Biphytane and stable carbon isotopic values of sugars and glycerol from ODP Hole 204-1245D and 201-1229A

The membrane lipids diglycosyl-glycerol dibiphytanyl glycerol tetraethers (2G-GDGTs) in marine subsurface sediments are believed to originate from uncultivated benthic archaea, yet the production of 2G-GDGTs from subseafloor samples has not been demonstrated in vitro. In order to validate sedimentary biosynthesis of 2G-GDGTs, we performed a stable carbon isotope probing experiment on a subseafloor sample with six different 13C-labelled substrates (bicarbonate, methane, acetate, leucine, glucose and Spirulina platensis biomass). After 468 days of anoxic incubation, only glucose and S. platensis resulted in label uptake in lipid moieties of 2G-GDGTs, indicating incorporation of carbon from these organic substrates. The hydrophobic moieties of 2G-GDGTs showed minimal label incorporation, with up to 4 per mil 13C enrichment detected in crenarchaeol-derived tricyclic biphytane from the S. platensis-supplemented slurries. The 2G-GDGT-derived glucose or glycerol moieties also showed 13C incorporation (Dd13C = 18 - 38 per mil) in the incubations with glucose or S. platensis, consistent with a lipid salvage mechanism utilized by marine benthic archaea to produce new 2G-GDGTs. The production rates were nevertheless rather slow, even when labile organic matter was supplied. The 2G-GDGT turnover times of 1700 - 20 500 years were much longer than those estimated for subseafloor microbial communities, implying that sedimentary 2G-GDGTs as biomarkers of benthic archaea are cumulative records of past and present generations.

Results of an in situ stable isotope labeling study in intertidal sediment

We report the results of an in situ tracer experiment in an intertidal sediment, where bacterial carbon was tagged with stable carbon-isotope label, after the injection of 13C-glucose. The appearance of label in bacteria (based on label incorporation in bacteria-specific, phospholipid-derived fatty acids) and subsequent transfer to meiobenthos (group level) and macrobenthos (species level) was followed for 36 days. The label dynamics of benthic taxa were either fitted with a simple-isotope model or evaluated against enrichment in bacteria, to derive the importance of bacterially derived carbon for the meiobenthos and macrobenthos. Although selective uptake of bacteria was evident, as 2.4 times more bacterial carbon was grazed as expected from indiscriminate feeding, bacterial carbon accounted on average for only 0.08 and 0.11 of the carbon requirements of meiobenthic and macrobenthic taxa, respectively. Additionally, the contribution of bacterial carbon to total carbon requirements did not depend on the living/feeding depth in the sediment or organism size (evaluated over a size range of four orders of magnitude). The observed overall low contribution of bacterial carbon implies that most intertidal benthic fauna depend primarily on other carbon resources that may assert a stronger control on the structure of intertidal-sediment communities.

Results of an isotope tracer experiment with arctic deep-sea nematodes

A stable isotope (13C)-labeling experiment was performed to quantify the importance of bacterial carbon as a food source for an Arctic deep-sea nematode community. Bacterial functional groups were isotopically enriched with 13C-glucose, 13C-acetate, 13C- bicarbonate, and 13C-amino acids injected into sediments collected from 1280 m depth at 79uN, 6uE, west of Svalbard. Incorporation of the 13C label into bacterial phospholipid-derived fatty acids (PLFAs) and nematodes in the top 5 cm of the sediment was monitored over a 7-d period. The 13C dynamics of nematodes was fitted with a simple isotope turnover model to derive the importance of the different bacterial functional groups as carbon sources for the nematodes. The different substrates clearly labeled different bacterial groups as evidenced by differential labeling of the PLFA patterns. The deep-sea nematode community incorporated a very limited amount of the label, and the isotope turnover model showed that the dynamics of the isotope transfer could not be attributed to bacterivory. The low enrichment of nematodes suggests a limited passive uptake of injected 13C-labeled substrates. The lack of accumulation suggests that the injected 13C-labeled dissolved organic carbon compounds are not important as carbon sources for deep-sea nematodes. Since earlier studies with isotopically enriched algae also found limited uptake by nematodes, the food sources of deep-sea nematodes remain unclear.