Natural 15N abundance of epiphytes depends on the position within the forest canopy: source signals and isotope fractionation

The natural ¹⁵N abundance (δ¹⁵N) of epiphytes and its N sources were studied in the canopy of a lowland rainforest in Costa Rica. Vascular and non‐vascular epiphytes and canopy soils were collected from four canopy zones and analysed for N contents and δ¹⁵N signals. In addition, the N concentrations and δ¹⁵N signatures of bulk precipitation, throughfall and stemflow were measured during the wet and the dry season. The δ¹⁵N values of epiphyte leaves decreased significantly from the lower zones (means of ?3·9 and ?4·3‰) to the upper zones (means of ?5·4 and ?6·1‰) of the canopy. In contrast, δ¹⁵N signatures of canopy soils (average ?0·3‰) differed little between the zones. Bulk deposition was enriched in ¹⁵N (+4·3‰) compared to all other potential N sources and was higher than throughfall and stemflow (+0·5 to ?1·3‰). δ¹⁵N values of atmospheric deposition were inversely related to those of the epiphyte leaves, whereas N isotopic composition of canopy soils did not vary significantly. Consequently, it is concluded that the variations in foliar N isotope composition of epiphytes were not simply caused by utilization of isotopically different N sources, but by different ¹⁵N discrimination during N acquisition.     

Australian mulga ecosystems –13C and 15N natural abundances of biota components and their ecophysiological significance

Samples of recently produced shoot material collected in winter/spring from common plant species of mulga vegetation in eastern and Western Australia were assayed for ¹³C and ¹⁵N natural abundance. ¹³C analyses showed only three of the 88 test species to exhibit C₄ metabolism and only one of seven succulent species to be in CAM mode. Non-succulent winter ephemeral C₃ species showed significantly lower mean δ¹³C values (– 28·0‰) than corresponding C₃-type herbaceous perennials, woody shrubs or trees (– 26·9, – 25·7 and – 26·2‰, respectively), suggesting lower water stress and poorer water use efficiency in carbon acquisition by the former than latter groups of taxa. Corresponding values for δ¹⁵N of the above growth and life forms lay within the range 7·5–15·5‰. δ¹⁵N of soil NH₄⁺ (mean 19·6‰) at a soft mulga site in Western Australia was considerably higher than that of NO₃^– (4·3‰). Shoot dry matter of Acacia spp. exhibited mean δ¹⁵N values (9·10 ± 0·6‰) identical to those of 37 companion non-N₂-fixing woody shrubs and trees (9·06 ± 0·5‰). These data, with no evidence of nodulation, suggested little or no input of fixed N₂ by the legumes in question. However, two acacias and two papilionoid legumes from a dune of wind-blown, heavily leached sand bordering a lake in mulga in Western Australia recorded δ¹⁵N values in the range 2·0–3·0‰ versus 6·4–10·7‰ for associated non-N₂-fixing taxa. These differences in δ¹⁵N, and prolific nodulation of the legumes, indicated symbiotic inputs of fixed N in this unusual situation. δ¹⁵N signals of lichens, termites, ants and grasshoppers from mulga of Western Australia provided evidence of N₂ fixation in certain termite colonies and by a cyanobacteria-containing species of lichen. Data are discussed in relation to earlier evidence of nitrophily and water availability constraints on nitrate utilization by mulga vegetation.     

Pan-European δ13C values of air and organic matter from forest ecosystems

We present carbon stable isotope, δ¹³C, results from air and organic matter samples collected during 98 individual field campaigns across a network of Carboeuroflux forest sites in 2001 (14 sites) and 2002 (16 sites). Using these data, we tested the hypothesis that δ¹³C values derived from large‐scale atmospheric measurements and models, which are routinely used to partition carbon fluxes between land and ocean, and potentially between respiration and photosynthesis on land, are consistent with directly measured ecosystem‐scale δ¹³C values. In this framework, we also tested the potential of δ¹³C in canopy air and plant organic matter to record regional‐scale ecophysiological patterns. Our network estimates for the mean δ¹³C of ecosystem respired CO₂ and the related ‘discrimination’ of ecosystem respiration, δ_er and Δ_er, respectively, were ?25.6±1.9‰ and 17.8 ±2.0‰ in 2001 and ?26.6±1.5‰ and 19.0±1.6‰ in 2002. The results were in close agreement with δ¹³C values derived from regional‐scale atmospheric measurement programs for 2001, but less so in 2002, which had an unusual precipitation pattern. This suggests that regional‐scale atmospheric sampling programs generally capture ecosystem δ¹³C signals over Europe, but may be limited in capturing some of the interannual variations. In 2001, but less so in 2002, there were discernable longitudinal and seasonal trends in δ_er. From west to east, across the network, there was a general enrichment in ¹³C (～3‰ and ～1‰ for the 2 years, respectively) consistent with increasing Gorczynski continentality index for warmer and drier conditions. In 2001 only, seasonal ¹³C enrichment between July and September, followed by depletion in November (from about ?26.0‰ to ?24.5‰ to ?30.0‰), was also observed. In 2001, July and August δ_er values across the network were significantly related to average daytime vapor pressure deficit (VPD), relative humidity (RH), and, to a lesser degree, air temperature (T_a), but not significantly with monthly average precipitation (P_m). In contrast, in 2002 (a much wetter peak season), δ_er was significantly related with T_a, but not significantly with VPD and RH. The important role of plant physiological processes on δ_er in 2001 was emphasized by a relatively rapid turnover (between 1 and 6 days) of assimilated carbon inferred from time‐lag analyses of δ_er vs. meteorological parameters. However, this was not evident in 2002. These analyses also noted corresponding diurnal cycles of δ_er and meteorological parameters in 2001, indicating a rapid transmission of daytime meteorology, via physiological responses, to the δ_er signal during this season. Organic matter δ¹³C results showed progressive ¹³C enrichment from leaves, through stems and roots to soil organic matter, which may be explained by ¹³C fractionation during respiration. This enrichment was species dependent and was prominent in angiosperms but not in gymnosperms. δ¹³C values of organic matter of any of the plant components did not well represent short‐term δ_er values during the seasonal cycle, and could not be used to partition ecosystem respiration into autotrophic and heterotrophic components.     

Bicarbonate stimulates non‐structural carbohydrate pools of Camptotheca acuminata

The role of root‐derived dissolved inorganic carbon (DIC) has been emphasized lately, as it can provide an alternative source of carbon for photosynthesis. The fate of newly fixed DIC and its effect on non‐structural carbohydrate (NSC) pools has not been thoroughly elucidated to date. To this end, we used ¹³C (NaHCO₃) as a substrate tracer to investigate the incorporation of newly fixed bicarbonate into the plant organs and NSC compounds of Camptotheca acuminata seedlings for 24 and 72 h. NSC levels across the organs were all markedly increased within 24 h of labeling treatment and afterward only decreased in stems at 72 h. The variation range of NSC concentrations in roots was considerably smaller than in the stem and leaves. As time passed, the δ¹³C in NSC compounds was significantly affected by ¹³C labeling and was more positive in the roots than in the stem and leaves. Starch was more ¹³C‐enriched than was soluble carbohydrate, and the δ¹³C of root starch was as high as ?4.70‰. Bicarbonate incorporation into newly formed NSC compounds contributed up to 0.24% of the root starch within 72 h. These data provided strong evidence that bicarbonate not only acted as a C source that contributed slightly to the NSC pools but also stimulated the increase in NSC pools. The present study expands our understanding of the rapid change of NSC pools across the organs in response to bicarbonate.     

Short-term variation in the isotopic composition of organic matter allocated from the leaves to the stem of Pinus sylvestris: effects of photosynthetic and postphotosynthetic carbon isotope fractionation

We aimed to quantify the separate effects of photosynthetic and postphotosynthetic carbon isotope discrimination on δ¹³C of the fast‐turn‐over carbon pool (water soluble organic carbon and CO₂ emitted from heterotrophic tissues), including their diel variation, along the pathway of carbon transport from the foliage to the base of the stem. For that purpose, we determined δ¹³C in total and water‐soluble organic matter of the foliage plus δ¹³C and δ¹⁸O in phloem organic matter of twigs and at three heights along the stem of Pinus sylvestris over a nine‐day period, including four measurements per day. These data were related to meteorological and photosynthesis parameters and to the δ¹³C of stem‐emitted CO₂. In the canopy (foliage and twigs), the δ¹³C of soluble organic matter varied diurnally with amplitudes of up to 1.9‰. The greatest ¹³C enrichment was recorded during the night/early morning, indicating a strong influence of starch storage and remobilization on the carbon isotope signatures of sugars exported from the leaves. ¹³C enrichment of soluble organic matter from the leaves to the twig phloem and further on to the phloem of the stem was supposed to be a result of carbon isotope fractionation associated with metabolic processes in the source and sink tissues. CO₂ emitted from the stem was enriched by 2.3–5.2‰ compared with phloem organic matter. When day‐to‐day variation was addressed, water‐soluble leaf δ¹³C and twig phloem δ¹⁸O were strongly influenced by c_i/c_a and stomatal conductance (G_s), respectively. These results show that both photosynthetic and postphotosynthetic carbon isotope fractionation influence δ¹³C of organic matter over time, and over the length of the basipetal transport pathway. Clearly, these influences on the δ¹³C of respired CO₂ must be considered when using the latter for partitioning of ecosystem CO₂ fluxes or when the assessment of δ¹³C in organic matter is applied to estimate environmental effects in c_i/c_a.     

Beyond CO2-fixation by Rubisco – an interpretation of 13C/12C variations in tree rings from novel intra-seasonal studies on broad-leaf trees

Evidence is presented for a very specific, seasonally recurring tri‐phase carbon isotope pattern in tree rings of broad‐leaf deciduous tree species. It is derived from highly resolved intra‐annual measurements of ¹³C/¹²C ratios of wood and cellulose from tree rings of Fagus sylvatica, Populus nigra, Quercus petraea and Morus alba. Investigations on δ¹³C from buds and leaves of Fagus sylvatica revealed a similar tri‐phase δ¹³C pattern. At the very beginning of a growing season, the δ¹³C trend of tree rings and foliage shows a marked increase of up to 5‰. The maximum δ¹³C‐value of each vegetation period always occurs in young heterotrophic leaves shortly after bud burst and persistently in the early wood of each tree ring, when growth depends on carbon reserves. Thereafter, δ¹³C profiles represent the autotrophic stage of the leaves, which show different patterns of variation, by and large characterized by a decline. The minimum δ¹³C‐value always shows up in the late wood of each tree ring. At the very end of each tree ring δ¹³C‐values start rising again. This increase in δ¹³C marks the gradual switch‐over to storage‐dependent growth and can also be observed in senescent leaves. Seasonal changes of more than 4‰ were measured, whereas contiguous δ¹³C values rarely differed from each other by more than 0.3‰. This tri‐phase pattern cannot be explained by the common model of carbon isotope fractionation during photosynthesis. It appears to be primarily an indication of seasonal changes in down‐stream processes of the carbohydrate metabolism. Environmental influences on the carbon isotope fractionation during photosynthesis are presumably of secondary importance and expressed by certain peculiarities showing up during the autotrophic phase, i.e. the mid‐section of the seasonal δ¹³C pattern.     

Differences in δ13C and δ15N stable isotopes in the pearly razorfish Xyrichtys novacula related to the sex,location and spawning period

In the present study, Xyrichtys novacula (Labridae) were sampled at five locations around the islands of Ibiza and Formentera (western Mediterranean Sea). Isotopic signatures of δ¹³C, δ¹⁵N and the C:N ratio were analysed in relation to locality, sex and size differences. δ¹³C and δ¹⁵N partitioning was also studied in the reproductive spawning period. There were significant differences in the δ¹³C signature between localities for both sexes, but not for δ¹⁵N. Sex differences were also found with a mean ±s.e . value of ?17·38 ± 0·06‰δ¹³C and 8·36 ± 0·05‰δ¹⁵N for females and ?17·17 ± 0·07‰δ¹³C and 8·80 ± 0·06‰δ¹⁵N for males. Increasing total length in both sexes was positively correlated with δ¹⁵N enrichment and a significant positive linear regression was established for both variables. During the reproductive spawning period, there were changes in δ¹³C fractioning with enrichment in postspawning females and males (with respect to prespawning and spawning periods) and δ¹⁵N impoverishment in postspawning females (with respect to prespawning and spawning periods). Xyrichtys novacula uses local food sources, as confirmed by δ¹³C and δ¹⁵N, and females and males use different food sources, thus avoiding intraspecific competition. This was confirmed by δ¹⁵N enrichment as size increased. Spawning leads to special requirements for gonad maturation, which is reflected in the isotopic signatures for both sexes.     

Temporal variability in stable isotope ratios of C and N in the vibrissa of captive and wild adult South American sea lions Otaria byronia: More than just diet shifts

We analyzed the δ¹³C and δ¹⁵N values in the vibrissae of captive adult breeding South American sea lions (Otaria byronia) fed at a constant diet and then used this information to analyze the change in stable isotope values along the vibrissae from wild individuals. The overall diet‐to‐vibrissa discrimination factor of the captive animals was 3.0‰ ± 0.1‰ for δ¹³C and 3.6‰ ± 0.1‰ for δ¹⁵N, but the stable isotope ratios fluctuated periodically despite constant diet. The δ¹³C and δ¹⁵N values of the captive male declined at the end of the breeding season, whereas the δ¹³C values of the female increased during the central part of pregnancy and the δ¹⁵N values peaked during lactation. The δ¹³C and δ¹⁵N values of adult wild specimens also fluctuated periodically and vibrissae growth rate (0.15 mm/d in both sexes) was slightly lower than in captivity (0.17 mm/d), assuming an annual periodicity for oscillations. Similarities in the amplitude of the cycles of captive and wild males suggested that fasting was probably the main source of periodic variability in the δ¹⁵N of wild males, whereas pregnancy and lactation were probably the main source of periodic variability for the δ¹³C of wild females.     

Stable‐isotope comparisons between embryos and mothers of a placentatrophic shark species

Stable nitrogen (δ¹⁵N) and carbon (δ¹³C) isotopes of Atlantic sharpnose shark Rhizoprionodon terraenovae embryos and mothers were analysed. Embryos were generally enriched in ¹⁵N in all studied tissue relative to their mothers' tissue, with mean differences between mother and embryo δ¹⁵N (i.e. Δδ¹⁵N) being 1·4‰ for muscle, 1·7‰ for liver and 1·1‰ for cartilage. Embryo muscle and liver were enriched in ¹³C (both Δδ¹³C means = 1·5‰) and embryo cartilage was depleted (Δδ¹³C mean = ?1·01‰) relative to corresponding maternal tissues. While differences in δ¹⁵N and δ¹³C between mothers and their embryos were significant, muscle δ¹⁵N values indicated embryos to be within the range of values expected if they occupied a similar trophic position as their respective mothers. Positive linear relationships existed between embryo total length (L_T) and Δδ¹⁵N for muscle and liver and embryo L_T and Δδ¹³C for muscle, with those associations possibly resulting from physiological differences between smaller and larger embryos or differences associated with the known embryonic nutrition shift (yolk feeding to placental feeding) that occurs during the gestation of this placentatrophic species. Together these results suggest that at birth, the δ¹⁵N and δ¹³C values of R. terraenovae are likely higher than somewhat older neonates whose postpartum feeding habits have restructured their isotope profiles to reflect their postembryonic diet.     

Diurnal variation in the stable isotope composition of water and dry matter in fruiting Lupinus angustifolius under field conditions

In this paper, we present an integrated account of the diurnal variation in the stable isotopes of water (δD and δ¹⁸O) and dry matter (δ¹⁵N, δ¹³C, and δ¹⁸O) in the long‐distance transport fluids (xylem sap and phloem sap), leaves, pod walls, and seeds of Lupinus angustifolius under field conditions in Western Australia. The δD and δ¹⁸O of leaf water showed a pronounced diurnal variation, ranging from early morning minima near 0‰ for both δD and δ¹⁸O to early afternoon maxima of 62 and 23‰, respectively. Xylem sap water showed no diurnal variation in isotopic composition and had mean values of ?13·2 and ?2·3‰ for δD and δ¹⁸O. Phloem sap water collected from pod tips was intermediate in isotopic composition between xylem sap and leaf water and exhibited only a moderate diurnal fluctuation. Isotopic compositions of pod wall and seed water were intermediate between those of phloem and xylem sap water. A model of average leaf water enrichment in the steady state (Craig & Gordon, pp. 9–130 in Proceedings of a Conference on Stable Isotopes in Oceanographic Studies and Palaeotemperatures, Lischi and Figli, Pisa, Italy, 1965; Dongmann et al., Radiation and Environmental Biophysics 11, 41–52, 1974; Farquhar & Lloyd, pp. 47–70 in Stable Isotopes and Plant Carbon–Water Relations, Academic Press, San Diego, CA, USA, 1993) agreed closely with observed leaf water enrichment in the morning and early afternoon, but poorly during the night. A modified model taking into account non‐steady‐state effects (Farquhar and Cernusak, unpublished) gave better predictions of observed leaf water enrichments over a full diurnal cycle. The δ¹⁵N, δ¹³C, and δ¹⁸O of dry matter varied appreciably among components. Dry matter δ¹⁵N was highest in xylem sap and lowest in leaves, whereas dry matter δ¹³C was lowest in leaves and highest in phloem sap and seeds, and dry matter δ¹⁸O was lowest in leaves and highest in pod walls. Phloem sap, leaf, and fruit dry matter δ¹⁸O varied diurnally, as did phloem sap dry matter δ¹³C. These results demonstrate the importance of considering the non‐steady‐state when modelling biological fractionation of stable isotopes in the natural environment.     

δ13C of CO2 respired in the dark in relation to δ13C of leaf metabolites: comparison between Nicotiana sylvestris and Helianthus annuus under drought

The variations of δ¹³C in leaf metabolites (lipids, organic acids, starch and soluble sugars), leaf organic matter and CO₂ respired in the dark from leaves of Nicotiana sylvestris and Helianthus annuus were investigated during a progressive drought. Under well‐watered conditions, CO₂ respired in the dark was ¹³C‐enriched compared to sucrose by about 4‰ in N. sylvestris and by about 3‰ and 6‰ in two different sets of experiments in H. annuus plants. In a previous work on cotyledonary leaves of Phaseolus vulgaris, we observed a constant ¹³C‐enrichment by about 6‰ in respired CO₂ compared to sucrose, suggesting a constant fractionation during dark respiration, whatever the leaf age and relative water content. In contrast, the ¹³C‐enrichment in respired CO₂ increased in dehydrated N. sylvestris and decreased in dehydrated H. annuus in comparison with control plants. We conclude that (i) carbon isotope fractionation during dark respiration is a widespread phenomenon occurring in C₃ plants, but that (ii) this fractionation is not constant and varies among species and (iii) it also varies with environmental conditions (water deficit in the present work) but differently among species. We also conclude that (iv) a discrimination during dark respiration processes occurred, releasing CO₂ enriched in ¹³C compared to several major leaf reserves (carbohydrates, lipids and organic acids) and whole leaf organic matter.     

Dietary analysis of the herbivorous hemiramphid Hyporhamphus regularis ardelio: an isotopic approach

The stable isotope values for a range of size classes of Hyporhamphus regularis ardelio from Moreton Bay, south‐east Australia were determined. There was a positive linear relationship between δ¹³C and standard length (L_S)(δ¹³C = 0·034 L_S ? 16·23; r² = 0·78). δ¹³C ranged from ?8·48 to ?17·29‰ with the smallest size class (50 mm L_S) being on average 1·04‰ enriched with respect to that of zooplankton (Temora turbinata) and 7·97‰ depleted compared to Zostera capricorni. δ¹³C was positively correlated with L_S(P < 0·01)(more enriched with increasing L_S) with those fish of the largest size class (225 mm L_S) being 9·86 and 0·84‰ enriched than T. turbinata and Z. capricorni, respectively. There was no detectable trend in δ¹⁵N values with L_S(P > 0·01) with δ¹⁵N, ranging from 9·18 to 11·00‰. Fish of all size classes were on average 2·32 and 7·63‰ more enriched than zooplankton and seagrass, respectively. Carbon isotope data indicate that H. r. ardelio commence life as carnivores and change to a diet in which seagrass is the primary carbon source. The dependence on animal matter, however, is always present. Due to the low percentage of nitrogen in Z. capricorni(2·5%) compared to zooplankton (9·1%) it appears that nitrogen from zooplankton is necessary throughout their life history with the carbon requirements for these fish coming chiefly from Z. capricorni.     

Trophic size‐structure of sailfish Istiophorus platypterus in eastern Taiwan estimated by stable isotope analysis

To examine trophic dynamics over different size classes, an isotopic study of sailfish Istiophorus platypterus life‐history stages was carried out. Samples were collected from eastern Taiwan and the South China Sea during April 2009 and February 2012. A total of 263 samples (111–245 cm, lower jaw fork length, L_LJFL) were examined for changes in trophic structure in relation to L_LJFL by using stable isotope analysis of carbon (δ¹³C) and nitrogen (δ¹⁵N). The δ¹⁵N values for I. platypterus ranged from 7·51 to 14·19‰ (mean ± s.d . = 12·06 ± 1·16‰) and the δ¹³C values ranged from ?22·04 to ?15·48‰ (mean ± s.d . = ?17·62 ± 1·10‰). The δ¹⁵N values were positively dependent on L_LJFL (r² = 0·377), whereas δ¹³C were negatively dependent on L_LJFL (r² = 0·063). There were significantly different seasonal changes in nitrogen and carbon isotopic concentration, but no significant differences in concentrations between eastern Taiwan and the South China Sea were reported. The trophic level (T_L) of each L_LJFL class was correlated, starting from 2·84 T_L for size class I (L_LJFL < 140 cm) and reaching 5·03 T_L for size class VI (L_LJFL > 221 cm). The mean ± s.d . T_L was 4·43 ± 0·19 for all samples. The results reveal that I. platypterus occupies a wide range of trophic levels and different size classes occupy different trophic positions in the pelagic ecosystem.     

Genetic parameters and QTL analysis of δ13C and ring width in maritime pine

Classical quantitative genetics and quantitative trait dissection analysis (QTL) approaches were used in order to investigate the genetic determinism of wood cellulose carbon isotope composition (δ¹³C, a time integrated estimate of water use efficiency) and of diameter growth and their relationship on adult trees (15 years) of a forest tree species (maritime pine). A half diallel experimental set‐up was used to (1) estimate heritabilities for δ¹³C and ring width and (2) to decompose the phenotypic δ¹³C/growth correlation into its genetic and environmental components. Considerable variation was found for δ¹³C (range of over 3‰) and for ring width (range of over 5 mm) and significant heritabilities (narrow sense 0·17/0·19 for δ¹³C and ring width, respectively, 100% additivity). The significant phenotypic correlation between δ¹³C and ring width was not determined by the genetic component, but was attributable to environmental components. Using a genetic linkage map of a full‐sib family, four significant and four suggestive QTLs were detected for δ¹³C, the first for δ¹³C in a forest tree species, as far as known to the authors. Two significant and four suggestive QTLs were found for ring width. No co‐location of QTLs was found between δ¹³C and growth.     

Diffusive fractionation complicates isotopic partitioning of autotrophic and heterotrophic sources of soil respiration

Carbon isotope ratios (δ¹³C) of heterotrophic and rhizospheric sources of soil respiration under deciduous trees were evaluated over two growing seasons. Fluxes and δ¹³C of soil respiratory CO₂ on trenched and untrenched plots were calculated from closed chambers, profiles of soil CO₂ mole fraction and δ¹³C and continuous open chambers. δ¹³C of respired CO₂ and bulk carbon were measured from excised leaves and roots and sieved soil cores. Large diel variations (>5‰) in δ¹³C of soil respiration were observed when diel flux variability was large relative to average daily fluxes, independent of trenching. Soil gas transport modelling supported the conclusion that diel surface flux δ¹³C variation was driven by non‐steady state gas transport effects. Active roots were associated with high summertime soil respiration rates and around 1‰ enrichment in the daily average δ¹³C of the soil surface CO₂ flux. Seasonal δ¹³C variability of about 4‰ (most enriched in summer) was observed on all plots and attributed to the heterotrophic CO₂ source.     

Environmental and physiological controls on the carbon isotope composition of CO2 respired by leaves and roots of a C3 woody legume (Prosopis velutina) and a C4 perennial grass (Sporobolus wrightii)

Accurate estimates of the δ¹³C value of CO₂ respired from roots (δ¹³C_{R_root}) and leaves (δ¹³C_{R_leaf}) are important for tracing and understanding changes in C fluxes at the ecosystem scale. Yet the mechanisms underlying temporal variation in these isotopic signals are not fully resolved. We measured δ¹³C_{R_leaf}, δ¹³C_{R_root}, and the δ¹³C values and concentrations of glucose and sucrose in leaves and roots in the C₄ grass Sporobolus wrightii and the C₃ tree Prosopis velutina in a savanna ecosystem in southeastern Arizona, USA. Night‐time variation in δ¹³C_{R_leaf} of up to 4.6 ± 0.6‰ in S. wrightii and 3.0 ± 0.6‰ in P. velutina were correlated with shifts in leaf sucrose concentration, but not with changes in δ¹³C values of these respiratory substrates. Strong positive correlations between δ¹³C_{R_root} and root glucose δ¹³C values in P. velutina suggest large diel changes in δ¹³C_{R_root} (were up to 3.9‰) influenced by short‐term changes in δ¹³C of leaf‐derived phloem C. No diel variation in δ¹³C_{R_root} was observed in S. wrightii. Our findings show that short‐term changes in δ¹³C_{R_leaf} and δ¹³C_{R_root} were both related to substrate isotope composition and concentration. Changes in substrate limitation or demand for biosynthesis may largely control short‐term variation in the δ¹³C of respired CO₂ in these species.     

Seasonal dynamics in the stable carbon isotope composition δ¹³C from non-leafy branch, trunk and coarse root CO₂ efflux of adult deciduous (Fagus sylvatica) and evergreen (Picea abies) trees

Respiration is a substantial driver of carbon (C) flux in forest ecosystems and stable C isotopes provide an excellent tool for its investigation. We studied seasonal dynamics in δ¹³C of CO₂ efflux (δ¹³C_E) from non‐leafy branches, upper and lower trunks and coarse roots of adult trees, comparing deciduous Fagus sylvatica (European beech) with evergreen Picea abies (Norway spruce). In both species, we observed strong and similar seasonal dynamics in the δ¹³C_E of above‐ground plant components, whereas δ¹³C_E of coarse roots was rather stable. During summer, δ¹³C_E of trunks was about ?28.2‰ (Beech) and ?26.8‰ (Spruce). During winter dormancy, δ¹³C_E increased by 5.6–9.1‰. The observed dynamics are likely related to a switch from growth to starch accumulation during fall and remobilization of starch, low TCA cycle activity and accumulation of malate by PEPc during winter. The seasonal δ¹³C_E pattern of branches of Beech and upper trunks of Spruce was less variable, probably because these organs were additionally supplied by winter photosynthesis. In view of our results and pervious studies, we conclude that the pronounced increases in δ¹³C_E of trunks during the winter results from interrupted access to recent photosynthates.     

Feeding ecology of pelagic fish larvae and juveniles in slope waters of the Gulf of Mexico

Stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) were used to investigate feeding patterns of larval and early juvenile pelagic fishes in slope waters of the Gulf of Mexico. Contribution of organic matter supplied to fishes and trophic position within this pelagic food web was estimated in 2007 and 2008 by comparing dietary signatures of the two main producers in this ecosystem: phytoplankton [based on particulate organic matter (POM)] and Sargassum spp. Stable isotope ratios of POM and pelagic Sargassum spp. were significantly different from one another with δ¹³C values of POM depleted by 3–6‰ and δ¹⁵N values enriched by 2 relative to Sargassum spp. Stable isotope ratios were significantly different among the five pelagic fishes examined: blue marlin Makaira nigricans, dolphinfish Coryphaena hippurus, pompano dolphinfish Coryphaena equiselis, sailfish Istiophorus platypterus and swordfish Xiphias gladius. Mean δ¹³C values ranged almost 2 among fishes and were most depleted in I. platypterus. In addition, mean δ¹⁵N values ranged 4–5 with highest mean values found for both C. hippurus and C. equiselis and the lowest mean value for M. nigricans during both years. Increasing δ¹³C or δ¹⁵N with standard length suggested that shifts in trophic position and diet occurred during early life for several species examined. Results of a two‐source mixing model suggest approximately an equal contribution of organic matter by both sources (POM = 55%; pelagic Sargassum spp. = 45%) to the early life stages of pelagic fishes examined. Contribution of organic matter, however, varied among species, and sensitivity analyses indicated that organic source estimates changed from 2 to 13% for a δ¹³C fractionation change of ±0·25‰ or a δ¹⁵N fractionation change of ± 1·0‰ relative to original fractionation values.     

Identifying critical habitat of the endangered vaquita (Phocoena sinus) with regional δ13C and δ15N isoscapes of the Upper Gulf of California,Mexico

The vaquita (Phocoena sinus) is the world's most endangered cetacean and has experienced a 60% reduction in the size of its population in the past decade. Knowledge of its basic ecology is essential for developing successful management plans to protect and conserve this species. In this study, we identified vaquita foraging areas by creating an isoscape of the Upper Gulf of California (UGC) based on sediment and zooplankton carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope values. Our results confirm that this species is confined to the western region of the UGC, which is characterized by relatively high δ¹⁵N values (sediments: 10.2‰ ± 2.0‰, zooplankton: 15.8‰ ± 1.3‰), higher sea surface temperatures (～16°C–25°C), higher concentrations of silt in sediments, and the highest turbidity. In contrast, the eastern region of the UGC had relatively low sediment (7.7‰ ± 2.4‰) and zooplankton (14.6‰ ± 1.0‰) δ¹⁵N values, and the highest concentrations of sand in sediments. Our approach is an effective use of marine isoscapes over a small spatial scale (<200 km) to identify the environmental characteristics that define the critical habitat for an extremely endangered marine mammal.     

Using stable isotope ratios as a tracer of feeding adaptation in released Japanese flounder Paralichthys olivaceus

Release‐recapture experiments were conducted to examine temporal changes of the carbon and nitrogen stable isotope (δ¹³C and δ¹⁵N) ratios in the muscle tissue of artificially produced Japanese flounder Paralichthys olivaceus, juveniles. About 9000 juveniles (mean ± s .d . 43·3 ± 5·2 mm in standard length and 1·07 ± 0·37 g, n = 15) were released in each of three coastal areas: Chojagasaki, Arasaki and Jogashima with different geographical conditions, along Sagami Bay, Pacific coast of central Japan. Recapture efforts were made on 4, 11, 18, 40 and 55 days after the release. The stable isotope ratios, RNA:DNA ratio, stomach content mass (per body mass M_sc) and condition factor (K) of recaptured individuals were measured. The mean ± s .d . δ¹³C and δ¹⁵N values (n = 15) were ?18·3 ± 0·2‰ and 12·2 ± 0·2‰, respectively at the release. Wild Japanese flounder juveniles were captured only in Chojagasaki, and the δ¹³C and δ¹⁵N values (n = 6) were ?14·0 ± 0·4‰ and 13·2 ± 0·7‰, respectively; these values were considered to represent the wild diet. Nutritional conditions of the released and recaptured juveniles as determined by the RNA : DNA ratio, M_SC and K were indicated to be the best in Chojagasaki, in which the stable isotope ratios gradually shifted towards and reached the wild values within 40 days. This result along with stomach content analyses suggested that the released juveniles had acquired a wild feeding habit. In Arasaki and Jogashima, nutritional conditions of the recaptured juveniles were poorer, with no clear changes in the stable isotope ratios. Greatly varied stable isotope ratio values were observed in the juveniles recaptured in Chojagasaki 11 days after the release, ranging from the release levels to the wild levels. The extent of changes in the stable isotope ratios had a positive correlation to the RNA : DNA ratio and K of these juveniles (r = 0·87, n = 10 and r = 0·83, n = 18, respectively). The analyses of stable isotope ratios coupled with nutritional condition were considered to be an effective tool to examine post‐release feeding adaptation of Japanese flounder juveniles.