Erik A. Hobbie

Global patterns of foliar nitrogen isotopes and their relationships with climate, mycorrhizal fungi, foliar nutrient concentrations, and nitrogen availability.

Ratios of nitrogen (N) isotopes in leaves could elucidate underlying patterns of N cycling across ecological gradients. To better understand global-scale patterns of N cycling, we compiled data on foliar N isotope ratios (delta(15)N), foliar N concentrations, mycorrhizal type and climate for over 11,000 plants worldwide. Arbuscular mycorrhizal, ectomycorrhizal, and ericoid mycorrhizal plants were depleted in foliar delta(15)N by 2 per thousand, 3.2 per thousand, 5.9 per thousand, respectively, relative to nonmycorrhizal plants. Foliar delta(15)N increased with decreasing mean annual precipitation and with increasing mean annual temperature (MAT) across sites with MAT >or= -0.5 degrees C, but was invariant with MAT across sites with MAT < -0.5 degrees C. In independent landscape-level to regional-level studies, foliar delta(15)N increased with increasing N availability; at the global scale, foliar delta(15)N increased with increasing foliar N concentrations and decreasing foliar phosphorus (P) concentrations. Together, these results suggest that warm, dry ecosystems have the highest N availability, while plants with high N concentrations, on average, occupy sites with higher N availability than plants with low N concentrations. Global-scale comparisons of other components of the N cycle are still required for better mechanistic understanding of the determinants of variation in foliar delta(15)N and ultimately global patterns in N cycling.

Correlations between foliar δ15N and nitrogen concentrations may indicate plant-mycorrhizal interactions.

Abstract Nitrogen isotope measurements may provide insights into changing interactions among plants, mycorrhizal fungi, and soil processes across environmental gradients. Here, we report changes in δ15N signatures due to shifts in species composition and nitrogen (N) dynamics. These changes were assessed by measuring fine root biomass, net N mineralization, and N concentrations and δ15N of foliage, fine roots, soil, and mineral N across six sites representing different post-deglaciation ages at Glacier Bay, Alaska. Foliar δ15N varied widely, between 0 and –2‰ for nitrogen-fixing species, between 0 and –7‰ for deciduous non-fixing species, and between 0 and –11‰ for coniferous species. Relatively constant δ15N values for ammonium and generally low levels of soil nitrate suggested that differences in ammonium or nitrate use were not important influences on plant δ15N differences among species at individual sites. In fact, the largest variation among plant δ15N values were observed at the youngest and oldest sites, where soil nitrate concentrations were low. Low mineral N concentrations and low N mineralization at these sites indicated low N availability. The most plausible mechanism to explain low δ15N values in plant foliage was a large isotopic fractionation during transfer of nitrogen from mycorrhizal fungi to plants. Except for N-fixing plants, the foliar δ15N signatures of individual species were generally lower at sites of low N availability, suggesting either an increased fraction of N obtained from mycorrhizal uptake (f), or a reduced proportion of mycorrhizal N transferred to vegetation (Tr). Foliar and fine root nitrogen concentrations were also lower at these sites. Foliar N concentrations were significantly correlated with δ15N in foliage of Populus, Salix, Picea, and Tsuga heterophylla, and also in fine roots. The correlation between δ15N and N concentration may reflect strong underlying relationships among N availability, the relative allocation of carbon to mycorrhizal fungi, and shifts in either f or Tr.

15N IN SYMBIOTIC FUNGI AND PLANTS ESTIMATES NITROGEN AND CARBON FLUX RATES IN ARCTIC TUNDRA

When soil nitrogen is in short supply, most terrestrial plants form symbioses with fungi (mycorrhizae): hyphae take up soil nitrogen, transport it into plant roots, and receive plant sugars in return. In ecosystems, the transfers within the pathway fractionate nitrogen isotopes so that the natural abundance of 15N in fungi differs from that in their host plants by as much as 12% per hundred. Here we present a new method to quantify carbon and nitrogen fluxes in the symbiosis based on the fractionation against 15N during transfer of nitrogen from fungi to plant roots. We tested this method, which is based on the mass balance of 15N, with data from arctic Alaska where the nitrogen cycle is well studied. Mycorrhizal fungi provided 61-86% of the nitrogen in plants; plants provided 8-17% of their photosynthetic carbon to the fungi for growth and respiration. This method of analysis avoids the disturbance of the soil-microbe-root relationship caused by collecting samples, mixing the soil, or changing substrate concentrations. This analytical technique also can be applied to other nitrogen-limited ecosystems, such as many temperate and boreal forests, to quantify the importance for terrestrial carbon and nitrogen cycling of nutrient transfers mediated by mycorrhizae at the plant-soil interface.

Redefining fine roots improves understanding of below-ground contributions to terrestrial biosphere processes.

Fine roots acquire essential soil resources and mediate biogeochemical cycling in terrestrial ecosystems. Estimates of carbon and nutrient allocation to build and maintain these structures remain uncertain because of the challenges of consistently measuring and interpreting fine-root systems. Traditionally, fine roots have been defined as all roots ≤ 2 mm in diameter, yet it is now recognized that this approach fails to capture the diversity of form and function observed among fine-root orders. Here, we demonstrate how order-based and functional classification frameworks improve our understanding of dynamic root processes in ecosystems dominated by perennial plants. In these frameworks, fine roots are either separated into individual root orders or functionally defined into a shorter-lived absorptive pool and a longer-lived transport fine-root pool. Using these frameworks, we estimate that fine-root production and turnover represent 22% of terrestrial net primary production globally - a c. 30% reduction from previous estimates assuming a single fine-root pool. Future work developing tools to rapidly differentiate functional fine-root classes, explicit incorporation of mycorrhizal fungi into fine-root studies, and wider adoption of a two-pool approach to model fine roots provide opportunities to better understand below-ground processes in the terrestrial biosphere.

Insights into nitrogen and carbon dynamics of ectomycorrhizal and saprotrophic fungi from isotopic evidence

Abstract The successful use of natural abundances of carbon (C) and nitrogen (N) isotopes in the study of ecosystem dynamics suggests that isotopic measurements could yield new insights into the role of fungi in nitrogen and carbon cycling. Sporocarps of mycorrhizal and saprotrophic fungi, vegetation, and soils were collected in young, deciduous-dominated sites and older, coniferous-dominated sites along a successional sequence at Glacier Bay National Park, Alaska. Mycorrhizal fungi had consistently higher δ15N and lower δ13C values than saprotrophic fungi. Foliar δ13C values were always isotopically depleted relative to both fungal types. Foliar δ15N values were usually, but not always, more depleted than those in saprotrophic fungi, and were consistently more depleted than in mycorrhizal fungi. We hypothesize that an apparent isotopic fractionation by mycorrhizal fungi during the transfer of nitrogen to plants may be attributed to enzymatic reactions within the fungi producing isotopically depleted amino acids, which are subsequently passed on to plant symbionts. An increasing difference between soil mineral nitrogen δ15N and foliar δ15N in later succession might therefore be a consequence of greater reliance on mycorrhizal symbionts for nitrogen supply under nitrogen-limited conditions. Carbon signatures of mycorrhizal fungi may be more enriched than those of foliage because the fungi use isotopically enriched photosynthate such as simple sugars, in contrast to the mixture of compounds present in leaves. In addition, some 13C fractionation may occur during transport processes from leaves to roots, and during fungal chitin biosynthesis. Stable isotopes have the potential to help clarify the role of fungi in ecosystem processes.

CARBON ALLOCATION TO ECTOMYCORRHIZAL FUNGI CORRELATES WITH BELOWGROUND ALLOCATION IN CULTURE STUDIES

Ectomycorrhizal fungi form symbioses with most temperate and boreal tree species, but difficulties in measuring carbon allocation to these symbionts have prevented the assessment of their importance in forest ecosystems. Here, I surveyed allocation patterns in 14 culture studies and five field studies of ectomycorrhizal plants. In culture studies, allocation to ectomycorrhizal fungi (NPPf) was linearly related to total belowground net primary production (NPPb) by the equation NPPf = 41.5% x NPPb - 11.3% (r2 = 0.55, P < 0.001) and ranged from 1% to 21% of total net primary production. As a percentage of NPP, allocation to ectomycorrhizal fungi was highest at lowest plant growth rates and lowest nutrient availabilities. Because total belowground allocation can be estimated using carbon balance techniques, these relationships should allow ecologists to incorporate mycorrhizal fungi into existing ecosystem models. In field studies, allocation to ectomycorrhizal fungi ranged from 0% to 22% of total allocation, but wide differences in measurement techniques made intercomparisons difficult. Techniques such as fungal in-growth cores, root branching-order studies, and isotopic analyses could refine our estimates of turnover rates of fine roots, mycorrhizae, and extraradical hyphae. Together with ecosystem modeling, such techniques could soon provide good estimates of the relative importance of root vs. fungal allocation in belowground carbon budgets.

Nitrogen isotopes link mycorrhizal fungi and plants to nitrogen dynamics.

In this review, we synthesize field and culture studies of the 15N/14N (expressed as δ15N) of autotrophic plants, mycoheterotrophic plants, parasitic plants, soil, and mycorrhizal fungi to assess the major controls of isotopic patterns. One major control for plants and fungi is the partitioning of nitrogen (N) into either 15N-depleted chitin, ammonia, or transfer compounds or 15N-enriched proteinaceous N. For example, parasitic plants and autotrophic hosts are similar in δ15N (with no partitioning between chitin and protein), mycoheterotrophic plants are higher in δ15 N than their fungal hosts, presumably with preferential assimilation of fungal protein, and autotrophic, mycorrhizal plants are lower in 15N than their fungal symbionts, with saprotrophic fungi intermediate, because mycorrhizal fungi transfer 15N-depleted ammonia or amino acids to plants. Similarly, nodules of N2-fixing bacteria transferring ammonia are often higher in δ15N than their plant hosts. N losses via denitrification greatly influence bulk soil δ15N, whereas δ15N patterns within soil profiles are influenced both by vertical patterns of N losses and by N transfers within the soil-plant system. Climate correlates poorly with soil δ15N; climate may primarily influence δ15N patterns in soils and plants by determining the primary loss mechanisms and which types of mycorrhizal fungi and associated vegetation dominate across climatic gradients.

Nitrogen isotopes in ectomycorrhizal sporocarps correspond to belowground exploration types

Nitrogen isotope values (δ15N) are higher in ectomycorrhizal fungi than in their plant hosts but the wide variability in δ15N among sporocarps of different fungal taxa is unexplained. We propose that fungal δ15N reflects sequestration of fungal nitrogen to build fungal biomass, and should accordingly reflect fungal exploration strategies and hyphal properties. To test this, we compared δ15N to exploration types, hyphal hydrophobicity, and the presence of rhizomorphs in ectomycorrhizal species from surveys at four sites in temperate and boreal coniferous forests. Fungi with exploration types of high biomass, such as long-distance (e.g., Suillus), medium-distance mat (e.g., Hydnellum), and medium-distance fringe (e.g., Cortinarius) were 4–7‰ more enriched in 15N than fungi with exploration types of low biomass [medium-distance smooth (e.g., Amanita), short-distance (e.g., Inocybe), and contact (e.g., Hygrophorus)]. High biomass types comprised 79% (Åheden, northern Sweden), 65% (Deer Park, Pacific Northwest, USA), 45% (Stadsskogen, central Sweden), and 39% (Hoh, Pacific Northwest, USA) of ectomycorrhizal species, with these types more prevalent at sites of lower nitrogen availability. Species with hydrophobic hyphae or with rhizomorphs were 3–4‰ more enriched in 15N than taxa with hydrophilic hyphae or without rhizomorphs. The consistency of these patterns suggest that δ15N measurements could provide insights into belowground functioning of poorly known taxa of ectomycorrhizal fungi and into relative fungal biomass across ectomycorrhizal communities.

Natural Abundance of 15N in Nitrogen-Limited Forests and Tundra Can Estimate Nitrogen Cycling Through Mycorrhizal Fungi: A Review

The hyphae of ectomycorrhizal and ericoid mycorrhizal fungi proliferate in nitrogen (N)-limited forests and tundra where the availability of inorganic N is low; under these conditions the most common fungal species are those capable of protein degradation that can supply their host plants with organic N. Although it is widely understood that these symbiotic fungi supply N to their host plants, the transfer is difficult to quantify in the field. A novel approach uses the natural 15N:14N ratios (expressed as δ15N values) in plants, soils, and mycorrhizal fungi to estimate the fraction of N in symbiotic trees and shrubs that enters through mycorrhizal fungi. This calculation is possible because mycorrhizal fungi discriminate against 15N when they create compounds for transfer to plants; host plants are depleted in 15N, whereas mycorrhizal fungi are enriched in 15N. The amount of carbon (C) supplied to these fungi can be stoichiometrically calculated from the fraction of plant N derived from the symbiosis, the N demand of the plants, the fungal C:N ratio, and the fraction of N retained in the fungi. Up to a third of C allocated belowground, or 20% of net primary production, is used to support ectomycorrhizal fungi. As anthropogenic N inputs increase, the C allocation to fungi decreases and plant δ15N increases. Careful analyses of δ15N patterns in systems dominated by ectomycorrhizal and ericoid mycorrhizal symbioses may reveal the ecosystem-scale effects of alterations in the plant–mycorrhizal symbioses caused by shifts in climate and N deposition.

Patterns of natural 15N in soils and plants from chemically and organically fertilized uplands

Diagnostic tests for organic production of crops would be useful. In this study, the difference in natural 15N abundances (δ15N) of soils and plants between fertilizer-applied upland (FU) and compost-applied upland (CU) fields was investigated to study using δ15N as a marker of organic produce. Twenty samples each of soils and plants were collected from each field in early summer after applying fertilizer or compost. The δ15N of fertilizers and composts was −1.6±1.5‰ (n=8) and 17.4±1.2‰ (n=10), respectively. The δ15N of total soil-N was significantly (P<0.05) higher in CU fields (8.8±2.0‰) than in FU fields (5.9±0.7‰) due to long-term continuous application of 15N-enriched compost, as indicated by a positive correlation (r=0.62) between N content and δ15N of total soil-N. The NO3− pool of CU soils (11.6±4.5‰) was also significantly (P<0.05) enriched in 15N compared to FU soils (4.7±1.1‰), while the 15N contents of NH4+ pool were not different between both soils. Compost application resulted in 15N enrichment of plants; the δ15N values were 14.6±3.3‰ for CU and 4.1±1.7‰ for FU fields. These results showed that long-term application of compost resulted in a significant 15N-enrichment of soils and plants relative to fertilizer. Therefore, this study suggested that δ15N could serve as promising indicators of organic fertilizers application when used with other independent evidence. However, further studies under many conditions should be conducted to prepare reliable δ15N guidelines for organic produce, since the δ15N of inorganic soil-N and plant-N are influenced by various factors such as soil type, plant species, the rate of N application, and processes such as mineralization, nitrification, and denitrifcation.