John F. Stolz

The role of microbes in accretion, lamination and early lithification of modern marine stromatolites.

For three billion years, before the Cambrian diversification of life, laminated carbonate build-ups called stromatolites were widespread in shallow marine seas. These ancient structures are generally thought to be microbial in origin and potentially preserve evidence of the Earths earliest biosphere. Despite their evolutionary significance, little is known about stromatolite formation, especially the relative roles of microbial and environmental factors in stromatolite accretion. Here we show that growth of modern marine stromatolites represents a dynamic balance between sedimentation and intermittent lithification of cyanobacterial mats. Periods of rapid sediment accretion, during which stromatolite surfaces are dominated by pioneer communities of gliding filamentous cyanobacteria, alternate with hiatal intervals. These discontinuities in sedimentation are characterized by development of surface films of exopolymer and subsequent heterotrophic bacterial decomposition, forming thin crusts of microcrystalline carbonate. During prolonged hiatal periods, climax communities develop, which include endolithic coccoid cyanobacteria. These coccoids modify the sediment, forming thicker lithified laminae. Preservation of lithified layers at depth creates millimetre-scale lamination. This simple model of modern marine stromatolite growth may be applicable to ancient stromatolites.

Bacillus arsenicoselenatis, sp. nov., and Bacillus selenitireducens, sp. nov.: Two haloalkaliphiles from Mono Lake, California that respire oxyanions of selenium and arsenic

Abstract Two gram-positive anaerobic bacteria (strains E1H and MLS10) were isolated from the anoxic muds of Mono Lake, California, an alkaline, hypersaline, arsenic-rich water body. Both grew by dissimilatory reduction of As(V) to As(III) with the concomitant oxidation of lactate to acetate plus CO2. Bacillus arsenicoselenatis (strain E1H) is a spore-forming rod that also grew by dissimilatory reduction of Se(VI) to Se(IV). Bacillus selenitireducens (strain MLS10) is a short, non-spore-forming rod that grew by dissimilatory reduction of Se(IV) to Se(0). When the two isolates were cocultured, a complete reduction of Se(VI) to Se(0) was achieved. Both isolates are alkaliphiles and had optimal specific growth rates in the pH range of 8.5–10. Strain E1H had a salinity optimum at 60 g l–1 NaCl, while strain MLS10 had optimal growth at lower salinities (24–60 g l–1 NaCl). Both strains have limited abilities to grow with electron donors and acceptors other than those given above. Strain MLS10 demonstrated weak growth as a microaerophile and was also capable of fermentative growth on glucose, while strain E1H is a strict anaerobe. Comparative 16S rRNA gene sequence analysis placed the two isolates with other Bacillus spp. in the low G+C gram-positive group of bacteria.

A Bacterium That Can Grow by Using Arsenic Instead of Phosphorus

Evidence is offered for arsenate replacing phosphate as a molecular building block in a Mono Lake, California, bacterium. Life is mostly composed of the elements carbon, hydrogen, nitrogen, oxygen, sulfur, and phosphorus. Although these six elements make up nucleic acids, proteins, and lipids and thus the bulk of living matter, it is theoretically possible that some other elements in the periodic table could serve the same functions. Here, we describe a bacterium, strain GFAJ-1 of the Halomonadaceae, isolated from Mono Lake, California, that is able to substitute arsenic for phosphorus to sustain its growth. Our data show evidence for arsenate in macromolecules that normally contain phosphate, most notably nucleic acids and proteins. Exchange of one of the major bio-elements may have profound evolutionary and geochemical importance.

Arsenic(III) fuels anoxygenic photosynthesis in hot spring biofilms from Mono Lake, California

Phylogenetic analysis indicates that microbial arsenic metabolism is ancient and probably extends back to the primordial Earth. In microbial biofilms growing on the rock surfaces of anoxic brine pools fed by hot springs containing arsenite and sulfide at high concentrations, we discovered light-dependent oxidation of arsenite [As(III)] to arsenate [As(V)] occurring under anoxic conditions. The communities were composed primarily of Ectothiorhodospira-like purple bacteria or Oscillatoria-like cyanobacteria. A pure culture of a photosynthetic bacterium grew as a photoautotroph when As(III) was used as the sole photosynthetic electron donor. The strain contained genes encoding a putative As(V) reductase but no detectable homologs of the As(III) oxidase genes of aerobic chemolithotrophs, suggesting a reverse functionality for the reductase. Production of As(V) by anoxygenic photosynthesis probably opened niches for primordial Earths first As(V)-respiring prokaryotes.

Dissimilatory arsenate reduction with sulfide as electron donor: experiments with mono lake water and Isolation of strain MLMS-1, a chemoautotrophic arsenate respirer.

ABSTRACT Anoxic bottom water from Mono Lake, California, can biologically reduce added arsenate without any addition of electron donors. Of the possible in situ inorganic electron donors present, only sulfide was sufficiently abundant to drive this reaction. We tested the ability of sulfide to serve as an electron donor for arsenate reduction in experiments with lake water. Reduction of arsenate to arsenite occurred simultaneously with the removal of sulfide. No loss of sulfide occurred in controls without arsenate or in sterilized samples containing both arsenate and sulfide. The rate of arsenate reduction in lake water was dependent on the amount of available arsenate. We enriched for a bacterium that could achieve growth with sulfide and arsenate in a defined, mineral medium and purified it by serial dilution. The isolate, strain MLMS-1, is a gram-negative, motile curved rod that grows by oxidizing sulfide to sulfate while reducing arsenate to arsenite. Chemoautotrophy was confirmed by the incorporation of H14CO3− into dark-incubated cells, but preliminary gene probing tests with primers for ribulose-1,5-biphosphate carboxylase/oxygenase did not yield PCR-amplified products. Alignment of 16S rRNA sequences indicated that strain MLMS-1 was in the δ-Proteobacteria, located near sulfate reducers like Desulfobulbus sp. (88 to 90% similarity) but more closely related (97%) to unidentified sequences amplified previously from Mono Lake. However, strain MLMS-1 does not grow with sulfate as its electron acceptor.

Sulfurospirillum barnesii sp. nov. and Sulfurospirillum arsenophilum sp. nov., new members of the Sulfurospirillum clade of the ε-Proteobacteria

Two strains of dissimilatory arsenate-reducing vibrio-shaped bacteria are assigned to the genus Sulfurospirillum. These two new species, Sulfurospirillum barnesii strain SES-3T and Sulfurospirillum arsenophilum strain MIT-13T, in addition to Sulfurospirillum sp. SM-5, two strains of Sulfurospirillum deleyianum, and Sulfurospirillum arcachonense, form a distinct clade within the epsilon subclass of the Proteobacteria based on 16S rRNA analysis.

The respiratory arsenate reductase from Bacillus selenitireducens strain MLS10

The respiratory arsenate reductase from the Gram-positive, haloalkaliphile, Bacillus selenitireducens strain MLS10 was purified and characterized. It is a membrane bound heterodimer (150 kDa) composed of two subunits ArrA (110 kDa) and ArrB (34 kDa), with an apparent K(m) for arsenate of 34 microM and V(max) of 2.5 micromol min(-1) mg(-1). Optimal activity occurred at pH 9.5 and 150 g l(-1) of NaCl. Metal analysis (inductively coupled plasma mass spectrometry) of the holoenzyme and sequence analysis of the catalytic subunit (ArrA; the gene for which was cloned and sequenced) indicate it is a member of the DMSO reductase family of molybdoproteins.

Evolution of Nitrate Reductase: Molecular and Structural Variations on a Common Function

The biological transformation of nitrogen oxyanions is widespread in nature and gives rise to a robust biogeochemical cycle. The first step in nitrate reduction is carried out by the enzyme nitrate reductase (NR). Although NR always catalyzes the same chemical reaction (conversion of nitrate into nitrite), its location in the cell, structure, and function are organism‐dependent. We use protein sequence data to determine phylogenetic relationships and to examine similarities in structure and function. Three distinct clades of NR are apparent: the eukaryotic assimilatory NR (Euk‐NR) clade, the membrane‐associated prokaryotic NR (Nar) clade, and a clade that includes both the periplasmic NR (Nap) and prokaryotic assimilatory NR (Nas). The high degree of sequence similarity and a phylogenetic distribution that follows taxonomic classification suggest a monophyletic origin for the Euk‐NR early on in the evolution of eukaryotic cells. In contrast, sequence conservation, phylogenetic analysis, and physiology suggest that both Nar and Nap were acquired by horizontal gene transfer. Nap and Nas share a lesser degree of similarity, with Nap a subclade of Nas. Nap from strict anaerobic bacteria such as Desulfovibrio desulfuricans is ancestral to facultative species and may provide an evolutionary link between Nap and Nas. We observed conserved binding sites for molybdenum and pterin cofactors in all four proteins. In pathways involving Euk‐NR, Nas, and Nar, for which ammonia is the end product, nitrite is reduced to ammonia by a siroheme nitrite reductase. Nap, however, is coupled to a pentaheme nitrite reductase. In denitrification, whether Nar or Nap is involved, nitrite is reduced to nitric oxide by either a cytochrome cd1 or a copper‐containing nitrite reductase. This complexity underscores the importance of nitrate reduction as a key biological process.

The Microbial Community in the Layered Sediments at Laguna Figueroa, Baja California, Mexico: Does it Have Precambrian Analogues?

Abstract In the hypersaline lagoon at Laguna Figueroa vertically stratified diverse communities of microorganisms thrive. The modern sediments of Baja California at Laguna Figueroa contain cyanobacterial communities and sedimentary structures produced by these blue greens that have already been studied by Horodyski and his colleagues. This paper provides an introduction to the complex microbial communities, primarily those that underlie the laminated Microcoleus mats. They are composed of anaerobic photosynthetic and heterotrophic bacteria. The following genera of cyanobacteria at least are components of these mat communities: Lyngbya, Microcoleus, Entophysalis, Phormidium, Pseudoanabaena, Anabaena and Schizothrix. Among the photosynthetic bacteria several species of Thiocapsa-like microbes formed major surface components of certain mats and scums; rhodospirilli, rhodopseudomonads, chromatis and others were seen. The following nonphotosynthetic bacteria were identified: Nocardia sp., three types of spirilli, two types of Spirochaeta sp., two types of Desulfovibria sp., a new strain of red Beneckea and four distinctive unidentified coccoid and filamentous bacteria. Reasons are given for believing several of the species are new to science and that the microbial diversity is far greater than the approximately twenty species reported here. Eukaryotes are extremely rare. Only one species of animal, a herpachtechoid copepod, was ever seen in the 8-km long microbial communities of the hypersaline basin. Dunaliella salina, a chlorophyte and Aspergillus sydowi, an ascomycetous fungus were the only eukaryotes that were observed to be regular components of mat communities. Ciliates, amoebae (including a chrysarchnion-like microbe) and diatom tests, mostly empty, were the only other eukaryotes observed. Attempts to enrich for eukaryotic microorganisms were not successful whereas attempts to enrich for bacteria, especially anaerobes led to such a profusion of forms that to continue detailed study of them was beyond our means. Unidentified small rods and cocci constituted the largest fraction of individuals in the subsurface community. The microbes isolated from mats are adapted for alternating dry and wet conditions as well as high concentrations of salt and low concentrations of oxygen.

Arsenic in the Evolution of Earth and Extraterrestrial Ecosystems

If you were asked to speculate about the form extra-terrestrial life on Mars might take, which geomicrobial phenomenon might you select as a model system, assuming that life on Mars would be ‘primitive’? Give your reasons. At the end of my senior year at Rensselaer Polytechnic Institute in 1968, I took Professor Ehrlichs final for his Geomicrobiology course. The above question beckoned to me like the Sirens to Odysseus, for if I answered, it would take so much time and thought that I would never get around to the exams other essay questions and consequently, would be “shipwrecked” by flunking the course. So, I passed it up. With this 41-year perspective in mind, this manuscript is now submitted to Professor Ehrlich for (belated) “extra-credit.” R.S. Oremland