Search

Abstract We applied stable carbon and nitrogen isotopic analyses to understand the faunivory of the four sympatric wild Paradoxurinae civet species in Borneo, which share similar ecological characteristics. We also employed compound-specific nitrogen isotope analysis of amino acids to estimate these species’ trophic positions (TPs). The bulk stable isotope analysis revealed distinctly lower nitrogen isotope ratios in binturongs than in the other three species. This suggests that binturongs exhibit the lowest degree of faunivory among the four species. Binturongs had the lowest TP estimated from the nitrogen isotope ratios of amino acids (2.0–2.1), followed by small-toothed palm civets (2.4–2.5), masked palm civets (2.7), and common palm civets (2.9). These results suggest that there is little faunivory in binturong and variations in faunivory in the other species. Although the number of samples measured for the nitrogen isotope ratios of amino acids is small (n = 2 for each species), our results suggest that the varying degree of consumption of animal food sources, such as insects, is the key mechanism of niche partitioning in these four Paradoxurinae civet species in Borneo. Such subtle but essential differences in closely related sympatric species would maintain high biodiversity in tropical regions.

Abstract Returned samples from the carbonaceous asteroid (162173) Ryugu provide pristine information on the original aqueous alteration history of the Solar System. Secondary precipitates, such as carbonates and phyllosilicates, reveal elemental partitioning of the major component ions linked to the primordial brine composition of the asteroid. Here, we report on the elemental partitioning and Mg isotopic composition (25Mg/24Mg) of breunnerite [(Mg, Fe, Mn)CO3] from the Ryugu C0002 sample and the A0106 and C0107 aggregates by sequential leaching extraction of salts, exchangeable ions, carbonates, and silicates. Breunnerite was the sample most enriched in light Mg isotopes, and the 25Mg/24Mg value of the fluid had shifted lower by ~0.38‰ than the initial value (set to 0‰) before dolomite precipitation. As a simple model, the Mg2+ first precipitated in phyllosilicates, followed by dolomite precipitation, at which time ~76−87% of Mg2+ had been removed from the primordial brine. A minor amount of phyllosilicate precipitation continued after dolomite precipitation. The element composition profiles of the latest solution that interacted with the cation exchange pool of Ryugu were predominantly Na-rich. Na+ acts as a bulk electrolyte and contributes to the stabilization of the negative surface charge of phyllosilicates and organic matter on Ryugu.

Abstract This paper is an attempt to generalize the nitrogen isotope information of the molecules that make up the cell from our point of view. Nitrogen in the cell exists as 20 proteinaceous amino acids, nucleobases, hemes, chlorophylls, and others, and their composition is similar among organisms. Based on a physiologically simple autonomous system that maintains a balance between inputs and outputs, it is theoretically predicted that 15N is distributed to each cellular compound with a certain regularity, and thus a specific relationship in nitrogen isotopic ratios among compounds. Previous studies essentially confirm this. The nitrogen isotopic ratio of compound i constituting the cell can be generalized as δ 15N i = δ 15Nplant + Δ i (TP − 1) + γ i , where TP represents the trophic position, and Δ i and γ i are the trophic discrimination factor and the intracellular 15N distribution of compound i, respectively. Knowing reliable values of Δ i and γ i will help us to better understand nitrogen dynamics in the biosphere and advance our understanding of the picture of the Earth’s environment through biogeochemistry.

Abstract We report primordial aqueous alteration signatures in water-soluble organic molecules from the carbonaceous asteroid (162173) Ryugu by the Hayabusa2 spacecraft of JAXA. Newly identified low-molecular-weight hydroxy acids (HO-R-COOH) and dicarboxylic acids (HOOC-R-COOH), such as glycolic acid, lactic acid, glyceric acid, oxalic acid, and succinic acid, are predominant in samples from the two touchdown locations at Ryugu. The quantitative and qualitative profiles for the hydrophilic molecules between the two sampling locations shows similar trends within the order of ppb (parts per billion) to ppm (parts per million). A wide variety of structural isomers, including α- and β-hydroxy acids, are observed among the hydrophilic molecules. We also identify pyruvic acid and dihydroxy and tricarboxylic acids, which are biochemically important intermediates relevant to molecular evolution, such as the primordial TCA (tricarboxylic acid) cycle. Here, we find evidence that the asteroid Ryugu samples underwent substantial aqueous alteration, as revealed by the presence of malonic acid during keto–enol tautomerism in the dicarboxylic acid profile. The comprehensive data suggest the presence of a series for water-soluble organic molecules in the regolith of Ryugu and evidence of signatures in coevolutionary aqueous alteration between water and organics in this carbonaceous asteroid.

The chirality of amino acids in extraterrestrial materials may provide an insight into the origin of the essential l-enantiopure amino acids in the terrestrial biosphere. In 2020, the Hayabusa2 mission succeeded in bringing back surface materials from the C-type asteroid (162173) Ryugu to the Earth. Amino acids were one of the targeted organic molecules to be studied in the Ryugu samples. To analyze the various structural isomers of amino acids, which were expected to be present, from the limited amount of the returned samples, the development of a highly-sensitive and selective analytical method was necessary. In the present study, a three-dimensional high-performance liquid chromatography (3D-HPLC) system has been developed for the enantioselective determination of five proteinogenic and three non-proteinogenic amino acids in the Ryugu samples, in which amino acids in the sample were separated by reversed-phase, anion-exchange and enantioselective columns after the fluorescence derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiozole. The applicability of the analytical system to the extraterrestrial samples was evaluated by analyzing several types of carbonaceous meteorites before applying the system to the Ryugu samples. In the analysis of the Ryugu samples, all of the target amino acids were successfully determined quantitatively. Non-proteinogenic amino acids including 2-amino-n-butyric acid, isovaline and norvaline, rarely present in the terrestrial environment, were found as almost racemic mixtures with 47.1 to 55.2%l.

Abstract Samples from the carbonaceous asteroid (162173) Ryugu provide information on the chemical evolution of organic molecules in the early solar system. Here we show the element partitioning of the major component ions by sequential extractions of salts, carbonates, and phyllosilicate-bearing fractions to reveal primordial brine composition of the primitive asteroid. Sodium is the dominant electrolyte of the salt fraction extract. Anions and NH4 + are more abundant in the salt fraction than in the carbonate and phyllosilicate fractions, with molar concentrations in the order SO4 2− > Cl− > S2O3 2− > NO3 − > NH4 +. The salt fraction extracts contain anionic soluble sulfur-bearing species such as S n -polythionic acids (n

Abstract We performed in-situ analysis on a ~ 1 mm-sized grain A0080 returned by the Hayabusa2 spacecraft from near-Earth asteroid (162173) Ryugu to investigate the relationship of soluble organic matter (SOM) to minerals. Desorption electrospray ionization-high resolution mass spectrometry (DESI-HRMS) imaging mapped more than 200 CHN, CHO, CHO–Na (sodium adducted), and CHNO soluble organic compounds. A heterogeneous spatial distribution was observed for different compound classes of SOM as well as among alkylated homologues on the sample surface. The A0080 sample showed mineralogy more like an Ivuna-type (CI) carbonaceous chondrite than other meteorites. It contained two different lithologies, which are either rich (lithology 1) or poor (lithology 2) in magnetite, pyrrhotite, and dolomite. CHN compounds were more concentrated in lithology 1 than in lithology 2; on the other hand, CHO, CHO–Na, and CHNO compounds were distributed in both lithologies. Such different spatial distribution of SOM is likely the result of interaction of the SOM with minerals, during precipitation of the SOM via fluid activity, or could be due to difference in transportation efficiencies of SOMs in aqueous fluid. Organic-related ions measured by time-of-flight secondary ion mass spectrometry (ToF–SIMS) did not coincide with the spatial distribution revealed by DESI-HRMS imaging. This result may be because the different ionization mechanism between DESI and SIMS, or indicate that the ToF–SIMS data would be mainly derived from methanol-insoluble organic matter in A0080. In the Orgueil meteorite, such relationship between altered minerals and SOM distributions was not observed by DESI-HRMS analysis and field-emission scanning electron microscopy, which would result from differences of SOM formation processes and sequent alteration process on the parent bodies or even on the Earth. Alkylated homologues of CHN compounds were identified in A0080 by DESI-HRMS imaging as observed in the Murchison meteorite, but not from the Orgueil meteorite. These compounds with a large C number were enriched in Murchison fragments with abundant carbonate grains. In contrast, such relationship was not observed in A0080, implying different formation or growth mechanisms for the alkylated CHN compounds by interaction with fluid and minerals on the Murchison parent body and asteroid Ryugu. Graphical Abstract

Uracil was identified in the sample returned from the asteroid Ryugu. Having been provided to the early Earth as a component in such asteroidal materials, these molecules might have played a role for prebiotic chemical evolution on the early Earth

Abstract Osaka Bay adjacent to the Kyoto–Osaka–Kobe metropolitan area was affected by severe metal pollution during the twentieth century; yet little is known about the trace metal sources and pre-industrial human activities. We have determined the elemental concentrations and zinc stable isotope ratios (δ66Zn) in bulk sediments and the trace metal concentrations in chemical fractions of a 9-m-long sediment core from Osaka Bay. Our goals were (1) to reconstruct the historical trace metal contamination, and (2) to identify anthropogenic Zn sources and the solid phases of anthropogenic trace metals. The core provided a continuous environmental record of the last 2300 years based on radiocarbon dating of molluscan shells. Copper, Zn, and Pb showed an initial enrichment from the 1670s AD, which could be caused by human activities due to an increasing population. In agreement with previous findings, the trace metal concentrations slightly increased from the 1870s, strongly increased from the beginning of the twentieth century, and peaked around 1960 before environmental pollution control laws were enacted. Increasing trace metal concentrations in the acid-labile and reducible fractions obtained by the Community Bureau of Reference (BCR) sequential extraction procedure toward the surface indicate carbonates and Mn oxyhydroxides were the primary fractions for anthropogenic trace metals. The δ66Zn values (1) were constant until the 1940s, suggesting that the average δ66Zn of industrial sources was indistinguishable from that value of the natural background, (2) showed a slight decrease from the 1950s and remained constant until the present, and (3) fell in a binary mixing process between a lithogenic (~ + 0.27‰) and an anthropogenic endmember (~ + 0.17‰), the latter likely representing a mixture of various Zn sources such as road dust, tire wear, industrial effluents, and effluents from wastewater treatment plants. We conclude the combination of Zn stable isotopes together with chemical fractions obtained by the BCR method represents a promising approach to assess the trace metal sources and their potential mobility in sediment cores from anthropogenically affected coastal areas.

Abstract We present the first data describing molecular evidence of planktonic coenzyme factor 430 (hereafter referred to as F430), a key signature of methanogenic archaea, obtained from a water column sample in a freshwater lake in Japan. We report concentrations of native F430 ranging from 6.8–35 × 102 femto mol g-wet−1 and 8.5–86 × 102 femto mol g-wet−1 in the water column and core-top sediment, respectively. Among these profiles, the epimer F430 of the deactivated chemical formula was very low overall in both planktonic and benthic environments. Small subunit (SSU) rRNA gene sequencing revealed that the planktonic microbial community was clearly different from that of the benthic sediment. Planktonic cyanobacteria (e.g., Microcystis) were predominant in the water column, whereas a wide variety of bacteria and archaea, including methanogens (e.g., Methanobacterium, Methanoregula, Methanothrix, Methanosarcina, and Methanocella), were observed in the core-top sediment. These results suggest that the planktonic cyanobacterial community may be rapidly degraded at the water–sediment interface driven by activated benthic microbial heterotrophic processes. Graphic abstract

Tracking migration of highly migratory marine fish using isotope analysis (iso-logging) has become a promising tool in recent years. However, application of this method is often hampered by the lack of essential information such as spatial variations in isotope ratios across habitats (isoscapes) and ontogenetic shifts of isotope ratios of target animals. Here, we test the utility of geostatistical analysis to generate isoscapes of δ13C and δ15N in the western Pacific and estimate the ontogenetic shifts in δ13C and δ15N values of a target species. We first measured δ13C and δ15N in the white muscle of juvenile (n = 210) and adult (n = 884) skipjack tuna Katsuwonus pelamis sampled across the northwest Pacific. Next we fitted a geostatistical model to account for the observed spatial variations in δ13C and δ15N of skipjack by fork length and other environmental variables with spatial random effects. We then used the best-fit models to predict the isoscapes of δ13C and δ15N in 2021. Furthermore, we measured δ15N of amino acids (δ15NAAs) of skipjack (n = 5) to determine whether the observed spatial variation of isotope ratios resulted from baseline shifts or differences in trophic position. The geostatistical model reasonably estimated both isoscapes and ontogenetic shifts from isotope ratios of skipjack, and the isoscapes showed that δ13C and δ15N can clearly distinguish the latitudinal migration of skipjack in the western Pacific. The δ15NAAs supported the results of the geostatistical model, that is, observed variations in skipjack δ15N were largely derived from a baseline shift rather than regional differences in trophic position. Thus, we showed that geostatistical analysis can provide essential basic information required for iso-logging without compound-specific isotope analysis.

Abstract In the high‐productivity subarctic ocean, phytoplankton growth is assumed to rely mainly on nitrate, based on snapshot observations conducted in spring or summer. However, the year‐round utilization of nitrate and ammonium by phytoplankton remains unexplored. Here, we show the seasonal transition of the dominant nitrogen source for phytoplankton by using the nitrogen isotopic composition (δ15N) of nitrate, ammonium, and sinking particles in the northwestern North Pacific. To decouple the combined effects of nitrogen sources for phytoplankton and subsequent trophic transfers in δ15N of sinking particles, we used compound‐specific δ15N analysis of amino acids (δ15NAA) in sinking particles. The seasonal change in δ15NAA revealed that most of the seasonal variation in δ15N of sinking particles arose from changes in the availability of nitrogenous nutrients in the euphotic layer. The δ15N of sinking particles suggested that the dominant nitrogen source for phytoplankton switches from nitrate to ammonium in autumn.

Abstract The southwestern Ryukyu Trench represents the ultimate sink of sediments shed from Taiwan into the Philippine Sea, which are mainly transported to the trench by turbidity currents via submarine canyons. Here, we present trench turbidites intercalated with hemipelagites in a gravity pilot core and a piston core acquired on the Ryukyu Trench floor at 6147 m water depth. We performed X-ray fluorescence core scans (ITRAX profiles), magnetic measurements, and organic geochemistry analyses to discriminate turbidites from hemipelagites. We identified 36 turbidites (0.9–4.2 cm thick) based on visual core descriptions and Ca/Fe ratios in the ITRAX profiles. Three of these turbidites show magnetic signatures indicating the presence of pyrrhotite and peaks in the magnetic susceptibility profile, suggesting that Taiwan-sourced sediments are transported to the Ryukyu Trench by long-runout turbidity currents. Pyrrhotite is also present in hemipelagites of the upper part of the retrieved cores, indicating a dominant sediment source in Taiwan over the last several thousand years. Ca/Fe and Zr/Rb ratios in the ITRAX profiles mark distal turbidites (about 1–3 cm thick), and Zr/Rb peaks mainly reflect grain size changes. Detailed analyses of a representative turbidite show good correlation between Ca/Fe and Zr/Rb peaks with upward-coarsening and upward-fining trends that delimit the turbidite. Sedimentary organic matter in hemipelagites is characterized by higher total organic carbon and total nitrogen contents and higher δ13C values than that in turbidites. Our multi-proxy approach employing high-resolution XRF core scans to differentiate turbidites from hemipelagites contributes to establishing a comprehensive view of modern trench sedimentation from Taiwan to the southwestern Ryukyu Trench.

Abstract Nitrogen isotope analysis of chloropigments provides information on the sources of nitrogenous nutrients assimilated by phytoplankton. The abundant, ubiquitous chlorophyll a records nitrogen isotopic compositions (δ15N) of eukaryotic phytoplankton and cyanobacteria, whereas more source‐specific chloropigments, such as the divinylchlorophylls exclusively possessed by the marine picocyanobacterium Prochlorococcus, can potentially resolve isotopic variability within the phytoplankton community. In this study, we analyzed the δ15N of both chlorophyll a and divinylchlorophyll a isolated from suspended particulate material collected at the subsurface chlorophyll maximum (SCM) along a meridional transect at 88°E in the oligotrophic eastern Indian Ocean. The nitrogen isotopic compositions of Prochlorococcus (δ15NPRO) and the combined biomass of eukaryotic phytoplankton and Synechococcus (δ15NEU+SYN) estimated from the δ15N of divinylchlorophyll a and chlorophyll a, respectively, revealed systematic variations that were not apparent from bulk isotope measurements. Whereas consistently low δ15NPRO indicated reliance of Prochlorococcus on regenerated nitrogen throughout the transect, elevation in δ15NEU+SYN values at several stations was interpreted to reflect assimilation of subsurface NO3− by eukaryotic phytoplankton. The δ15N distributions revealed subtle differences in NO3− availability at the SCM along the transect, which were consistently explained by the occurrence of mesoscale eddies in the Bay of Bengal, deepening of the mixed layer induced by a seasonal Wyrtki Jet in the equatorial region, and substantial deepening of the nutricline in the South Indian Ocean gyre. Our results highlight the utility of compound‐specific isotopic measurements of multiple species of chlorophylls in obtaining essential non‐incubation‐based biogeochemical constraints on the primary production in the ocean.

Abstract Stable nitrogen (15N/14N) and carbon (13C/12C) isotopic compositions of amino acids in organisms have widely been employed as a powerful tool to evaluate resource utilization and trophic connection among organisms in diverse ecosystems. However, little is known about the physiological factors or mechanisms responsible for determining the isotopic discrimination (particularly for carbon) within amino acids of organisms. In the present study, we investigated the inter-trophic discrimination of nitrogen and carbon isotopes within amino acids (Δδ 15NAA and Δδ 13CAA, respectively) using four consumer–diet pairs. Each pairing illustrates a metabolic perspective of isotopic fractionation of amino acids. The Δδ 15NAA values in these combinations reveal a trend consistent with those observed in many other combinations in previous studies. This further validates a standard scenario: the deamination preferentially removes 14N amino group from diet-derived amino acids, leaving behind the 15N-enriched amino acids in consumer biomass. The Δδ 15NAA values thus mirror the activity of amino acid deamination in consumers. In contrast, the trends in the Δδ 13CAA value suggest a different metabolic fate for the amino acid carbon isotope. Based on our results, we predict the following scenario: decarboxylation preferentially removes 12C α-carbon (i.e., carbonyl-carbon) from pyruvic acid in glycolysis, and from α-ketoglutaric acid in the tricarboxylic acid cycle, leaving behind the 13C-enriched both pyruvic and α-ketoglutaric acids. The 13C is then transferred to amino acids that are synthesized from the 13C-enriched precursor molecules within consumers. The Δδ 13CAA values therefore mirror the pathways of de novo amino acid synthesis in consumers. The proposed link between nitrogen and carbon isotopes can refine our knowledge of the potential processes affecting the isotopic fractionation within diet and consumer compartments, as well as environmental samples. Graphical abstract

Fish migration has always played an essential role in marine conservation and fisheries management. However, migration patterns are changing globally alongside changing ocean conditions. This affects the spatial scale of required governance and, consequently, our food supply. Technological advances in the bio-tracer approach using isotopes would make it possible to monitor fish migration differently from the conventional bio-logging method. Eye lenses of fish are incrementally grown and metabolically inert. Therefore, stable isotope analysis of eye lenses can reconstruct the geographic and trophic histories of fish. However, it is difficult to distinguish between the two pieces of information: migration and changing trophic relationships with the conventional bulk stable isotope approach. In this study, we measured the nitrogen isotope ratios of individual amino acids in eye lenses of chub mackerel (n = 3) from the western North Pacific to test whether the two pieces of information could be distinguished. A maximum of 34 growth layers was obtained from a single eye lens. For assigning the isotopic trends observed in lens tissues to specific life stages or size classes, we established a relationship between lens size and fork length (FL) as a reference framework. The reconstructed nitrogen isotope chorology of chub mackerel showed an increase in trophic position of about 1 unit from juvenile (FL ∼30 mm) to adult stages (FL ∼300 mm). The variation in nitrogen isotope ratios of phenylalanine (a source amino acid) in the eye lenses was –3 to 5‰. This baseline variability could be used for estimating fish movements. This study proposed compound-specific nitrogen isotope analysis of amino acids in eye lenses as a new tool capable of reconstructing the geographic and trophic histories of migratory fish.

Nitrogen (N) availability influences patterns of terrestrial productivity and global carbon cycling, imparting strong but poorly resolved feedbacks on Earth's climate system. Central questions concern the timescale of N cycle response to elevated CO2 concentration in the atmosphere and whether availability of this limiting nutrient increases or decreases with climate change. Nitrogen isotopic composition of bulk plant leaves provides information on large-scale patterns of N availability in the modern environment. Here we examine the utility of chlorins, degradation products of chlorophyll, hypothesized to persist in soil subsequent to plant decay, as proxies for reconstructing past plant δ15N. Specifically, we test the hypothesis that δ15N of plant leaves (δ15Nleaf) is recorded in δ15N of pheophytin a (δ15Npheo) along the leaf–litter–soil continuum across an array of ecosystem climate conditions and plant functional types (C3, C4, legumes, and woody plants). The δ15N of live foliage and bulk soil display marked declines with increasing rainfall, consistent with past studies in Hawaii and patterns worldwide. We find measurable chlorin concentrations along soil–depth profiles at all sites, with pheophytin a present in amounts required for isotopic analysis (>10 nmol). δ15Npheo in leaves, litter, and soil track δ15Nleaf of plant leaves. We find potential for δ15Npheo records from soil to provide proxy information on δ15Nleaf.

To study the influence of different ocean currents on the trophic spectra found in a sympatric fish community, we analyzed the radiocarbon contents (Δ14C) and stable carbon (δ13C) and nitrogen (δ15N) isotopic compositions in their bulk muscle tissues and the δ15N of individual amino acids in 26 species of demersal fish collected from off Tohoku (Pacific coast), northeastern Japan. The Δ14C values of the fish varied from –42 to +41‰, consistent with the Δ14C of the dissolved inorganic carbon in the cold Oyashio Current (typically about –50‰) and the warm Kuroshio Current (typically about +50‰). The trophic positions (TPs) of the fish estimated from the δ15N values of amino acids increased from 3.1 to 4.5 with increasing snout length. A negative correlation was observed between Δ14C and TP in gadiform fish and flatfish, suggesting that the Oyashio Current delivers a high TP diet to these fish groups. These results suggest that the trophic ecology of marine fish in the coastal western North Pacific is primarily controlled by the two major surface water currents, but is also significantly influenced by a combination of species-specific feeding and migration strategies.

Density stratification between freshwater and brine is periodically formed during massive evaporation events, which often associates deposition of organic-rich sediments. Here, we investigated phototrophic communities and nitrogen cycle during the deposition of two organic-rich shale beds of gypsum–shale alternation, representing the initial stage of the Messinian salinity crisis (Vena del Gesso, Northern Apennines, Italy). The structural distributions and the carbon and nitrogen isotopic compositions of geoporphyrins show a common pattern in the two shales, indicating the predominance of a particular phototrophic community under freshwater–brine stratified conditions. The ∼6‰ difference in δ13C of total organic carbon between PLG 4 and 5 shales was associated with similar shift in δ13C of the porphyrins derived from chlorophyll c, suggesting that the eukaryotic algae producing chlorophyll c were the major constituent of the phototrophic community. Importantly, these porphyrins show δ15N values (-7.6–-4.7‰) indicative of N2-fixation. We suggest that nitrate-depletion in the photic zone induced the predominance of diazotrophic cyanobacteria, which supplied new nitrogen for the chlorophyll c-producing eukaryotic algae. The large difference in the δ13C values of porphyrins and total organic carbon between PLG 4 and 5 shales are interpreted to reflect the depth of the chemocline, which fluctuates in response to changes in the regional evaporation–precipitation balance. Such variation in the chemocline depth may have dynamically changed the mode of the nitrogen cycle (i.e., nitrification–denitrification–N2-fixation coupling vs. phototrophic assimilation of ammonium) in the density-stratified marginal basins during the Messinian salinity crisis.

Periphytic algae attached to a streambed substrate (periphyton) are an important primary producer in stream ecosystems. We determined the isotopic composition of chlorophyll a in periphyton collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, a pure aquatic primary producer (Cladophora sp.) and a terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Epeorus latifolium). Periphyton Δ14Cchl values (−258 ‰ in April and −190 ‰ in October) were slightly lower than Δ14Cbulk values (−228 ‰ in April and −179 ‰ in October) but were close to the Δ14C value for dissolved inorganic carbon (DIC; −217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = −1000 ‰), CO2 derived from aquatic and terrestrial organic matters (variable Δ14C) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (−215 ‰ in April and −199 ‰ in October) and Cladophora sp. (−210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

Chloropigments and their derivative pheopigments preserved in sediments can directly be linked to photosynthesis. Their carbon and nitrogen stable isotopic compositions have been shown to be a good recorder of recent and past surface ocean environmental conditions tracing the carbon and nitrogen sources and dominant assimilation processes of the phytoplanktonic community. In this study we report results from combined compound-specific radiocarbon and stable carbon and nitrogen isotope analysis to examine the time-scales of synthesis and fate of chlorophyll-a and its degradation products pheophytin-a, pyropheophytin-a, and 132,173-cyclopheophorbide-a-enol until burial in Black Sea core-top sediments. The pigments are mainly of marine phytoplanktonic origin as implied by their stable isotopic compositions. Pigment δ15N values indicate nitrate as the major uptake substrate but 15N-depletion towards the open marine setting indicates either contribution from N2-fixation or direct uptake of ammonium from deeper waters. Radiocarbon concentrations translate into minimum and maximum pigment ages of approximately 40 to 1200 years. This implies that protective mechanisms against decomposition such as association with minerals, storage in deltaic anoxic environments, or eutrophication-induced hypoxia and light limitation are much more efficient than previously thought. Moreover, seasonal variations of nutrient source, growth period, and habitat and their associated isotopic variability are likely at least as strong as long-term trends. Combined triple isotope analysis of sedimentary chlorophyll and its primary derivatives is a powerful tool to delineate biogeochemical and diagenetic processes in the surface water and sediments, and to assess their precise time-scales.

We determined both the nitrogen and carbon isotopic compositions of various vanadyl alkylporphyrins isolated from siliceous marine sediments of the Onnagawa Formation (middle Miocene, northeastern Japan) to investigate the biogeochemistry and ecology of photoautotrophs living in the paleo-ocean. The distinctive isotopic signals support the interpretations of previous works that the origin of 17-nor-deoxophylloerythroetioporphyrin (DPEP) is chlorophylls-c1-3, whereas 8-nor-DPEP may have originated from chlorophylls-a2 or b2 or bacteriochlorophyll-a. Although DPEP and cycloheptanoDPEP are presumably derived from common precursory pigments, their isotopic compositions differed in the present study, suggesting that the latter represents a specific population within the photoautotrophic community. The average δ15N value for the entire photoautotrophic community is estimated to be –2 to +1‰ from the δ15N values of DPEP (–6.9 to –3.6‰; n=7), considering that the empirical isotopic relationships that the tetrapyrrole nuclei of chloropigments are depleted in 15N by ~4.8‰ and enriched in 13C by ~1.8‰ relative to the whole cells. This finding suggests that nitrogen utilized in the primary production was supplied mainly through N2-fixation by diazotrophic cyanobacteria. Based on the δ13C values of DPEP (–17.9 to –15.6‰; n=7), we estimated isotopic fractionation associated with photosynthetic carbon fixation to be 8–14‰. This range suggests the importance of β-carboxylation and/or active transport of the carbon substrate, indicating in turn the substantial contribution of diazotrophic cyanobacteria to primary production. Based on the δ15N values of 17-nor-DPEP (–7.4 to –2.4‰ n=7), the δ15N range of chlorophylls-c-producing algae was estimated to be –3 to +3‰. This relative depletion in sup>15N suggests that these algae mainly utilized nitrogen regenerated from diazotrophic cyanobacteria. Given that diatoms are likely to have constituted the chlorophylls-c-producing algae within the biogenic-silica-rich Onnagawa Formation, cyanobacteria-hosting diatoms may have been important contributors to primary production.

In Livello Bonarelli black shale deposited during Cretaceous Oceanic Anoxic Event 2 (OAE-2, ca. 94 Ma), nitrogen isotopic compositions of bulk sediments are mostly in a narrow range from –2.7 to –0.7‰. We also determined molecular distribution and nitrogen isotopic compositions of geoporphyrins extracted from the black shale. The nitrogen isotopic compositions of C32 Ni deoxophylloerythroetioporphyrin (DPEP) and total Ni porphyrins are –3.5 and –3.3‰, respectively, leading us to the estimation that the mean nitrogen isotopic composition of photoautotrophic cells were around +1‰ during the formation of Bonarelli black shale. This value is suggestive of N2-fixation, a dominant process for these photoautotrophs when assimilating nitrogen. Furthermore, Ni-chelated C32 DPEP, derived mainly from chlorophyll a had the highest concentration. Based on this evidence, we conclude that diazotrophic cyanobacteria were major primary producers during that time. Cyanobacteria may be key photoautotrophs during the formation of black shale type sediments intermittently observed throughout the later half of the Earth's history, and hence may have played a crucial role in the evolution of geochemical cycles even in the later half of the Earth's history.