▪ Abstract The use of stable isotope techniques in plant ecological research has grown steadily during the past two decades. This trend will continue as investigators realize that stable isotopes can serve as valuable nonradioactive tracers and nondestructive integrators of how plants today and in the past have interacted with and responded to their abiotic and biotic environments. At the center of nearly all plant ecological research which has made use of stable isotope methods are the notions of interactions and the resources that mediate or influence them. Our review, therefore, highlights recent advances in plant ecology that have embraced these notions, particularly at different spatial and temporal scales. Specifically, we review how isotope measurements associated with the critical plant resources carbon, water, and nitrogen have helped deepen our understanding of plant-resource acquisition, plant interactions with other organisms, and the role of plants in ecosystem studies. Where possible we also...

Stable Isotopes in Plant Ecology

Equilibrium and kinetic isotope fractionations during incomplete reactions result in minute differences in the ratio between the two stable N isotopes, 15N and 14N, in various N pools. In ecosystems such variations (usually expressed in per mil [δ15N] deviations from the standard atmospheric N2) depend on isotopic signatures of inputs and outputs, the input–output balance, N transformations and their specific isotope effects, and compartmentation of N within the system. Products along a sequence of reactions, e.g. the N mineralization–N uptake pathway, should, if fractionation factors were equal for the different reactions, become progressively depleted. However, fractionation factors vary. For example, because nitrification discriminates against 15N in the substrate more than does N mineralization, NH4+ can become isotopically heavier than the organic N from which it is derived.Levels of isotopic enrichment depend dynamically on the stoichiometry of reactions, as well as on specific abiotic and biotic conditions. Thus, the δ15N of a specific N pool is not a constant, and δ15N of a N compound added to the system is not a conservative, unchanging tracer. This fact, together with analytical problems of measuring δ15N in small and dynamic pools of N in the soil–plant system, and the complexity of the N cycle itself (for instance the abundance of reversible reactions), limit the possibilities of making inferences based on observations of 15N abundance in one or a few pools of N in a system. Nevertheless, measurements of δ15N might offer the advantage of giving insights into the N cycle without disturbing the system by adding 15N tracer.Such attempts require, however, that the complex factors affecting δ15N in plants be taken into account, viz. (i) the source(s) of N (soil, precipitation, NOx, NH3, N2-fixation), (ii) the depth(s) in soil from which N is taken up, (iii) the form(s) of soil-N used (organic N, NH4+, NO3−), (iv) influences of mycorrhizal symbioses and fractionations during and after N uptake by plants, and (v) interactions between these factors and plant phenology. Because of this complexity, data on δ15N can only be used alone when certain requirements are met, e.g. when a clearly discrete N source in terms of amount and isotopic signature is studied. For example, it is recommended that N in non-N2-fixing species should differ more than 5‰ from N derived by N2-fixation, and that several non-N2-fixing references are used, when data on δ15N are used to estimate N2-fixation in poorly described ecosystems.As well as giving information on N source effects, δ15N can give insights into N cycle rates. For example, high levels of N deposition onto previously N-limited systems leads to increased nitrification, which produces 15N-enriched NH4+ and 15N-depleted NO3−. As many forest plants prefer NH4+ they become enriched in 15N in such circumstances. This change in plant δ15N will subsequently also occur in the soil surface horizon after litter-fall, and might be a useful indicator of N saturation, especially since there is usually an increase in δ15N with depth in soils of N-limited forests.Generally, interpretation of 15N measurements requires additional independent data and modelling, and benefits from a controlled experimental setting. Modelling will be greatly assisted by the development of methods to measure the δ15N of small dynamic pools of N in soils. Direct comparisons with parallel low tracer level 15N studies will be necessary to further develop the interpretation of variations in δ15N in soil–plant systems. Another promising approach is to study ratios of 15N[ratio ]14N together with other pairs of stable isotopes, e.g. 13C[ratio ]12C or 18O[ratio ]16O, in the same ion or molecules. This approach can help to tackle the challenge of distinguishing isotopic source effects from fractionations within the system studied.

Tansley Review No. 95 15N natural abundance in soil-plant systems

https://www.cell.com/trends/plant-science/pdf/S1360-1385(01)01889-1.pdf

Physiological mechanisms influencing plant nitrogen isotope composition.

We assembled a globally-derived data set for site-averaged foliar delta(15)N, the delta(15)N of whole surface mineral soil and corresponding site factors (mean annual rainfall and temperature, latitude, altitude and soil pH). The delta(15)N of whole soil was related to all of the site variables (including foliar delta(15)N) except altitude and, when regressed on latitude and rainfall, provided the best model of these data, accounting for 49% of the variation in whole soil delta(15)N. As single linear regressions, site-averaged foliar delta(15)N was more strongly related to rainfall than was whole soil delta(15)N. A smaller data set showed similar, negative correlations between whole soil delta(15)N, site-averaged foliar delta(15)N and soil moisture variations during a single growing season. The negative correlation between water availability (measured here by rainfall and temperature) and soil or plant delta(15)N fails at the landscape scale, where wet spots are delta(15)N-enriched relative to their drier surroundings. Here we present global and seasonal data, postulate a proximate mechanism for the overall relationship between water availability and ecosystem delta(15)N and, newly, a mechanism accounting for the highly delta(15)N-depleted values found in the foliage and soils of many wet/cold ecosystems. These hypotheses are complemented by documentation of the present gaps in knowledge, suggesting lines of research which will provide new insights into terrestrial N-cycling. Our conclusions are consistent with those of Austin and Vitousek (1998) that foliar (and soil) delta(15)N appear to be related to the residence time of whole ecosystem N.

The 15N natural abundance (δ15N) of ecosystem samples reflects measures of water availability

Contents
 	Summary	367	
I.	Introduction	367	
II.	Background on isotopes	368	
III.	Patterns of soil δ15N	370	
IV.	Patterns of fungal δ15N	372	
V.	Biochemical basis for the influence of fungi on δ15N patterns in plant–soil systems	373	
VI.	Patterns of δ15N in plant and fungal culture studies	374	
VII.	Mycoheterotrophic and parasitic plants	375	
VIII.	Patterns of foliar δ15N in autotrophic plants	376	
IX.	Controls over plant δ15N	377	
X.	Conclusions and research needs	378	
 	Acknowledgements	379	
 	References	379	


Summary
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 δ15N 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.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1469-8137.2012.04300.x

Nitrogen isotopes link mycorrhizal fungi and plants to nitrogen dynamics

SUMMARY
The 15N natural abundance and N concentrations of fruit bodies from 70 species (23 genera) of ectomycorrhizal fungi found in boreal forests are presented. Large intraspecific and intrageneric differences were found, e.g. 8.3‰15N in the species Dermocybe crocea and 12.6‰ in the genus Cortinarius. In addition, significant differences in both δ15N and %N were found between different parts of fruit bodies, with cap material giving consistently higher values. Proteins and amino acids were enriched by 9.7±0.4‰ (mean ± 1 SE) relative to chitin, irrespective of the part of the fruit body examined. Chitin had δ15N values similar to that of plant hosts. The higher δ15N and %N values of the caps than of the stipes probably reflect a higher portion of proteins and amino acids in the caps. The δ15N of mycorrhizal fungi can be a function of the N species used (organic N, NH4+, NO3−), the depth of soil at which the mycelium occurs, and metabolic fractionations. The metabolic fractionations, e.g. potential transaminations during the flux of N from the soil through the fungus to the plant, make it difficult, at present, to make inferences about sources of N based on δ15N values alone. No effect of sample drying temperature on δ15values of fungal material was detected.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1046/j.1469-8137.1997.00788.x

Natural 15N abundance in fruit bodies of ectomycorrhizal fungi from boreal forests

Summary
Mucor hiemalis, Zygorhynchus heterogamous, Aspergillus niger and Penicillium frequentans were incubated in media containing either [methyl-14C] paraquat or [ethylene-14C] diquat. Aspergillus niger liberated 14CO2 from both herbicides and Penicillium frequentans only from paraquat. Mucor hiemalis and Zygorhynchus heterogamous were unable to release 14CO2 from either. Those species able to degrade paraquat or diquat have correspondingly low intracellular levels of the herbicides and are able to tolerate high concentrations in the medium. By contrast, species unable to break them down are characterized by high intracellular levels and by their inability to grow at low concentrations. Tolerance to these herbicides appears to be the result of an ability to degrade them.

https://www.jstor.org/stable/pdfplus/2433482.pdf

The breakdown of paraquat and diquat by soil fungi

Summary
Paraquat and diquat cause rapid loss of potassium and phosphate from Aspergillus niger, Penicillium frequentans, Mucor hiemalis and Zygorrhynchus heterogamus. The rate of loss is greater at higher concentrations of herbicide, particularly from Zygorrhynchus and Mucor. A brief incubation in paraquat or diquat is followed by sustained loss of potassium when colonies of the four species are transferred to water. Sodium azide also stimulates loss of potassium but thisceases when colonies are transferred to water. Paraquat enhances the level of malondialdehyde, a breakdown product of membrane fatty acids, in Zygorrhynchus and Mucor. These results are discussed with reference to the possible mode of action of bipyridyl herbicides in fungi.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1469-8137.1981.tb02321.x

The effect of bipyridyl herbicides on the loss of nutrients from fungi

/pdf/keeping-bloody-relics-liquid-3wijxkfvt2.pdf

Anthony J. E. Lyon

Papers

Natural 15N abundance in fruit bodies of ectomycorrhizal fungi from boreal forests

The breakdown of paraquat and diquat by soil fungi

The effect of bipyridyl herbicides on the loss of nutrients from fungi

Keeping bloody relics liquid