modules encoding the injection machinery and receptor binding proteins (RBPs) of bacteriophages are variable genomic units predisposed to mutation and recombination to maintain infectivity toward co-evolving bacterial hosts. In this study, we reveal howAlteromonas mediterraneaschitovirus A5 shares its host recognition module, including tail fiber (TF) and cognate chaperone, with phages from distantly related families includingAlteromonasmyovirus, V22. While the chaperone of V22 is essential for producing active TFs, here we demonstrate production of functional A5 TFs regardless of chaperone co-expression. AlphaFold-generated models of TF and chaperone pairs from phages A5, V22, and otherAlteromonasphages reveal how amino acid insertions within both A5-like proteins results in a knob domain duplication in the TF and a β-hairpin “tentacle” extension of the chaperone. These structural modifications are linked to chaperone dependency differences between the A5 and V22 TFs. Structural similarity between the chaperones and intramolecular chaperone domains of other phage RBPs suggests an additional function of these chaperones as transient TF “caps”. Finally, our identification of homologous host recognition modules used by morphologically distinct phages implies that HGT and recombination events between unrelated phages may be a more common process than previously thought amongCaudoviricetesphages.

The first microbial rhodopsin, a light-driven proton pump bacteriorhodopsin from Halobacterium salinarum (HsBR), was discovered in 1971. Since then, this seven-α-helical protein, comprising a retinal molecule as a cofactor, became a major driver of groundbreaking developments in membrane protein research. However, until 1999 only a few archaeal rhodopsins, acting as light-driven proton and chloride pumps and also photosensors, were known. A new microbial rhodopsin era started in 2000 when the first bacterial rhodopsin, a proton pump, was discovered. Later it became clear that there are unexpectedly many rhodopsins, and they are present in all the domains of life and even in viruses. It turned out that they execute such a diversity of functions while being "nearly the same." The incredible evolution of the research area of rhodopsins and the scientific and technological potential of the proteins is described in the review with a focus on their function-structure relationships.

Most microbial groups have not been cultivated yet, and the only way to approach the enormous diversity of rhodopsins that they contain in a sensible timeframe is through the analysis of their genomes. High-throughput sequencing technologies have allowed the release of community genomics (metagenomics) of many habitats in the photic zones of the ocean and lakes. Already the harvest is impressive and included from the first bacterial rhodopsin (proteorhodopsin) to the recent discovery of heliorhodopsin by functional metagenomics. However, the search continues using bioinformatic or biochemical routes.

The long-term fate of plastics in the ocean and their interactions with marine microorganisms remain poorly understood. In particular, the role of sinking plastic particles as a transport vector for surface microbes towards the deep sea has not been investigated. Here, we present the first data on the composition of microbial communities on floating and suspended plastic particles recovered from the surface to the bathypelagic water column (0-2000 m water depth) of the North Pacific Subtropical Gyre. Microbial community composition of suspended plastic particles differed from that of plastic particles afloat at the sea surface. However, in both compartments, a diversity of hydrocarbon-degrading bacteria was identified. These findings indicate that microbial community members initially present on floating plastics are quickly replaced by microorganisms acquired from deeper water layers, thus suggesting a limited efficiency of sinking plastic particles to vertically transport microorganisms in the North Pacific Subtropical Gyre. HN, AV were financed through the European Research Council (ERC-CoG Grant Nr 772923, project VORTEX). PDM was supported by NWO (VI.Veni.212.040).

The first microbial rhodopsin, a light-driven proton pump bacteriorhodopsin from Halobacterium salinarum (HsBR), was discovered in 1971. Since then, this seven-α-helical protein, comprising a retinal molecule as a cofactor, became a major driver of groundbreaking developments in membrane protein research. However, until 1999 only a few archaeal rhodopsins, acting as light-driven proton and chloride pumps and also photosensors, were known. A new microbial rhodopsin era started in 2000 when the first bacterial rhodopsin, a proton pump, was discovered. Later it became clear that there are unexpectedly many rhodopsins, and they are present in all the domains of life and even in viruses. It turned out that they execute such a diversity of functions while being “nearly the same.” The incredible evolution of the research area of rhodopsins and the scientific and technological potential of the proteins is described in the review with a focus on their function–structure relationships.

A next-generation sequencing (NGS)-based taxonomic analysis was carried out on airborne bacteria sampled at ground level in two periods (May and September) and two opposite locations on the north–south axis of the island of Sardinia. Located in a central position of the Mediterranean basin, Sardinia constitutes a suitable outpost for revealing possible immigration of bacterial taxa during transcontinental particle discharge between Africa and Europe. With the aim of verifying relative effects of dust outbreaks, sampling period, and sampling site on the airborne bacterial community composition, we compared air collected during dust-carrying meteorological events to that coming from wind regimes not associated with long-distance particle lifting. Results indicated that (a) higher microbial diversity and richness (118 vs. 65 orders) and increased community evenness were observed in the campaign carried out in September in comparison to the one in May, irrespective of the place of collection and of the presence or absence of dust outbreaks; (b) during the period of standard wind regimes without transcontinental outbreaks, a synchronous concerted turnover of bacterial communities across distant locations of the same island, accompanied as mentioned by a parallel rise in bacterial diversity and community evenness, appears to have occurred; (c) changes in wind provenance could transiently change community composition in the locality placed on the coast facing the incoming wind but not in the one located at the opposite side of the island, and for this reason the community changes brought from dust outbreaks of African origin are observed only in the sampling station exposed to the south; (d) the same winds, once proceeding over land, appear to uplift bacteria belonging to a common core already present over the region, which dilute or replace those that were associated with the air coming from the sea or conveyed by the dust particulates, explaining the two prior points; and (e) the hierarchy of the variables tested in determining bacterial assemblages composition results is as follows: sampling period≫ongoing meteorological events>sampling location within the island.

An NGS-based taxonomic analysis was carried out on airborne bacteria sampled at ground level in two periods (May and September) and two opposite localities on the North-South axis of the Sardinia Island. Located in a central position of the Mediterranean basin, Sardinia constitutes a suitable outpost to reveal possible immigration of bacterial taxa during transcontinental particle discharge between Africa and Europe. With the aim of verifying relative effects of dust outbreaks, sampling period and sampling site, on the airborne bacterial community composition, we compared air collected during dust-carrying meteorological events to that coming from wind regimes not associated to long-distance particle lifting. Results indicated that: (a) a higher microbial diversity (118 orders vs 65) and increased community evenness were observed in the campaign carried out in September in comparison to the one in May, irrespective of the place of collection and of the presence or absence of dust outbreaks. (b) During the period of standard wind regimes without transcontinental outbreaks a synchronous, concerted succession of bacterial communities across distant locations of the same island, accompanied as mentioned by a parallel rise in bacterial diversity and community evenness appears to have occurred. (c) changes in wind provenance could transiently change community composition in the locality placed on the coast facing the incoming wind, but not in the one located at the opposite side of the island; for this reason the community changes brought from dust outbreaks of African origin are observed only in the sampling station exposed to south; (d) the same winds, once proceeding over land appear to uplift bacteria belonging to a common core already present over the region, which dilute or replace those that were associated with the air coming from the sea or conveyed by the dust particulate, explaining the two prior points. (e) the hierarchy of the variables tested in determining bacterial assemblages composition results: sampling period >> ongoing meteorological events > sampling location within the island.

BackgroundLake Baikal is the largest body of liquid freshwater on Earth. Previous studies have described the microbial composition of this habitat, but the viral communities from this ecosystem have not been characterized in detail.ResultsHere, we describe the viral diversity of this habitat across depth and seasonal gradients. We discovered 19,475 bona fide viral sequences, which are derived from viruses predicted to infect abundant and ecologically important taxa that reside in Lake Baikal, such as Nitrospirota, Methylophilaceae, and Crenarchaeota. Diversity analysis revealed significant changes in viral community composition between epipelagic and bathypelagic zones. Analysis of the gene content of individual viral populations allowed us to describe one of the first bacteriophages that infect Nitrospirota, and their extensive repertoire of auxiliary metabolic genes that might enhance carbon fixation through the reductive TCA cycle. We also described bacteriophages of methylotrophic bacteria with the potential to enhance methanol oxidation and the S-adenosyl-L-methionine cycle.ConclusionsThese findings unraveled new ways by which viruses influence the carbon cycle in freshwater ecosystems, namely, by using auxiliary metabolic genes that act upon metabolisms of dark carbon fixation and methylotrophy. Therefore, our results shed light on the processes through which viruses can impact biogeochemical cycles of major ecological relevance.

Phytoplankton is the base of the marine food chain, oxygen, carbon cycle playing a global role in climate and ecology. Nucleocytoplasmic Large DNA Viruses regulating the dynamics of phytoplankton comprise genes of rhodopsins of two distinct families. We present a function-structure characterization of two homologous proteins representatives of family 1 of viral rhodopsins, OLPVR1 and VirChR1. VirChR1 is a highly selective, Ca2+-dependent, Na+/K+- conducting channel and, in contrast to known cation channelrhodopsins (ChRs), is impermeable to Ca2+ ions. In human neuroblastoma cells, upon illumination, VirChR1 depolarizes the cell membrane to a level sufficient to fire neurons. It suggests its unique optogenetic potential. 1.4 Å resolution structure of OLPVR1 reveals their remarkable difference from the known channelrhodopsins and a unique ion-conducting pathway. The data suggest that viral channelrhodopsins mediate phototaxis of algae enhancing the host anabolic processes to support virus reproduction, and therefore, their key role in global phytoplankton dynamics.

Marine phages play a variety of critical roles in regulating the microbial composition of our oceans. Despite constituting the majority of genetic diversity within these environments, there are relatively few isolates with complete genome sequences or in-depth analyses of their host interaction mechanisms, such as characterization of their receptor binding proteins (RBPs). Here, we present the 92,760-bp genome of the Alteromonas-targeting phage V22. Genomic and morphological analyses identify V22 as a myovirus; however, due to a lack of sequence similarity to any other known myoviruses, we propose that V22 be classified as the type phage of a new Myoalterovirus genus within the Myoviridae family. V22 shows gene homology and synteny with two different subfamilies of phages infecting enterobacteria, specifically within the structural region of its genome. To improve our understanding of the V22 adsorption process, we identified putative RBPs (gp23, gp24, and gp26) and tested their ability to decorate the V22 propagation strain, Alteromonas mediterranea PT11, as recombinant green fluorescent protein (GFP)-tagged constructs. Only GFP-gp26 was capable of bacterial recognition and identified as the V22 RBP. Interestingly, production of functional GFP-gp26 required coexpression with the downstream protein gp27. GFP-gp26 could be expressed alone but was incapable of host recognition. By combining size-exclusion chromatography with fluorescence microscopy, we reveal how gp27 is not a component of the final RBP complex but instead is identified as a new type of phage-encoded intermolecular chaperone that is essential for maturation of the gp26 RBP., mSystems, 5 (3), ISSN:2379-5077

Members of the SAR11 clade, despite their high abundance, are often poorly represented by metagenome‐assembled genomes. This fact has hampered our knowledge about their ecology and genetic diversity. Here we examined 175 SAR11 genomes, including 47 new single‐amplified genomes. The presence of the first genomes associated with subclade IV suggests that, in the same way as subclade V, they might be outside the proposed Pelagibacterales order. An expanded phylogenomic classification together with patterns of metagenomic recruitment at a global scale have allowed us to define new ecogenomic units of classification (genomospecies), appearing at different, and sometimes restricted, metagenomic data sets. We detected greater microdiversity across the water column at a single location than in samples collected from similar depth across the global ocean, suggesting little influence of biogeography. In addition, pangenome analysis revealed that the flexible genome was essential to shape genomospecies distribution. In one genomospecies preferentially found within the Mediterranean, a set of genes involved in phosphonate utilization was detected. While another, with a more cosmopolitan distribution, was unique in having an aerobic purine degradation pathway. Together, these results provide a glimpse of the enormous genomic diversity within this clade at a finer resolution than the currently defined clades. This work was supported by grant ‘VIREVO’ CGL2016-76273-P [AEI/FEDER, EU] (cofounded with FEDER funds) from the Spanish Ministerio de Economía, Industria y Competitividad to FRV, and grants CGL2013-40564-R and SAF2013-49267-EXP from the Spanish Ministerio de Economía, Industria y Competitividad, grant ACIF/2015/332 from Generalitat Valenciana and grant 5334 from the Betty Moore Foundation to MMG. FRV was also a beneficiary of the 5top100-program of the Ministry for Science and Education of Russia. JHM was supported by a Ph.D. fellowship from the Spanish Ministerio de Economía y Competitividad (BES-2014-067828). MLP was supported by a postdoctoral fellowship from the Spanish Ministerio de Economía, Industria y Competitividad (IJCI-2017-34002).

Viruses are extremely abundant and diverse biological entities that contribute to the functioning of marine ecosystems. Despite their recognized importance no studies have addressed trends of micro-diversity in marine viral communities across depth gradients. To fill this gap we obtained metagenomes from both the cellular and viral fractions of Mediterranean seawater samples spanning the epipelagic to the bathypelagic zone at 15, 45, 60 and 2000 meters deep. The majority of viral genomic sequences obtained were derived from bacteriophages of the order Caudovirales, and putative host assignments suggested that they infect some of the most abundant bacteria in marine ecosystems such as Pelagibacter, Puniceispirillum and Prochlorococcus. We evaluated micro-diversity patterns by measuring the accumulation of synonymous and non-synonymous mutations in viral genes. Our results demonstrated that the degree of micro-diversity differs among genes encoding metabolic, structural, and replication proteins and that the degree of micro-diversity increased with depth. These trends of micro-diversity were linked to the changes in environmental conditions observed throughout the depth gradient, such as energy availability, host densities and proportion of actively replicating viruses. These observations allowed us to generate hypotheses regarding the selective pressures acting upon marine viruses from the epipelagic to the bathypelagic zones.

We present two genomes of widespread freshwater picocyanobacteria isolated by extinction dilution from a Spanish oligotrophic reservoir. Based on microscopy and genomic properties, both picocyanobacteria were tentatively designated Synechococcus lacustris Tous, formerly described as a metagenome assembled genome (MAG) from the same habitat, and Cyanobium usitatum Tous, described here for the first time. Both strains were purified in unicyanobacterial cultures, and their genomes were sequenced. They are broadly distributed in freshwater systems; the first seems to be a specialist on temperate reservoirs (Tous, Amadorio, Dexter, Lake Lanier, Sparkling), and the second appears to also be abundant in cold environments including ice-covered lakes such as Lake Baikal, Lake Erie or the brackish Baltic Sea. Having complete genomes provided access to the flexible genome that does not assemble in MAGs. We found several genomic islands in both genomes, within which there were genes for nitrogen acquisition, transporters for a wide set of compounds and biosynthesis of phycobilisomes in both strains. Some of these regions of low coverage in metagenomes also included antimicrobial compounds, transposases and phage defence systems, including a novel type III CRISPR-Cas phage defence system that was only detected in Synechococcus lacustris Tous.

The root nodules of two wild legume species endemic to Algeria, Hedysarum naudinianum and He. perrauderianum, were investigated with regard to their anatomy and histology, and the identity of the associated bacteria. Both plants were found to form root nodules with regular features and well infected by rod-shaped bacteria. The culturable fraction of bacteria that could be obtained from surface-sterilized nodules included a prevailing presence of Enterobacteriaceae having 100 % 16S rDNA sequence identity with both Enterobacter cloacae and E. ludwigii. In H. perrauderianum, this taxon was the sole cultured isolate, while from H. naudinianum we also found Bacillus, Lactobacillus, Staphylococcus, Rothia, and isolates that were 100 % identical to Corynebacterium pseudodiphthericum, which is known to be an agent of respiratory and cardiac infections in humans. Whereas no culturable rhizobia and alike could be obtained on plates, PCR-based culture-independent approaches revealed in both plants the presence of a Mesorhizobium sp., which in H. perrauderianum was identical to isolates nodulating other legumes from Africa, European Mediterranean countries, and Asia, while in H. naudinianum it bore a single nucleotide polymorphism which is so far unique for any observed mesorhizobia. Data from the microsymbionts appear to suggest interesting clues to interpret the evolutionary ecology of their host plants.

Background: Haloquadratum walsbyi dominates saturated thalassic lakes worldwide where they can constitute up to 80-90% of the total prokaryotic community. Despite the abundance of the enigmatic square-flattened cells, only 7 isolates are currently known with 2 genomes fully sequenced and annotated due to difficulties to grow them under laboratory conditions. We have performed a transcriptomic analysis of one of these isolates, the Spanish strain HBSQ001 in order to investigate gene transcription under light and dark conditions. Results: Despite a potential advantage for light as additional source of energy, no significant differences were found between light and dark expressed genes. Constitutive high gene expression was observed in genes encoding surface glycoproteins, light mediated proton pumping by bacteriorhodopsin, several nutrient uptake systems, buoyancy and storage of excess carbon. Two low expressed regions of the genome were characterized by a lower codon adaptation index, low GC content and high incidence of hypothetical genes. Conclusions: Under the extant cultivation conditions, the square hyperhalophile devoted most of its transcriptome towards processes maintaining cell integrity and exploiting solar energy. Surface glycoproteins are essential for maintaining the large surface to volume ratio that facilitates light and organic nutrient harvesting whereas constitutive expression of bacteriorhodopsin warrants an immediate source of energy when light becomes available The research leading to these results has received funding from the European Union Seventh Framework Program (FP7/2007-2013- MaCuMBA project) under grant agreement n° 311,975 (HB and FRV; salary (in part), consumables and sequencing costs) Salaries of RR and ABMC were partly payed by the projects “VIREVO” CGL2016-76273-P [AEI/FEDER, UE], from Agencia Estatal de Investigación (AEI) y por la Unión Europea a través del Fondo Europeo de Desarrollo Regional - FEDER – “Una manera de hacer Europa” Acciones de dinamización “REDES DE EXCELENCIA” CONSOLIDER- CGL2015-71523-REDC from the Spanish Ministerio de Economía, Industria y Competitividad PROMETEO II/2014/012 “AQUAMET” from Conselleria d’Educació, Cultura i Esport. Direcció General d’Universitat, Estudis Superiors i Ciència. Generalitat Valenciana

Assessing the response of microbial communities to nutrient inputs in man-managed soils is of primary importance to understand the impact on ecosystem services provided by the soil microbiome. In this study, a low-nutrient soil was supplemented with seven different innovative fertilizers including matrixes of plant, animal, fungal or synthetic origin, and dosed to deliver the same amount of nitrogen. Growth of a potted grass crop (Cynodon dactylon) was recorded and the fertilizers were scored by the plant yield obtained in a greenhouse study. Soil was sampled at 9 and 58 days after the addition and bacterial community composition was analyzed after soil DNA extraction through pyrosequencing of 16S rDNA gene amplicons. Over 900 bacterial genera were detected, belonging to 21 described and 19 candidate phyla. In spite of the equal dose of nitrogen delivered, specific groups were fostered by given fertilizers, in particular marked effects on some phyla were displayed by a yeast-based fertilizer, which was also most effective in plant productivity. The main shifts were observed shortly after the fertilizer application, followed by a gradual stabilization of the equilibrium and by a rise in community evenness.

Organic fertilizers are of high interest in agriculture as they promise a retarded release of the nitrogen to soil, which improves uptake efficiency by plants and reduces negative impacts like the formation of nitrate by nitrifiers or N2O by denitrifying microorganisms. In this study we tested the effects of seven different commercially available organic fertilizers of plant-, animal-, or microbial origin in a two-month greenhouse trial, using the perennial grass Cynodon dactylon in pots and monitoring effects on plant growth as well as on the abundance of prokaryotic nitrifiers and denitrifiers by realtime PCR. In most cases a single application of the fertilizers induced plant growth but did not increase the abundance of nitrifiers and denitrifiers. In contrast a repeated application stimulated, in addition to plant growth, also the increase of the two functional groups studied and a faster mobilization of nitrogen from the different fertilizers. Upon analyzing total bacterial DNA extracted from soil nirK abundance was found responsive to plant presence. Bacterial amoA and nosZ gene copies were significantly positively correlated with plant growth and cumulative dry weight at harvest. The latter was responsive as early as 9 days after fertilizer supplementation.

Kopf, Anna ... et. al.-- 5 pages, 1 figure.-- This manuscript is NOAA-GLERL contribution number 1763, Ocean Sampling Day was initiated by the EU-funded Micro B3 (Marine Microbial Biodiversity, Bioinformatics, Biotechnology) project to obtain a snapshot of the marine microbial biodiversity and function of the world’s oceans. It is a simultaneous global mega-sequencing campaign aiming to generate the largest standardized microbial data set in a single day. This will be achievable only through the coordinated efforts of an Ocean Sampling Day Consortium, supportive partnerships and networks between sites. This commentary outlines the establishment, function and aims of the Consortium and describes our vision for a sustainable study of marine microbial communities and their embedded functional traits, This work was supported by the Micro B3 project, which is funded from the European Union’s Seventh Framework Programme (FP7; Joint Call OCEAN.2011‐2: Marine microbial diversity – new insights into marine ecosystems functioning and its biotechnological potential) under the grant agreement no 287589

Dust particles lifting and discharge from Africa to Europe is a recurring phenomenon linked to air circulation conditions. The possibility that microorganisms are conveyed across distances entails important consequences in terms of biosafety and pathogens spread. Using culture independent DNA-based analyses via next generation sequencing of the 16 S genes from the airborne metagenome, the atmospheric microbial community was characterized and the hypothesis was tested that shifts in species diversity could be recorded in relation to dust discharge. As sampling ground the island of Sardinia was chosen, being an ideal cornerstone within the Mediterranean and a crossroad of wind circulation amidst Europe and Africa. Samples were collected in two opposite coastal sites and in two different weather conditions comparing dust-conveying winds from Africa with a control situation with winds from Europe. A major conserved core microbiome was evidenced but increases in species richness and presence of specific taxa were nevertheless observed in relation to each wind regime. Taxa which can feature strains with clinical implications were also detected. The approach is reported as a recommended model monitoring procedure for early warning alerts in frameworks of biosafety against natural spread of clinical microbiota across countries as well as to prevent bacteriological warfare.

The complex world of interactive patterns between microorganisms and plants includes relationships that span from soil, to rhizosphere, to phyllosphere and culminate inside the plant tissues in the endophytic condition. The nature of these interactions can be beneficial, neutral or pathogenic with a number of subtle differences and some overlaps between these conditions. The availability of metagenomics and NGS technologies has opened new perspectives and powerful ways to investigate this scenario. The questions that are central to such field involve: the identity and diversity of microorganisms relating with plants, the array of genes that are functional to this lifestyle, the relative effects of host genotype vs. environment of source in determining which taxa can become established as plant-associated microbiota; which genes are expressed when on or inside plants and which proteins are abundant in the different stages of the process. Metagenomics can in this respect be efficiently coupled to other—omics to provide a multiphasic picture of the ongoing phenomena that exploits the growing potentialities of high throughput investigations. The practical applications of these studies, besides the clearer insight into environmental biodiversity and ecology of microorganisms, can also involve valuable strategies for the improvement of crop productivity and their defense against plant pathogens.