Nanako O. Ogawa

Geographical origin of polished rice based on multiple element and stable isotope analyses

We determined carbon and nitrogen contents (C and N contents) and stable carbon, nitrogen, and oxygen isotopic compositions (δ(13)C, δ(15)N, and δ(18)O) of polished rice in order to develop a simple method to discriminate its geographical origin. As a first attempt, we examined a single cultivar, Koshihikari rice, from 14 different cultivation areas including Australia (n=1), Japan (n=12), and USA (n=1). For all rice samples, C and N contents and the isotopic compositions are consistent with those of general plant materials, being 37.2-40.0% (C content), 0.8-1.4% (N content), -27.1 to -25.4% (δ(13)C), +0.4 to +9.0% (δ(15)N), and +18.8 to +22.9% (δ(18)O). However, its cultivated area is clearly distinguished by a pentagonal radar plot based on the elemental and isotopic compositions. Thus, the comparison of C and N contents and δ(13)C, δ(15)N, and δ(18)O values would potentially be useful for rapid and routine discrimination of geographical origin of the polished rice.

15N/14N ratios of amino acids as a tool for studying terrestrial food webs: a case study of terrestrial insects (bees, wasps, and hornets)

Compound-specific stable isotope analysis (CSIA) of amino acids is a new method that enables estimates of trophic position for consumers in food webs. We examined the nitrogen isotopic composition (δ15N) of amino acids of Japanese social insects (three bee, three wasp, and four hornet species) to evaluate the potential of CSIA of amino acids in studies of terrestrial food webs. For wasps, we also examined samples at different growth stages (ranging from egg to adult) to assess the effect of metamorphosis on CSIA estimates of trophic position. The δ15N values of bulk tissues for Japanese social insects are only weakly correlated with the biologically expected trophic positions. In contrast, the trophic positions estimated from the δ15N values of amino acids (yielding values of between 2.0 and 2.3 for bees, between 2.8 and 3.3 for wasps, and between 3.5 and 4.1 for hornets) are consistent with the biologically expected trophic positions for these insects (i.e., 2.0 for bees, 3.0 for wasps, and 3.0–4.0 for hornets). Although large variability is observed among the δ15N values of individual amino acids (e.g., ranging from 3.0 to 14.9‰ for phenylalanine), no significant change is observed in the trophic position during wasp metamorphosis. Thus, the CSIA of amino acids is a powerful tool for investigating not only aquatic food webs but also terrestrial food webs with predatory insects.

A low trophic position of Japanese eel larvae indicates feeding on marine snow.

What eel larvae feed on in the surface layer of the ocean has remained mysterious. Gut contents and bulk nitrogen stable isotope studies suggested that these unusual larvae, called leptocephali, feed at a low level in the oceanic food web, whereas other types of evidence have suggested that small zooplankton are eaten. In this study, we determined the nitrogen isotopic composition of amino acids of both natural larvae and laboratory-reared larvae of the Japanese eel to estimate the trophic position (TP) of leptocephali. We observed a mean TP of 2.4 for natural leptocephali, which is consistent with feeding on particulate organic matter (POM) such as marine snow and discarded appendicularian houses containing bacteria, protozoans and other biological materials. The nitrogen isotope enrichment values of the reared larvae confirm that the primary food source of natural larvae is consistent only with POM. This shows that leptocephali feed on readily available particulate material originating from various sources closely linked to ocean primary production and that leptocephali are a previously unrecognized part of oceanic POM cycling.

Evidence of Global Chlorophyll d

Although analyses of chlorophyll d (Chl d)‐dominated oxygenic photosystems have been conducted since their discovery 12 years ago, Chl d distribution in the environment and quantitative importance for aquatic photosynthesis remain to be investigated. We analyzed the pigment compositions of surface sediments and detected Chl d and its derivatives from diverse aquatic environments. Our data show that the viable habitat for Chl d‐producing phototrophs extends across salinities of 0 to 50 practical salinity units and temperatures of 1� to 40�C, suggesting that Chl d production can be ubiquitously observed in aquatic environments that receive near-infrared light. The relative abundances of Chl d derivatives over that of Chl a derivatives in the studied samples are up to 4%, further suggesting that Chl d‐based photosynthesis plays a quantitatively important role in the aquatic photosynthesis.

Biochemical and physiological bases for the use of carbon and nitrogen isotopes in environmental and ecological studies

We review the biochemical and physiological bases of the use of carbon and nitrogen isotopic compositions as an approach for environmental and ecological studies. Biochemical processes commonly observed in the biosphere, including the decarboxylation and deamination of amino acids, are the key factors in this isotopic approach. The principles drawn from the isotopic distributions disentangle the complex dynamics of the biosphere and allow the interactions between the geosphere and biosphere to be analyzed in detail. We also summarize two recently examined topics with new datasets: the isotopic compositions of individual biosynthetic products (chlorophylls and amino acids) and those of animal organs for further pursuing the basis of the methodology. As a tool for investigating complex systems, compound-specific isotopic analysis compensates the intrinsic disadvantages of bulk isotopic signatures. Chlorophylls provide information about the particular processes of various photoautotrophs, whereas amino acids provide a precise measure of the trophic positions of heterotrophs. The isotopic distributions of carbon and nitrogen in a single organism as well as in the whole biosphere are strongly regulated, so that their major components such as amino acids are coordinated appropriately rather than controlled separately.

New organic reference materials for carbon- and nitrogen-stable isotope ratio measurements provided by Center for Ecological Research, Kyoto University, and Institute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology

We determined both carbon and nitrogen isotope ratios of ten organic reference materials (CERKU-01 to CERKU-10) in the Center for Ecological Research (CER), Kyoto University, and three organic reference materials (BG-A, BG-P, and BG-T) in the Institute of Biogeosciences (BioGeos), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), using an internationally recommended calibration method of two-point anchoring. The reference materials cover δ13CVPDB range of −34.92 to −9.45‰ and δ15NAir range of −5.22 to 22.71‰ and can be used to measure isotope ratios of naturally occurring substances.

Tracking aquatic change using chlorin‐specific carbon and nitrogen isotopes: The last glacial‐interglacial transition at Lake Suigetsu, Japan

Joint carbon and nitrogen isotope measurements were made from chlorins (chlorophyll a, phaeophytin a and pyrophaeophytin a) extracted from the last glacial-interglacial transition sediments of Lake Suigetsu, central Japan. These data highlight both the potential and difficulty of using chlorin-specific isotopes to track aquatic change from lake sediments. δ13C and δ15N of the three chlorins show coherent patterns with time, supporting the theory that phaeophytin a and pyrophaeophytin a are early diagenetic products of chlorophyll a and that despite this transition, their isotopic signatures remain intact. However, our data suggest that the isotopic composition of phaeophytin a and pyrophaeophytin a can be imprecise proxies for the isotope composition of chlorophyll a, possibly owing to the complex array of factors which affect the synthesis, transformation and sedimentation of these phaeopigments in nature. The total accumulation of organic matter in Lake Suigetsu appears to be controlled by the balance of allocthonous and authocthonous material as reflected by the C/N ratio. However, both bulk organic and chlorin-specific δ13C show similar changes, suggesting that the first order variability in bulk organic δ13C reflects aquatic change. By contrast, there is no similarity between chlorin and bulk δ15N, suggesting that interpretation of bulk δ15N in this setting is compromised by diagenetic alteration. The isotopic composition of chlorins are interpreted to reflect the response of aquatic primary productivity to post-glacial environmental change. However, further research into the synthesis and transformation of chlorins in the modern environment is required in order to facilitate a more rigorous approach to interpreting isotope ratios in chlorins extracted from sediments.

Compound-Specific Nitrogen Isotope Analysis of d-Alanine, l-Alanine, and Valine: Application of Diastereomer Separation to δ15N and Microbial Peptidoglycan Studies

We have developed an analytical method to determine the compound-specific nitrogen isotope compositions of individual amino acid enantiomers using gas chromatography/combustion/isotope ratio mass spectrometry. A novel derivatization of amino acid diastereomers by optically active (R)-(-)-2-butanol or (S)-(+)-2-butanol offers two advantages for nitrogen isotope analysis. First, chromatographic chiral separation can be achieved without the use of chiral stationary-phase columns. Second, the elution order of these compounds on the chromatogram can be switched by a designated esterification reaction. We applied the method to the compound-specific nitrogen isotope analysis of D- and L-alanine in a peptidoglycan derived from the cell walls of cultured bacteria (Firmicutes and Actinobacteria; Enterococcus faecalis, Staphylococcus aureus, Staphylococcus staphylolyticus, Lactobacillus acidophilus, Bacillus subtilis, Micrococcus luteus, and Streptomyces sp.), natural whole bacterial cells (Bacillus subtilis var. natto), (pseudo)-peptidoglycan from archaea (Methanobacterium sp.), and cell wall from eukaryota (Saccharomyces cerevisiae). We observed statistically significant differences in nitrogen isotopic compositions; e.g., delta15N ( per thousand vs air) in Staphylococcus staphylolyticus for d-alanine (19.2 +/- 0.5 per thousand, n = 4) and L-alanine (21.3 +/- 0.8 per thousand, n = 4) and in Bacillus subtilis for D-alanine (6.2 +/- 0.2 per thousand, n = 3) and L-alanine (8.2 +/- 0.4 per thousand, n = 3). These results suggest that enzymatic reaction pathways, including the alanine racemase reaction, produce a nitrogen isotopic difference in amino acid enantiomers, resulting in 15N-depleted D-alanine. This method is expected to facilitate compound-specific nitrogen isotope studies of amino acid stereoisomers.

Quantitative analysis of coenzyme F430 in environmental samples: a new diagnostic tool for methanogenesis and anaerobic methane oxidation.

Coenzyme F430 is a nickel hydrocorphinoid and is the prosthetic group of methyl-coenzyme M reductase that catalyzes the last step of the methanogenic reaction sequence and its reversed reaction for anaerobic methane oxidation by ANME. As such, function-specific compound analysis has the potential to reveal the microbial distribution and activity associated with methane production and consumption in natural environments and, in particular, in deep subsurface sediments where microbiological and geochemical techniques are restricted. Herein, we report the development of a technique for high-sensitivity analysis of F430 in environmental samples, including paddy soils, marine sediments, microbial mats, and an anaerobic groundwater. The lower detection limit of F430 analysis by liquid chromatography/mass spectrometry is 0.1 femto mol, which corresponds to 6 × 10(2) to 1 × 10(4) cells of methanogens. F430 concentrations in these natural environmental samples range from 63 × 10(-6) to 44 nmol g(-1) and are consistent with the methanogenic archaeal biomass estimated by microbiological analyses.

Implications for chloro- and pheopigment synthesis and preservation from combined compound-specific δ 13 C, δ 15 N, and Δ 14 C analysis

Chloropigments and their derivative pheopig- ments preserved in sediments can directly be linked to pho- tosynthesis. Their carbon and nitrogen stable isotopic com- positions 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 pro- cesses of the phytoplanktonic community. In this study we report results from combined compound-specific radiocar- bon and stable carbon and nitrogen isotope analysis to exam- ine the time-scales of synthesis and fate of chlorophyll-a and its degradation products pheophytin-a, pyropheophytin-a, and 13 2 ,17 3 -cyclopheophorbide-a-enol until burial in Black Sea core-top sediments. The pigments are mainly of ma- rine phytoplanktonic origin as implied by their stable isotopic compositions. Pigment 15 N values indicate nitrate as the major uptake substrate but 15 N-depletion towards the open marine setting indicates either contribution from N 2-fixation or direct uptake of ammonium from deeper waters. Ra- diocarbon concentrations translate into minimum and max- imum 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.