Shoot δ15N Research Articles

Symbiotic N nutrition, carbon accumulation, and water-use efficiency in species of the genus Aspalathus native to the Cape fynbos, South Africa

Low soil moisture and N in the fynbos are a major constraint to plant growth and N2 fixation in legumes, including Aspalathus species. The 15N and 13C natural abundance techniques were used to study N nutrition and plant water relations in different Aspalathus species sampled from 13 study sites in the fynbos. The results showed that A. acuminata shoots recorded lower δ15N values at Kalbaskraal, Worcester Langerug and Roundebosch, resulting in higher %Ndfa at those sites relative to Elim, which registered greater δ15N values. However, A. hispida shoots showed the lowest δ15N at Elim and Dutoitskloof than Yzerfontein, and Silwerstroomstrand, resulting in high %Ndfa at Elim. Of A. cordata, A. astroites, A. hispida, and A. wildenowiana collected from Dutoitskloof differed in δ15N and %Ndfa. A. cordata and A. astroites shoots showed lower δ15N, leading to higher %Ndfa than A. wildenowiana, which exhibited greater shoot δ15N. Of four Aspalathus species (A. cordata, A. carnosa, A. capensis and A. juniperina) sampled from Silvermine, A. cordata revealed the lowest δ15N and hence the highest %Ndfa. Conversely, A. carnosa recorded the highest shoot δ15N and the lowest %Ndfa. At Kokrivier, shoot δ15N was much greater in A. zeyheri and lowest in A. incurvifolia, resulting in higher %Ndfa in the latter than the former. The Aspalathus species differed in water-use efficiency or shoot δ13C values, which ranged from −30.35‰ to −26.93‰. Rainfall, soil moisture deficit and altitude were the major determinants of shoot δ13C values, but how these factors influence shoot δ13C of plants remains to be determined.

Effects of copper contamination on N cycling microbial guilds and plant performance in two contrasting grassland soils

Heavy metal pollution has important effects on ecosystem nitrogen (N) cycling, but factors driving differences between the direction, onset and intensity of responses are poorly understood. We used two contrasting grassland soils to examine the effects of copper (Cu) on the abundance and activity of N cycling microbial guilds and plant responses, including plant δ15N as an integrator of the N cycle. A low pH sandy soil and a high pH sandy loam soil were aged two years in outdoor mesocosms with copper (Cu) treatments of background, 200, 400 or 1000 mg kg−1 Cu. After two years, increased Cu treatments resulted in significantly lower abundances of ammonia oxidizing archaea, Nitrospira nitrite oxidizing bacteria (NOB), potential ammonia oxidation rates and plant biomass in both soils. Plants possessed significantly increased N content and enriched shoot δ15N in with higher Cu in both soils. While abundances of ammonia oxidizing bacteria were unaffected by Cu, the response among Nitrobacter NOB and denitrifiers and plant δ13C differed between the two soils. In contrast to plants, differences in the intensity and direction of microbial guild responses were not explained by increasing soluble Cu but rather shaped by soil type. This indicates that the two soils differed in metal bioavailability to plants, as well as harbored microbial communities with inherent differences in metal sensitivity. Furthermore, effects of increasing Cu on microbial N-cycling guilds became more apparent with longer incubation time, emphasizing the importance of long-term studies to assess important ecosystem effects of Cu contamination. Taken together, we conclude that a combination of plant and microbial responses can give better insights on how Cu is affecting the N cycle in polluted soils.

Relationships between stable isotope natural abundances (δ13C and δ15N) and water use efficiency in rice under alternate wetting and drying irrigation in soils with high clay contents.

Natural abundance of the stable isotope (δ13C and δ15N) in plants is widely used to indicate water use efficiency (WUE). However, soil water and texture properties may affect this relationship, which remains largely elusive. Therefore, the purpose of this study was to evaluate δ13C as affected by different combinations of alternate wetting and drying irrigation (AWD) with varied soil clay contents in different organs and whole plant and assess the feasibility of using δ13C and δ15N as a physiological indicator of whole-plant water use efficiency (WUEwhole-plant). Three AWD regimes, I100 (30 mm flooded when soil reached 100% saturation), I90 (30 mm flooded when reached 90% saturation) and I70 (30 mm flooded when reached 70% saturation) and three soil clay contents, 40% (S40), 50% (S50), and 60% (S60), were included. Observed variations in WUEwhole-plant did not conform to theoretical expectations of the organs δ13C (δ13Corgans) of plant biomass based on pooled data from all treatments. However, a positive relationship between δ13Cleaf and WUEET (dry biomass/evapotranspiration) was observed under I90 regime, whereas there were no significant relationships between δ13Corgans and WUEET under I100 or I70 regimes. Under I100, weak relationships between δ13Corgans and WUEET could be explained by (i) variation in C allocation patterns under different clay content, and (ii) relatively higher rate of panicle water loss, which was independent of stomatal regulation and photosynthesis. Under I70, weak relationships between δ13Corgans and WUEET could be ascribed to (i) bigger cracks induced by water-limited irrigation regime and high clay content soil, and (ii) damage caused by severe drought. In addition, a negative relationship was observed between WUEwhole-plant and shoot δ15N (δ15Nshoot) across the three irrigation treatments, indicating that WUEwhole-plant is tightly associated with N metabolism and N isotope discrimination in rice. Therefore, δ13C should be used cautiously as an indicator of rice WUEwhole-plant at different AWD regimes with high clay content, whereas δ15N could be considered an effective indicator of WUEwhole-plant.

Black Seedcoat Pigmentation Is a Marker for Enhanced Nodulation and N2 Fixation in Bambara Groundnut (Vigna Subterranea L. Verdc.) Landraces

Field studies on the effect of differing seedcoat colour on plant growth and symbiotic performance of three Bambara groundnut landraces using the 15N natural abundance technique and ureide analysis in the xylem stream and petiole extracts revealed a consistent pattern in nodule function between ureide levels and shoot δ15N values, as well as between ureide levels and percent N derived from fixation at Gbalahi, Kpalisogu, and Manga. At those three sites, where shoot δ15N was low and percent N derived from fixation high, ureide concentrations in the xylem sap and petiole extracts were also high, indicating that the two techniques were comparatively quite robust in estimating N2 fixation at a higher precision in the test Bambara groundnut landraces. A comparison of the effect of differing seedcoat pigmentation on the concentration of phenolics released by imbibed seeds showed that the Black landrace contained higher levels of flavonoids and anthocyanins, followed by the Red, and lowest in the Cream landrace. The Black landrace also recorded much higher nodule number and nodule fresh weight per plant, greater shoot biomass, lower shoot δ15N values, greater N derived from fixation, higher symbiotic N contribution, and increased water-use efficiency across all the five study sites, followed by the Red landrace, which produced the highest grain yield. Black seedcoat pigmentation in Bambara groundnut is therefore a marker for increased nodulation, N2 fixation, and plant growth that can be tapped in breeding programs for developing high N2-fixing, water-use efficient grain legumes for use by farmers in Africa.

Direct and indirect trophic interactions of soil nematodes impact chickpea and oat nutrition

Root lesion nematodes (RLN) have negative impacts on legume-grass systems. These impacts might be moderated by bacterial feeding nematodes (BFN) presence. It remains unknown how these trophic groups of nematodes interactively impact plant productivity and dynamics of carbon (C) and nitrogen (N) in grass-legume mixtures. We addressed this research gap using inter-kingdom interactions in a model system. Chickpea and oat were grown in mono- and mixed cultures, and RLN and BFN were applied alone and in combination. Plant biomass, shoot C and N content and isotopic composition, and mineral N in soil were measured. RLN presence reduced root biomass across treatments. This reduction was stronger in the grass than the legume and was not modified by BFN. Nematodes increased plant shoot N concentrations. RLN and BFN had interactive effects on shoot nutrient concentration resulting in reduced shoot C concentration when both trophic groups were combined. Shoot δ15N data revealed transfer of symbiotically fixed N from chickpea to oat in the presence of RLN. However, this N transfer did not result in improved oat growth. Interactive effects of soil organisms can cascade aboveground, influencing C and N dynamics and ecosystem productivity.

Low 15N Natural Abundance in Shoot Tissue of Brachiaria humidicola Is an Indicator of Reduced N Losses Due to Biological Nitrification Inhibition (BNI).

The tropical forage grass Brachiaria humidicola (Bh) suppresses the activity of soil nitrifiers through biological nitrification inhibition (BNI). As a result, nitrate () formation and leaching are reduced which is also expected to tighten the soil nitrogen (N) cycle. However, the beneficial relationship between reduced losses and enhanced N uptake due to BNI has not been experimentally demonstrated yet. Nitrification discriminates against the 15N isotope and leads to 15N depleted , but 15N enriched in soils. Leaching of 15N depleted enriches the residual N pool in the soil with 15N. We hypothesized that altered nitrification and leaching due to diverging BNI magnitudes in contrasting Bh genotypes influence soil 15N natural abundance (δ15N), which in turn is reflected in distinct δ15N in Bh shoot biomass. Consequently, high BNI was expected to be reflected in low plant δ15N of Bh. It was our objective to investigate under controlled conditions the link between shoot value of δ15N in several Bh genotypes and leached amounts and shoot N uptake. Additionally, plant 15N and N% was monitored among a wide range of Bh genotypes with contrasting BNI potentials in field plots for 3 years. We measured leaf δ15N of young leaves (regrown after cutback) of Bh and combined it with nitrification rates (NRs) of incubated soil to test whether there is a direct relationship between plant δ15N and BNI. Increased leached was positively correlated with higher δ15N in Bh, whereas the correlation between shoot N uptake and shoot δ15N was inverse. Field cultivation of a wide range of Bh genotypes over 3 years decreased NRs in incubated soil, while shoot δ15N declined and shoot N% increased over time. Leaf δ15N of Bh genotypes correlated positively with NRs of incubated soil. It was concluded that decreasing plant δ15N of Bh genotypes over time reflects the long-term effect of BNI as linked to lower formation and reduced leaching. Accordingly, a low δ15N in Bh shoot tissue verified its potential as indicator of high BNI activity of Bh genotypes.

Quantifying N-loss by root abscission: consequences for wheat N budgets and δ15N values

Lower plant δ15N values relative to source δ15N are commonly attributed to 15N efflux. We determined the extent to which root abscission contributes to plant N-loss and consequences for plant δ15N. Wheat (Triticum aestivum L. cv. SST015) was grown in hydroponics with direct aeration, aeration constrained within a pipe and circulation of nutrient solution through sand, representing three levels of stability for root growth. The δ15N of nutrient solutions and root fragments were periodically determined, as well as root and shoot δ15N. Plants in solution had significantly more negative δ15N (−8.9 and −9.2‰) than plants in sand (−6.9‰), suggesting greater 15N-loss; root fragments were major biomass- (six-fold greater than root dry weight) and N-loss (two-fold greater than plant net N uptake) pathways in solution. These plants had more ephemeral roots and two-fold more root tips than the sand treatment. We estimated that root fragment loss decreased plant δ15N by at least −3.7, −2.6 and −1.0‰ in the direct, pipe and sand treatments, respectively. Positive nutrient solution δ15N in all treatments relative to the source δ15N suggests that plant N, probably derived from efflux, was present in solution. Despite this, root abscission and root turnover are also important N-loss pathways in plants, while plant δ15N values are probably influenced by a combination of root abscission and N efflux.

N2-fixation and N contribution by grain legumes under different soil fertility status and cropping systems in the Guinea savanna of northern Ghana

Continuous cereal-based cropping has led to a rapid decline in soil fertility in the Guinea savanna agro-ecological zone of northern Ghana with corresponding low crop yields. We evaluated the effects of cropping system and soil fertility status on grain yields and N2-fixation by grain legumes and net N contribution to soil fertility improvement in contrasting sites in this agro-ecological zone. Maize was intercropped with cowpea, soybean and groundnut within a row, with a maize stand alternated with two equally spaced cowpea or groundnut stands and in the maize-soybean system, four equally spaced soybean stands. These intercrops were compared with sole crops of maize, cowpea, soybean and groundnut in fertile and poorly fertile fields at sites in the southern (SGS) and the northern (NGS) Guinea savanna. The proportion of N derived from N2-fixation (%Ndfa) was comparable between intercrops and sole crops. However, the amount of N2-fixed was significantly larger in sole crops due to a greater biomass accumulation. Legumes in poorly fertile fields had significantly smaller shoot δ15N enrichment (−2.8 to +0.7‰) and a larger %Ndfa (55–94%) than those in fertile fields (−0.8 to +2.2‰; 23–85%). The N2-fixed however was larger in fertile fields (16–145kgNha−1) than in poorly fertile fields (15–123kgNha−1) due to greater shoot dry matter and N yields. The legumes grown in the NGS obtained more of their N requirements from atmospheric N2-fixation (73–88%) than legumes grown in the SGS (41–69%). The partial soil N balance (inkgha−1) was comparable between intercrops (−14 to 21) and sole legumes (−8 to 23) but smaller than that of sole maize receiving N fertiliser (+7 to +34). With other N inputs (aerial deposition) and outputs (leaching and gaseous losses) unaccounted for, there is uncertainty surrounding the actual amount of soil N balances of the cropping systems, indicating that partial N balances are not reliable indicators of the sustainability of cropping systems. Nevertheless, the systems with legumes seem more attractive due to several non-N benefits. Our results suggest that soybean could be targeted in the SGS and cowpea in the NGS for greater productivity while groundnut is suited to both environments. Grain legumes grown in poorly fertile fields contributed more net N to the soil but growing legumes in fertile fields seems more lucrative due to greater grain and stover yields and non-N benefits.

Contrasting grass nitrogen strategies reflect interspecific trade-offs between nitrogen acquisition and use in a semi-arid temperate grassland

Nitrogen (N) niche differentiations play an important role in community structure and biogeochemical cycling in terrestrial ecosystems. However, very few studies have examined how plant N strategies specialize via trade-offs between N acquisition and use abilities under natural field conditions. A field experiment was conducted to investigate N strategies by coexisting grass species using the in-situ stable isotope labeling technique. We injected 15N–labeled nitrate, 15N–labeled ammonium, and 13C-15N-labeled glycine solutions in early and late vegetative growing seasons. Shoot δ15N and N concentration were measured to determine the N uptake and use abilities of 4 common species. All plant species preferred to take up nitrate (the dominant N form) over ammonium and glycine, and N-acquisition capacity varied with temporal variation of soil N. However, the dominant species was more N-conservative than less-dominant species and had lower overall N uptake rates (shoot 15N excess 48 hours after injection) and higher N use efficiencies (aboveground biomass : N ratio). The different N strategies may reflect mixed effects of environmental filtering and interspecific competition and have significant implications for species coexistence as well as for ecosystem functions such as nutrient cycling.

Effects of walnut trees on biological nitrogen fixation and yield of intercropped alfalfa in a Mediterranean agroforestry system

While intercropping annual non nitrogen-fixing crops with deciduous hardwood species is now well documented, there is a need to investigate if nitrogen-fixing intercrops may succeed in agroforestry systems. Intercropping with trees usually leads to a decline in crop yield, and could in addition possibly reduce the biological N fixation (BNF) over time due to the competition for resources. In a Mediterranean experimental site, 17 year-old hybrid walnut trees (Juglans nigra x Juglans regia L.) planted in East-West oriented lines were intercropped with alfalfa (Medicago sativa L.) to assess the impact of competition for light and water on alfalfa yield and BNF. Alfalfa yield and shoot δ15N values (a proxy for the proportion of N derived from the air, %Ndfa) were measured during one year at different distances from the tree row in two directions (north and south). Alfalfa yield was reduced close to the tree row (−28% and −22% on the northern and southern sides respectively), but less than the reduction of irradiation (−59% and −33% respectively). Shading improved by 35% the apparent light use efficiency (LUE: aboveground biomass produced per unit of global radiation) of alfalfa, indicating that alfalfa was shade tolerant at this Mediterranean site. Alfalfa shoot δ15N values were lower close to the tree rows than at mid inter-row: BNF was stimulated close to the trees. Compensative and facilitative mechanisms between trees and alfalfa plants led to a rise in LUE and%Ndfa in shaded areas. These results contradict the frequent assumption that N fixation is reduced in the shade of trees. Appropriate tree canopy management may help maintain light competition between trees and alfalfa to a level that still enhance complementary, which would further improve the sustainability of the use of alfalfa as an intercrop in Mediterranean regions.

Are seagrass beds indicators of anthropogenic nutrient stress in the rocky intertidal?

It is well established that anthropogenic nutrient inputs harm estuarine seagrasses, but the influence of nutrients in rocky intertidal ecosystems is less clear. In this study, we investigated the effect of anthropogenic nutrient loading on Phyllospadix spp., a rocky intertidal seagrass, at local and regional scales. At sites along California, Washington, and Oregon, we demonstrated a significant, negative correlation of urban development and Phyllospadix bed thickness. These results were echoed locally along an urban gradient on the central California coast, where Phyllospadix shoot δ15N was negatively associated with Phyllospadix bed thickness, and experimentally, where nutrient additions in mesocosms reduced Phyllospadix shoot formation and increased epiphytic cover on Phyllospadix shoots. These findings provide evidence that coastal development can threaten rocky intertidal seagrasses through increased epiphytism. Considering that seagrasses provide vital ecosystem services, mitigating eutrophication and other factors associated with development in the rocky intertidal coastal zone should be a management priority.

Interspecific variation in leaf–root differences in δ15N among three tree species grown with either nitrate or ammonium

Interspecific variation in nitrogen isotope composition of roots and leaves of tree seedlings grown in a steady-state nitrogen environment reflects known variation in sites of assimilation and nitrogen source preference in three tree species. As a time-integrated measure of nitrogen use, discrimination against the heavier stable isotope (15N) during the uptake and assimilation of inorganic nitrogen has the potential to provide information on interspecific differences in inorganic nitrogen source preference. Here, nitrogen isotope composition (δ15N) at natural abundance was measured for the roots and shoots from seedlings of three forest tree species: Populus tremuloides (aspen), Pinus contorta var. latifolia (pine) and Picea glauca (spruce). The seedlings were grown hydroponically with low (0.1 mM) or high (1.5 mM) concentrations of NO3 − or NH4 +, or in sand with NO3 −, NH4 + or an equal mix of NO3 − and NH4 + (0.1 mM). Whole-plant nitrogen isotope discrimination was observed in hydroponically grown seedlings but not in sand culture. Differences in δ15N between shoots and roots were greater in aspen when grown with NO3 − (3.02 ‰) than with NH4 + (1.27 ‰). There were no significant differences between the δ15N of leaves and roots for pine and spruce on either source. Although whole-plant nitrogen isotope discrimination was not observed in sand culture, shoot δ15N was, again, greater than root δ15N for NO3 −-grown aspen. Interspecific variation in nitrogen isotope discrimination was observed in both hydroponics and sand culture. The differences in nitrogen isotope composition under steady-state conditions indicate that interspecific differences in nitrogen source preference are consistent with previous experiments using alternative methods to identify differences in nitrogen uptake and assimilation in the same tree species.

Evaluating N2 fixation by food grain legumes in farmers’ fields in three agro-ecological zones of Zambia, using 15N natural abundance

A survey of N2 fixation in farmers’ fields of Northern (>1,000 mm rainfall), Central (800–1,000 mm rainfall), and Southern (<800 mm rainfall) Zambia revealed some significant differences in plant growth and symbiotic performance of different food grain legumes. Of the three grain legumes (i.e., Bambara groundnut (Vigna subterranea L. Verdc.), cowpea (Vigna unguiculata L. Walp.), and groundnut (Arachis hypogaea L.)) grown in Southern Zambia, cowpea showed greater shoot biomass and significantly lower shoot δ15N values than groundnut and Bambara groundnut. The lower shoot δ15N resulted in greater %Ndfa (59%) in shoots and higher amounts of N-fixed, whether per square meters (6,394.0 mg N), per plant (650.8 mg N), or per hectare (63.9 kg N) relative to groundnut and Bambara groundnut, even though the number of cowpea plants per square meter was significantly lower than that of groundnut or Bambara groundnut. Although the shoot δ15N values of cowpea, Bambara groundnut and common bean (Phaseolus vulgaris L.) were significantly lower than those of groundnut in Central Zambia and their %Ndfa values, therefore, greater, the higher number of groundnut plants per square meter resulted in significantly greater shoot N content, as well as N-fixed per square meter and per hectare relative to the other species. Despite having similar plant density as cowpea in Central Zambia, common bean could fix only 6.0 kg N ha−1 compared with 35.4 kg N ha−1 by cowpea. In Northern Zambia, Bambara groundnut showed the lowest mean shoot δ15N value (0.54 ± 0.3‰), followed by groundnut (1.59 ± 1.0‰), and then common bean (the three grain legumes grown in that region). As a result, %Ndfa and N-fixed were significantly greater in groundnut (69.7% and 566.0 mg N per plant) and Bambara groundnut (62.9% and 440.1 mg N per plant) than in common bean (2.6% and 2.4 mg N per plant). In Northern Zambia, groundnut, Bambara groundnut and common bean fixed 78.7, 67.6, and 0.9 kg N ha−1, respectively, even though the plant density per square meter of common bean (which fixed the lowest amount of N per hectare) was twice that of groundnut and Bambara groundnut. A species × site analysis showed that cowpea fixed relatively greater amounts of N per plant, per square meter, and per hectare in Southern than Central Zambia. Bambara groundnut and common bean also had significantly lower δ15N values and higher %Ndfa in Central than Northern Zambia.

Symbiotic N2 fixation in 30 field-grown cowpea (Vigna unguiculata L. Walp.) genotypes in the Upper West Region of Ghana measured using 15N natural abundance

In this study, 30 cowpea genotypes were assessed for symbiotic N2 fixation in 2005, and 15 of them were re-evaluated in 2006 using the 15N natural abundance technique. Shoot dry matter yield of cowpea genotypes increased significantly in cvs. Vuli-1, Glenda, IT93K-2045-29, IT90K-59, Omondaw, Apagbaala, and IT84S-2246 in 2005 producing about 3.0 to 3.6-fold more biomass relative to cv. Vallenga. In 2006, seven out of the 15 cowpea genotypes tested (namely, IT97K-499-39, TVu11424, Botswana White, IT84S-2246, Sanzie, Brown Eye, and Glenda) also produced more dry matter than cv. CH14. Shoot δ15N values ranged from −0.58‰ to 1.49‰ in 2005, and −1.51‰ to 1.40‰ in 2006, and these resulted in %Ndfa values of 63.5–86.7% and 56.2–96.3%, respectively. The amount of N-fixed was 49–178 kg N ha−1 in 2005 and 62–198 kg N ha−1 in 2006. Furthermore, there was a direct relationship between the level of symbiotic N nutrition and plant growth, and between grain yield and amount of N-fixed in 2005 and 2006. As a result, genotypes that fixed the most N also produced the largest biomass and the greatest amount of grain yield. The observed relationship between N2 fixation and biomass confirmed our view that cowpea (and other grain legumes) can be concurrently selected for higher N2 fixation, superior plant growth, and greater grain yield. The high levels of N-fixed by many of the cowpea genotypes in this study suggest that they can contribute large amounts of N to cropping systems in African agriculture.

Shoot δ15N gives a better indication than ion concentration or Δ13C of genotypic differences in the response of durum wheat to salinity.

We compared the performance of different physiological traits that reveal genotypic variations in tolerance to salinity in durum wheat. A set of 114 genotypes was grown in hydroponics for over 3 months. Three conditions: control, moderate (12 dS m-1) and severe (17 dS m-1) salinity, were maintained for nearly 8 weeks before harvest. The genotype biomass in control conditions correlated with the biomass at the two salinity levels. Subsequently, two subsets of 10 genotypes each were selected on the basis of extreme differences in biomass at the two salinity levels while showing relatively similar biomass in control conditions. Carbon isotope discrimination (Δ13C), nitrogen isotope composition (δ15N), and the concentration of nitrogen, phosphorus and several ions (K+, Na+, Ca2+, Mg2+) were analysed in the two subsets for the three treatments. At 12 dS m-1, K+ concentration, K+/Na+ ratio, Δ13C and δ15N correlated positively and Na+ correlated negatively with shoot biomass. Under control conditions and at 17 dS m-1 no correlation was observed. However, the trait that correlated best with genotypic differences in biomass was δ15N at 12 dS m-1. This trait was the first variable chosen at each of the two salinity levels in a stepwise analysis. We consider the possible mechanisms relating δ15N to biomass and the use of this isotopic signature as a selection trait.

Plant isotopic composition provides insight into mechanisms underlying growth stimulation by AM fungi in a semiarid environment.

We hypothesised that improved plant water status and enhanced transpiration are key mechanisms involved in plant growth stimulation by native arbuscular mycorrhizal fungi (AMF) in semiarid calcareous soils. Seedlings of the dryland shrubs Pistacia lentiscus L. and Retama sphaerocarpa L. were pre-inoculated with a mixture of eight native Glomus spp. fungi, or left un-inoculated, before transplanting into a degraded site in south-eastern Spain. Pre-inoculated Pistacia and Retama shrubs grew faster after transplanting, despite spontaneous colonisation of control plants by local AMF. Pre-inoculation enhanced shoot water content and shoot δ15N in both shrub species. Increased potassium uptake and improved water relations were key mechanisms behind growth stimulation by native AMF in Pistacia. Shoot δ18O (a proxy measure of stomatal conductance) was significantly less negative in AMF-inoculated than in control Pistacia seedlings, indicating enhanced cumulative transpiration in the former. In contrast, shoot δ18O was unaffected by AMF inoculation in Retama, a leafless leguminous shrub with photosynthetic stems. Growth stimulation by native AMF in Retama was attributed to increased phosphorus uptake, enhanced atmospheric nitrogen fixation and a largely nutrient-mediated improvement of plant water status. Shoot δ13C was not significantly influenced by AMF inoculation in either shrub species, thus suggesting roughly parallel upshifts in photosynthetic and transpiration rates which did not affect plant water use efficiency.

Shoot δ15N and δ13C values of non-host Brassica rapa change when exposed to ±Glomus etunicatum inoculum and three levels of phosphorus and nitrogen

Glasshouse experiments were conducted to study the response of non-host Brassica rapa and host Sorghum bicolor to inoculation with the arbuscular mycorrhizal fungus (AMF) Glomus etunicatum when given different levels of N (0.9mmol kg(-1) sand, 2.7mmol kg(-1) sand, 8.1mmol kg(-1) sand) and P (3.6µmol kg(-1) sand, 10.7µmol kg(-1) sand, 32.0µmol kg(-1) sand) fertiliser. On both plant species, the presence of G. etunicatum inoculum (+AMF) was associated with significant changes of shoot δ(15)N values, with +AMF plants having larger average δ(15)N values than uninoculated plants (-AMF). These values are the largest average differences in shoot δ(15)N yet recorded for AMF and nutrient effects. B. rapa shoot δ(15)N average differences ranged from 1.67‰ to 2.70‰, while for S. bicolor they range between 2.07‰ and 4.40‰. For shoot δ(13)C only the non-host B. rapa responded to ±AMF and added N. Although the harvested dry weight biomass (-35.2% B. rapa; +39.8% S. bicolor) of both plant species responded to AMF inoculation, no direct relationship was observed between isotopic discrimination and growth inhibition for the non-host B. rapa. In this paper we discuss some implications regarding AMF inocula on the basis of our findings and current literature.