Foliar δ13C response patterns along a moisture gradient arising from genetic variation and phenotypic plasticity in grassland species of Inner Mongolia

Plants depend upon both genetic differences and phenotypic plasticity to cope with environmental variation over different timescales. The spatial variation in foliar δ¹³C levels along a moisture gradient represents an overlay of genetic and plastic responses. We hypothesized that such a spatial variation would be more obvious than the variation arising purely from a plastic response to moisture change. Leymus chinensis and Stipa spp. were sampled from Inner Mongolia along a dry‐wet transect, and some of these species were transplanted to an area with a moisture gradient. For Stipa spp., the slope of foliar δ¹³C and mean annual precipitation along the transect was significantly steeper than that of foliar δ¹³C and mean annual precipitation after the watering treatment. For L. chinensis, there was a general decreasing trend in foliar δ¹³C under the different (increasing) watering levels; however, its populations showed an irregular relationship between foliar δ¹³C and moisture origin. Therefore, support for our hypothesis was obtained from Stipa spp., but not from L. chinensis.     

Effects of sampling method on foliar δ13C of Leymus chinensis at different scales

Stable carbon isotope composition (δ¹³C) usually shows a negative relationship with precipitation at a large scale. We hypothesized that sampling method affects foliar δ¹³C and its response pattern to precipitation. We selected 11 sites along a precipitation gradient in Inner Mongolia and collected leaves of Leymus chinensis with five or six replications repeatedly in each site from 2009 to 2011. Additionally, we collected leaves of L. chinensis separately from two types of grassland (grazed and fenced) in 2011. Foliar δ¹³C values of all samples were measured. We compared the patterns that foliar δ¹³C to precipitation among different years or different sample sizes, the differences of foliar δ¹³C between grazed and fenced grassland. Whether actual annual precipitation (AAP) or mean annual precipitation (MAP), it was strongly correlated with foliar δ¹³C every year. Significant difference was found between the slopes of foliar δ¹³C to AAP and MAP every year, among the slopes of foliar δ¹³C to AAP from 2009 to 2011. The more samples used at each site the lower and convergent P‐values of the linear regression test between foliar δ¹³C and precipitation. Furthermore, there was significant lower foliar δ¹³C value in presence of grazed type than fenced type grassland. These findings provide evidence that there is significant effect of sampling method to foliar δ¹³C and its response pattern to precipitation of L. chinensis. Our results have valuable implications in methodology for future field sampling studies.     

Carbon isotopes of C3 herbs correlate with temperature on removing the influence of precipitation across a temperature transect in the agro‐pastoral ecotone of northern China

Plant δ¹³C–temperature (δ‐T) relation has been established in many systems and is often used as paleotemperature transfer function. However, it is still unclear about the exact contributions of temperature variation to plant ¹³C discrimination because of covariation between temperature and precipitation (aridity), which reduces confidence in reconstruction of paleoclimate. In this study, we measured carbon isotope composition (δ¹³C) of 173 samples of C3 perennial herbs from 22 sites across a temperature gradient along the 400 mm isohyet in the farming‐pastoral zone of North China. The results showed that precipitation obviously affected the correlations of temperatures and foliar δ¹³C. After removing the influence of precipitation by analysis of covariance (ANCOVA), a more strongly positive relationship was obtained between site‐mean foliar δ¹³C and annual mean temperature (AMT), with a regression coefficient of 0.1636‰/°C (p = .0024). For widespread species, Artemisia lavandulaefolia and Artemisia capillaries, the slopes (or coefficients) of foliar δ¹³C and AMT were significantly steeper (larger) than those of foliar δ¹³C and AMT where the precipitation influence was not excluded, whereas the δ‐T coefficients of Polygonum persicaria and Leymus chinensis showed little change across the transect after deducting the precipitation effect. Moreover, the positive relationship between temperature and δ¹³C over the transect could be explained by soil moisture availability related to temperature. Our results may afford new opportunities for investigating the nature of past climate variability.     

Thyme travels: 15N isoscapes of Thymus vulgaris L. invasion in lightly grazed pastoral communities

Alterations to ecosystem nitrogen (N) cycling by introduced plant species may increase the invasibility of habitat providing a positive feedback for the introduced species to become invasive. Spatial patterns of foliar and soil δ¹⁵N ratios reflect variation in rates and process of N‐cycling across invaded landscapes and provide insight into N‐source uptake and utilization strategies of invasive plant species. To evaluate invasion‐associated changes in soil and foliar δ¹⁵N at different scales: regional (among different sites), local (between north‐ and south‐facing aspect at the same site), and microsite (within populations in the same community), we measured foliar and soil δ¹⁵ N, animal faeces cover (as a proxy for grazing intensity) and N₂‐fixing species cover from inside to outside Thymus vulgaris L. (thyme)‐invaded lightly grazed pastoral communities in Central Otago, southern South Island, New Zealand. Mean thyme foliar δ¹⁵N were near‐zero across the invaded landscape, and did not change across the advancing edge of invasion or with aspect. There was no evidence that associations with N₂‐fixing species provide a potential N source. Soil δ¹⁵N was lower inside of thyme compared to at the edge or outside of thyme and was varied between aspects at some sites. Animal faeces cover as a proxy for grazing intensity explained only 23% of this observed variation of soil δ¹⁵N. Thyme invasion may result in lowered soil δ¹⁵N reflecting alterations to N dynamics. Associated invasion‐related impacts of animal grazing may also impact soil δ¹⁵N. Further studies are required to distinguish the underlying mechanism responsible for the observed patterns of foliar and soil δ¹⁵N values across thyme‐invaded Central Otago landscapes.     

Carbon (δ13C) and nitrogen (δ15N) stable isotope composition in plant and soil in Southern Patagonia's native forests

Stable isotope natural abundance measurements integrate across several biogeochemical processes in ecosystem N and C dynamics. Here, we report trends in natural isotope abundance (δ¹³C and δ¹⁵N in plant and soil) along a climosequence of 33 Nothofagus forest stands located within Patagonia, Southern Argentina. We measured 28 different abiotic variables (both climatic variables and soil properties) to characterize environmental conditions at each of the 33 sites. Foliar δ¹³C values ranged from ?35.4‰ to ?27.7‰, and correlated positively with foliar δ¹⁵N values, ranging from ?3.7‰ to 5.2‰. Soil δ¹³C and δ¹⁵N values reflected the isotopic trends of the foliar tissues and ranged from ?29.8‰ to ?25.3‰, and ?4.8‰ to 6.4‰, respectively, with no significant differences between Nothofagus species (Nothofagus pumilio, Nothofagus antarctica, Nothofagus betuloides). Principal component analysis and multiple regressions suggested that mainly water availability variables (mean annual precipitation), but not soil properties, explained between 42% and 79% of the variations in foliar and soil δ¹³C and δ¹⁵N natural abundance, which declined with increased moisture supply. We conclude that a decline in water use efficiency at wetter sites promotes both the depletion of heavy C and N isotopes in soil and plant biomass. Soil δ¹³C values were higher than those of the plant tissues and this difference increased as annual precipitation increased. No such differences were apparent when δ¹⁵N values in soil and plant were compared, which indicates that climatic differences contributed more to the overall C balance than to the overall N balance in these forest ecosystems.     

Environmental and species‐specific controls on δ13C and δ15N in dominant woody plants from central‐western Argentinian drylands

Spatial variation in mean annual precipitation is the principal driver of plant water and nitrogen status in drylands. The natural abundance of carbon stable isotopes (δ¹³C) in photosynthetic tissues of C3 plants is an indicator of time‐integrated behaviour of stomatal conductance; while that of nitrogen stable isotopes (δ¹⁵N) is an indicator of the main source of plant N (soil N vs. atmospheric N₂). Previous studies in drylands have documented that plant δ¹³C and δ¹⁵N values increase with decreasing mean annual precipitation due to reductions in stomatal conductance, and soil enriched in ¹⁵N, respectively. However, evidence for this comes from studies focused on stable isotopes measurements integrated at the plant community level or on dominant plants at the site level, but little effort has been made to study C and N isotope variations within a species growing along rainfall gradients. We analysed plant δ¹³C, δ¹⁵N and C/N values of three woody species having different phenological leaf traits (deciduous, perennial and aphyllous) along a regional mean annual precipitation gradient from the central‐western Argentinian drylands. Noticeably, plant δ¹³C and δ¹⁵N values in the three woody species did not increase towards sites with low precipitation or at the start of the growing season (drier period), as we expected. These results suggest that environmental factors other than mean annual precipitation may be affecting plant δ¹³C and δ¹⁵N. The short‐term environmental conditions may interact with species‐specific plant traits related to water and nitrogen use strategies and override the predictive influence of the mean annual precipitation on plant δ¹³C and δ¹⁵N widely reported in drylands.     

Stable isotope analysis indicates trophic differences among forest floor carabids in Japan

Differences in trophic niches among carabid beetles (Coleoptera: Carabidae) co‐occurring on the forest floors of warm temperate forests in central Japan were studied using carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope analyses. Different carabid species showed similar δ¹⁵N values, which were higher than those of their possible invertebrate prey (herbivores and detritivores) collected from the litter layer, indicating that these species were consumers in the same trophic level. In contrast, δ¹³C values differed among carabid species, indicating interspecific differences in prey animals. The variation in the δ¹³C value was larger in summer than in autumn. In summer, δ¹³C values indicated that some carabids depended highly on either grazing (low δ¹³C values) or detrital sources (high δ¹³C values) within the food chain [Chlaenius posticalis Motschulsky and Haplochlaenius costiger (Chaudoir), respectively], although other species with intermediate δ¹³C values likely depended on both. The latter group of species comprised mostly two dominant genera (Carabus and Synuchus). Although congeners might have similar feeding habits, the stable isotope ratios indicated trophic niche differences between adults of different species and between adults and larvae of the same genus.     

Hair of grazing cattle provides an integrated measure of the effects of site conditions and interannual weather variability on δ13C of temperate humid grassland

The carbon isotope composition (δ¹³C) of C3 ecosystems is sensitive to water availability, and provides important information for the assessment of terrestrial carbon (C) sink/source activity. Here, we report the effects of plant available soil water (PAW) on community ¹³C signatures of temperate humid grassland. The 5‐year study was conducted on pastures exhibiting a large range of PAW capacity that were located on two site types: peat and mineral soils. The data set included the centennial drought year 2003, and data from wet years (2000 and 2002). Seasonal variation of PAW was modeled using PAW capacity of each pasture, precipitation inputs and evapotranspiration estimates. Community ¹³C signatures were derived from the δ¹³C of vegetation and segments of tail switch hair of cattle grown while grazing pastures. Hair ¹³C signatures provided an assimilation‐weighted ¹³C signal that integrated both spatial (paddock‐scale) and temporal (grazing season) variation of ¹³C signatures on a pasture. The δ¹³C of hair and vegetation increased with decreasing modeled PAW in the same way on mineral and peat soils. But, at a given PAW, the δ¹³C of hair was 2.6‰ less negative than that of vegetation, reflecting the diet‐hair isotopic shift. Furthermore, the δ¹³C of hair and vegetation on peat soil pastures was 0.5‰ more negative than on pastures situated on mineral soil. This may have resulted from a ～10 ppm CO₂ enrichment of canopy air derived from ongoing peat mineralization. Community‐scale season‐mean ¹³C discrimination (Δ) exhibited a saturation‐type response towards season‐mean modeled PAW (r²=0.78), and ranged between 19.8‰ on soils with low PAW capacity during the drought year of 2003, and 21.4‰ on soils with high PAW capacity in a wet year. This indicated relatively small variation in season‐mean assimilation‐weighted p_i/p_a (0.68–0.75) between contrasting sites and years. However, this range is similar to that reported in other studies, which encompass the range from subtropical arid to humid temperate grassland. Furthermore, the tight relationship between season‐mean Δ and modeled mean PAW suggests that PAW may be used as proxy for Δ.     

Site‐specific responses to short‐term environmental variation are reflected in leaf and phloem‐sap carbon isotopic abundance of field grown Eucalyptus globulus

The carbon isotopic composition (δ¹³C) of plant material has been used extensively as an indirect measure of carbon fixation per volume of water used. More recently, the δ¹³C of phloem sap (δ¹³C_phl) has been used as a surrogate measure of short‐term, canopy scale δ¹³C. Using a combination of δ¹³C physiological, structural and chemical indices from leaves and phloem sap of Eucalyptus globulus at sites of contrasting water availability, we sought to identify short‐term, canopy scale resource limitations. Results illustrate that δ¹³C_phl offers valid reflections of short‐term, canopy scale values of leaf δ¹³C and tree water status. Under conditions limited by water, leaf and phloem sap photoassimilates differ in ¹³C abundance of a magnitude large enough to significantly influence predictions of water use efficiency. This pattern was not detected among trees with adequate water supply indicating fractionation into heterotrophic tissues that may be sensitive to plant water status. Trees employed a range of physiological, biochemical and structural adaptations to acclimate to resource limitation that differed among sites providing a useful context upon which to interpret patterns in δ¹³C. Our results highlight that such easily characterized properties are ideal for use as minimally invasive tools to monitor growth and resilience of plants to variations in resource availability.     

Variation in δ13C among species and sexes in the family Restionaceae along a fine‐scale hydrological gradient

Consistent, repeatable segregation of plant species along hydrological gradients is an established phenomenon that must in some way reflect a trade‐off between plants' abilities to tolerate the opposing constraints of drought and waterlogging. In C₃ species tissue carbon isotope discrimination (δ¹³C) is known to vary sensitively in response to stomatal behaviour, reflecting stomatal limitation of photosynthesis during the period of active growth. However, this has not been studied at fine‐spatial scale in natural communities. We tested how δ¹³C varied between species and sexes of individuals in the family Restionaceae growing along a monitored hydrological gradient. Twenty Restionaceae species were investigated using species‐level phylogeny at two sites in the Cape Floristic Region, a biodiversity hotspot. A spatial overlap analysis showed the Restionaceae species segregated significantly (P < 0.001) at both sites. Moreover, there were significant differences in δ¹³C values among the Restionaceae species (P < 0.001) and between male and female individuals of each species (P < 0.01). However, after accounting for phylogeny, species δ¹³C values did not show any significant correlation with the hydrological gradient. We suggest that some other variable (e.g. plant phenology) could be responsible for masking a simple response to water availability.     

Stipa tenacissima Does not Affect the Foliar δ13C and δ15N of Introduced Shrub Seedlings in a Mediterranean Semi-arid Steppe

Recent studies have shown that the tussock grass Stipa tenacissima L. facilitates the establishment of late-successional shrubs, in what constitutes the first documented case of facilitation of woody plants by grasses. With the aim of increasing our knowledge of this interaction, in the present study we investigated the effects of S. tenacissima on the foliar δ13C, δ15N, nitrogen concentration, and carbon ： nitrogen ratio of introduced seedlings of Pistacia lentiscus L., Quercus coccifera L., and Medicago arborea L. in a semi-arid Mediterranean steppe. Six months after planting, the values of δ13C ranged between -26.9‰ and -29.6‰, whereas those of δ15N ranged between -1.9‰ and 2.7‰. The foliar C ： N ratio ranged between 10.7 and 53.5, and the nitrogen concentration ranged between 1.0% and 4.4%. We found no significant effect of the microsite provided by S. tenacissima on these variables in any of the species evaluated. The values of δ13C were negatively correlated with predawn water potentials in M. arborea and were positively correlated with relative growth rate in Q. coccifera. The values of δ15N were positively correlated with the biomass allocation to roots in the latter species. The present results suggest that the modification of environmental conditions in the are surrounding S. tenacissima was not strong enough to modify the foliar isotopic and nitrogen concentration of shrubs during the early stages after planting.     

δ 13C variation of C3 and C4 plants across an Asian monsoon rainfall gradient in arid northwestern China

We have investigated carbon isotopic compositions of four plant genus/species, Bothriochloa ischaemum (C₄), Stipa bungeana (C₃), Lespedeza sp. (C₃) and Heteropappus less (C₃), along a precipitation gradient in northwest China in order to assess the impact of water availability on the carbon isotopic discrimination against ¹³C during carbon assimilation in this area. This information is necessary for reconstruction of paleovegetation, particularly paleo‐C₃/C₄ plant ratios using δ¹³C value of organic matter in loess and paleosols in the Chinese Loess Plateau. The δ¹³C of C₃ plants, as a group, exhibits a negative correlation with the annual precipitation amount with a total change and sensitivity of 5‰ and ?1.1‰/100 mm, respectively, for the precipitation range from 200 to 700 mm. The C₄ grass, B. ischaemum responds to aridity by decreasing 1.7‰ for over the precipitation range from 350 to 700 mm; the plant δ¹³C is significantly correlated with annual precipitation with a slope ?0.61‰/100 mm. This result implies that without considering the effect of water availability on the plant δ¹³C values, reconstruction of percent C₄ vegetation during the last glaciation can be overestimated by about a factor of two.     

Increased snowfall weakens complementarity of summer water use by different plant functional groups

Winter snowfall is an important water source for plants during summer in semiarid regions. Snow, rain, soil water, and plant water were sampled for hydrogen and oxygen stable isotopes analyses under control and increased snowfall conditions in the temperate steppe of Inner Mongolia, China. Our study showed that the snowfall contribution to plant water uptake continued throughout the growing season and was detectable even in the late growing season. Snowfall versus rainfall accounted for 30% and 70%, respectively, of the water source for plants, on the basis of hydrogen stable isotope signature (δD) analysis, and accounted for 12% and 88%, respectively, on the basis of oxygen stable isotope signature (δ¹⁸O) analysis. Water use partitioning between topsoil and subsoil was found among species with different rooting depths. Increased snowfall weakened complementarity of plant water use during summer. Our study provides insights into the relationships between precipitation regimes and species interactions in semiarid regions.     

Carbon isotope composition of fossil charcoal reveals aridity changes in the NW Mediterranean Basin

Although several proxies for the inference of precipitation have been proposed, evidence of changes in aridity during the Holocene is scarce, and most is only qualitative. Moreover, precipitation regimes show relatively poor spatial correlations and can exhibit contrasting responses to global climate trends in different areas. Thus, there is a need to concentrate efforts at the local scale in order to increase the spatial resolution of palaeoclimate records, especially regarding water availability in semiarid zones. We propose the analysis of carbon isotope composition (δ¹³C) in fossil charcoal (routinely recovered from archaeological sites) to quantify changes in water availability in the past. We applied this approach to reconstruct variations in aridity during the last four millennia in the Ebro Depression (NE Iberian Peninsula). First, we studied the effect of carbonization over a range of temperatures (300–500°C) on the δ¹³C of Aleppo pine (Pinus halepensis Mill.) wood cores, collected from nine locations in NE Iberian Peninsula with distinct water availability. Despite significant changes in δ¹³C caused by carbonization, the original climatic signal of wood δ¹³C was well preserved. Moreover, δ¹³C shifts induced by this process were successfully corrected by accounting for variation in charcoal carbon concentration (%C). After removing the effect of carbonization, we estimated annual precipitation (P) and the ratio between annual precipitation and evapotranspiration (P/E) from the δ¹³C of fossil charcoal. In general, estimated water availability in the past was higher than present values, indicating that latter‐day (semiarid) conditions are mostly due to recent climate changes. The good agreement between our findings and other evidence indicates that the analysis of δ¹³C in charcoal may be useful to expand current palaeoclimate records as it provides a complementary (and quantitative) source of information to assess climate dynamics.     

Foliar and soil δ15N values reveal increased nitrogen partitioning among species in diverse grassland communities

Plant and soil nitrogen isotope ratios (δ¹⁵N) were studied in experimental grassland plots of varying species richness. We hypothesized that partitioning of different sources of soil nitrogen among four plant functional groups (legumes, grasses, small herbs, tall herbs) should increase with diversity. Four years after sowing, all soils were depleted in ¹⁵N in the top 5 cm whereas in non‐legume plots soils were enriched in ¹⁵N at 5–25 cm depth. Decreasing foliar δ¹⁵N and Δδ¹⁵N (= foliar δ¹⁵N ? soil δ¹⁵N) values in legumes indicated increasing symbiotic N₂ fixation with increasing diversity. In grasses, foliar Δδ¹⁵N also decreased with increasing diversity suggesting enhanced uptake of N depleted in ¹⁵N. Foliar Δδ¹⁵N values of small and tall herbs were unaffected by diversity. Foliar Δδ¹⁵N values of grasses were also reduced in plots containing legumes, indicating direct use of legume‐derived N depleted in ¹⁵N. Increased foliar N concentrations of tall and small herbs in plots containing legumes without reduced foliar δ¹⁵N indicated that these species obtained additional mineral soil N that was not consumed by legumes. These functional group and species specific shifts in the uptake of different N sources with increasing diversity indicate complementary resource use in diverse communities.     

Temporal variation in body composition (C : N) helps explain seasonal patterns of zooplankton δ13C

1. The stable carbon isotope ratio δ¹³C is a useful tracer of energy flow in lake food webs, and the zooplankton signature is commonly used to establish a baseline for the pelagic habitat. However, sources of temporal variability in the δ¹³C of different zooplankton taxa are rarely considered. 2. Here, we investigate to what extent temporal variation in the δ¹³C of particulate organic matter (POM) (<41 μm) and the C : N of zooplankton can explain the temporal variability in δ¹³C of freshwater zooplankton. We compare temporal patterns of δ¹³C and C : N for Daphnia, Hesperodiaptomus franciscanus and Leptodiaptomus tyrelli over a 6‐month period at four sites in two oligotrophic lakes. 3. In all three taxa, seasonal variation in zooplankton C : N explained more of the variation in zooplankton δ¹³C than did the δ¹³C of POM. This suggests that variation in the lipid content of zooplankton can strongly influence temporal variation of δ¹³C in zooplankton. 4. Using these data, we evaluate procedures that estimate the δ¹³C of only the non‐lipid component of zooplankton. If zooplankton lipids are primarily dietary in origin, than extracting lipids or ‘normalising’δ¹³C based on C : N will exclude a major dietary source, and therefore may be inappropriate. 5. We conclude that temporal variation in body composition (C : N) of zooplankton can significantly influence the temporal variation of zooplankton δ¹³C signatures.     

Carbon and nitrogen parasitism by a xylem‐tapping mistletoe (Tapinanthus oleifolius) along the Kalahari Transect: a stable isotope study

The present study explores the xylem‐tapping parasitism by mistletoe (Tapinanthus oleifolius) on native tree species along the Kalahari Transect (KT) using the stable isotopes of carbon and nitrogen. Mistletoe‐host pairs were collected at three geographical locations along the KT rainfall gradient in the 2005 and 2006 wet seasons. Foliar total carbon, total nitrogen and their stable isotope compositions (δ¹³C and δ¹⁵N) were measured. Heterotrophy (H) was calculated using foliar δ¹³C values of mistletoes and their hosts as an indicator of proportion of carbon in the mistletoes derived from host photosynthate. Based on the mistletoe H‐value and relationship between the mistletoe foliar δ¹⁵N and their host foliar δ¹⁵N, the results showed that mistletoes along the KT derived both nitrogen and carbon from their hosts. Mistletoes may regulate water use in relation to nitrogen supply. The proportion of carbon in the mistletoes derived from host photosynthate was between 35% and 78%, and the degree of heterotrophy was species‐specific with only limited annual variation. The study emphasizes the importance of incorporating parasitic associations in future studies on studying carbon, water and nutrient cycling along the Kalahari.