Fungal communities from geothermal soils in Yellowstone National Park

Geothermal soils offer unique insight into the way extreme environmental factors shape communities of organisms. However, little is known about the fungi growing in these environments and in particular how localized steep abiotic gradients affect fungal diversity. We used metabarcoding to characterize soil fungi surrounding a hot spring-fed thermal creek with water up to ~85 C and pH ~10 in Yellowstone National Park. No soil variable we measured determined fungal community composition. However, soils with pH >8 had lower fungal richness and different fungal assemblages when compared to less extreme soils. Saprotrophic fungi community profile followed more closely overall community patterns while ectomycorrhizal fungi did not, highlighting potential differences in the factors that structure these different fungal trophic guilds. In addition, in vitro growth experiments in four target fungal species revealed a wide range of tolerances to pH levels but not to heat. Overall, our results documenting fungal communities within a few hundred meters suggest stronger statistical power and wider sampling are needed to untangle so many co-varying environmental factors affecting such diverse species communities.

Draft genome of a novel methanotrophic Methylobacter sp. from the volcanic soils of Pantelleria Island

The genus Methylobacter is considered an important and often dominant group of aerobic methane-oxidizing bacteria in many oxic ecosystems, where members of this genus contribute to the reduction of CH4 emissions. Metagenomic studies of the upper oxic layers of geothermal soils of the Favara Grande, Pantelleria, Italy, revealed the presence of various methane-oxidizing bacteria, and resulted in a near complete metagenome assembled genome (MAG) of an aerobic methanotroph, which was classified as a Methylobacter species. In this study, the Methylobacter sp. B2 MAG was used to investigate its metabolic potential and phylogenetic affiliation. The MAG has a size of 4,086,539 bp, consists of 134 contigs and 3955 genes were found, of which 3902 were protein coding genes. All genes for CH4 oxidation to CO2 were detected, including pmoCAB encoding particulate methane monooxygenase (pMMO) and xoxF encoding a methanol dehydrogenase. No gene encoding a formaldehyde dehydrogenase was present and the formaldehyde to formate conversion follows the tetrahydromethanopterin (H4MPT) pathway. “Ca. Methylobacter favarea” B2 uses the Ribulose-Mono-Phosphate (RuMP) pathway for carbon fixation. Analysis of the MAG indicates that Na+/H+ antiporters and the urease system might be important in the maintenance of pH homeostasis of this strain to cope with acidic conditions. So far, thermoacidophilic Methylobacter species have not been isolated, however this study indicates that members of the genus Methylobacter can be found in distinct ecosystems and their presence is not restricted to freshwater or marine sediments.

A Special Issue on Microorganisms from Extreme Environments in Memory of Luigi Michaud (1974–2014)

A special issue (SI) titled “Microbial Diversity in Extreme Environments: Implications for Ecological and Applicative Perspectives” has been launched with the aim of showcasing the diversity and biotechnological potential of extremophilic microorganisms. The issue includes 10 research papers and four reviews that mainly address prokaryotes inhabiting hyperarid, hypercold, hyperalkaline and hypersaline (or polyextreme) environments, spanning from deserts to meromictic and glacier lakes around the globe. Thermophilic prokaryotes from shallow hydrothermal vents and Antarctic geothermal soils are also treated. The ecology and biotechnological perspectives of eukaryotes are discussed in two review papers and one research paper. This special issue serves as a memorial to Dr. Luigi Michaud (1974–2014), who dramatically passed away in Antarctica during underwater sampling activities.

Methanotrophic activity and diversity of methanotrophs in volcanic geothermal soils at Pantelleria (Italy)

Volcanic and geothermal systems emit endogenous gases by widespread degassing from soils, including CH4, a greenhouse gas twenty-five times as potent as CO2. Recently, it has been demonstrated that volcanic or geothermal soils are not only a source of methane, but are also sites of methanotrophic activity. Methanotrophs are able to consume 10–40 Tg of CH4 a−1 and to trap more than 50% of the methane degassing through the soils. We report on methane microbial oxidation in the geothermally most active site of Pantelleria (Italy), Favara Grande, whose total methane emission was previously estimated at about 2.5 Mg a−1 (t a−1). Laboratory incubation experiments with three top-soil samples from Favara Grande indicated methane consumption values of up to 59.2 nmol g−1 soil d.w. h−1. One of the three sites, FAV2, where the highest oxidation rate was detected, was further analysed on a vertical soil profile, the maximum methane consumption was measured in the top-soil layer, and values greater than 6.23 nmol g−1 h−1 were still detected up to a depth of 13 cm. The highest consumption rate was measured at 37 °C, but a still detectable consumption at 80 °C (> 1.25 nmol g−1 h−1) was recorded. The soil total DNA extracted from the three samples was probed by Polymerase Chain Reaction (PCR) using standard proteobacterial primers and newly designed verrucomicrobial primers, targeting the unique methane monooxygenase gene pmoA; the presence of methanotrophs was detected at sites FAV2 and FAV3, but not at FAV1, where harsher chemical–physical conditions and negligible methane oxidation were detected. The pmoA gene libraries from the most active site (FAV2) pointed to a high diversity of gammaproteobacterial methanotrophs, distantly related to Methylocaldum-Metylococcus genera, and the presence of the newly discovered acido-thermophilic Verrucomicrobia methanotrophs. Alphaproteobacteria of the genus Methylocystis were isolated from enrichment cultures under a methane-containing atmosphere at 37 °C. The isolates grow at a pH range of 3.5 to 8 and temperatures of 18–45 °C, and consume 160 nmol of CH4 h−1 mL−1 of culture. Soils from Favara Grande showed the largest diversity of methanotrophic bacteria detected until now in a geothermal soil. While methanotrophic Verrucomicrobia are reported as dominating highly acidic geothermal sites, our results suggest that slightly acidic soils, in high-enthalpy geothermal systems, host a more diverse group of both culturable and uncultivated methanotrophs.

Methanotrophic activity and bacterial diversity in volcanic-geothermal soils at Pantelleria island (Italy)

Volcanic and geothermal systems emit endogenous gases by widespread degassing from soils, including CH4, a greenhouse gas twenty-five times as potent as CO2. Recently, it has been demonstrated that volcanic/geothermal soils are source of methane, but also sites of methanotrophic activity. Methanotrophs are able to consume 10–40 Tg of CH4 a−1 and to trap more than 50% of the methane degassing through the soils. We report on methane microbial oxidation in the geothermally most active site of Pantelleria island (Italy), Favara Grande, whose total methane emission was previously estimated in about 2.5 t a−1. Laboratory incubation experiments with three top-soil samples from Favara Grande indicated methane consumption values up to 950 ng g−1 dry soil h−1. One of the three sites, FAV2, where the highest oxidation rate was detected, was further analysed on a vertical soil profile and the maximum methane consumption was measured in the top-soil layer but values > 100 ng g−1 h−1 were maintained up to a depth of 15 cm. The highest consumption rate was measured at 37 °C, but a still recognizable consumption at 80 °C (> 20 ng g−1 h−1) was recorded. In order to estimate the bacterial diversity, total soil DNA was extracted from Favara Grande and analysed using a Temporal Temperature Gradient gel Electrophoresis (TTGE) analysis of the amplified bacterial 16S rRNA gene. The three soil samples were probed by PCR using standard proteobacterial primers and newly designed verrucomicrobial primers targeting the unique methane monooxygenase gene pmoA; the presence of methanotrophs was detected in sites FAV2 and FAV3, but not in FAV1, where harsher chemical-physical conditions and negligible methane oxidation were detected. The pmoA gene libraries from the most active site FAV2 pointed out a high diversity of gammaproteobacterial methanotrophs distantly related to Methylococcus/Methylothermus genera and the presence of the newly discovered acido-thermophilic methanotrophs Verrucomicrobia. Alphaproteobacteria of the genus Methylocystis were isolated from enrichment cultures, under a methane containing atmosphere at 37 °C. The isolates grow at pH 3.5–8 and temperatures of 18–45 °C, and show a methane oxidation rate of ~ 450 μmol mol−1 h−1. Soils from Favara Grande showed the largest diversity of methanotrophic bacteria until now detected in a geothermal soil. While methanotrophic Verrucomicrobia are reported to dominate highly acidic geothermal sites, our results suggest that slightly acidic soils, in high enthalpy geothermal systems, host a more diverse group of both culturable and uncultivated methanotrophs.

Thermogemmatispora onikobensis gen. nov., sp. nov. and Thermogemmatispora foliorum sp. nov., isolated from fallen leaves on geothermal soils, and description of Thermogemmatisporaceae fam. nov. and Thermogemmatisporales ord. nov. within the class Ktedonobacteria

Two thermophilic, Gram-stain-positive, sporulating bacterial strains, which formed branched vegetative and aerial mycelia, were isolated from fallen leaves sampled from geothermal soils and designated ONI-1T and ONI-5T. Strain ONI-1T grew at 50–74 °C, with optimum growth at 60–65 °C, and strain ONI-5T grew at 45–74 °C, with optimum growth at 60–65 °C. The pH range for growth of the strains was pH 4.6–8.0, with optimum growth at pH 7.0. The DNA G+C contents of strains ONI-1T and ONI-5T were 60.2 and 58.1 mol%, respectively. The major fatty acid was iso-C17 : 0 and the major menaquinone was MK-9(H2). The cell walls of the strains contained glutamic acid, serine, glycine, histidine, alanine and ornithine. The polar lipids consisted of phosphatidylinositol, phosphatidylglycerol and a glycolipid. The cell-wall sugar was rhamnose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that the strains belong to the class Ktedonobacteria and that strains ONI-1T and ONI-5T are most closely related to Thermosporothrix hazakensis SK20-1T (85.3 and 84.5 % sequence similarity, respectively). 16S rRNA gene sequence similarity between the two strains was 96.6 %. Based on the phenotypic features and phylogenetic position, we propose that strains ONI-1T and ONI-5T constitute a novel genus containing two novel species, for which we propose the names Thermogemmatispora onikobensis gen. nov., sp. nov. (the type species; type strain ONI-1T = JCM 16817T = KCTC 19768T) and Thermogemmatispora foliorum sp. nov. (type strain ONI-5T = JCM 16818T = KCTC 19767T), within the new family Thermogemmatisporaceae fam. nov. and order Thermogemmatisporales ord. nov.