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1.

Background and aims

Crop residues and soil types play an important role in soil C and N storage. The objectives of this study were to quantify the effects of crop residue quality and interactions with soil type on soil C and N, in the short- and medium-term, and to determine the responses related to the priming effect (PE).

Methods

Residues of vetch (Vicia sativa L.), pea (Pisum sativum L.) and wheat (Triticum aestivum L.) crops with different chemical compositions and labelled with 13C and 15N were left to decompose on the surface of either a sandy-loam soil or a clay soil incubated under laboratory conditions at 25 °C for 360 days. We measured the total CO2-C and CO2-13C emitted during decomposition, the soil mineral N content and the amounts of 13C and 15N remaining in both the surface residue particles and the bulk soil.

Results

Over the short-term, the vetch residues decomposed faster than those of wheat and pea on the soil surface due to their more favourable chemical composition for biodegradation; after one year, however, this difference disappeared. We observed extra soil C mineralization in all cases, i.e., the PE was positive for all treatments and was directly related to the water-soluble (vetch > pea > wheat) and soil C contents (clay soil > sandy-loam soil). Conversely, the fate of the added 15N and net N mineralization differed considerably between the three residues and was strongly related to the initial N content of the residue.

Conclusions

Crop residue quality and soil type affected the soil PE and soil C balance but not the fate of crop residue-C after one year. Net soil N mineralization was observed in all crop residues, with large early differences (vetch > pea > wheat), which were maintained on a medium-term basis. Our results emphasize the need to jointly consider C and N dynamics as well as short- and medium-term effects to manage agricultural and environmental services provided by the recycling of crop residues to agricultural soils.
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2.

Introduction

Root-mediated changes in soil organic matter (SOM) decomposition, termed rhizosphere priming effects (RPE), play crucial roles in the global carbon (C) cycle, but their mechanisms and field relevance remain ambiguous. We hypothesize that nitrogen (N) shortages may intensify SOM decomposition in the rhizosphere because of increase of fine roots and rhizodeposition.

Methods

RPE and their dependence on N-fertilization were studied using a C3-to-C4 vegetation change. N-fertilized and unfertilized soil cores, with and without maize, were incubated in the field for 50 days. Soil CO2 efflux was measured, partitioned for SOM- and root-derived CO2, and RPE was calculated. Plant biomass, microbial biomass C (MBC) and N (MBN), and enzyme activities (β-1,4-glucosidase; N-acetylglucosaminidase; L-leucine aminopeptidase) were analyzed.

Results

Roots enhanced SOM mineralization by 35 % and 126 % with and without N, respectively. This was accompanied by higher specific root-derived CO2 in unfertilized soils. MBC, MBN and enzyme activities increased in planted soils, indicating microbial activation, causing positive RPE. N-fertilization had minor effects on MBC and MBN, but it reduced β-1,4-glucosidase and L-leucine aminopeptidase activities under maize through lower root-exudation. In contrast, N-acetylglucosaminidase activity increased with N-fertilization in planted and unplanted soils.

Conclusions

This study showed the field relevance of RPE and confirmed that, despite higher root biomass, N availability reduces RPE by lowering root and microbial activity.
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3.

Introduction

The aphid Rhopalosiphum padi L. is a vector of Barley yellow dwarf virus (BYDV) in wheat and other economically important cereal crops. Increased atmospheric CO2 has been shown to alter plant growth and metabolism, enhancing BYDV disease in wheat. However, the biochemical influences on aphid metabolism are not known.

Objectives

This work aims to determine whether altered host-plant quality, influenced by virus infection and elevated CO2, impacts aphid weight and metabolism.

Methods

Untargeted 1H NMR metabolomics coupled with multivariate statistics were employed to profile the metabolism of R. padi reared on virus-infected and non-infected (sham-inoculated) wheat grown under ambient CO2 (aCO2, 400 µmol mol?1) and future, predicted elevated CO2 (eCO2, 650 µmol mol?1) concentrations. Un-colonised wheat was also profiled to observe changes to host-plant quality (i.e., amino acids and sugars).

Results

The direct impacts of virus or eCO2 were compared. Virus presence increased aphid weight under aCO2 but decreased weight under eCO2; whilst eCO2 increased non-viruliferous (sham) aphid weight but decreased viruliferous aphid weight. Discriminatory metabolites due to eCO2 were succinate and sucrose (in sham wheat), glucose, choline and betaine (in infected wheat), and threonine, lactate, alanine, GABA, glutamine, glutamate and asparagine (in aphids), irrespective of virus presence. Discriminatory metabolites due to virus presence were alanine, GABA, succinate and betaine (in wheat) and threonine and lactate (in aphids), irrespective of CO2 treatment.

Conclusion

This study confirms that virus and eCO2 alter host-plant quality, and these differences are reflected by aphid weight and metabolism.
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4.

Background and aims

Variations in root-associated fungal communities contribute to the so-called ‘crop rotation benefit’ on soil productivity. We assessed the effects of chickpea, lentil, and pea in wheat-based rotations, as compared to wheat monoculture, on the structure of root-associated fungal communities, and described the legacy of pulses on a following wheat crop.

Methods

The internal transcribed spacer (ITS) and 18S rRNA gene markers, and 454 amplicon pyrosequencing were used to describe the fungal communities of crop roots and rhizosphere soil in a field experiment and agronomic data were collected.

Results

Pulses influenced only the structure of the non-mycorrhizal fungal community of roots. Fusarium tricinctum, Clonostachys rosea, Fusarium redolens, and Cryptococcus sp. were specific to certain crops. Despite the absence of selective effects of pulses on their associated arbuscular mycorrhizal (AM) fungal community, pea had a legacy effect on the structure of the AM fungal community associated with the roots of the following wheat crop, in one of the two year/sites examined. Species of Mortierella, Cryptococcus, and Paraglomus in wheat rhizosphere soil may benefit yield, whereas species of Fusarium, Davidiella, Lachnum, Sistotrema and Podospora may reduce yield.

Conclusion

The effect of pulse crops on root fungal communities varied with rotation crop species. Pulses had various effects on the physiology of the following wheat crop, including increased productivity.
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5.

Background and aims

Positive below-ground interactions (facilitation) should be more pronounced when resources limit crop growth, according to the stress-gradient hypothesis. Our aim was to test this hypothesis for intercropped durum wheat and faba bean along a P-fertilizer gradient.

Methods

A field experiment was conducted in a long-term P-fertilizer trial with three rates of P-fertilization (No, Low and High P). Microbial biomass was assessed by chloroform fumigation-extraction. Quantitative PCR was applied to evaluate the abundance of relevant microbial groups.

Results

Phosphorus availability and microbial biomass systematically increased in the rhizosphere compared to bulk soil. P-fertilization resulted in higher abundance of targeted bacterial phyla, whole bacterial and fungal communities, and depressed mycorrhizal colonization of durum wheat, but not faba bean. Microbial biomass carbon significantly increased in the rhizosphere only in P-fertilized treatments, pointing to P limitation of microbial communities. Intercropping yielded a significant effect on rhizosphere microbial properties only at High P. Microbial biomass P increased in the rhizosphere of intercropped faba bean only at No P level, and was thus the sole finding supporting the stress-gradient hypothesis.

Conclusions

P-fertilization was the main driver of microbial communities in this field trial, and P-fertilizer application modulated the species-specific effect in the intercrop. Plant performance did not validate the stress-gradient hypothesis as positive plant-plant interactions occurred regardless of the level of P-fertilization.
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6.

Background and aims

In Malawi, strategies are being sought to boost maize production through improvements in soil fertility. This study assessed the impact of intercropping maize (Zea mays) with pigeon pea (Cajanus cajan) in Lixisols of Malawi on yield, biological N fixation, soil aggregation, and P forms within soil aggregates.

Methods

Maize and pigeon pea were grown intercropped in pots, with varying degrees of root interaction in order to understand the relative importance of biochemical versus physical rhizospheric interactions. Following harvest, soils were separated into aggregate fractions using wet-sieving, and the nutrient content of all fractions was assessed.

Results

The proportion of macroaggregates and microaggregates increased by 52 and 111%, respectively, in the intercropping treatment compared to sole maize, which significantly increased organic P storage in the microaggregates of intercropped compared to sole maize (84 versus 29 mg P kg?1, respectively). Biologically fixed N increased from 89% in the sole pigeon pea to 96% in the intercropped system.

Conclusions

Intercropping maize with pigeon pea can have a significant and positive impact on soil structure as well as nutrient storage in these high P-sorbing soils. This is caused primarily by physical root contact and to a lesser degree by biochemical activities.
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7.

Aims

A better understanding of how plant growth, N nutrition and symbiotic nitrogen fixation (SNF) are influenced by soil inorganic N availability, for a wide range of legume species, is crucial to optimise legume productivity, N2 fixation, while limiting environmental risks such as N leaching.

Methods

A comparative analysis was performed for ten legume crops, grown in a field experiment and supplied with four N fertiliser rates. Dry matter, N concentration and SNF were measured. In parallel, root elongation rates were studied in a greenhouse experiment.

Results

For most species, N fertilisation had little effect on plant growth and N accumulation. SNF was reduced by soil inorganic N available at sowing but with large differences in the magnitude of the response among species. The response varied according to plant N requirements for growth and plant ability to retrieve inorganic N. Accordingly, root lateral expansion rate measured in RhizoTubes was highly correlated with plant ability to retrieve inorganic N measured in the field experiment.

Conclusion

Combining SNF response to soil inorganic N, shoot N and plant ability to retrieve inorganic N, allowed a robust evaluation of differential response to soil inorganic N among a wide range of legume species.
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8.

Aims

The study aimed to find soil parameters that are best related to Se plant uptake for low Se soils with predominantly organic Se, and to explore the mechanisms that control Se bioavailability in the soils under study.

Methods

A pot experiment using nineteen soil samples taken from different fields of arable land (potato fields) in the Netherlands was conducted on summer wheat (Triticum aestivum L.). Selenium in wheat shoots and soil parameters, including basic soil properties, C:N ratio, inorganic selenite content, and Se and organic C in different soil extractions (0.01 M CaCl2, 0.43 M HNO3, hot water, ammonium oxalate, aqua regia) were analysed. Regression analysis was performed to identify soil parameters that determine Se content in wheat shoots.

Results

The regression model shows that Se:DOC ratio in 0.01 M CaCl2 soil extraction explained about 88 % of the variability of Se uptake in wheat shoots. Selenium uptake increased with Se:DOC ratio in CaCl2 extraction, which can be interpreted as a measure of the content of soluble Se-rich organic molecules. Selenium:DOC ratio in CaCl2 extraction and Se uptake increased towards higher soil pH and lower soil C:N ratio. The soil C:N ratio is also negatively correlated to Se:organic C ratio in other extractions (0.43 M HNO3, hot water, ammonium oxalate, aqua regia), indicating that at low soil C:N ratio soil organic matter is richer in Se. Contrarily, the soil pH is positively and strongly correlated to Se:organic C ratio in CaCl2 and hot water extractions, but only weakly correlated to Se:organic C ratio in other extractions.

Conclusions

Selenium-rich dissolved organic matter is the source of bioavailable Se in low Se soils with predominantly organic Se. The soil pH and quality of soil organic matter (i.e. soil C:N ratio) are the main soil properties determining Se bioavailability in these soil types.
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9.

Aim

To investigate the effects of biochar on biological and chemical phosphorus (P) processes and identify potential interactive effects between P fertilizer and biochar on P bioavailability in the rhizosphere of maize.

Methods

We conducted a pot-experiment with maize in a sandy loam soil with two fertilizer levels (0 and 100 mg P kg ?1) and three biochars produced from soft wood (SW), rice husk (RH) and oil seed rape (OSR). Sequential P fractionation was performed on biochar, bulk soil, and rhizosphere soil samples. Acid and alkaline phosphatase activity and root exudates of citrate, glucose, fructose, and sucrose in the rhizosphere were determined.

Results

RH and OSR increased readily available soil P, whereas SW had no effect. However, over time available P from the biochars moved to less available P pools (Al-P and Fe-P). There were no interactive effects between P fertilizer and biochar on P bioavailability. Exudates of glucose and fructose were strongly affected by especially RH, whereas sucrose was mostly affected by P fertilizer. Alkaline phosphatase activity was positively correlated with pH, and citrate was positively correlated with readily available P.

Conclusion

Biochar effects on biological and chemical P processes in the rhizosphere are driven by biochar properties.
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10.

Background and aims

Symbiotic N2 fixation is essential in the development of sustainable agriculture, but the nodulation of legumes is usually inhibited by N fertilization. Here, the intercropping of maize and pea in strips under various N managements was used as a means to alleviate the inhibitory effect of mineral N on pea nodulation and N2 fixation and to improve system performance.

Methods

N natural abundance (δ 15N) analysis was employed to quantify N2 fixation in the 3 years (2012 to 2014) of field experiment in Hexi Corridor of Northwestern China. Four N management systems with N rate of 0 kg N ha?1 (the control), 90?+?45 kg N ha?1 (base N plus topdressing N), 90?+?90 kg N ha?1, and 90?+?135 kg N ha?1 were implemented in the maize/pea strip intercropping to form different ratios of base N to topdressing N.

Results

Intercropped pea improved nodule biomass per plant by 99 %, increased nitrogen derived from the atmosphere (Ndfa) by 35 %, and promoted aboveground plant tissue N accumulation by 35 % as compared with sole pea, averaged across the four N treatments. Compared to the highest N fertilizer treatment, a reduction of topdressing to 45 kg N ha?1 increased the nodule biomass of intercropped pea by 116 %, Ndfa by 35 %, and grain yield by 6 %.

Conclusions

Adaptation of suitable N management in cereal/legume intercropping systems will allow an effective conversion of atmospheric N2 into crop available N and thus maximizing the system productivity.
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11.

Aims

The aim of our study was to clarify whether inoculating a soil with Pseudomonas sp. RU47 (RU47) bacteria would stimulate the enzymatic cleavage of organic P compounds in the rhizosphere and bulk soil, promoting plant growth. Adding either viable or heat treated RU47 cells made it possible to separate direct from indirect effects of the inoculum on P cycling in soil and plants.

Methods

We performed a rhizobox experiment in the greenhouse with tomato plants (Solanum lycopersicum) under low P soil conditions. Three inoculation treatments were conducted, using unselectively grown soil bacteria (bacterial mix), heat treated (HT-RU47) and viable RU47 (RU47) cells, and one not inoculated, optimally P-fertilized treatment. We verified plant growth, nutrient availability, enzyme activities and microbial community structure in soil.

Results

A plant growth promotion effect with improved P uptake was observed in both RU47 treatments. Inoculations of RU47 cells increased microbial phosphatase activity (PA) in the rhizosphere.

Conclusions

Plant growth promotion by RU47 cells is primarily associated with increased microbial PA in soil, while promotion of indigenous Pseudomonads as well as phytohormonal effects appear to be the dominant mechanisms when adding HT-RU47 cells. Thus, using RU47 offers a promising approach for more efficient P fertilization in agriculture.
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12.

Background and Aims

Litter decomposition serves an important role in maintaining nitrogen (N) availability within forest ecosystems. However, the interactive effects of exogenous N, drought, and litter quality and mixing on N immobilization during decomposition remain unclear. The aim of this study was to assess the effects of litter quality, reduced precipitation, N addition, and their interactions on litter mass loss and N immobilization.

Methods

This field study analyzed the effects of N addition and decreased precipitation on the decomposition rates and associated N immobilization of four types of litter: Quercus mongolica (QM), Tilia amurensis (TA), Pinus koraiensis (PK), and a mixture (MIX) of all three. The chemical quality of the MIX was prepared in a 4:3:3 (mass) ratio of PK, TA, and QM litters. Litterbags were placed in an N addition and precipitation manipulation forest field and collected after 92, 154, 365, 457, and 874 days. Decomposing litter residues were characterized for mass loss and N content to assess N immobilization.

Results

The addition of N had no significant effect on litter decomposition under both precipitation conditions, but a reduction in precipitation significantly depressed litter decomposition. The increases in N immobilization with N addition depended on the litter type and decomposition period. Precipitation reduction had significant effects on N immobilization and enhanced the magnitude and duration of N immobilization in decomposing litter, and both of which can be increased by N addition. The results indicate that the litter species is the major regulator that controls mass loss and N immobilization. Furthermore, the MIX treatment did not show non-additive effects on mass loss but did exhibit some weak synergistic effects on N immobilization.

Conclusions

Our results suggest that decomposing litters could help to sequester N depending on the litter identity and water regime in temperate forest ecosystems.
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13.

Aims

Our goals were (1) to determine whether tree species diversity affects nutrient (N, P and K) cycling, and (2) to assess whether there is competition for these nutrients between microbial biomass and trees.

Methods

We measured nutrient resorption efficiency by trees, nutrient contents in leaf litterfall, decomposition rates of leaf litter, nutrient turnover in decomposing leaf litter, and plant-available nutrients in the soil in mono-species stands of beech, oak, hornbeam and lime and in mixed-species stands, each consisting of three of these species.

Results

Cycling of nutrients through leaf litter input and decomposition were influenced by the types of tree species and not simply by tree species diversity. Trees and microbial biomass were competing strongly for P, less for K and only marginally for N. Such competition was most pronounced in mono-species stands of beech and oak, which had low nutrient turnover in their slow decomposing leaf litter, and less in mono-species stands of hornbeam and lime, which had high nutrient turnover in their fast decomposing leaf litter.

Conclusions

The low soil P and K availability in beech stands, which limit the growth of beech at Hainich, Germany, were alleviated by mixing beech with hornbeam and lime. These species-specific effects on nutrient cycling and soil nutrient availability can aid forest management in improving productivity and soil fertility.
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14.

Background

Zinc (Zn) deficiency is one of the most important micronutrient disorders affecting human health. Wheat is the staple food for 35% of the world’s population and is inherently low in Zn, which increases the incidence of Zn deficiency in humans. Major wheat-based cropping systems viz. rice–wheat, cotton–wheat and maize–wheat are prone to Zn deficiency due to the high Zn demand of these crops.

Methods

This review highlights the role of Zn in plant biology and its effect on wheat-based cropping systems. Agronomic, breeding and molecular approaches to improve Zn nutrition and biofortification of wheat grain are discussed.

Results

Zinc is most often applied to crops through soil and foliar methods. The application of Zn through seed treatments has improved grain yield and grain Zn status in wheat. In cropping systems where legumes are cultivated in rotation with wheat, microorganisms can improve the available Zn pool in soil for the wheat crop. Breeding and molecular approaches have been used to develop wheat genotypes with high grain Zn density.

Conclusions

Options for improving grain yield and grain Zn concentration in wheat include screening wheat genotypes for higher root Zn uptake and grain translocation efficiency, the inclusion of these Zn-efficient genotypes in breeding programs, and Zn fertilization through soil, foliar and seed treatments.
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15.

Background and aims

Given the worldwide effort to improve the nitrogen (N) economy of crops, it is critical to understand the mechanisms of improved N uptake which have resulted from selection pressure for grain yield in Australian wheat (Triticum aestivum L.). Changes in root system traits and N uptake were examined in nine Australian wheat varieties released between 1958 and 2007.

Methods

Wheat varieties were grown in rhizo-boxes in a glasshouse. We measured nitrogen uptake and mapped root growth and proliferation to quantify root length density (RLD), root length per plant, root biomass, specific root length, and plant nitrogen uptake per unit root length.

Results

Selection for yield reduced total RLD and total root length, and increased N uptake per unit root length that overrode the reduction in root system size, effectively explaining the increase in N uptake. Importantly, N uptake in our experiment under controlled conditions matched field measurements, reinforcing the agronomic significance of the present study.

Conclusions

Wheat varieties released in Australia between 1958 and 2007 increased their N uptake, not because of increasing their root length and RLD, but for progressively increasing the efficiency of their root system in capturing N. Our collection of varieties is therefore an interesting model to probe for variation in the affinity of the root system for nitrate.
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16.

Aims

Understanding the linkage of soil respiration (Rs) with forest development is essential for long-term C cycle models. We estimated the variation and temperature sensitivity (Q10 value) of Rs and its hetero-, (Rh) and autotrophic (Ra) components in relation to abiotic and biotic factors in Norway spruce stands of different ages, and the effect of trenching on microbial and soil characteristics.

Methods

Trenching method was used to partition Rs into Rh and Ra. Ingrowth core method was used to estimate fine root production. Soil microbial biomass was measured using manometric respirometers.

Results

Rs varied in differently aged stands demonstrating non-linear response to development stage. The variation of Rs was explained by changes in biotic factors rather than by changes in soil microclimate. Rh was more sensitive to Ts than Rs or Ra. After 4 years of trenching soil pH, N, SOM and dehydrogenase activity were significantly changed in trenched plots compared to control plots.

Conclusions

Different Q10 values of Rh and Ra in stands of different ages indicate the importance of Rs partitioning. Trenching should be used during a limited number of years because of the possible changes in chemical characteristics of soil and in the activity of soil microbial community.
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17.

Background and aims

Soil nutrient dynamics are affected by root-microbe interactions and plant development. We investigated the influence of plant growth stage and arbuscular mycorrhiza fungi (AMF) on carbon (C) and nitrogen (N) rhizodeposition and the transfer into the microbial biomass (MB).

Methods

Pea varieties (Pisum sativum L.) with (Frisson) and without mycorrhiza (P2) were 13C-15N-labelled and harvested at 45, 63, 71, and 95 days after sowing. Mycorrhization, MB, total C, N, 13C, 15N were determined in plant and soil compartments to calculate C and N derived from rhizodeposition (CdfR, NdfR).

Results

Total CdfR increased until pea maturity, NdfR until end of flowering. Their relative contribution steadily decreased over time, accounting for 4–10% of total plant C and N at harvest. Rhizodeposition contributed between 1 and 6% to MB C and N, although 20% of the rhizodeposits were discovered in the MB. Frisson released more NdfR than P2 but it was not possible to accurately estimate AMF effects on C and N due to differences in biomass partitioning.

Conclusions

CdfR followed an even flow from early growth until senescence. NdfR flow ceased after flowering possibly due to N relocation within the plant. Rhizodeposits contribute very little to MB in our study.
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18.

Aims

The objective of this study was to investigate how plants maintain productivity under a limited supply of water and N along the topographical soil water and N gradients in semi-arid forests.

Methods

We investigated forest structure and productivity, N cycling, and water and N use by plants at three different slope positions in a forested area near an arid boundary on a loess plateau in China.

Results

Net primary production (NPP) and aboveground N uptake decreased as soil water and/or N availability decreased on upper slopes; however, NPP and aboveground N uptake were only slightly lower than those of more humid forest ecosystems. Water use efficiency (WUE), N use efficiency (NUE), and fine root biomass increased as soil water and/or N supply decreased with altitude. High NUE was linked to higher N mean residence time, caused by higher N resorption efficiency rather than increasing N productivity.

Conclusions

Our results suggest that NPP and N uptake can be maintained by increasing WUE and NUE and increasing fine root biomass in water and N co-limited semi-arid forest ecosystems near arid boundaries. Such changes in resource use and acquisition strategy can affect production and N cycling via plant-soil feedback systems.
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19.

Aims

Hydro-biogeochemical processes in the rhizosphere regulate nutrient and water availability, and thus ecosystem productivity. We hypothesized that two such processes often neglected in rhizosphere models — diel plant water use and competitive cation exchange — could interact to enhance availability of K+ and NH4 +, both high-demand nutrients.

Methods

A rhizosphere model with competitive cation exchange was used to investigate how diel plant water use (i.e., daytime transpiration coupled with no nighttime water use, with nighttime root water release, and with nighttime transpiration) affects competitive ion interactions and availability of K+ and NH4 +.

Results

Competitive cation exchange enabled low-demand cations that accumulate against roots (Ca2+, Mg2+, Na+) to desorb NH4 + and K+ from soil, generating non-monotonic dissolved concentration profiles (i.e. ‘hotspots’ 0.1–1 cm from the root). Cation accumulation and competitive desorption increased with net root water uptake. Daytime transpiration rate controlled diel variation in NH4 + and K+ aqueous mass, nighttime water use controlled spatial locations of ‘hotspots’, and day-to-night differences in water use controlled diel differences in ‘hotspot’ concentrations.

Conclusions

Diel plant water use and competitive cation exchange enhanced NH4 + and K+ availability and influenced rhizosphere concentration dynamics. Demonstrated responses have implications for understanding rhizosphere nutrient cycling and plant nutrient uptake.
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20.

Background

Nitrous oxide (N2O), a long-standing anesthetic, is also neurotoxic by interfering with the bioavailability of vitamin B12 if abused. A few case studies have reported the neurological and psychiatric complications of N2O.

Case presentation

Here, we reported a patient of N2O induced subacute combined degeneration (SCD) with longitudinally extensive myelopathy with inverted V-sign exhibiting progressive limb paresthesia and unsteady gait.

Conclusions

This case raises the awareness of an important mechanism of neural toxicity of N2O, and clinical physicians should be well recognized this in the field of substance-related disorders.
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