Interpreting tree responses to thinning and fertilization using tree-ring stable isotopes

? Carbon sequestration has focused renewed interest in understanding how forest management affects forest carbon gain over timescales of decades, and yet details of the physiological mechanisms over decades are often lacking for understanding long-term growth responses to management. ? Here, we examined tree-ring growth patterns and stable isotopes of cellulose (δ(13)C(cell) and δ(18)O(cell)) in a thinning and fertilization controlled experiment where growth increased substantially in response to treatments to elucidate physiological data and to test the dual isotope approach for uses in other locations. ? δ(13)C(cell) and δ(18)O(cell) results indicated that fertilization caused an increase in intrinsic water-use efficiency through increases in photosynthesis (A) for the first 3 yr. The combination treatment caused a much larger increase in A and water-use efficiency. Only the thinning treatments showed consistent significant increases in δ(18)O(cell) above controls. Changes in canopy microclimate are the likely drivers for δ(18)O(cell) increases with decreases in relative humidity and increases in leaf temperature associated with thinning being the most probable causes. ? Tree-ring isotopic records, particularly δ(13)C(cell), remain a viable way to reconstruct long-term physiological mechanisms affecting tree carbon gain in response to management and climate fluctuations.     

Effects of sample preparation on stable isotope ratios of carbon and nitrogen in marine invertebrates: implications for food web studies using stable isotopes

Trophic ecology has benefitted from the use of stable isotopes for the last three decades. However, during the last 10 years, there has been a growing awareness of the isotopic biases associated with some pre-analytical procedures that can seriously hamper the interpretation of food webs. We have assessed the extent of such biases by: (1) reviewing the literature on the topic, and (2) compiling C and N isotopic values of marine invertebrates reported in the literature with the associated sample preparation protocols. The factors considered were: acid-washing, distilled water rinsing (DWR), sample type (whole individuals or pieces of soft tissues), lipid content, and gut contents. Two-level ANOVA revealed overall large and highly significant effects of acidification for both delta(13)C values (up to 0.9 per thousand decrease) and delta(15) N values (up to 2.1 per thousand decrease in whole individual samples, and up to 1.1 per thousand increase in tissue samples). DWR showed a weak overall effect with delta(13)C increments of 0.6 per thousand (for the entire data set) or decrements of 0.7 per thousand in delta(15) N values (for tissue samples). Gut contents showed no overall significant effect, whereas lipid extraction resulted in the greatest biases in both isotopic signatures (delta(13)C, up to -2.0 per thousand in whole individuals; delta(15)N, up to +4.3 per thousand in tissue samples). The study analyzed separately the effects of the various factors in different taxonomic groups and revealed a very high diversity in the extent and direction of the effects. Maxillopoda, Gastropoda, and Polychaeta were the classes that showed the largest isotopic shifts associated with sample preparation. Guidelines for the standardization of sample preparation protocols for isotopic analysis are proposed both for large and small marine invertebrates. Broadly, these guidelines recommend: (1) avoiding both acid washing and DWR, and (2) performing lipid extraction and gut evacuation in most cases.     

Estimating the timing of diet shifts using stable isotopes

Stable isotope analysis has become an important tool in studies of trophic food webs and animal feeding patterns. When animals undergo rapid dietary shifts due to migration, metamorphosis, or other reasons, the isotopic composition of their tissues begins changing to reflect that of their diet. This can occur both as a result of growth and metabolic turnover of existing tissue. Tissues vary in their rate of isotopic change, with high turnover tissues such as liver changing rapidly, while relatively low turnover tissues such as bone change more slowly. A model is outlined that uses the varying isotopic changes in multiple tissues as a chemical clock to estimate the time elapsed since a diet shift, and the magnitude of the isotopic shift in the tissues at the new equilibrium. This model was tested using published results from controlled feeding experiments on a bird and a mammal. For the model to be effective, the tissues utilized must be sufficiently different in their turnover rates. The model did a reasonable job of estimating elapsed time and equilibrial isotopic changes, except when the time since the diet shift was less than a small fraction of the half-life of the slowest turnover tissue or greater than 5–10 half-lives of the slowest turnover tissue. Sensitivity analyses independently corroborated that model estimates became unstable at extremely short and long sample times due to the effect of random measurement error. Subject to some limitations, the model may be useful for studying the movement and behavior of animals changing isotopic environments, such as anadromous fish, migratory birds, animals undergoing metamorphosis, or animals changing diets because of shifts in food abundance or competitive interactions.     

Measuring the trophic ecology of ants using stable isotopes

Ants are prominent components of most terrestrial arthropod food webs, yet due to their highly variable diet, the role ants play in arthropod communities can be difficult to resolve. Stable isotope analysis is a promising method for determining the dietary history of an organism, and has the potential to advance our understanding of the food web ecology of social insects. However, some unique characteristics of eusocial organisms can complicate the application of this technique to the study of their trophic ecology. Using stable isotopes of N and C, we investigated levels of intraspecific variation both within and among colonies. We also examined the effect of a common preservation technique on δ¹⁵N and δ¹³C values. We discuss the implications of our results on experimental design and sampling methods for studies using stable isotopes to investigate the trophic ecology of social insects. Received 4 February 2005; revised 23 June 2005; accepted 4 July 2005.     

利用氮、氧稳定同位素识别水体硝酸盐污染源研究进展

水体硝酸盐污染已经成为一个相当普遍且重要的环境问题.为了保证人类的身体健康、水环境的良性演化,有效识别水体中硝酸盐污染的来源就显得尤为重要.水体中不同来源的硝酸盐具有不同的氮、氧稳定同位素组成,因此,可以利用氮、氧稳定同位素对水体中的硝酸盐污染进行源识别.本文介绍了氮、氧稳定同位素在氮循环主要过程中的分馏系数和主要硝酸盐来源的氮、氧稳定同位素组成,对比了5种硝酸盐氮、氧同位素分析预处理方法的优缺点,综述了国内外学者在该方向的研究进展并划分为3个阶段:单独使用氮稳定同位素;同时使用氮、氧稳定同位素;结合数学模型的应用.最后,对该领域今后的研究方向进行了展望.     

Determination of food sources and trophic position in Malaysian tropical highland streams using carbon and nitrogen stable isotopes

Stable isotope analysis has been extensively used as an effective tool in determination of trophic relationship in ecosystems. In freshwater ecosystem, aquatic invertebrates represent main component of a river food web. This study was carried out to determine potential food sources of freshwater organism together with pattern of trophic position along the river food web. In this study, rivers of Belum-Temengor Forest Complex (BTFC) has been selected as sampling site as it is a pristine area that contains high diversity and abundance of organisms and can be a benchmark for other rivers in Malaysia. Stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) were applied to estimate trophic position and food web paradigm. Analysis of stable isotopes based on organic material collected from the study area revealed that the highest δ¹³C value was reported from filamentous algae (? 22.68 ± 0.126⁰/₀₀) and the lowest δ¹³C was in allocthonous leaf packs (? 31.58 ± 0.187⁰/₀₀). Meanwhile the highest δ¹⁵N value was in fish (8.45 ± 0.177⁰/₀₀) and the lowest value of δ¹⁵N was in autochthonous aquatic macrophyte (2.00 ± 1.234⁰/₀₀). Based on the δ¹⁵N results, there are three trophic levels in the study river and it is suggested that the trophic chain begins with organic matter followed by group of insects and ends with fish (organic matter < insects < fish).     

Determination of root biomasses of three species grown in a mixture using stable isotopes of carbon and nitrogen

A method is evaluated that employs variation in stable C and N isotopes from fractionations in C and N acquisition and growth to predict root biomasses of three plant species in mixtures. Celtis laevigata Willd. (C₃), Prosopis glandulosa Torr. (C₃, legume) and Schizachyrium scoparium (Michx.) Nash (C₄), or Gossypium hirsutum L. (C₃), Glycine max (L.) Merr. (C₃ legume), and Sorghum bicolor (L.) Moench (C₄) were grown together in separate, three-species combinations. Surface roots (0–10 cm depth) of each species from each of the two combinations were mixed in various proportions, and the relative abundances of ¹⁵N and ¹⁴N and ¹³C and ¹²C in prepared mixtures, surface roots of single species, and roots extracted from the 80-cm soil profile in which each species combination was grown were analyzed by mass spectrometry. An algebraic determination which employed the δ ¹³C, % ¹⁵N, and C and N concentrations of root subsamples of individual species accounted for more than 95% of the variance in biomass of each species in prepared mixtures with G. max, G. hirsutum, and S. bicolor. A similar analysis demonstrated species-specific differences in rooting patterns. Root biomasses of the C₄ monocots in each combination, S. scoparium and S. bicolor, were concentrated in the upper 20 cm of soil, while those of G. hirsutum and the woody P. glandulosa were largest in lower soil strata. Analyses of stable C and N isotopes can effectively be used to distinguish roots of species which differ in ratios of ¹⁵N to ¹⁴N and ¹³C to ¹²C and thus to study belowground competition between or rooting patterns of associated species with different C and N isotope signatures. The method evaluated can be extended to quantify aboveground and belowground biomasses of component species in mixtures with isotopes of other elements or element concentrations that differ consistently among plants of interest.     

The 'lost years' of green turtles: using stable isotopes to study cryptic lifestages

Ignorance of the location or inaccessible locations of lifestages can impede the study and management of species. We used stable isotopes of carbon and nitrogen to identify the habitats and diets and to estimate the duration of a 'missing' lifestage: the early juvenile stage of the green turtle, Chelonia mydas. Stable isotopes in scute from young herbivorous green turtles in shallow-water habitats revealed that they spend 3-5 years as carnivores in oceanic habitats before making a rapid ontogenetic shift in diet and habitat. Stable isotopes in persistent and continuously growing tissues, such as some fish scales, bird bills and claws and mammal hair and claws, can be used to evaluate the ecology of inaccessible lifestages.     

Tracing origins and migration of wildlife using stable isotopes: a review

To understand the ecology of migratory animals it is important to link geographic regions used by individuals including breeding, wintering, and intermediate stopover sites. Previous conventional approaches used to track animal movements have relied on extrinsic markers and typically the subsequent recovery of individuals. This approach has generally been inappropriate for most small, or non-game animals. The use of intrinsic markers such as fatty acid profiles, molecular DNA analyses, and the measurement of naturally occurring stable isotopes in animal tissues offer alternative approaches. This paper reviews the use of stable isotope analyses (primarily δ¹³C, δ¹⁵N, δ³⁴S, δD, δ⁸⁷Sr) to trace nutritional origin and migration in animals. This approach relies on the fact that foodweb isotopic signatures are reflected in the tissues of organisms and that such signatures can vary spatially based on a variety of biogeochemical processes. Organisms moving between isotopically distinct foodwebs can carry with them information on the location of previous feeding. Such an approach has been used to track animal use of inshore versus offshore, marine versus freshwater, terrestrial C₃ versus marine, terrestrial mesic versus xeric, and C₃ versus C₄ or Crassulacean acid metabolism foodwebs. More recently, the use of stable hydrogen isotope analyses (δD) to link organisms to broad geographic origin in North America is based on large-scale isotopic contours of growing-season average δD values in precipitation. This technique, especially when combined with the assay of other stable isotopes, will be extremely useful in helping to track migration and movement of a wide range of animals from insects to birds and mammals. Future research to refine our understanding of natural and anthropogenic-induced isotopic gradients in nature, and to explore the use of stable isotopes of other elements, is recommended. Received: 1 July 1998 / Accepted: 9 December 1998     

Determining biological tissue turnover using stable isotopes: the reaction progress variable

The reaction progress variable is applied to stable isotope turnover of biological tissues. This approach has the advantage of readily determining whether more than one isotope turnover pool is present; in addition, the normalization process inherent to the model means that multiple experiments can be considered together although the initial and final isotope compositions are different. Consideration of multiple isotope turnover pools allows calculation of diet histories of animals using a time sequence of isotope measurements along with isotope turnover pools. The delayed release of blood cells from bone marrow during a diet turnover experiment can be quantified using this approach. Turnover pools can also be corrected for increasing mass during an experiment, such as when the animals are actively growing. Previous growth models have been for exponential growth; the approach here can be used for several different growth models. Electronic Supplementary Material The online version of this article () contains supplementary material, which is available to authorized users.     

Variations and controls of nitrogen stable isotopes in particulate organic matter of lakes

Nitrogen stable isotope (δ¹⁵N) data of particulate organic matter (POM) from the literature were analyzed to provide an understanding of the variations and controls of δ¹⁵N_POM in lakes at the global scale. The δ¹⁵N_POM variability characterized by seasonal mean, minimum, maximum, and amplitude (defined as δ¹⁵N_POM maximum − δ¹⁵N_POM minimum) from 36 lakes with seasonal data did not change systematically with latitude, but was significantly lower in small lakes than in large lakes. The seasonal mean δ¹⁵N_POM increased from oligotrophic lakes to eutrophic lakes despite large variations that are attributed to the occurrences of nitrogen fixation across the trophic gradient and the differences in δ¹⁵N of dissolved inorganic nitrogen (DIN) in individual lakes. Seasonal mean δ¹⁵N_POM was significantly correlated with DIN concentration and δ¹⁵N_DIN in two subsets of lakes. Seasonal minimum δ¹⁵N_POM in individual lakes is influenced by nitrogen fixation and δ¹⁵N_DIN while seasonal maximum δ¹⁵N_POM is influenced by lake trophic state and δ¹⁵N_DIN. As a result of the dominance of non-living POM in the unproductive surface waters, seasonal δ¹⁵N_POM amplitude was small (mean = 4.2‰) in oligotrophic lakes of all latitudes. On the other hand, seasonal δ¹⁵N_POM amplitude in eutrophic lakes was large (mean = 10.3‰), and increased from low to high latitudes, suggesting that the seasonal variability of δ¹⁵N in the phytoplankton-dominated POM pool was elevated by the greater spans of solar radiation and thermal regimes at high latitudes. The δ¹⁵N_POM from 42 lakes with no seasonal data revealed no consistent patterns along latitude, lake area, and trophic gradients, and a greater than 2‰ depletion compared to the lakes with seasonal data. Along with the large seasonal variability of δ¹⁵N_POM within lakes, these results provide insightful information on sampling design for the studies of food web baseline in lakes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effects of nutritional restriction on nitrogen and carbon stable isotopes in growing seabirds

When using stable isotopes as dietary tracers it is essential to consider effects of nutritional state on isotopic fractionation. While starvation is known to induce enrichment of ¹⁵N in body tissues, effects of moderate food restriction on isotope signatures have rarely been tested. We conducted two experiments to investigate effects of a 50–55% reduction in food intake on δ¹⁵N and δ¹³C values in blood cells and whole blood of tufted puffin chicks, a species that exhibits a variety of adaptive responses to nutritional deficits. We found that blood from puffin chicks fed ad libitum became enriched in ¹⁵N and ¹³C compared to food-restricted chicks. Our results show that ¹⁵N enrichment is not always associated with food deprivation and argue effects of growth on diet–tissue fractionation of nitrogen stable isotopes (Δ¹⁵N) need to be considered in stable isotope studies. The decrease in δ¹³C of whole blood and blood cells in restricted birds is likely due to incorporation of carbon from ¹³C-depleted lipids into proteins. Effects of nutritional restriction on δ¹⁵N and δ¹³C values were relatively small in both experiments (δ¹⁵N: 0.77 and 0.41‰, δ¹³C: 0.20 and 0.25‰) compared to effects of ecological processes, indicating physiological effects do not preclude the use of carbon and nitrogen stable isotopes in studies of seabird ecology. Nevertheless, our results demonstrate that physiological processes affect nitrogen and carbon stable isotopes in growing birds and we caution isotope ecologists to consider these effects to avoid drawing spurious conclusions.     

Early plant defence against insect attack: involvement of reactive oxygen species in plant responses to insect egg deposition

Planta - Pinus sylvestris responds to insect egg deposition by ROS accumulation linked with reduced activity of the ROS scavenger catalase. Egg mortality in needles with hypersensitive response...     

Carbon and nitrogen in a temperate agroforestry system: Using stable isotopes as a tool to understand soil dynamics

Natural exchanges of carbon (C) between the atmosphere, the oceans, and terrestrial ecosystems are currently being modified through human activities as a result of fossil fuel burning and the conversion of tropical forests to agricultural land. These activities have led to a steady increase of atmospheric carbon dioxide (CO₂) over the last two Centuries. The goal of this study was to determine the potential of temperate agroforestry systems to sequester C in soil. Therefore, changes in the soil organic C (SOC) and nitrogen (N) pools were quantified and the δ¹³C and δ¹⁵N stable isotope technique was applied to assess soil C and N dynamics in a 13-year old hybrid poplar alley cropping system in Southern Canada. Results from this study showed that after 13 years of alley cropping the SOC and N pools did not differ significantly (p = 0.01) with distance from the tree row, although a trend of a larger SOC and N pool near the tree row could be observed. Soil organic C after 13 years of alley cropping, was 19 mg C g⁻¹ compared to 11 mg C g⁻¹ upon initiation of agroforestry. Soil organic C and N were not evenly distributed throughout the plow layer. The largest C and N pool occurred in the top 20 cm, which is due to the accumulation of organic material in the upper horizons as a result of no-till cultivation. The entire soil, to a 40 cm depth, showed a δ¹³C shift to that of C₃ residue. This shift reflects the greater input of residues from C₃ plants such as that derived from beans, wheat, and hybrid poplar leaf litterfall. The proportion of C derived from a C₃ source ranged from 64 to 69% to a 40 cm depth. The soil δ¹⁵N signature of this study is similar to that of mineral soil, and reflect values characteristic of N mineralization processes. However, the entire soil shows a positive shift in δ¹⁵N as a result of historical additions of manure and current use of mineral fertilizers, and ongoing processes of denitrification and nitrate leaching, which leads to an enrichment of the soil.     

Trophic structure of macro- and meso-invertebrates in Japanese coniferous forest: Carbon and nitrogen stable isotopes analyses

Few studies have been published on the feeding ecology of Japanese soil fauna based on stable isotope analysis. Therefore, the present work aims to use this technique for studying the trophic structure of Japanese soil fauna at two coniferous forests. Significant differences were observed between investigated sites (Arahama and Gamo) in genus richness and abundance, while for Shannon diversity indexes the difference was non-significant. The isotopic signatures (δ¹³C and δ¹⁵N) of the invertebrates collected at Arahama ranged from 0.3 to 6.3‰ for δ¹⁵N and from −27.3 to −23.3‰ for δ¹³C. At Gamo, invertebrates δ¹³C values ranged from −26.1 to −23.5‰ and δ¹⁵N values ranged from 1.6 to 6.8‰. At both sites, invertebrates formed two distinct groups on the basis of combined C and N stable isotope ratios. The locations of these groups related to δ¹³C values. The less enriched group (δ¹³C < −25‰) and the more enriched one (δ¹³C > −25‰). The range of δ¹⁵N for the present animals exceeded two trophic levels. While, the gradual ¹⁵N enrichment within the invertebrates species may indicate the dominance of omnivory in soil food webs. The differences between sites in δ¹⁵N confirm the importance of studying the trophic structure of soil fauna locally.