Oxygenic photosynthesis is widely accepted as the most important bioenergetic process happening in Earth's surface environment. It is thought to have evolved within the cyanobacterial lineage, but it has been difficult to determine when it began. Evidence based on the occurrence and appearance of stromatolites and microfossils indicates that phototrophy occurred as long ago as 3,465 Myr although no definite physiological inferences can be made from these objects. Carbon isotopes and other geological phenomena provide clues but are also equivocal. Biomarkers are potentially useful because the three domains of extant life-Bacteria, Archaea and Eukarya-have signature membrane lipids with recalcitrant carbon skeletons. These lipids turn into hydrocarbons in sediments and can be found wherever the record is sufficiently well preserved. Here we show that 2-methyl-bacteriohopanepolyols occur in a high proportion of cultured cyanobacteria and cyanobacterial mats. Their 2-methylhopane hydrocarbon derivatives are abundant in organic-rich sediments as old as 2,500 Myr. These biomarkers may help constrain the age of the oldest cyanobacteria and the advent of oxygenic photosynthesis. They could also be used to quantify the ecological importance of cyanobacteria through geological time.

There is a close connection between modern-day biosynthesis of particular triterpenoid biomarkers and presence of molecular oxygen in the environment. Thus, the detection of steroid and triterpenoid hydrocarbons far back in Earth history has been used to infer the antiquity of oxygenic photosynthesis. This prompts the question: were these compounds produced similarly in the past? In this paper, we address this question with a review of the current state of knowledge surrounding the oxygen requirement for steroid biosynthesis and phylogenetic patterns in the distribution of steroid and triterpenoid biosynthetic pathways.The hopanoid and steroid biosynthetic pathways are very highly conserved within the bacterial and eukaryotic domains, respectively. Bacteriohopanepolyols are produced by a wide range of bacteria, and are methylated in significant abundance at the C2 position by oxygen-producing cyanobacteria. On the other hand, sterol biosynthesis is sparsely distributed in distantly related bacterial taxa and the pathways do not produce the wide range of products that characterize eukaryotes. In particular, evidence for sterol biosynthesis by cyanobacteria appears flawed. Our experiments show that cyanobacterial cultures are easily contaminated by sterol-producing rust fungi, which can be eliminated by treatment with cycloheximide affording sterol-free samples.Sterols are ubiquitous features of eukaryotic membranes, and it appears likely that the initial steps in sterol biosynthesis were present in their modern form in the last common ancestor of eukaryotes. Eleven molecules of O 2 are required by four enzymes to produce one molecule of cholesterol. Thermodynamic arguments, optimization of function and parsimony all indicate that an ancestral anaerobic pathway is highly unlikely.The known geological record of molecular fossils, especially steranes and triterpanes, is notable for the limited number of structural motifs that have been observed. With a few exceptions, the carbon skeletons are the same as those found in the lipids of extant organisms and no demonstrably extinct structures have been reported. Furthermore, their patterns of occurrence over billion year time-scales correlate strongly with environments of deposition. Accordingly, biomarkers are excellent indicators of environmental conditions even though the taxonomic affinities of all biomarkers cannot be precisely specified. Biomarkers are ultimately tied to biochemicals with very specific functional properties, and interpretations of the biomarker record will benefit from increased understanding of the biological roles of geologically durable molecules.

No abstract

The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C 20:1 and cy-C 21 fatty acids dominated all of the Aquificales, while the alkyl glycerol ethers were mainly C 18:0 . These Aquificales biomarkers were major constituents of the lipid extracts of two Octopus Spring samples, a biofilm associated with the siliceous vent walls, and the well-known pink streamer community (PSC). Both the biofilm and the PSC contained mono-and dialkyl glycerol ethers in which C 18 and C 20 alkyl groups were prevalent. Phospholipid fatty acids included both the Aquificales n-C 20:1 and cy-C 21 , plus a series of isobranched fatty acids (i-C 15:0 to i-C 21:0 ), indicating an additional bacterial component. Biomass and lipids from the PSC were depleted in 13 C relative to source water CO 2 by 10.9 and 17.2‰, respectively. The C 20-21 fatty acids of the PSC were less depleted than the iso-branched fatty acids, 18.4 and 22.6‰, respectively. The biomass of T. ruber grown on CO 2 was depleted in 13 C by only 3.3‰ relative to C source. In contrast, biomass was depleted by 19.7‰ when formate was the C source. Independent of carbon source, T. ruber lipids were heavier than biomass (؉1.3‰). The depletion in the C 20-21 fatty acids from the PSC indicates that Thermocrinis biomass must be similarly depleted and too light to be explained by growth on CO 2 . Accordingly, Thermocrinis in the PSC is likely to have utilized formate, presumably generated in the spring source region.Based on phylogenetic analysis of small-subunit rRNA sequences, hyperthermophilic organisms proliferate in the deepest branches of the Bacterial and Archaeal domains. The branch lengths of these hyperthermophilic lineages tend to be short, which further suggests that such organisms are the closest known extant descendants of the last common ancestor and retain many ancestral phenotypic properties (49). The recent discovery of filamentous microfossils preserved in a 3,235-million-year-old submarine volcanogenic deposit lends considerable weight to the theory that hydrothermal vent organisms have had a very long history on Earth (41). Hyperthermophilic microbes are also attracting astrobiological and biogeochemical interest because of their potential role in the formation of many kinds of mineral deposits and the generation of rock textures and mineral assemblages that may be diagnostic for extant or extinct life beyond Earth (5).A well-known example of a hyperthermophilic chemolithotrophic ecosystem is the pink filamentous streamers found at Octopus Spring in Yellowstone National Park (YNP), United States that were described by Brock in 1965 (3, 4). Similar streamer communities were first reported b...

No abstract

Symbiotic associations between microbes and invertebrates have resulted in some of the most unusual physiological and morphological adaptations that have evolved in the animal world. We document a new symbiosis between marine polychaetes of the genus Osedax and members of the bacterial group Oceanospirillales, known for heterotrophic degradation of complex organic compounds. These organisms were discovered living on the carcass of a grey whale at 2891 m depth in Monterey Canyon, off the coast of California. The mouthless and gutless worms are unique in their morphological specializations used to obtain nutrition from decomposing mammalian bones. Adult worms possess elaborate posterior root-like extensions that invade whale bone and contain bacteriocytes that house intracellular symbionts. Stable isotopes and fatty acid analyses suggest that these unusual endosymbionts are likely responsible for the nutrition of this locally abundant and reproductively prolific deep-sea worm.

No abstract

The molecular and isotopic compositions of lipid biomarkers from cultured filamentous cyanobacteria (Phormidium, also known as Leptolyngbya) have been used to investigate the community and trophic structure of photosynthetic mats from alkaline hot springs of the Lower Geyser Basin at Yellowstone National Park. We studied a shallow‐water coniform mat from Octopus Spring (OS) and a submerged, tufted mat from Fountain Paint Pots (FPP) and found that 2‐methylhopanepolyols and mid‐chain branched methylalkanes were diagnostic for cyanobacteria, whereas abundant wax esters were representative of the green non‐sulphur bacterial population. The biomarker composition of cultured Phormidium‐isolates varied, but was generally representative of the bulk mat composition. The carbon isotopic fractionation for biomass relative to dissolved inorganic carbon (DIC; ɛCO2) for cultures grown with 1% CO2 ranged from 21.4 to 26.1 and was attenuated by diffusion limitation associated with filament aggregation (i.e. cell clumping). Isotopic differences between biomass and lipid biomarkers, and between lipid classes, depended on the cyanobacterial strain, but was positively correlated with overall fractionation. Acetogenic lipids (alkanes and fatty acids) were generally more depleted than isoprenoids (phytol and hopanoids). The δ13CTOC for OS and FPP mats were somewhat heavier than for cultures (−16.9 and −23.6, respectively), which presumably reflects the lower availability of DIC in the natural environment. The isotopic dispersions among cyanobacterial biomarkers, biomass and DIC reflected those established for culture experiments. The 7‐methyl‐ and 7,11‐dimethylheptadecanes were from 9 to 11 depleted relative to the bulk organic carbon, whereas 2‐methylhopanols derived from the oxidation‐reduction of bacteriohopanepolyol were enriched relative to branched alkanes by approximately 5–7. These isotopic relationships survived with depth and indicated that the relatively heavy isotopic composition of the OS mat resulted from diffusion limitation. This study supports the suggestion that culture studies can establish valid isotopic relationships for interpretation of trophic structure in modern and ancient microbial ecosystems.