当前期刊: Environmental Microbiology Go to current issue    加入关注   
显示样式:        排序: 导出
  • Catabolism of biogenic amines in Pseudomonas species
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-14
    José M. Luengo; Elías R. Olivera

    Biogenic amines (BAs; 2‐phenylethylamine, tyramine, dopamine, epinephrine, norepinephrine, octopamine, histamine, tryptamine, serotonin, agmatine, cadaverine, putrescine, spermidine, spermine and certain aliphatic amines) are widely distributed organic molecules that play basic physiological functions in animals, plants and microorganisms. Pseudomonas species can grow in media containing different BAs as carbon and energy sources, a reason why these bacteria are excellent models for studying such catabolic pathways. In this review, we analyse most of the routes used by different species of Pseudomonas (P. putida, P. aeruginosa, P. entomophila and P. fluorescens) to degrade BAs. Analysis of these pathways has led to the identification of a huge number of genes, catabolic enzymes, transport systems and regulators, as well as to understanding of their hierarchy and functional evolution. Knowledge of these pathways has allowed the design and collection of genetically manipulated microbes useful for eliminating BAs from different sources, highlighting the biotechnological applications of these studies.

  • Who is eating fructose within the Aedes albopictus gut microbiota?
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Morgane Guégan; Van Tran Van; Edwige Martin; Guillaume Minard; Florence‐Hélène Tran; Benjamin Fel; Anne‐Emmanuelle Hay; Laurent Simon; Mohamed Barakat; Patrick Potier; Feth el Zahar Haichar; Claire Valiente Moro

    The Asian tiger mosquito Aedes albopictus is a major public health concern because of its invasive success and its ability to transmit pathogens. Given the low availability of treatments against mosquito‐borne diseases, vector control remains the most suitable strategy. The methods used thus far are becoming less effective, but recent strategies have emerged from the study of mosquito‐associated microorganisms. Although the role of the microbiota in insect biology does not require further proof, much remains to be deciphered in mosquitoes, especially the contribution of the microbiota to host nutrient metabolism. Mosquitoes feed on plant nectar, composed of mostly fructose. We used stable isotope probing to identify bacteria and fungi assimilating fructose within the gut of Ae. albopictus. Mosquitoes were fed a 13C‐labelled fructose solution for 24 h. Differences in the active microbial community according to the sex of mosquitoes were highlighted. The bacterium Lelliottia and the fungi Cladosporium and Aspergillus dominated the active microbiota in males, whereas the bacterium Ampullimonas and the yeast Cyberlindnera were the most active in females. This study is the first to investigate trophic interactions between Ae. albopictus and its microbiota, thus underscoring the importance of the microbial component in nectar feeding in mosquitoes.

  • Thiomicrorhabdus streamers and sulfur cycling in perennial hypersaline cold springs in the Canadian high Arctic
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Elisse Magnuson; Nadia C.S. Mykytczuk; Andre Pellerin; Jacqueline Goordial; Susan M. Twine; Boswell Wing; Simon J. Foote; Kelly Fulton; Lyle G. Whyte

    The Gypsum Hill (GH) springs on Axel Heiberg Island in the Canadian high Arctic are host to chemolithoautotrophic, sulfur‐oxidizing streamers that flourish in the high Arctic winter in water temperatures from −1.3‐7°C with ~8% salinity in a high Arctic winter environment with air temperatures commonly less than −40°C and an average annual air temperature of −15°C. Metagenome sequencing and binning of streamer samples produced a 96% complete Thiomicrorhabdus sp. metagenome‐assembled genome representing a possible new species or subspecies. This is the most cold‐ and salt‐extreme source environment for a Thiomicrorhabdus genome yet described. Metaproteomic and metatranscriptomic analysis attributed nearly all gene expression in the streamers to the Thiomicrorhabdus sp. and suggested that it is active in CO2 fixation and oxidation of sulfide to elemental sulfur. In situ geochemical and isotopic analyses of the fractionation of multiple sulfur isotopes determined the biogeochemical transformation of sulfur from its source in Carboniferous evaporites to biotic processes occurring in the sediment and streamers. These complementary molecular tools provided a functional link between the geochemical substrates and the collective traits and activity that define the microbial community's interactions within a unique polar saline habitat where Thiomicrorhabdus‐dominated streamers form and flourish.

  • Symbiodiniaceae‐bacteria interactions: rethinking metabolite exchange in reef‐building corals as multi‐partner metabolic networks.
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Jennifer L. Matthews; Jean‐Baptiste Raina; Tim Kahlke; Justin R. Seymour; Madeleine J. H. van Oppen; David J. Suggett

    The intimate relationship between scleractinian corals and their associated microorganisms is fundamental to healthy coral reef ecosystems. Coral‐associated microbes (Symbiodiniaceae and other protists, bacteria, archaea, fungi, and viruses) support coral health and resilience through metabolite transfer, inter‐partner signalling, and genetic exchange. However, much of our understanding of the coral holobiont relationship has come from studies that have investigated either coral‐Symbiodiniaceae or coral‐bacteria interactions in isolation, while relatively little research has focused on other ecological and metabolic interactions potentially occurring within the coral multi‐partner symbiotic network. Recent evidences of intimate coupling between phytoplankton and bacteria have demonstrated that obligate resource exchange between partners fundamentally drives their ecological success. Here, we posit that similar associations with bacterial consortia regulate Symbiodiniaceae productivity and are in turn central to the health of corals. Indeed, we propose that this bacteria‐Symbiodiniaceae‐coral relationship underpins the coral holobiont's nutrition, stress tolerance, and potentially influences the future survival of coral reef ecosystems under changing environmental conditions. Resolving Symbiodiniaceae‐bacteria associations is therefore a logical next step towards understanding the complex multi‐partner interactions occurring in the coral holobiont.

  • Microbial communities from Arctic marine sediments respond slowly to methane addition during ex situ enrichments
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Scott Klasek; Marta E. Torres; Douglas H. Bartlett; Madeline Tyler; Wei‐Li Hong; Frederick Colwell

    Anaerobic methanotrophic archaea (ANME) consume methane in marine sediments, limiting its release to the water column, but their responses to changes in methane and sulfate supplies remain poorly constrained. To address how methane exposure may affect microbial communities and methane‐ and sulfur‐cycling gene abundances in Arctic marine sediments, we collected sediments from offshore Svalbard that represent geochemical horizons where anaerobic methanotrophy is expected to be active, previously active, and long‐inactive based on reaction‐transport biogeochemical modelling of porewater sulfate profiles. Sediment slurries were incubated at in situ temperature and pressure with different added methane concentrations. Sediments from an active area of seepage began to reduce sulfate in a methane‐dependent manner within months, preceding increased relative abundances of anaerobic methanotrophs ANME‐1 within communities. In previously active and long‐inactive sediments, sulfur‐cycling Deltaproteobacteria became more dominant after 30 days, though these communities showed no evidence of methanotrophy after nearly 8 months of enrichment. Overall, enrichment conditions, but not methane, broadly altered microbial community structure across different enrichment times and sediment types. These results suggest that active ANME populations may require years to develop, and consequently microbial community composition may affect methanotrophic responses to potential large‐scale seafloor methane releases in ways that provide insight for future modelling studies.

  • Deciphering proteolysis pathways for the error‐prone DNA polymerase in cyanobacteria
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Haojie Jin; Rick Kim; Devaki Bhaya

    Protein quality control pathways require AAA+ proteases, such as Clp and Lon. Lon protease maintains UmuD, an important component of the error‐prone DNA repair polymerase (Pol V), at very low levels in E. coli. Most members of the phylum Cyanobacteria lack Lon (including the model cyanobacterium, Synechocystis sp. PCC6803), so maintenance of UmuD at low levels must employ different proteases. We demonstrate that the first 19 residues from the N‐terminus of UmuD (Sug1‐19) fused to a reporter protein are adequate to trigger complete proteolysis and that mutation of a single leucine residue (L6) to aspartic acid inhibits proteolysis. This process appears to follow the N‐end rule and is mediated by ClpA/P protease and the ClpS adaptor. Additionally, mutations of arginine residues in the Sug1‐19 tag suggest that the ClpX/P pathway also plays a role in proteolysis. We propose that there is a dual degron at the N‐terminus of the UmuD protein in Synechocystis sp. PCC6803, which is distinct from the degron required for degradation of UmuD in E. coli. The use of two proteolysis pathways to tune levels of UmuD might reflect how a photosynthetic organism responds to multiple environmental stressors.

  • A diverse uncultivated microbial community is responsible for organic matter degradation in the Black Sea sulphidic zone
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-13
    Saara Suominen; Nina Dombrowski; Jaap S. Sinninghe Damsté; Laura Villanueva

    Organic matter degradation in marine environments is essential for the recycling of nutrients, especially under conditions of anoxia where organic matter tends to accumulate. However, little is known about the diversity of the microbial communities responsible for the mineralization of organic matter in the absence of oxygen, as well as the factors controlling their activities. Here, we determined the active heterotrophic prokaryotic community in the sulphidic water column of the Black Sea, an ideal model system, where a tight coupling between carbon, nitrogen and sulphur cycles is expected. Active microorganisms degrading both dissolved organic matter (DOM) and protein extracts were determined using quantitative DNA stable isotope probing incubation experiments. These results were compared with the metabolic potential of metagenome‐assembled genomes obtained from the water column. Organic matter incubations showed that groups like Cloacimonetes and Marinimicrobia are generalists degrading DOM. Based on metagenomic profiles the degradation proceeds in a potential interaction with members of the Deltaproteobacteria and Chloroflexi Dehalococcoidia. On the other hand, microbes with small genomes like the bacterial phyla Parcubacteria, Omnitrophica and of the archaeal phylum Woesearchaeota, were the most active, especially in protein‐amended incubations, revealing the potential advantage of streamlined microorganisms in highly reduced conditions.

  • Extremely Halophilic Archaeal Communities are Resilient to Short‐Term Entombment in Halite
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-09
    Tom J C Huby; Dave R Clark; Boyd A McKew; Terry J McGenity

    Some haloarchaea avoid the harsh conditions present in evaporating brines by entombment in brine inclusions within forming halite crystals, where a subset of haloarchaea survive over geological time. However, shifts in the community structure of halite‐entombed archaeal communities remain poorly understood. Therefore, we analysed archaeal communities from in‐situ hypersaline brines collected from Trapani saltern (Sicily) and their successional changes in brines versus laboratory‐grown halite over 21 weeks, using high‐throughput sequencing. Haloarchaea were dominant, comprising >95% of the archaeal community. Unexpectedly, the OTU richness of the communities after 21 weeks was indistinguishable from the parent brine and overall archaeal abundance in halite showed no clear temporal trends. Furthermore, the duration of entombment was less important than the parent brine from which the halite derived in determining the community composition and relative abundances of most genera in halite‐entombed communities. These results show that halite‐entombed archaeal communities are resilient to entombment durations of up to 21 weeks, and that entombment in halite may be an effective survival strategy for near complete communities of haloarchaea. Additionally, the dominance of ‘halite specialists” observed in ancient halite must occur over periods of years, rather than months, hinting at long‐term successional dynamics in this environment.

  • Anammox and beyond
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-08
    J. Gijs Kuenen

    When looking back and wonder how we did it, I became even more aware of how my wanderings in microbiology are all linked, from the start of my PhD with Hans Veldkamp on sulphur‐oxidizing bacteria in chemostats. My interests broadened from obligate chemolithoautotrophic bacteria to facultative organisms and the question about the ecological niches of these different metabolic types. The sulphide oxidizing bacteria also may be used to produce elemental sulphur, which can easily be removed from wastewater. This fitted in a long‐standing collaboration with Dimitry Sorokin on the ecophysiology and application of alkaliphilic sulphur bacteria. Then came the denitrifying sulphur‐oxidizing bacteria and their application to remove sulphide from wastewater, which lead to our interest in nitrate, nitrite and ammonium removal in general. The big surprise was the serendipitous discovery of the ‘anammox’‐process, whereby ammonium is anaerobically oxidized to dinitrogen gas with nitrite as electron acceptor. The early days of our anammox research are the main focus of this article, which describes the struggle of growing and identifying the most peculiar bacteria we ever came across. A specialized organelle, the anammoxosome was shown to be responsible for the key ammonium oxidation, whereby a rocket fuel, hydrazine, turned out to be an intermediate. Soon after we became aware that anammox is everywhere and in the marine environment makes up a major portion of the nitrogen cycle. The intense scientific collaboration with Mike Jetten and Mark van Loosdrecht and colleagues led to our further understanding and application of this fascinating process, which is briefly summarized in this article. My broader interest in environmental microbiology and microbial ecology has been a regularly returning theme, taking me all over the world to great collaborations lasting to this very day.

  • Environmental conditions steer phenotypic switching in acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus, affecting PirAVP/PirBVP toxins production
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-08
    Vikash Kumar; Suvra Roy; Kartik Baruah; Delphi Van Haver; Francis Impens; Peter Bossier

    Bacteria in nature are widely exposed to differential fluid shears which are often a trigger for phenotypic switches. The latter mediates transcriptional and translation remodelling of cellular metabolism impacting among others virulence, antimicrobial resistance and stress resistance. In this study, we evaluated the role of fluid shear on phenotypic switch in an acute hepatopancreatic necrosis disease (AHPND)‐causing Vibrio parahaemolyticus M0904 strain under both in vitro and in vivo conditions. The results showed that V. parahaemolyticus M0904 grown at lower shaking speed (110 rpm constant agitation, M0904/110), causing low fluid shear, develop cellular aggregates or floccules. These cells increased levan production (as verified by concanavalin binding) and developed differentially stained colonies on Congo red agar plates and resistance to antibiotics. In addition, the phenotypic switch causes a major shift in the protein secretome. At 120 rpm (M0904/120), PirAVP/PirBVP toxins are mainly produced, while at 110 rpm PirAVP/PirBVP toxins production is stopped and an alkaline phosphatase (ALP) PhoX becomes the dominant protein in the protein secretome. These observations are matched with a very strong reduction in virulence of M0904/110 towards two crustacean larvae, namely, Artemia and Macrobrachium. Taken together, our study provides substantial evidence for the existence of two phenotypic forms in AHPND V. parahaemolyticus strain displaying differential phenotypes. Moreover, as aerators and pumping devices are frequently used in shrimp aquaculture facilities, they can inflict fluid shear to the standing microbial agents. Hence, our study could provide a basis to understand the behaviour of AHPND‐causing V. parahaemolyticus in aquaculture settings and open the possibility to monitor and control AHPND by steering phenotypes.

  • Colonies of the fungus Aspergillus niger are highly differentiated to adapt to local carbon source variation
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-06
    Paul Daly; Mao Peng; Hugh D. Mitchell; Young‐Mo Kim; Charles Ansong; Heather Brewer; Peter de Gijsel; Mary S. Lipton; Lye Meng Markillie; Carrie D. Nicora; Galya Orr; Ad Wiebenga; Kristiina S. Hildén; Mirjam A. Kabel; Scott E. Baker; Miia R. Mäkelä; Ronald P. de Vries

    Saprobic fungi, such as Aspergillus niger, grow as colonies consisting of a network of branching and fusing hyphae that are often considered to be relatively uniform entities in which nutrients can freely move through the hyphae. In nature, different parts of a colony are often exposed to different nutrients. We have investigated, using a multi‐omics approach, adaptation of A. niger colonies to spatially separated and compositionally different plant biomass substrates. This demonstrated a high level of intra‐colony differentiation, which closely matched the locally available substrate. The part of the colony exposed to pectin‐rich sugar beet pulp and to xylan‐rich wheat bran showed high pectinolytic and high xylanolytic transcript and protein levels respectively. This study therefore exemplifies the high ability of fungal colonies to differentiate and adapt to local conditions, ensuring efficient use of the available nutrients, rather than maintaining a uniform physiology throughout the colony.

  • Sinorhizobium fredii HH103 syrM inactivation affects the expression of a large number of genes, impairs nodulation with soybean and extends the host‐range to Lotus japonicus
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-05
    Sebastián Acosta‐Jurado; Cynthia Alias‐Villegas; Pilar Navarro‐Gómez; Andrés Almozara; Miguel A. Rodríguez‐Carvajal; Carlos Medina; José‐María Vinardell

    Sinorhizobium fredii HH103 RifR is a broad host‐range rhizobial strain able to nodulate with soybean and Lotus burttii, but it is ineffective with L. japonicus. Here, we study the role of the HH103 RifR SyrM protein in the regulation of gene expression and its relevance in symbiosis with those three legumes. RNAseq analyses show that HH103 SyrM is an important transcriptional regulator not only in the presence of inducer flavonoids but also in its absence. Lack of SyrM increases Nod factors production and decreases genistein‐mediated repression of exopolysaccharide production in HH103. In symbiosis, mutation of syrM partially impaired interaction with soybean but improves effectiveness with L. burttii and extends the host‐rage to L. japonicus Gifu. In addition, HH103 syrM mutants enter in both Lotus species by infection threads, whereas HH103 uses the more primitive intercellular infection to enter into L. burttii roots These symbiotic phenotypes were previously observed in two other HH103 mutants affected in symbiotic regulators, nodD2 and nolR, revealing that in S. fredii HH103 numerous transcriptional regulators finely modulate symbiotic gene expression.

  • Small RNA coaR contributes to intestinal colonization in Vibrio cholerae via the two‐component system EnvZ/OmpR
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-05
    Daoyi Xi; Yujia Li; Junxiang Yan; Yuehua Li; Xiaochen Wang; Boyang Cao

    Vibrio cholerae is a waterborne bacterium responsible for worldwide outbreaks of acute and fatal cholera. Recently, small regulatory RNAs (sRNAs) have become increasingly recognized as important regulators of virulence gene expression in response to environmental signals. In this study, we determined that two‐component system EnvZ/OmpR was required for intestinal colonization in V. cholerae O1 EI Tor strain E12382. Analysis of the characteristics of OmpR revealed a potential binding site in the intergenic region between vc1470 and vc1471, and qRT‐PCR showed that expression of the intergenic region increased 5.3‐fold in the small intestine compared to LB medium. Race and northern blot assays were performed and demonstrated a new sRNA, coaR (cholerae osmolarity and acidity related regulatory RNA). A ΔcoaR mutant showed a deficient colonization ability in small intestine with CI of 0.15. We identified a target of coaR, tcpI, a negative regulator of the major pilin subunit of TcpA. The ΔtcpI mutant has an increased colonization with CI of 3.16. The expression of coaR increased 2.8‐fold and 3.3‐fold under relative acidic and hypertonic condition. In summary, coaR was induced under the condition of high osmolarity and acid stress via EnvZ/OmpR and explained that tcpI relieves pH‐mediated repression of toxin co‐regulated pilus synthesis.

  • Biological inhibition of soil nitrification by forest tree species affects Nitrobacter populations
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-05
    Amandine Laffite; Alessandro Florio; Kasaina Sitraka Andrianarisoa; Charline Creuze des Chatelliers; Brigitte Schloter‐Hai; Sidy M. Ndaw; Charlotte Periot; Michael Schloter; Bernd Zeller; Franck Poly; Xavier Le Roux

    Some temperate tree species are associated with very low soil nitrification rates, with important implications for forest N dynamics, presumably due to their potential for biological nitrification inhibition (BNI). However, evidence for BNI in forest ecosystems is scarce so far and the nitrifier groups controlled by BNI‐tree species have not been identified. Here, we evaluated how some tree species can control soil nitrification by providing direct evidence of BNI and identifying the nitrifier group(s) affected. First, by comparing 28 year‐old monocultures of several tree species, we showed that nitrification rates correlated strongly with the abundance of the nitrite oxidizers Nitrobacter (50‐ to 1000‐fold changes between tree monocultures) and only weakly with the abundance of ammonia oxidizing archaea (AOA). Second, using reciprocal transplantation of soil cores between low and high nitrification stands, we demonstrated that nitrification changed 16 months after transplantation and was correlated with changes in the abundance of Nitrobacter, not AOA. Third, extracts of litter or soil collected from the low nitrification stands of Picea abies and Abies nordmanniana inhibited the growth of Nitrobacter hamburgensis X14. Our results provide for the first time direct evidence of BNI by tree species directly affecting the abundance of Nitrobacter.

  • Viral SPP1 DNA is infectious in naturally competent Bacillus subtilis cells: inter‐ and intramolecular recombination pathways
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-03
    Ester Serrano; Cristina Ramos; Silvia Ayora; Juan C. Alonso

    A proteolyzed bacteriophage (phage) might release its DNA into the environment. Here, we define the recombination functions required to resurrect an infective lytic phage from inactive environmental viral DNA in naturally competent Bacillus subtilis cells. Using phage SPP1 DNA, a model that accounts for the obtained data is proposed (i) the DNA uptake apparatus takes up environmental SPP1 DNA, fragments it, and incorporates into the cytosol different linear single‐stranded (ss) DNA molecules shorter than genome‐length; (ii) the SsbA‐DprA mediator loads RecA onto any fragmented linear SPP1 ssDNA, but negative modulators (RecX and RecU) promote a net RecA disassembly from these ssDNAs not homologous to the host genome; (iii) single strand annealing (SSA) proteins, DprA and RecO, anneal the SsbA‐ or SsbB‐coated complementary strands, yielding tailed SPP1 duplex intermediates; (iv) RecA polymerized on these tailed intermediates invades a homologous region in another incomplete molecule, and in concert with RecD2 helicase, reconstitutes a complete linear phage genome with redundant regions at the ends of the molecule; and (v) DprA, RecO or viral G35P SSA, may catalyze the annealing of these terminally redundant regions, alone or with the help of an exonuclease, to produce a circular unit‐length duplex viral genome ready to initiate replication.

  • Chromobacterium violaceum delivers violacein, a hydrophobic antibiotic, to other microbes in membrane vesicles
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-02
    Seong Yeol Choi; Sungbin Lim; Gayoung Cho; Jisoo Kwon; Wonsik Mun; Hansol Im; Robert J. Mitchell

    This study describes Chromobacterium violaceum's use of extracellular membrane vesicles (MVs) to both solubilize and transport violacein to other microorganisms. Violacein is a hydrophobic bisindole with known antibiotic activities against other microorganisms. Characterization of the MVs found they carried more violacein than protein (1.37 ± 0.19‐fold), suggesting they may act as a reservoir for this compound. However, MVs are not produced in response to violacein – a ΔvioA isogenic mutant, which is incapable of making violacein, actually produced significantly more MVs (3.2‐fold) than the wild‐type strain. Although violacein is insoluble in water (Log Poctanol:water = 3.34), 79.5% remained in the aqueous phase when it was present within the C. violaceum MVs, an increase in solubility of 1740‐fold. Moreover, tests with a strain of Staphylococcus aureus showed MV‐associated violacein is bactericidal, with 3.1 mg/l killing 90% of S. aureus in 6 h. Tests with the ΔvioA MVs found no loss in the S. aureus viability, even when its MVs were added at much higher concentrations, demonstrating violacein is the active component within the wild‐type MVs. In conclusion, our study clearly demonstrates C. violaceum produces MVs and uses them as vehicles to solubilize violacein and transport this hydrophobic antibiotic to other microbes.

  • Comparison of sulfide‐oxidizing Sulfurimonas strains reveals a new mode of thiosulfate formation in subsurface environments
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-02
    Sven Lahme; Cameron M. Callbeck; Lucy E. Eland; Anil Wipat; Dennis Enning; Ian M. Head; Casey R.J. Hubert

    Sulfur‐oxidizing Sulfurimonas spp. are widespread in sediments, hydrothermal vent fields, aquifers and subsurface environments such as oil reservoirs where they play an important role in the sulfur cycle. We determined the genome sequence of the oil field isolate Sulfurimonas sp. strain CVO and compared its gene expression during nitrate‐dependent sulfide oxidation to the coastal sediment isolate Sulfurimonas denitrificans. Formation of elemental sulfur (S0) and high expression of sulfide quinone oxidoreductase (SQR) genes indicates that sulfide oxidation in both strains is mediated by SQR. Subsequent oxidation of S0 was achieved by the sulfur oxidation enzyme complex (SOX). In the coastal S. denitrificans, the genes are arranged and expressed as two clusters: soxXY1Z1AB and soxCDY2Z2H, and sulfate was the sole metabolic end product. By contrast, the oil field strain CVO has only the soxCDY2Z2H cluster and not soxXY1Z1AB. Despite the absence of the soxXY1Z1AB cluster, strain CVO oxidized S0 to thiosulfate and sulfate, demonstrating that soxCDY2Z2H genes alone are sufficient for S0 oxidation in Sulfurimonas spp. and that thiosulfate is an additional metabolic end product. Screening of publicly available metagenomes revealed that Sulfurimonas spp. with only the soxCDY2Z2H cluster are widespread suggesting this mechanism of thiosulfate formation is environmentally significant.

  • Methane oxidation in anoxic lake water stimulated by nitrate and sulfate addition
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-01
    Sigrid van Grinsven; Jaap S. Sinninghe Damsté; Alejandro Abdala Asbun; Julia C. Engelmann; John Harrison; Laura Villanueva

    Methanotrophic bacteria play a key role in limiting methane emissions from lakes. It is generally assumed that methanotrophic bacteria are mostly active at the oxic‐anoxic transition zone in stratified lakes, where they use oxygen to oxidize methane. Here, we describe a methanotroph of the genera Methylobacter that is performing high‐rate (up to 72 μM day−1) methane oxidation in the anoxic hypolimnion of the temperate Lacamas Lake (Washington, USA), stimulated by both nitrate and sulfate addition. Oxic and anoxic incubations both showed active methane oxidation by a Methylobacter species, with anoxic rates being threefold higher. In anoxic incubations, Methylobacter cell numbers increased almost two orders of magnitude within 3 days, suggesting that this specific Methylobacter species is a facultative anaerobe with a rapid response capability. Genomic analysis revealed adaptations to oxygen‐limitation as well as pathways for mixed‐acid fermentation and H2 production. The denitrification pathway was incomplete, lacking the genes narG/napA and nosZ, allowing only for methane oxidation coupled to nitrite‐reduction. Our data suggest that Methylobacter can be an important driver of the conversion of methane in oxygen‐limited lake systems and potentially use alternative electron acceptors or fermentation to remain active under oxygen‐depleted conditions.

  • The fungal mitochondrial membrane protein, BbOhmm, antagonistically controls hypoxia tolerance
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-01
    Zhangjiang He; Xin Zhao; Yifei Gao; Nemat O Keyhani; Huifang Wang; Juan Deng; Zhuoyue Lu; Yanze Kan; Zhibing Luo; Yongjun Zhang

    Adaptation to low‐oxygen environment in host tissues is crucial for microbial pathogens, particuarlly fungi, to successfully infect target hosts. However, the underlying mechanisms responsible for hypoxia tolerance in most pathogens are poorly understood. A mitochondrial protein, BbOhmm, is demonstrated to limit oxidative stress resistance and virulence in the insect fungal pathogen, Beauveria bassiana. Here, we found that BbOhmm negatively affected hypoxic adaptation in the insect haemocoel while regulating respiration‐related events, heme synthesis and mitochondrial iron homeostasis. A homologue of the mammalian sterol regulatory element‐binding proteins (SREBPs), BbSre1, was shown to be involved in BbOhmm‐mediated low‐oxygen adaptation. Inactivation of BbSre1 resulted in a significant increase in sensitivity to hypoxic and oxidative stress. Similar to ΔBbOhmm, ΔBbSre1 or the ΔBbOhmmΔBbSre1 double mutant accumulated high levels of heme and mitochondrial iron, regulating the similar pathways during hypoxic stress. BbSre1 transcriptional activity and nuclear import were repressed in ΔBbOhmm cells, and affected by intracellular ROS and oxygen levels. These findings have led to a new model in which BbOhmm affects ROS homeostasis in combination with available oxygen to control the transcriptional activity of BbSre1, which in turn mediates low‐oxygen adaptation by regulating mitochondrial iron homeostasis, heme synthesis and respiration‐implicated genes.

  • Spatio‐temporal study of microbiology in the stratified oxic‐hypoxic‐euxinic, freshwater‐to‐hypersaline Ursu Lake
    Environ. Microbiol. (IF 5.147) Pub Date : 2020-01-01
    Andreea Baricz; Cecilia Maria Chiriac; Adrian‐Ștefan Andrei; Paul Adrian Bulzu; Erika Andrea Levei; Oana Cadar; Karina Paula Battes; Mirela Cîmpean; Marin ȘenilĂ; Adorján Cristea; Vasile Muntean; Mircea Alexe; Cristian Coman; Edina Kriszta Szekeres; Cosmin Ionel Sicora; Artur Ionescu; David Blain; William Kenneth O'Neill; Jessica Edwards; John Edward Hallsworth; Horia Leonard Banciu

    Ursu Lake is located in the Middle Miocene salt deposit of Central Romania. It is stratified, and the water column has three distinct water masses: an upper, freshwater‐to‐moderately saline stratum (0–3 m), an intermediate stratum exhibiting a steep halocline (3–3.5 m), and a lower, hypersaline stratum (4 m and below) that is euxinic (i.e. anoxic and sulphidic). Recent studies have characterised the lake's microbial taxonomy, and given rise to intriguing ecological questions. Here, we explore whether the communities are dynamic or stable in relation to taxonomic composition, geochemistry and biophysics, and ecophysiological functions during the annual cycle. We found: (i) seasonally fluctuating, light‐dependent communities in the upper layer (≥0.987–0.990 water‐activity), a stable but phylogenetically diverse population of heterotrophs in the hypersaline stratum (water activities down to 0.762), and a persistent plate of green sulphur bacteria that connects these two (0.958–0.956 water activity) at 3–3.5 m; (ii) communities which might be be involved in carbon‐ and sulphur cycling between and within the lake's three main water masses; (iii) uncultured lineages including Acetothermia (OP1), Candidate Phyla Radiation, Cloacimonetes (WWE1), Marinimicrobia (SAR406), Omnitrophicaeota (OP3), Parcubacteria (OD1), and SR1, in the hypersaline stratum (likely involved in the anaerobic steps of carbon‐ and sulphur cycling); and (iv) that species richness and habitat stability are associated with high redox‐potentials. Ursu Lake has a unique and complex ecology, exhibiting both dynamic fluctuations and stability, and can be used as a comparator system for other stratified hypersaline systems and a modern analogue for ancient euxinic water bodies.

  • Unravelling interspecies interactions across heterogeneities in complex biofilm communities
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-10
    Henriette L. Røder; Nanna M. C. Olsen; Marvin Whiteley; Mette Burmølle

    The importance of microbial biofilms has been well‐recognized for several decades, and focus is now shifting towards investigating multispecies biofilm communities rather than mono‐ or dual‐species biofilms. Therefore, the demand for techniques that provide a sufficient amount of information at adequate resolution is increasing. One major challenge for multispecies studies is that diversity and spatial organization often lead to a high degree of spatial and chemical heterogeneity. Many current approaches do not account for such heterogeneity and therefore only provide average information (−omics techniques in particular), which could obscure important information about the community. Here, we bring attention to the issues of heterogeneity when analysing synthetic multi‐species biofilms, in vitro, and the importance of multi‐scale approaches. We provide an overview of current and newer approaches that can be applied to biofilm communities, in order to elucidate interactions at the appropriate scale.

  • A common mechanism for efficient N2O reduction in diverse isolates of nodule‐forming bradyrhizobia
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-07-18
    Daniel Mania; Kedir Woliy; Tulu Degefu; åsa Frostegård

    Bradyrhizobia are abundant soil bacteria, which can form nitrogen‐fixing symbioses with leguminous plants, including important crops such as soybean, cowpea and peanut. Many bradyrhizobia can denitrify, but studies have hitherto focused on a few model organisms. We screened 39 diverse Bradyrhizobium strains, isolated from legume nodules. Half of them were unable to reduce N2O, making them sources of this greenhouse gas. Most others could denitrify NO3− to N2. Time‐resolved gas kinetics and transcription analyses during transition to anaerobic respiration revealed a common regulation of nirK, norCB and nosZ (encoding NO2−, NO and N2O reductases), and differing regulation of napAB (encoding periplasmic NO3− reductase). A prominent feature in all N2‐producing strains was a virtually complete hampering of NO3− reduction in the presence of N2O. In‐depth analyses suggest that this was due to a competition between electron transport pathways, strongly favouring N2O over NO3− reduction. In a natural context, bacteria with this feature would preferentially reduce available N2O, produced by themselves or other soil bacteria, making them powerful sinks for this greenhouse gas. One way to augment such populations in agricultural soils is to develop inoculants for legume crops with dual capabilities of efficient N2‐fixation and efficient N2O reduction.

  • Unravelling plasmidome distribution and interaction with its hosting microbiome
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-15
    Aya Brown Kav; Roye Rozov; David Bogumil; Søren Johannes Sørensen; Lars Hestbjerg Hansen; Itai Benhar; Eran Halperin; Ron Shamir; Itzhak Mizrahi

    Horizontal gene transfer via plasmids plays a pivotal role in microbial evolution. The forces that shape plasmidomes functionality and distribution in natural environments are insufficiently understood. Here, we present a comparative study of plasmidomes across adjacent microbial environments present in different individual rumen microbiomes. Our findings show that the rumen plasmidome displays enormous unknown functional potential currently unannotated in available databases. Nevertheless, this unknown functionality is conserved and shared with published rat gut plasmidome data. Moreover, the rumen plasmidome is highly diverse compared with the microbiome that hosts these plasmids, across both similar and different rumen habitats. Our analysis demonstrates that its structure is shaped more by stochasticity than selection. Nevertheless, the plasmidome is an active partner in its intricate relationship with the host microbiome with both interacting with and responding to their environment.

  • Mismatch repair hierarchy of Pseudomonas putida revealed by mutagenic ssDNA recombineering of the pyrF gene
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-07
    Tomas Aparicio; Akos Nyerges; István Nagy; Csaba Pal; Esteban Martínez‐García; Víctor de Lorenzo

    The mismatch repair (MMR) system is one of the key molecular devices that prokaryotic cells have for ensuring fidelity of DNA replication. While the canonical MMR of E. coli involves 3 proteins (encoded by mutS, mutL and mutH), the soil bacterium Pseudomonads putida has only 2 bona fide homologues (mutS and mutL) and the sensitivity of this abridged system to different types of mismatches is unknown. In this background, sensitivity to MMR of this bacterium was inspected through single stranded (ss) DNA recombineering of the pyrF gene (the prokaryotic equivalent to yeast's URA3) with mutagenic oligos representative of every possible mispairing under either wild‐type conditions, permanent deletion of mutS or transient loss of mutL activity (brought about by the thermoinducible dominant negative allele mutLE36K). Analysis of single nucleotide mutations borne by clones resistant to fluoroorotic acid (5FOA, the target of wild type PyrF) pinpointed prohibited and tolerated single‐nucleotide replacements and exposed a clear grading of mismatch recognition. The resulting data unequivocally established the hierarchy A:G < C:C < G:A < C:A, A:A, G:G, T:T, T:G, A:C, C:T < G:T, T:C as the one prevalent in Pseudomonas putida. This information is vital for enabling recombineering strategies aimed at single‐nucleotide changes in this biotechnologically important species.

  • Skeleton bones in museum indoor environments offer niches for fungi and are affected by weathering and deposition of secondary minerals
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-28
    Flavia Pinzari; Lorraine Cornish; Anne D. Jungblut

    Large skeleton specimens are often featured as iconic open displays in Natural History Museums, for example, the blue whale ‘Hope’ at the Natural History Museum, London. A study on Hope's bone surface was performed to assess the biodeterioration potential of fungi. Fungi were isolated, and a fungal internal transcribed spacer (ITS) clone library survey was performed on dust and bone material. Mineral particles derived from bone and dust were analysed using energy dispersive X‐ray spectroscopy, variable pressure scanning electron microscopy (SEM) and high vacuum SEM. Results showed that bone material, although mainly mineral in nature, and therefore less susceptible than organic materials to biodeterioration phenomena in the indoor environments, offers niches for specialized fungi and is affected by unusual and yet not so well‐documented mechanisms of alteration. Areas of bone surface were covered with a dense biofilm mostly composed of fungal hyphae, which produced tunnelling and extensive deposition of calcium and iron‐containing secondary minerals. Airborne halophilic and xerophilic fungi including taxa grouping into Ascomycota and Basidiomycota, capable of displacing salts and overcome little water availability, were found to dominate the microbiome of the bone surface.

  • The Neurospora RNA polymerase II kinase CTK negatively regulates catalase expression in a chromatin context‐dependent manner
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-10-21
    Jiabin Duan; Qingqing Liu; Sodgerel Su; Joonseok Cha; Yike Zhou; Ruiqi Tang; Xiao Liu; Ying Wang; Yi Liu; Qun He

    Clearance and adaptation to reactive oxygen species (ROS) are crucial for cell survival. As in other eukaryotes, the Neurospora catalases are the main enzymes responsible for ROS clearance and their expression are tightly regulated by the growth and environmental conditions. The RNA polymerase II carboxyl terminal domain (RNAPII CTD) kinase complex (CTK complex) is known as a positive elongation factor for many inducible genes by releasing paused RNAPII near the transcription start site and promoting transcription elongation. However, here we show that deletion of CTK complex components in Neurospora led to high CAT‐3 expression level and resistance to H2O2‐induced ROS stress. The catalytic activity of CTK‐1 is required for such a response. On the other hand, CTK‐1 overexpression led to decreased expression of CAT‐3. ChIP assays shows that CTK‐1 phosphorylates the RNAPII CTD at Ser2 residues in the cat‐3 ORF region during transcription elongation and deletion of CTK‐1 led to dramatic decreases of SET‐2 recruitment and H3K36me3 modification. As a result, histones at the cat‐3 locus become hyperacetylated to promote its transcription. Together, these results demonstrate that the CTK complex is negative regulator of cat‐3 expression by affecting its chromatin structure.

  • Identification of a complete dibenzothiophene biodesulfurization operon and its regulator by functional metagenomics
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-10-21
    Guadalupe Martín‐Cabello; Laura Terrón‐González; Manuel Ferrer; Eduardo Santero

    Functional screening for aromatic ring oxygenases of an oil contaminated soil metagenome identified 25 different clones bearing monooxygenases coding genes. One fosmid bore an operon containing four tightly linked genes coding for a complete dibenzothiophene biodesulfurization pathway, which included the predicted monooxygenases DszC and DszA, the desulfinase DszB, and an FMN‐oxidoreductase designated DszE. The dszEABC operon provided Escherichia coli with the ability to use dibenzothiophene as the only sulfur source. Transcription of the operon is driven from a σN‐dependent promoter and regulated by an activator that was designated dszR. DszR has been purified and characterized in vitro and shown to be a constitutively active σN‐dependent activator of the group IV, which binds to two contiguous sequences located upstream of the promoter. The dsz promoter and dszE and dszR genes have apparently been recruited from an aliphatic sulfonate biodegradation pathway. If transcribed from a heterologous upstream promoter, the σN‐dependent promoter region functions as an ‘insulator’ that prevents translation of dszE, by binding with its ribosome binding site. Translational coupling, in turn, prevents translation of the downstream dszABC genes. The silencer combined with translational coupling thus represents an effective way of preventing expression of operons when spuriously transcribed from upstream promoters.

  • In Ganoderma lucidum, Glsnf1 regulates cellulose degradation by inhibiting GlCreA during the utilization of cellulose
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-10-28
    Yanru Hu; Wenzhao Xu; Shishan Hu; Lingdan Lian; Jing Zhu; Liang Shi; Ang Ren; MingWen Zhao

    Cellulose is a by‐product of agricultural production and an abundant waste. As a carbon source, cellulose can be degraded and utilized by fungi. Carbon sources, which act as nutrients, not only provide energy but also serve as regulators of gene expression, metabolism and growth, through various signalling networks that enable cells to sense and adapt to varying environmental conditions. Nutrient‐sensing pathways prioritize the use of preferred carbon sources and regulate the production of cellulose‐degrading enzymes when necessary. Understanding the regulation of the fungal cellulolytic response will become increasingly important because we strive to increase the efficiency of the utilization of these renewable energy sources. Here, we show that Glsnf1, a sucrose‐nonfermenting serine–threonine‐protein kinase 1 (Snf1)/AMP‐activated protein kinase homologue in medicinal macro basidiomycete Ganoderma lucidum, actively responds to carbon alterations and positively regulates cellulase activity and cellulase‐related gene transcription. The carbon catabolite repressor CreA, a zinc binuclear cluster transcription factor that mediates the sensing of nutrients and suppression of the transcription of a number of genes necessary for the consumption of a less preferred carbon source, participates in the Glsnf1‐mediated regulation of cellulases. Glsnf1 not only negatively regulates the transcription level of the CreA gene but also hinders its localization in the nucleus. Overall, our findings reveal a key nutrient‐sensing mechanism that is critical for the modulation of carbon source adaptation in G. lucidum.

  • At the nexus of three kingdoms: the genome of the mycorrhizal fungus Gigaspora margarita provides insights into plant, endobacterial and fungal interactions
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-10-31
    Francesco Venice; Stefano Ghignone; Alessandra Salvioli di Fossalunga; Joëlle Amselem; Mara Novero; Xie Xianan; Kinga Sędzielewska Toro; Emmanuelle Morin; Anna Lipzen; Igor V. Grigoriev; Bernard Henrissat; Francis M. Martin; Paola Bonfante

    As members of the plant microbiota, arbuscular mycorrhizal fungi (AMF, Glomeromycotina) symbiotically colonize plant roots. AMF also possess their own microbiota, hosting some uncultivable endobacteria. Ongoing research has revealed the genetics underlying plant responses to colonization by AMF, but the fungal side of the relationship remains in the dark. Here, we sequenced the genome of Gigaspora margarita, a member of the Gigasporaceae in an early diverging group of the Glomeromycotina. In contrast to other AMF, G. margarita may host distinct endobacterial populations and possesses the largest fungal genome so far annotated (773.104 Mbp), with more than 64% transposable elements. Other unique traits of the G. margarita genome include the expansion of genes for inorganic phosphate metabolism, the presence of genes for production of secondary metabolites and a considerable number of potential horizontal gene transfer events. The sequencing of G. margarita genome reveals the importance of its immune system, shedding light on the evolutionary pathways that allowed early diverging fungi to interact with both plants and bacteria.

  • A crosstalk between c‐di‐GMP and cAMP in regulating transcription of GcsA, a diguanylate cyclase involved in swimming motility in Pseudomonas putida
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-06
    Yujie Xiao; Huizhong Liu; Meina He; Liang Nie; Hailing Nie; Wenli Chen; Qiaoyun Huang

    The ubiquitous bacterial second messenger c‐di‐GMP is synthesized by diguanylate cyclase (DGC) and degraded by phosphodiesterase (PDE). Pseudomonas putida has dozens of DGC/PDE‐encoding genes in its genome, but the phenotypical–genotypical correlation and transcriptional regulation of these genes are largely unknown. Herein, we characterize function and transcriptional regulation of a P. putida c‐di‐GMP‐metabolizing enzyme, GcsA. GcsA consists of two per‐ARNT‐sim (PAS) domains, followed by a canonical conserved central sequence pattern (GGDEF) domain and a truncated EAL domain. In vitro analysis confirmed the DGC activity of GcsA. The phenotypic observation revealed that GcsA inhibited swimming motility in an FlgZ‐dependent manner. In terms of transcriptional regulation, gcsA was found to be cooperatively regulated by c‐di‐GMP and cAMP via their effectors, FleQ and Crp respectively. The transcription of gcsA was promoted by c‐di‐GMP and inhibited by cAMP. In vitro binding analysis revealed that FleQ indirectly regulated the transcription of gcsA, while Crp directly regulated the transcription of gcsA by binding to its promoter. Besides, an inverse relationship between the cellular c‐di‐GMP and cAMP levels in P. putida was confirmed. These findings provide basic knowledge regarding the function and transcriptional regulation of GcsA and demonstrate a crosstalk between c‐di‐GMP and cAMP in the regulation of the expression of GcsA in P. putida.

  • ICETh1 and ICETh2, two interdependent mobile genetic elements in Thermus thermophilus transjugation
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-02
    Ignacio Baquedano; Mario Mencía; Alba Blesa; Vincent Burrus; José Berenguerfi

    Cell to cell DNA transfer between Thermus thermophilus, or transjugation, requires the natural competence apparatus (NCA) of the recipient cell and a DNA donation machinery in the donor. In T. thermophilus HB27, two mobile genetic elements with functional similarities to Integrative and Conjugative Elements (ICEs) coexist, ICETh1 encoding the DNA transfer apparatus and ICETh2, encoding a putative replication module. Here, we demonstrate that excision and integration of both elements depend on a single tyrosine recombinase encoded by ICETh2, and that excision is not required but improves the transfer of these elements to a recipient cell. These findings along with previous results suggest that ICETh1 and ICETh2 depend on each other for spreading among T. thermophilus by transjugation.

  • Bacillus subtilis builds structurally and functionally different spores in response to the temperature of growth
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-24
    Rachele Isticato; Mariamichela Lanzilli; Claudia Petrillo; Giuliana Donadio; Loredana Baccigalupi; Ezio Ricca

    Bacterial spores are commonly isolated from a variety of different environments, including extreme habitats. Although it is well established that such ubiquitous distribution reflects the spore resistance properties, it is not clear whether the growing conditions affect the spore structure and function. We used Bacillus subtilis spores of similar age but produced at 25, 37, or 42°C to compare their surface structures and functional properties. Spores produced at the 25°C were more hydrophobic while those produced at 42°C contained more dipicolinic acid, and were more resistant to heat or lysozyme treatments. Electron microscopy analysis showed that while 25°C spores had a coat with a compact outer coat, not tightly attached to the inner coat, 42°C spores had a granular, not compact outer coat, reminiscent of the coat produced at 37°C by mutant spores lacking the protein CotG. Indeed, CotH and a series of CotH‐dependent coat proteins including CotG were more abundantly extracted from the coat of 25 or 37°C than 42°C spores. Our data indicated that CotH is a heat‐labile protein with a major regulatory role on coat formation when sporulation occurs at low temperatures, suggesting that B. subtilis builds structurally and functionally different spores in response to the external conditions.

  • Acclimation to low ultraviolet‐B radiation increases photosystem I abundance and cyclic electron transfer with enhanced photosynthesis and growth in the cyanobacterium Nostoc sphaeroides
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-10
    Zhen Chen; Hai‐Bo Jiang; Kunshan Gao; Bao‐Sheng Qiu

    Ultraviolet‐B radiation is known to harm most photosynthetic organisms with the exception of several studies of photosynthetic eukaryotes in which UV‐B showed positive effects. In this study, we investigated the effect of acclimation to low UV‐B radiation on growth and photosynthesis of the cyanobacterium Nostoc sphaeroides. Exposure to 0.08 W m−2 UV‐B plus low visible light for 14 d significantly increased the growth rate and biomass production by 16% and 30%, respectively, compared with those under visible light alone. The UV‐B acclimated cells showed an approximately 50% increase in photosynthetic efficiency (α) and photosynthetic capacity (Pmax), a higher PSI/PSII fluorescence ratio, an increase in PSI content and consequently enhanced cyclic electron flow, relative to those of non‐acclimated cells. Both the primary quinone‐type acceptor and plastoquinone pool re‐oxidation were up‐regulated in the UV‐B acclimated cells. In parallel, the UV‐B acclimated colonies maintained a higher rate of D1 protein synthesis following exposure to elevated intensity of UV‐B or visible light, thus functionally mitigating photoinhibition. The present data provide novel insight into photosynthetic acclimation to low UV‐B radiation and suggest that UV‐B may act as a positive ecological factor for the productivity of some photosynthetic prokaryotes, especially during twilight periods or in shaded environments.

  • 100‐year‐old enigma solved: identification, genomic characterization and biogeography of the yet uncultured Planctomyces bekefii
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-11
    Svetlana N. Dedysh; Petra Henke; Anastasia A. Ivanova; Irina S. Kulichevskaya; Dmitriy A. Philippov; Jan P. Meier‐Kolthoff; Markus Göker; Sixing Huang; Jörg Overmann

    The first representative of the phylum Planctomycetes, Planctomyces bekefii, was described nearly one century ago. This morphologically conspicuous freshwater bacterium is a rare example of as‐yet‐uncultivated prokaryotes with validly published names and unknown identity. We report the results of molecular identification of this elusive bacterium, which was detected in a eutrophic boreal lake in Northern Russia. By using high‐performance cell sorting, P. bekefii‐like cell rosettes were selectively enriched from lake water. The retrieved 16S rRNA gene sequence was nearly identical to those in dozens of metagenomes assembled from freshwater lakes during cyanobacterial blooms and was phylogenetically placed within a large group of environmental sequences originating from various freshwater habitats worldwide. In contrast, 16S rRNA gene sequence similarity to all currently described members of the order Planctomycetales was only 83%–92%. The metagenome assembled for P. bekefii reached 43% genome coverage and showed the potential for degradation of peptides, pectins, and sulfated polysaccharides. Tracing the seasonal dynamics of P. bekefii by Illumina paired‐end sequencing of 16S rRNA gene fragments and by fluorescence in situ hybridization revealed that these bacteria only transiently surpass the detection limit, with a characteristic population peak of up to 104 cells ml−1 following cyanobacterial blooms.

  • A single Thaumarchaeon drives nitrification in deep oligotrophic Lake Constance
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-12
    Janina Herber; Franziska Klotz; Benjamin Frommeyer; Severin Weis; Dietmar Straile; Allison Kolar; Johannes Sikorski; Markus Egert; Michael Dannenmann; Michael Pester

    Ammonia released during organic matter mineralization is converted during nitrification to nitrate. We followed spatiotemporal dynamics of the nitrifying microbial community in deep oligotrophic Lake Constance. Depth‐dependent decrease of total ammonium (0.01–0.84 μM) indicated the hypolimnion as the major place of nitrification with 15N‐isotope dilution measurements indicating a threefold daily turnover of hypolimnetic total ammonium. This was mirrored by a strong increase of ammonia‐oxidizing Thaumarchaeota towards the hypolimnion (13%–21% of bacterioplankton) throughout spring to autumn as revealed by amplicon sequencing and quantitative polymerase chain reaction. Ammonia‐oxidizing bacteria were typically two orders of magnitude less abundant and completely ammonia‐oxidizing (comammox) bacteria were not detected. Both, 16S rRNA gene and amoA (encoding ammonia monooxygenase subunit B) analyses identified only one major species‐level operational taxonomic unit (OTU) of Thaumarchaeota (99% of all ammonia oxidizers in the hypolimnion), which was affiliated to Nitrosopumilus spp. The relative abundance distribution of the single Thaumarchaeon strongly correlated to an equally abundant Chloroflexi clade CL500‐11 OTU and a Nitrospira OTU that was one order of magnitude less abundant. The latter dominated among recognized nitrite oxidizers. This extremely low diversity of nitrifiers shows how vulnerable the ecosystem process of nitrification may be in Lake Constance as Central Europe's third largest lake.

  • GATA‐type transcriptional factor Gat1 regulates nitrogen uptake and polymalic acid biosynthesis in polyextremotolerant fungus Aureobasidium pullulans
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-10
    Xiaodan Song; Yongkang Wang; Pan Wang; Guihong Pu; Xiang Zou

    Polymalic acid (PMA) is a novel biopolymer produced by the polyextremotolerant fungus Aureobasidium pullulans. In this study, a GATA‐family transcriptional factor, Gat1, which regulates nitrogen uptake and PMA biosynthesis, was investigated. PMA production increased to 11.2% in the mutant overexpressing gat1 but decreased to 49.1% of the PMA titre when gat1 was knocked out from the genome of A. pullulans. Comparative transcriptome analysis of wild‐type and mutant strains (∆gat1 and OE::gat1) revealed that 23 common differentially expressed genes were related to oxidative phosphorylation, ribosome biogenesis, and nitrogen metabolism. Under nitrogen‐limited stress, regardless of the preferred nitrogen (glutamine, Gln) or non‐preferred nitrogen (proline, Pro), 70% of Gat1 in the cells was located in the nucleus–cytoplasm, which resulted in an increase in nitrogen uptake and PMA biosynthesis regulation. Quantitative RT‐PCR revealed that glucosekinase (GLK) in the glycolytic pathway and malate synthase (MLS) in the glyoxylate shunt pathway may be cross‐regulated by Gat1 and nitrogen concentration (Gln or Pro), Therefore, glk was overexpressed in mutant strain (OE::gat1), which resulted in an increased PMA titre and yield of 12.6% and 13.0% respectively. These findings indicate that Gat1 may play an important role in the dual regulation of the nitrogen and carbon metabolisms in PMA biosynthesis.

  • Evidence for the coexistence of direct and riboflavin‐mediated interspecies electron transfer in Geobacter co‐culture
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-10
    Lingyan Huang; Xing Liu; Yin Ye; Man Chen; Shungui Zhou

    Geobacter species can secrete free redox‐active flavins, but the role of these flavins in the interspecies electron transfer (IET) of Geobacter direct interspecies electron transfer (DIET) co‐culture is unknown. Here, we report the presence of a new riboflavin‐mediated interspecies electron transfer (RMIET) process in a traditional Geobacter DIET co‐culture; in this process, riboflavin contributes to IET by acting as a free‐form electron shuttle between free Geobacter species and serving as a bound cofactor of some cytochromes in Geobacter co‐culture aggregates. Multiple lines of evidence indicate that RMIET facilitates the primary initiation of syntrophic growth between Geobacter species before establishing the DIET co‐culture and provides additional ways alongside the DIET to transfer electrons to achieve electric syntrophy between Geobacter species. Redox kinetic analysis of riboflavin on either Geobacter species demonstrated that the Gmet_2896 cytochrome acts as the key riboflavin reduction site, while riboflavin oxidation by Geobacter sulfurreducens is the rate‐limiting step in RMIET, and the RMIET makes only a minor contribution to IET in Geobacter DIET co‐culture. The discovery of a new RMIET process in Geobacter DIET co‐culture suggests the complexity of IET in syntrophic bacterial communities and provides suggestions for the careful examination of the IET of other syntrophic co‐cultures.

  • High‐quality genome‐scale metabolic modelling of Pseudomonas putida highlights its broad metabolic capabilities
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-11
    Juan Nogales; Joshua Mueller; Steinn Gudmundsson; Francisco J. Canalejo; Estrella Duque; Jonathan Monk; Adam M. Feist; Juan Luis Ramos; Wei Niu; Bernhard O. Palsson

    Genome‐scale reconstructions of metabolism are computational species‐specific knowledge bases able to compute systemic metabolic properties. We present a comprehensive and validated reconstruction of the biotechnologically relevant bacterium Pseudomonas putida KT2440 that greatly expands computable predictions of its metabolic states. The reconstruction represents a significant reactome expansion over available reconstructed bacterial metabolic networks. Specifically, iJN1462 (i) incorporates several hundred additional genes and associated reactions resulting in new predictive capabilities, including new nutrients supporting growth; (ii) was validated by in vivo growth screens that included previously untested carbon (48) and nitrogen (41) sources; (iii) yielded gene essentiality predictions showing large accuracy when compared with a knock‐out library and Bar‐seq data; and (iv) allowed mapping of its network to 82 P. putida sequenced strains revealing functional core that reflect the large metabolic versatility of this species, including aromatic compounds derived from lignin. Thus, this study provides a thoroughly updated metabolic reconstruction and new computable phenotypes for P. putida, which can be leveraged as a first step toward understanding the pan metabolic capabilities of Pseudomonas.

  • Sequence, structure and function‐based classification of the broadly conserved FAH superfamily reveals two distinct fumarylpyruvate hydrolase subfamilies
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-10
    Hwaseok Hong; Hogyun Seo; Woojin Park; Kyung‐Jin Kim

    Fumarylacetoacetate hydrolase (FAH) superfamily proteins are found ubiquitously in microbial pathways involved in the catabolism of aromatic substances. Although extensive bioinformatic data on these proteins have been acquired, confusion caused by problems with the annotation of these proteins hinders research into determining their physiological functions. Here we classify 606 FAH superfamily proteins using a maximum likelihood (ML) phylogenetic tree, comparative gene‐neighbourhood patterns and in vitro enzyme assays. The FAH superfamily proteins used for the analyses are divided into five distinct subfamilies, and two of them, FPH‐A and FPH‐B, contain the majority of the proteins of undefined function. These subfamilies include clusters designated FPH‐I and FPH‐II, respectively, which include two distinct types of fumarylpyruvate hydrolase (FPH), an enzyme involved in the final step of the gentisate pathway. We determined the crystal structures of these FPH enzymes at 2.0 Å resolutions and investigate the substrate binding mode by which these types of enzymes can accommodate fumarylpyruvate as a substrate. Consequentially, we identify the molecular signatures of the two types of FPH enzymes among the broadly conserved FAH superfamily proteins. Our studies allowed us to predict the relationship of unknown FAH superfamily proteins using their sequence information.

  • Preferential catabolism of the (S)‐enantiomer of the herbicide napropamide mediated by the enantioselective amidohydrolase SnaH and the dioxygenase Snpd in Sphingobium sp. strain B2
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-14
    Junwei Huang; Dian Chen; Jiandong Jiang

    The (R)‐ and (S)‐enantiomers of the chiral herbicide napropamide (NAP) show different biological activities and ecotoxicities. These two enantiomers behave differently in the environment due to enantioselective catabolism by microorganisms. However, the molecular mechanisms underlying this enantioselective catabolism remain largely unknown. In this study, the genes (snaH and snpd) involved in the catabolism of NAP were cloned from Sphingobium sp. B2, which was capable of catabolizing both NAP enantiomers. Compared with (R)‐NAP, (S)‐NAP was much more rapidly transformed by the amidase SnaH, which initially cleaved the amide bonds of (S)/(R)‐NAP to form (S)/(R)‐2‐(1‐naphthalenyloxy)‐propanoic acid [(S)/(R)‐NP] and diethylamine. The α‐ketoglutarate‐dependent dioxygenase Snpd, showing strict stereoselectivity for (S)‐NP, further transformed (S)‐NP to 1‐naphthol and pyruvate. Molecular docking and site‐directed mutagenesis analyses revealed that when the (S)‐enantiomers of NAP and NP occupied the active sites, the distance between the ligand molecule and the coordination atom was shorter than that when the (R)‐enantiomers occupied the active sites, which facilitated formation of the transition state complex. This study enhances our understanding of the preferential catabolism of the (S)‐enantiomer of NAP on the molecular level.

  • Regional community assembly drivers and microbial environmental sources shaping bacterioplankton in an alpine lacustrine district (Pyrenees, Spain)
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-14
    Rüdiger Ortiz‐Álvarez; Joan Cáliz; Lluís Camarero; Emilio O. Casamayor

    Microbial communities in natural ecosystems are subject to strong ecological rules. The study of local communities along a regional metacommunity can reveal patterns of community assembly, and disentangle the underlying ecological processes. In particular, we seek drivers of community assembly at the regional scale using a large lacustrine dataset (>300 lakes) along the geographical, limnological and physico‐chemical gradients in the Pyrenees. By using high throughput amplicon sequencing of the 16S rRNA gene, and inferring environmental sources of bacterial immigrants, we showed that surface aquatic bacterial assemblages were strongly influenced by terrestrial populations from soil, biofilms or sediments, and primarily selected by a pH‐alkalinity gradient. Indeed, source proportions explained 27% of the community variation, and chemistry 15% of the total variation, half of it shared with the sources. Major taxonomic groups such as Verrucomicrobia, Actinobacteria and Bacteroidetes showed higher aquatic affinities than Parcubacteria, Gammaproteobacteria, Alphaproteobacteria or Betaproteobacteria, which may be recruited and selected through different hydrographic habitats. A regional fingerprint was observed with lower alpha diversity and higher beta diversity in the central Pyrenees than in both ends. We suggest an ecological succession process, likely influenced by complex interactions of environmental source dispersal and environmental filtering along the mountain range geography.

  • Short bZIP homologue of sulfur regulator Met4 from Ogataea parapolymorpha does not depend on DNA‐binding cofactors for activating genes in sulfur starvation
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-19
    Su Jin Yoo; Min Jeong Sohn; Da Min Jeong; Hyun Ah Kang

    The acquisition of sulfur from environment and its assimilation is essential for fungal growth and activities. Here, we describe novel features of the regulatory network of sulfur metabolism in Ogataea parapolymorpha, a thermotolerant methylotrophic yeast with high resistance to harsh environmental conditions. A short bZIP protein (OpMet4p) of O. parapolymorpha, displaying the combined structural characteristics of yeast and filamentous fungal Met4 homologues, plays a key role as a master regulator of cell homeostasis during sulfur limitation, but also its function is required for the tolerance of various stresses. Domain swapping analysis, combined with deletion analysis of the regulatory domains and genes encoding OpCbf1p, OpMet28p, and OpMet32p, indicated that OpMet4p does not require the interaction with these DNA‐binding cofactors to induce the expression of sulfur genes, unlike the Saccharomyces cerevisiae Met4p. ChIP analysis confirmed the notion that OpMet4p, which contains a canonical bZIP domain, can bind the target DNA in the absence of cofactors, similar to homologues in other filamentous fungi. Collectively, the identified unique features of the O. parapolymorpha regulatory network, as the first report on the sulfur regulation by a short yeast Met4 homologue, provide insights into conservation and divergence of the sulfur regulatory networks among diverse ascomycetous fungi.

  • Evidences of aromatic degradation dominantly via the phenylacetic acid pathway in marine benthic Thermoprofundales
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-22
    Wei‐Wei Liu; Jie Pan; Xiaoyuan Feng; Meng Li; Ying Xu; Fengping Wang; Ning‐Yi Zhou

    Thermoprofundales (Marine Benthic Group D archaea, MBG‐D) is a newly proposed archaeal order and widely distributed in global marine sediment, and the members in the order may play a vital role in carbon cycling. However, the lack of pure cultures of these oeganisms has hampered the recognition of their catabolic roles. Here, by constructing high‐quality metagenome‐assembled genomes (MAGs) of two new subgroups of Thermoprofundales from hydrothermal sediment and predicting their catabolic pathways, we here provide genomic evidences that Thermoprofundales are capable of degrading aromatics via the phenylacetic acid (PAA) pathway. Then, the gene sequences of phenylacetyl‐CoA ligase (PCL), a key enzyme for the PAA pathway, were searched in reference genomes. The widespread distribution of PCL genes among 14.9% of archaea and 75.9% of Thermoprofundales further supports the importance of the PAA pathway in archaea, particularly in Thermoprofundales where no ring‐cleavage dioxygenases were found. Two PCLs from Thermoprofundales MAGs, PCLM8‐3 and PCLM10‐15, were able to convert PAA to phenylacetyl‐CoA (PA‐CoA) in vitro, demonstrating the involvement of Thermoprofundales in aromatics degradation through PAA via CoA activation. Their acid tolerance (pH 5–7), high‐optimum temperatures (60°C and 80°C), thermostability (stable at 60°C and 50°C for 48 h) and broad substrate spectra imply that Thermoprofundales are capable of transforming aromatics under extreme conditions. Together with the evidence of in situ transcriptional activities for most genes related to the aromatics pathway in Thermoprofundales, these genomic, and biochemical evidences highlight the essential role of this ubiquitous and abundant archaeal order in the carbon cycle of marine sediments.

  • Energy flux controls tetraether lipid cyclization in Sulfolobus acidocaldarius
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-25
    Alice Zhou; Yuki Weber; Beverly K. Chiu; Felix J. Elling; Alec B. Cobban; Ann Pearson; William D. Leavitt

    Microorganisms regulate the composition of their membranes in response to environmental cues. Many Archaea maintain the fluidity and permeability of their membranes by adjusting the number of cyclic moieties within the cores of their glycerol dibiphytanyl glycerol tetraether (GDGT) lipids. Cyclized GDGTs increase membrane packing and stability, which has been shown to help cells survive shifts in temperature and pH. However, the extent of this cyclization also varies with growth phase and electron acceptor or donor limitation. These observations indicate a relationship between energy metabolism and membrane composition. Here we show that the average degree of GDGT cyclization increases with doubling time in continuous cultures of the thermoacidophile Sulfolobus acidocaldarius (DSM 639). This is consistent with the behavior of a mesoneutrophile, Nitrosopumilus maritimus SCM1. Together, these results demonstrate that archaeal GDGT distributions can shift in response to electron donor flux and energy availability, independent of pH or temperature. Paleoenvironmental reconstructions based on GDGTs thus capture the energy available to microbes, which encompasses fluctuations in temperature and pH, as well as electron donor and acceptor availability. The ability of Archaea to adjust membrane composition and packing may be an important strategy that enables survival during episodes of energy stress.

  • Diet shapes cold‐water corals bacterial communities
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-01
    Pierre E. Galand; Marine Remize; Anne‐Leila Meistertzheim; Audrey M. Pruski; Erwan Peru; Tim Jesper Suhrhoff; Nadine Le Bris; Gilles Vétion; Franck Lartaud

    Different cold‐water coral (CWC) species harbour distinct microbial communities and the community composition is thought to be linked to the ecological strategies of the host. Here we test whether diet shapes the composition of bacterial communities associated with CWC. We compared the microbiomes of two common CWC species in aquaria, Lophelia pertusa and Madrepora oculata, when they were either starved, or fed respectively with a carnivorous diet, two different herbivorous diets, or a mix of the 3. We targeted both the standing stock (16S rDNA) and the active fraction (16S rRNA) of the bacterial communities and showed that in both species, the corals' microbiome was specific to the given diet. A part of the microbiome remained, however, species‐specific, which indicates that the microbiome's plasticity is framed by the identity of the host. In addition, the storage lipid content of the coral tissue showed that different diets had different effects on the corals' metabolisms. The combined results suggest that L. pertusa may be preying preferentially on zooplankton while M. oculata may in addition use phytoplankton and detritus. The results cast a new light on coral microbiomes as they indicate that a portion of the CWC's bacterial community could represent a food influenced microbiome.

  • Recurrent seasonal changes in bacterial growth efficiency, metabolism and community composition in coastal waters
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-24
    Zuriñe Baña; Naiara Abad; Ainhoa Uranga; Iñigo Azúa; Itxaso Artolozaga; Marian Unanue; Juan Iriberri; Jesus M. Arrieta; Begoña Ayo

    The microbial response to environmental changes in coastal waters of the eastern Cantabrian Sea was explored for four years by analysing a broad set of environmental variables along with bacterial community metabolism and composition. A recurrent seasonal cycle emerged, consisting of two stable periods, characterized by low bacterial metabolic activity (winter) from October to March, and high bacterial metabolic activity (summer) from May to August. These two contrasting periods were linked by short transition periods in April (TA) and September (TS). The phylogenetic groups Alphaproteobacteria and Bacteroidetes were dominant during winter and summer respectively, and their recurrent alternation was mainly driven by the bloom of eukaryotic phytoplankton before TA and the bloom of prokaryotic phytoplankton before TS. Bacterial growth efficiency remained high and stable during the winter and summer periods but dropped during the two short transition periods. Our results suggest that bacterial growth efficiency should be considered a very resilient property that reflects different stages in the adaptation of the bacterial community composition to the environmental changes occurring throughout the seasonal cycle in this coastal ecosystem.

  • Transcriptional patterns of Emiliania huxleyi in the North Pacific Subtropical Gyre reveal the daily rhythms of its metabolic potential
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-27
    María D. Hernández Limón; Gwenn M. M. Hennon; Matthew J. Harke; Kyle R. Frischkorn; Sheean T. Haley; Sonya T. Dyhrman

    Emiliania huxleyi is a calcifying haptophyte, contributing to both the organic and inorganic marine carbon cycles. In marine ecosystems, light is a major driver of phytoplankton physiology and ultimately carbon flow through the ecosystem. Here, we analysed a Lagrangian time‐series of metatranscriptomes collected in the North Pacific Subtropical Gyre (NPSG) to examine how in situ populations of E. huxleyi modulate gene expression over day–night transitions. Many E. huxleyi contigs had a diel expression pattern, with 61% of contigs clustering into modules with statistically significant diel periodicity. Contigs involved in processes that build up energy stores, like carbon fixation and lipid synthesis, peaked around dawn. In contrast, contigs involved in processes that released energy stores, like respiration and lipid degradation, peaked mid‐day and towards dusk. These patterns suggest an orchestrated cycle of building, then consuming energy stores in E. huxleyi populations in the NPSG. Selected contigs related to the cell cycle also exhibited significant diel periodicity consistent with phased modulations of division observed in culture. Overall, these patterns of gene expression suggest a daily metabolic cascade that could contribute to both organic and inorganic carbon flow in this nutrient depleted ecosystem.

  • Mixed‐mode bacterial transmission in the common brooding coral Pocillopora acuta
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-25
    Katarina Damjanovic; Patricia Menéndez; Linda L. Blackall; Madeleine J. H. van Oppen

    Reef‐building corals form associations with a huge diversity of microorganisms, which are essential for the survival and well‐being of their host. While the acquisition patterns of Symbiodiniaceae microalgal endosymbionts are strongly linked to the coral's reproductive strategy, few studies have investigated the transmission mode of bacteria, especially in brooding species. Here, we relied on 16S rRNA gene and Internal Transcribed Spacer 2 marker metabarcoding in conjunction with fluorescence in situ hybridisation microscopy to describe the onset of microbial associations in the common brooding coral Pocillopora acuta. We analysed the bacterial and Symbiodiniaceae community composition in five adult colonies, their larvae, and 4‐day old recruits. Larvae and recruits inherited Symbiodiniaceae, as well as a small number of bacterial strains, from their parents. Rhodobacteraceae and Endozoicomonas were among the most abundant taxa that were likely maternally transmitted to the offspring. The presence of bacterial aggregates in newly released larvae was observed with confocal microscopy, confirming the occurrence of vertical transmission of bacteria in P. acuta. We concluded that host factors, as well as the environmental bacterial pool influenced the microbiome of P. acuta.

  • Emergence of new variants of antibiotic resistance genomic islands among multidrug‐resistant Salmonella enterica in poultry
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-25
    Emiliano Cohen; Maya Davidovich; Assaf Rokney; Lea Valinsky; Galia Rahav; Ohad Gal‐Mor

    Non‐typhoidal Salmonella enterica (NTS) are diverse and important bacterial pathogens consisting of more than 2600 different serovars, with varying host‐specificity. Here, we characterized the poultry‐associated serovars in Israel, analysed their resistome and illuminated the molecular mechanisms underlying common multidrug resistance (MDR) patterns. We show that at least four serovars including Infantis, Muenchen, Newport and Virchow present a strong epidemiological association between their temporal trends in poultry and humans. Worrisomely, 60% from all of the poultry isolates tested (n = 188) were multidrug resistant, mediated by chromosomal SNPs and different mobile genetics elements. A novel streptomycin‐azithromycin resistance island and previously uncharacterized versions of the mobilized Salmonella genomic island 1 (SGI1) were identified and characterized in S. Blockley and S. Kentucky isolates respectively. Moreover, we demonstrate that the acquisition of SGI1 does not impose fitness cost during growth under nutrient‐limited conditions or in the context of Salmonella infection in the mouse model. Overall, our data emphasize the role of the poultry production as a pool of specific epidemic MDR strains and autonomous genetic elements, which confer resistance to heavy metals and medically relevant antibiotics. These are likely to disseminate to humans via the food chain and fuel the increasing global antibiotic resistance crisis.

  • Microbial communities of the Laurentian Great Lakes reflect connectivity and local biogeochemistry
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-02
    Sara F. Paver; Ryan J. Newton; Maureen L. Coleman

    The Laurentian Great Lakes are a vast, interconnected freshwater system spanning strong physicochemical gradients, thus constituting a powerful natural laboratory for addressing fundamental questions about microbial ecology and evolution. We present a comparative analysis of pelagic microbial communities across all five Laurentian Great Lakes, focusing on Bacterial and Archaeal picoplankton characterized via 16S rRNA amplicon sequencing. We collected samples throughout the water column from the major basins of each lake in spring and summer over 2 years. Two oligotypes, classified as LD12 (Alphaproteobacteria) and acI‐B1 (Actinobacteria), were among the most abundant in every sample. At the same time, microbial communities showed distinct patterns with depth during summer stratification. Deep hypolimnion samples were frequently dominated by a Chloroflexi oligotype that reached up to 19% relative abundance. Stratified surface communities differed between the colder, less productive upper lakes (Superior, Michigan, Huron) and warmer, more productive lower lakes (Erie, Ontario), in part due to an Actinobacteria oligotype (acI‐C2) that averaged 7.7% of sequences in the lower lakes but <0.2% in the upper lakes. Together, our findings suggest that both hydrologic connectivity and local selective pressures shape microbial communities in the Great Lakes and establish a framework for future investigations.

  • Subpopulations of hyphae secrete proteins or resist heat stress in Aspergillus oryzae colonies
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-11-24
    Martin Tegelaar; Robert‐Jan Bleichrodt; Benjamin Nitsche; Arthur F. J. Ram; Han A. B. Wösten

    Hyphae at the outer part of colonies of Aspergillus niger and Aspergillus oryzae are heterogeneous with respect to transcriptional and translational activity. This heterogeneity is maintained by Woronin body mediated closure of septal pores that block interhyphal mixing of cytoplasm. Indeed, heterogeneity between hyphae is abolished in ΔhexA strains that lack Woronin bodies. The subpopulation of hyphae with high transcriptional and translational activity secretes enzymes that degrade the substrate resulting in breakdown products that serve as nutrients. The role of hyphae with low transcriptional and translational activity was not yet known. Here, we show that this subpopulation is more resistant to environmental stress in A. oryzae, in particular to temperature stress, when compared to hyphae with high transcriptional and translational activity. Notably, all hyphae of the ΔhexA strain of A. oryzae were sensitive to heat stress explained by the reduced heterogeneity in this strain. Together, we show that different subpopulations of hypha secrete proteins and resist heat stress showing the complexity of a fungal mycelium.

  • Sediment cooling triggers germination and sulfate reduction by heat‐resistant thermophilic spore‐forming bacteria
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-02
    Emma Bell; Angela Sherry; Giovanni Pilloni; Ana Suárez‐Suárez; Margaret A. Cramm; Guillermo Cueto; Ian M. Head; Casey R. J. Hubert

    Thermophilic endospores are widespread in cold marine sediments where the temperature is too low to support growth and activity of thermophiles in situ. These endospores are likely expelled from warm subsurface environments and subsequently dispersed by ocean currents. The endospore upper temperature limit for survival is 140°C, which can be tolerated in repeated short exposures, potentially enabling transit through hot crustal fluids. Longer‐term thermal tolerance of endospores, and how long they could persist in an environment hotter than their maximum growth temperature, is less understood. To test whether thermophilic endospores can survive prolonged exposure to high temperatures, sediments were incubated at 80–90°C for 6, 12 or 463 days. Sediments were then cooled by 10–40°C, mimicking the cooling in subsurface oil reservoirs subjected to seawater injection. Cooling the sediments induced sulfate reduction, coinciding with an enrichment of endospore‐forming Clostridia. Different Desulfofundulus, Desulfohalotomaculum, Desulfallas, Desulfotomaculum and Desulfofarcimen demonstrated different thermal tolerances, with some Desulfofundulus strains surviving for >1 year at 80°C. In an oil reservoir context, heat‐resistant endospore‐forming sulfate‐reducing bacteria have a survival advantage if they are introduced to, or are resident in, an oil reservoir normally too hot for germination and growth, explaining observations of reservoir souring following cold seawater injection.

  • CmVVD is involved in fruiting body development and carotenoid production and the transcriptional linkage among three blue‐light receptors in edible fungus Cordyceps militaris
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-02
    Jiaojiao Zhang; Fen Wang; Ying Yang; Ying Wang; Caihong Dong

    Fruiting body development and carotenoid production are light‐induced in Cordyceps militaris. Our previous studies have shown that two blue‐light receptors, CmWC‐1 and CmCRY‐DASH, regulate fruiting body development and secondary metabolism. However, the photosensory system of C. militaris remains unclear. Here, gene deletion of Cmvvd, coding for another blue‐light receptor, resulted in reduced conidiation level and significant promotion of carotenoid content. Cmvvd transcription levels at fruiting body stages were higher than at other stages, and fruiting bodies could not develop normally in ΔCmvvd strains, indicating that Cmvvd might play an important role in fruiting body development. Rhythm loops were not affected in ΔCmvvd strains but were regulated by Cmwc‐1, and the expression of the rhythm regulator gene Cmfrq was dependent on CmWC‐1. Chromatin immunoprecipitation assay confirmed that Cmvvd is the direct target of CmWC‐1 in this fungus. Our results also revealed interdependent transcriptional relationships between Cmwc‐1 and Cmvvd, and between Cmwc‐1 and Cmcry‐DASH. Cmcry‐DASH expression was affected by Cmvvd, and the function‐loss of Cmcry‐DASH might be compensated by the high transcription of Cmvvd. This is the first report of the transcriptional linkage among the three blue‐light receptors in edible fungi and will be helpful for studies of multicellular development in this fungus.

  • Genome analysis of sponge symbiont ‘Candidatus Halichondribacter symbioticus’ shows genomic adaptation to a host‐dependent lifestyle
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-08
    Stephen Knobloch; Ragnar Jóhannsson; Viggó Þór Marteinsson

    The marine sponge Halichondria panicea inhabits coastal areas around the globe and is a widely studied sponge species in terms of its biology, yet the ecological functions of its dominant bacterial symbiont ‘Candidatus Halichondribacter symbioticus’ remain unknown. Here, we present the draft genome of ‘Ca. H. symbioticus’ HS1 (2.8 Mbp, ca. 87.6% genome coverage) recovered from the sponge metagenome of H. panicea in order to study functions and symbiotic interactions at the genome level. Functional genome comparison of HS1 against closely related free‐living seawater bacteria revealed a reduction of genes associated with carbohydrate transport and transcription regulation, pointing towards a limited carbohydrate metabolism, and static transcriptional dynamics reminiscent of other bacterial symbionts. In addition, HS1 was enriched in sponge symbiont specific gene families related to host–symbiont interactions and defence. Similarity in the functional gene repertoire between HS1 and a phylogenetically more distant symbiont in the marine sponge Aplysina aerophoba, based on COG category distribution, suggest a convergent evolution of symbiont specific traits and general metabolic features. This warrants further investigation into convergent genomic evolution of symbionts across different sponge species and habitats.

  • Microbial signatures of protected and impacted Northern Caribbean reefs: changes from Cuba to the Florida Keys
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-11
    Laura Weber; Patricia González‐Díaz; Maickel Armenteros; Víctor M. Ferrer; Fernando Bretos; Erich Bartels; Alyson E. Santoro; Amy Apprill

    There are a few baseline reef‐systems available for understanding the microbiology of healthy coral reefs and their surrounding seawater. Here, we examined the seawater microbial ecology of 25 Northern Caribbean reefs varying in human impact and protection in Cuba and the Florida Keys, USA, by measuring nutrient concentrations, microbial abundances, and respiration rates as well as sequencing bacterial and archaeal amplicons and community functional genes. Overall, seawater microbial composition and biogeochemistry were influenced by reef location and hydrogeography. Seawater from the highly protected ‘crown jewel’ offshore reefs in Jardines de la Reina, Cuba had low concentrations of nutrients and organic carbon, abundant Prochlorococcus, and high microbial community alpha diversity. Seawater from the less protected system of Los Canarreos, Cuba had elevated microbial community beta‐diversity whereas waters from the most impacted nearshore reefs in the Florida Keys contained high organic carbon and nitrogen concentrations and potential microbial functions characteristic of microbialized reefs. Each reef system had distinct microbial signatures and within this context, we propose that the protection and offshore nature of Jardines de la Reina may preserve the oligotrophic paradigm and the metabolic dependence of the community on primary production by picocyanobacteria.

  • DsrL mediates electron transfer between NADH and rDsrAB in Allochromatium vinosum
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-29
    Maria Löffler; Julia Feldhues; Sofia S. Venceslau; Lydia Kammler; Fabian Grein; Inês A. C. Pereira; Christiane Dahl

    Dissimilatory sulphite reductase DsrAB occurs in sulphate/sulphite‐reducing prokaryotes, in sulphur disproportionators and also in sulphur oxidizers, where it functions in reverse. Predictions of physiological traits in metagenomic studies relying on the presence of dsrAB, other dsr genes or combinations thereof suffer from the lack of information on crucial Dsr proteins. The iron–sulphur flavoprotein DsrL is an example of this group. It has a documented essential function during sulphur oxidation and was recently also found in some metagenomes of probable sulphate and sulphite reducers. Here, we show that DsrL and reverse acting rDsrAB can form a complex and are copurified from the phototrophic sulphur oxidizer Allochromatium vinosum. Recombinant DsrL exhibits NAD(P)H:acceptor oxidoreductase activity with a strong preference for NADH over NADPH. In vitro, the rDsrABL complex effectively catalyses NADH‐dependent sulphite reduction, which is strongly enhanced by the sulphur‐binding protein DsrC. Our work reveals NAD+ as suitable in vivo electron acceptor for sulphur oxidation in organisms operating the rDsr pathway and points to reduced nicotinamide adenine dinucleotides as electron donors for sulphite reduction in sulphate/sulphite‐reducing prokaryotes that contain DsrL. In addition, dsrL cannot be used as a marker distinguishing sulphate/sulphite reducers and sulphur oxidizers in metagenomic studies without further analysis.

  • 2‐Methylcitrate cycle: a well‐regulated controller of Bacillus sporulation
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-28
    Cao Zheng; Zhaoqing Yu; Cuiying Du; Yujing Gong; Wen Yin; Xinfeng Li; Zhou Li; Ute Römling; Shan‐Ho Chou; Jin He

    Bacillus thuringiensis is the most widely used eco‐friendly biopesticide, containing two primary determinants of biocontrol, endospore and insecticidal crystal proteins (ICPs). The 2‐methylcitrate cycle is a widespread carbon metabolic pathway playing a crucial role in channelling propionyl‐CoA, but with poorly understood metabolic regulatory mechanisms. Here, we dissect the transcriptional regulation of the 2‐methylcitrate cycle operon prpCDB and report its unprecedented role in controlling the sporulation process of B. thuringiensis. We found that the transcriptional activity of the prp operon encoding the three critical enzymes PrpC, PrpD, and PrpB in the 2‐methylcitrate cycle was negatively regulated by the two global transcription factors CcpA and AbrB, while positively regulated by the LysR family regulator CcpC, which jointly account for the fact that the 2‐methylcitrate cycle is specifically and highly active in the stationary phase of growth. We also found that the prpD mutant accumulated 2‐methylcitrate, the intermediate metabolite of the 2‐methylcitrate cycle, which delayed and inhibited sporulation at the early stage. Thus, our results not only revealed sophisticated transcriptional regulatory mechanisms for the metabolic 2‐methylcitrate cycle but also identified 2‐methylcitrate as a novel regulator of sporulation in B. thuringiensis.

  • Genomic mosaicism underlies the adaptation of marine Synechococcus ecotypes to distinct oceanic iron niches
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-27
    Nathan A. Ahlgren; Bernard Shafer Belisle; Michael D. Lee

    Phytoplankton are limited by iron (Fe) in ~40% of the world's oceans including high‐nutrient low‐chlorophyll (HNLC) regions. While low‐Fe adaptation has been well‐studied in large eukaryotic diatoms, less is known for small, prokaryotic marine picocyanobacteria. This study reveals key physiological and genomic differences underlying Fe adaptation in marine picocyanobacteria. HNLC ecotype CRD1 strains have greater physiological tolerance to low Fe congruent with their expanded repertoire of Fe transporter, storage and regulatory genes compared to other ecotypes. From metagenomic analysis, genes encoding ferritin, flavodoxin, Fe transporters and siderophore uptake genes were more abundant in low‐Fe waters, mirroring paradigms of low‐Fe adaptation in diatoms. Distinct Fe‐related gene repertories of HNLC ecotypes CRD1 and CRD2 also highlight how coexisting ecotypes have evolved independent approaches to life in low‐Fe habitats. Synechococcus and Prochlorococcus HNLC ecotypes likewise exhibit independent, genome‐wide reductions of predicted Fe‐requiring genes. HNLC ecotype CRD1 interestingly was most similar to coastal ecotype I in Fe physiology and Fe‐related gene content, suggesting populations from these different biomes experience similar Fe‐selective conditions. This work supports an improved perspective that phytoplankton are shaped by more nuanced Fe niches in the oceans than previously implied from mostly binary comparisons of low‐ versus high‐Fe habitats and populations.

  • Eco‐holobiont: A new concept to identify drivers of host‐associated microorganisms
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-26
    Brajesh K. Singh; Hongwei Liu; Pankaj Trivedi

    Host microbiomes play a critical role in host fitness and health. Whilst the current 'holobiont' concept framework has greatly expanded eco‐evolutionary and functional understanding of host‐microbiome interactions, the important role of biotic interactions and microbial loop (compositional linkage between soil, plant and animal) in shaping host‐microbiome are poorly understood. We proposed an 'eco‐holobiont' concept to fill the knowledge gap.

  • Spatial heterogeneity stabilizes predator–prey interactions at the microscale while patch connectivity controls their outcome
    Environ. Microbiol. (IF 5.147) Pub Date : 2019-12-26
    Margarita Petrenko; Shmuel P. Friedman; Ronen Fluss; Zohar Pasternak; Amit Huppert; Edouard Jurkevitch

    Natural landscapes are both fragmented and heterogeneous, affecting the distribution of organisms, and their interactions. While predation in homogeneous environments increases the probability of population extinction, fragmentation/heterogeneity promotes coexistence and enhances community stability as shown by experimentation with animals and microorganisms, and supported by theory. Patch connectivity can modulate such effects but how microbial predatory interactions are affected by water‐driven connectivity is unknown. In soil, patch habitability by microorganisms, and their connectivity depend upon the water saturation degree (SD). Here, using the obligate bacterial predator Bdellovibrio bacteriovorus, and a Burkholderia prey, we show that soil spatial heterogeneity profoundly affects predatory dynamics, enhancing long‐term co‐existence of predator and prey in a SD‐threshold dependent‐manner. However, as patches and connectors cannot be distinguished in these soil matrices, metapopulations cannot be invoked to explain the dynamics of increased persistence. Using a set of experiments combined with statistical and physical models we demonstrate and quantify how under full connectivity, predation is independent of water content but depends on soil microstructure characteristics. In contrast, the SD below which predation is largely impaired corresponds to a threshold below which the water network collapses and water connectivity breaks down, preventing the bacteria to move within the soil matrix.

Contents have been reproduced by permission of the publishers.