Cover crop root morphology rather than quality controls the fate of root and rhizodeposition C into distinct soil C pools.

Cover crops increase carbon (C) inputs to agricultural soils, and thus have the potential to mitigate climate change through enhanced soil organic carbon (SOC) storage. However, few studies have explored the fate of belowground C inputs associated with varying root traits into the distinct SOC pools of mineral-associated organic carbon (MAOC) particulate organic carbon (POC). Therefore, a packed 0.5 m column trial was established with 0.25 m topsoil and 0.25 m subsoil with four cover crops species (winter rye, oilseed radish, chicory, and hairy vetch) known to differ in C:N ratio and root morphology. Cover crops were 14 CO2 -labeled for 3 months, and then, half of the columns were sampled to quantify root and rhizodeposition C. In the remaining columns, plant shoots were harvested and the undisturbed soil and roots were left for incubation. Bulk soil from both sampling times was subjected to a simple fractionation scheme, where 14 C in the <50 and >50 µm fraction was assumed to represent MAOC and POC, respectively. The fast-growing rye and radish produced the highest root C. The percentage loss of C via rhizodeposition (%ClvR) showed a distinct pattern, with 22% for the more branched roots (rye and vetch) and 6%-8% for the less branched roots (radish and chicory). This suggests that root morphology plays a key role in determining rhizodeposition C. After 1 year of incubation at room temperature, the remaining MAOC and POC were positively correlated with belowground inputs in absolute terms. However, topsoil MAOC formation efficiencies (cover crop-derived MAOC remaining as a share of belowground inputs) were higher for vetch and rye (21% and 15%, respectively) than for chicory and radish (9% and 10%, respectively), suggesting a greater importance of rhizodeposition (or indirectly, root morphology) than solely substrate C:N ratio for longer term C stabilization.

Robust Species Distribution Mapping of Crop Mixtures Using Color Images and Convolutional Neural Networks.

Crop mixtures are often beneficial in crop rotations to enhance resource utilization and yield stability. While targeted management, dependent on the local species composition, has the potential to increase the crop value, it comes at a higher expense in terms of field surveys. As fine-grained species distribution mapping of within-field variation is typically unfeasible, the potential of targeted management remains an open research area. In this work, we propose a new method for determining the biomass species composition from high resolution color images using a DeepLabv3+ based convolutional neural network. Data collection has been performed at four separate experimental plot trial sites over three growing seasons. The method is thoroughly evaluated by predicting the biomass composition of different grass clover mixtures using only an image of the canopy. With a relative biomass clover content prediction of R2 = 0.91, we present new state-of-the-art results across the largely varying sites. Combining the algorithm with an all terrain vehicle (ATV)-mounted image acquisition system, we demonstrate a feasible method for robust coverage and species distribution mapping of 225 ha of mixed crops at a median capacity of 17 ha per hour at 173 images per hectare.

ESTABLISHMENT OF ACUTE-PHASE PROTEIN AND SERUM PROTEIN ELECTROPHORESIS PRELIMINARY REFERENCE VALUES FOR PRONGHORN (ANTILOCAPRA AMERICANA).

Pronghorn (Antilocapra americana) are native to western North America and are found in 24 Association of Zoos and Aquariums (AZA)-accredited institutions. Acute-phase proteins (APP) are a broad class of proteins that are stimulated in response to inflammation and have been shown to be a sensitive measure of inflammation in equids and ruminants. In this study, blood samples from clinically normal free-ranging and captive populations of pronghorn were analyzed using assays for protein electrophoresis (EPH) and APP, including serum amyloid A (SAA) and haptoglobin (HP), to develop preliminary ranges to gauge potential differences between these populations. Additional samples were taken from clinically abnormal captive pronghorn with facial abscesses. By EPH measurements, albumin: globulin ratio mean and SE were significantly different (P <0.05) with 1.02 (0.08) for captive populations and 1.91 (0.05) for free-ranging populations. Total protein mean and SE were significantly different (P <0.05) for captive and free-ranging populations, respectively 5.6 (0.3) g/dl and 6.9 (0.1) g/dl. Mean and SD of SAA for captive pronghorn were 1.4 (3.2) mg/L, and were significantly different from the free-ranging population, which was below the limits of detection for (P <0.05). There was no difference in HP levels between these groups. In a case study of a pronghorn with facial abscesses, elevated levels of HP, but not SAA, suggested that HP maybe useful in certain disease states. Future studies should explore the use of these biomarkers as tools to monitor general health, prognosis, and subclinical disease.

Newly depolymerized large organic N contributes directly to amino acid uptake in young maize plants.

The contribution of large molecular size organic nitrogen (N) to plant N uptake is unclear. Soils with and without maize, at three pH levels, were treated with (carbon-14 and -13 (14 C, 13 C), 15 N) triple-labelled > 100 kDa organic N. After 48 h, soil and maize were sampled for bulk and compound specific isotope analysis to study the turnover in soil and plant 13 C and 15 N uptake. Mineralization of > 100 kDa organic N increased with higher pH only in soil without maize. The > 100 kDa organic N disappeared rapidly in soils with and without maize, but surprisingly more > 100 kDa organic N derived amino acids remained in soil with than without maize - most likely in the microbial biomass. Total 15 N uptake in maize increased with higher soil pH. The organic N uptake was estimated to account for 20-30% of the total 15 N uptake. Organic N uptake was confirmed by the presence of 13 C-labelled amino acids in maize roots. The study suggests that the importance of plant organic N uptake increases when N is derived from complex molecules such as proteins compared to studies using single amino acids as N source, and that rhizosphere microorganisms increase anabolic utilization of organic N compared to microorganisms in the bulk soil.

The influence of hydrolysis and derivatization on the determination of amino acid content and isotopic ratios in dual-labeled (13 C, 15 N) white clover.

RATIONALE: The cycling of peptide- and protein-bound amino acids (AAs) is important for studying the rate-limiting steps in soil nitrogen (N) turnover. A strong tool is stable C and N isotopes used in combination with compound-specific isotope analysis (CSIA), where a prerequisite for analysis is appropriate methods for peptide and protein hydrolysis and appropriate methods for derivatization of AAs for analysis by gas chromatography (GC). METHODS: We examined the efficiency of a standard acidic hydrolysis (6 M HCl, 20 h at 110°C) and a fast acidic hydrolysis (6 M HCl, 70 min at 150°C) on the recovery of AAs from a protein standard (bovine serum albumin). The best methods were used on dual-labeled (13 C and 15 N) clover shoot and root juice, divided into four molecular weight (Mw) size fractions. We used NAIP (N-acetyl isopropyl esterification) derivatization for GC/combustion-isotope ratio mass spectrometry (C-IRMS) analysis of AA standards. RESULTS: The NAIP derivatization gave very low limits of detection (LODs) (< 2 pmol) and limits of quantification (LOQs) ranging from 0.55 to 4.89 pmol. Comparing the concentrations of individual AAs in hydrolyzed versus unhydrolyzed clover juice samples of the low Mw size fraction (<1 kDa) showed a significant decline in concentration (p <0.03) for seven AAs after hydrolysis. Despite the decline in AA concentration, we found a linear connection between the obtained atomic fraction (13 C/total carbon and 15 N/total nitrogen) for individual AAs of hydrolyzed versus unhydrolyzed samples. CONCLUSIONS: The methodology distinguished differences in atomic fractions across AAs, in individual AAs in Mw size fractions, and between shoot and root samples of experimentally labeled white clover. Specifically, the method separated L-glutamate (Glu) and glutamine (Gln). Thus, for a broader use in plant and soil ecology, we present an optimized methodology for GC/C-IRMS analysis of AAs from organic nitrogen samples enriched with 13 C and 15 N - AA stable isotope probing (SIP).

Intercropping of Narrow-Leafed Lupin (Lupinus angustifolius L.) and Barley (Hordeum vulgare L.) Affects the Flavonoid Composition of Both Crops.

Barley (Hordeum vulgare L.) is a common cereal crop in agricultural production and is often included in legume-cereal intercropping. Flavonoids, a major class of secondary metabolites found in barley, are involved in plant defense and protection. However, the effect of intercropping on barley flavonoids remains unknown. Herein, an intercropping system involving barley and lupin (Lupinus angustifolius L.) was studied. Intercropping increased the level of luteolin in lupin roots. Lupin-barley intercropping considerably increased genistein, rutin, and apigenin in barley shoots. Genistein and apigenin were also detected in intercropped barley roots and rhizosphere soil. The three flavonoids have been reported as defense compounds, suggesting that lupin triggers a defense response in barley to strengthen its survival ability.

Data on growth, uptake and N2 fixation of grass-clover leys fertilized with mineral N fertilizer and cattle slurry.

This article presents the data obtained from a field experiment in which grass-clover leys were fertilized with increasing N rates applied in either mineral N fertilizer and/or cattle slurry forms. The leys were composed of a 2-species mixture of white clover (Trifolium repens L.) and ryegrass (Lolium perenne L.) and a 4-species mixture of white clover, red clover (Trifolium pratense L.), festulolium (Festulolium braunii) and ryegrass. In total, eighty fields were established at two farm sites in the western part of Denmark on sandy soils and monitored for two herbage seasons (2018-2019). Dry matter yield, botanical composition, N concentration and the proportion of N derived from the atmosphere using the 15 N dilution method were recorded in the harvestable biomass after each cut. Furthermore, the specific growth, N uptake and quantitative biological N fixation of the species were determined. The dataset can be used to establish the N balance, to calculate the optimal economic fertilization rate based on grass-clover composition and to predict N leaching and residual effect. The data presented were thoroughly used and discussed in the research article "Contrasting effects of slurry and mineral fertilizer on N2 fixation in grass-clover mixtures".

Deep-rooted perennial crops differ in capacity to stabilize C inputs in deep soil layers.

Comprehensive climate change mitigation necessitates soil carbon (C) storage in cultivated terrestrial ecosystems. Deep-rooted perennial crops may help to turn agricultural soils into efficient C sinks, especially in deeper soil layers. Here, we compared C allocation and potential stabilization to 150 cm depth from two functionally distinct deep-rooted perennials, i.e., lucerne (Medicago sativa L.) and intermediate wheatgrass (kernza; Thinopyrum intermedium), representing legume and non-legume crops, respectively. Belowground C input and stabilization was decoupled from nitrogen (N) fertilizer rate in kernza (100 and 200 kg mineral N ha-1), with no direct link between increasing mineral N fertilization, rhizodeposited C, and microbial C stabilization. Further, both crops displayed a high ability to bring C to deeper soil layers and remarkably, the N2-fixing lucerne showed greater potential to induce microbial C stabilization than the non-legume kernza. Lucerne stimulated greater microbial biomass and abundance of N cycling genes in rhizosphere soil, likely linked to greater amino acid rhizodeposition, hence underlining the importance of coupled C and N for microbial C stabilization efficiency. Inclusion of legumes in perennial cropping systems is not only key for improved productivity at low fertilizer N inputs, but also appears critical for enhancing soil C stabilization, in particular in N limited deep subsoils.

A novel box for aerosol and droplet guarding and evacuation in respiratory infection (BADGER) for COVID-19 and future outbreaks.

The coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has infected millions and killed more than 1.7 million people worldwide as of December 2020. Healthcare providers are at increased risk of infection when caring for patients with COVID-19. The mechanism of transmission of SARS-CoV-2 is beginning to emerge as airborne spread in addition to direct droplet and indirect contact as main routes of transmission. Here, we report on the design, construction, and testing of the BADGER (Box for Aerosol and Droplet Guarding and Evacuation in Respiratory Infection), an affordable, scalable device that contains droplets and aerosol particles, thus minimizing the risk of infection to healthcare providers. A semi-sealed environment is created inside the BADGER, which is placed over the head of the patient and maintains at least 12-air changes per hour using in-wall vacuum suction. Multiple hand-ports enable healthcare providers to perform essential tasks on a patient's airway and head. Overall, the BADGER has the potential to contain large droplets and small airborne particles as demonstrated by simulated qualitative and quantitative assessments to provide an additional layer of protection for healthcare providers treating COVID-19 and future respiratory contagions.

Gram-positive bacteria control the rapid anabolism of protein-sized soil organic nitrogen compounds questioning the present paradigm.

The cycling of especially large size organic nitrogen (N) from plants into stable microbial derived soil organic carbon (C) and N pools is understudied, in spite of organic N composing 90% of soil N and the intimate link between organic N and soil C stabilization. We investigated the fate of peptide-size and protein-size organic N fractions in soils from two long-term field experiments markedly differing in conditions for microorganisms. We combined amino acid stable isotope probing (AA-SIP) fingerprinting with PLFA-SIP to trace organic N into the soil microbial biomass. Contrary to the present paradigm, we found for both soils that greater molecular size did not protect against decomposition of these compounds neither did protection via strong sorption to the soil mineral phase. Instead, we found strong evidence that gram-positive bacteria are the key actors in the decomposition of protein-sized nitrogen compounds and that amino acids bound in large organic nitrogen compounds directly contribute to the build-up of bacterial tissue. We conclude that when large organic nitrogen compounds are dissolved, turnover occurs rapidly, irrespective of molecular size, and the bacterial incorporation of these rapid cycling compounds makes an important contribution to soil organic matter formation.

Forbs enhance productivity of unfertilised grass-clover leys and support low-carbon bioenergy.

Intensively managed grasslands are dominated by highly productive grass-clover mixtures. Increasing crop diversity by inclusion of competitive forbs may enhance biomass production and sustainable biofuel production. Here we examined if one or all of three forbs (chicory, Cichorium intybus L.; caraway, Carum carvi L.; plantain, Plantago lanceolata L.) included in ryegrass-red clover mixtures enhanced above- and below-ground productivity, and assessed their biofuel potentials, based on a three-year experiment with and without fertilisation as cattle slurry. We determined herbage yield, standing root biomass, and estimated methane energy output and greenhouse gas (GHG) emissions per energy unit using life cycle assessment. Results showed that plantain-containing grass-clover mixtures significantly increased herbage yield, while chicory- or caraway-containing mixtures maintained similar yields to the grass-clover mixture. Standing root biomass of the grass-clover mixture was enhanced by inclusion of caraway and plantain, with that of plantain further enhanced by fertilisation. The highest methane energy output was achieved in plantain-containing grass-clover mixtures. All unfertilised mixtures achieved the 60% reduction in GHG emissions compared to fossil fuel, whereas all fertilised mixtures did not meet the 60% reduction target. These findings suggest that including competitive forbs such as plantain in grass-clover mixtures enhances productivity, supporting low-carbon footprint bioenergy production.

Spatial variability in the mineralisation of the phenylurea herbicide linuron within a Danish agricultural field: multivariate correlation to simple soil parameters.

The spatial variability in the mineralisation rate of linuron [N-(3,4-dichlorophenyl)-N'-methoxy-N'-methylurea] was studied within a previously treated Danish agricultural field by sampling soils from eleven different plots randomly distributed across an area of 20 x 20 m. The soils were characterised with respect to different abiotic and biotic properties including moisture content, organic matter content, pH, nutrient content, bacterial biomass, potential for mineralisation of MCPA [(4-chloro-2-methylphenoxy)acetic acid] and linuron. Five soils had a potential for mineralisation of linuron, with 5-15% of the added [ring-U-14C]linuron metabolised to 14CO2 within 60 days at 10 degrees C, while no extensive mineralisation of linuron was observed in the six remaining soils within this period. A TLC analysis of the methanol-extractable residues showed no development of 14C-labelled metabolites from linuron in any of the samples. Multivariate analysis was conducted to elucidate relationships between the intrinsic properties of single soil samples and initial rate of linuron mineralisation. The analysis indicated that important soil parameters in determining the spatial heterogeneity included the C(total)/N(total) ratio, pH and the water-extractable potassium contents, with the first of these highly negatively correlated and the last two highly positively correlated to the initial linuron mineralisation rate. This study shows that enhanced biodegradation of linuron may develop with successive field treatments, but that considerable in-field spatial heterogeneity in the degradation rate still exists. Combined with a parallel enrichment study focused on the underlying microbial processes, the present results suggest that intrinsic soil properties affect the linuron-metabolising bacterial population and thereby determine the spatial variability in the linuron mineralisation activity.

Method for rapid screening of pesticide mineralization in soil.

A method has been developed for the analysis of (14)CO(2) evolution from the mineralization of (14)C-labelled organic compounds in soil samples. The new method is less space demanding and substantially cuts down laborious manual work compared to the traditional incubation bottle method used. Furthermore, the use of scintillation cocktail is largely reduced with the new method. In the new method, (14)CO(2) is trapped in filter paper held in the lid of a 20 ml glass vial by surface tension. The trapping solution used is Ca(OH)(2), which fixates CO(2) in the filter paper and the analysis of trapped (14)CO(2) is done using the Cyclone trade mark Storage Phosphor system. The lids are placed in a 32 well holder and exposed to a phosphor screen prior to scanning in a Cyclone trade mark scanner. The new filter method has been tested and compared to results obtained using the traditional method. The results show good agreement but due to a smaller capacity for CO(2) with the filter method compared to the traditional method, the interval between sampling has to be shorter using the filter method when the CO(2) development is high. The detection limits for the filter method is higher compared to the traditional method. With the filter method, the level of radioactivity has to exceed 300 dpm before detection is possible, while the same limit for the traditional method is around 30 dpm. On the other hand, the gas trapping faster and the efficiency is higher with the filter method.

Variation of MCPA, metribuzine, methyltriazine-amine and glyphosate degradation, sorption, mineralization and leaching in different soil horizons.

Pesticide mineralization and sorption were determined in 75 soil samples from 15 individually drilled holes through the vadose zone along a 28km long transect of the Danish outwash plain. Mineralization of the phenoxyacetic acid herbicide MCPA was high both in topsoils and in most subsoils, while metribuzine and methyltriazine-amine was always low. Organic matter and soil pH was shown to be responsible for sorption of MCPA and metribuzine in the topsoils. The sorption of methyltriazine-amine in topsoil was positively correlated with clay and negatively correlated with the pH of the soil. Sorption of glyphosate was tested also high in the subsoils. One-dimensional MACRO modeling of the concentration of MCPA, metribuzine and methyltriazine-amine at 2m depth calculated that the average concentration of MCPA and methyltriazine-amine in the groundwater was below the administrative limit of 0.1mug/l in all tested profiles while metribuzine always exceeded the 0.1mug/l threshold value.

Elucidating the key member of a linuron-mineralizing bacterial community by PCR and reverse transcription-PCR denaturing gradient gel electrophoresis 16S rRNA gene fingerprinting and cultivation.

A bacterial community from Danish agricultural soil was enriched with linuron [N-(3,4-dichlorophenyl)-N'-methoxy-N'-methylurea] as the sole carbon and nitrogen source. The community mineralized [ring-U-14C]linuron completely to 14CO2 and 14C-biomass. Denaturing gradient gel electrophoresis analysis and cultivation revealed that a Variovorax sp. was responsible for the mineralization activity.

Sorption-controlled degradation kinetics of MCPA in soil.

The relationship between sorption strength and degradation kinetics has been studied for the pesticide MCPA in a sandy top- and subsoil. After adding two types of sorbents (crushed peat and activated carbon) in various amounts to the sandy soils, sorption, desorption, and mineralization of 14C-MCPA were measured. The obtained Freundlich constants (KF) varied between 0.7 and 27.2 mg(1-nF) x L(nF)/kg, and the first-order mineralization rate constants varied between 0.001 and 0.128 d(-1). The results showed an inverse relationship between sorption strength and mineralization. A higher KF value corresponded to a smaller mineralization rate and less mineralization. A correlation coefficient of r2 = 0.934 between the log-transformed Freundlich desorption coefficient (K(F,des)) and the log-transformed mineralization rate constant (k) was obtained. After 7, 14, 22, and 35 days of incubation, soil samples were consecutively extracted by water, methanol, and 5 M NaOH to separate the remaining 14C into 3 different pools. The extractions showed that the mineralization only proceeded from the water extractable pool of MCPA. Thin-layer chromatography revealed a formation of small amounts of metabolites; <7% of initially added 14C was present as other compounds than 14C-MCPA in the water and methanol extractable pools. The study showed mineralization to be strongly correlated with sorption, represented by the desorption coefficient, and hence stresses the significance of bonding strength for estimating pesticide degradation in soil.