Phospholipid Fatty Acid Research Articles

Long‐term successive biochar application increases plant lignin and microbial necromass accumulation but decreases their contributions to soil organic carbon in rice–wheat cropping system

AbstractBiochar application is widely recognized as an effective approach for increasing soil organic carbon (SOC) and mitigating climate change in agroecosystems. However, the effects of biochar application on net accumulations and relative contributions of different SOC sources remain unclear. Here, we explored the effects of biochar application on plant‐derived (PDC) and microbial necromass C (MNC) in a 10‐year experimental rice–wheat rotation field receiving four different intensities of biochar application (0, 2.25, 11.5, and 22.5 t ha−1 for each crop season), using phospholipid fatty acids (PLFAs), lignin phenols and amino sugars as biomarkers of microbial biomass, PDC and MNC, respectively. Our results showed that biochar application increased SOC content and stock by 32.6%–203% and 26.4%–145%, respectively. Higher biochar application (11.5 and 22.5 t ha−1) increased soil pH, total nitrogen (TN), total phosphorus (TP), SOC/TN, and root biomass. In addition, higher biochar application enhanced bacterial, fungal, and total microbial biomass. Plant lignin phenols and MNC contents significantly increased, whereas their contributions to SOC significantly decreased with the increase in biochar application rates due to the disproportionate increase in PDC and MNC, and SOC. Fungal necromass had a greater contribution to SOC than bacterial necromass. The fungal/bacterial necromass decreased from 2.56 to 2.26 with increasing biochar application rates, because of the higher abundances of bacteria than that of fungi as indicated by PLFAs under higher biochar application rates. Random forest analyses revealed that pH, TP, and SOC/TN were the main factors controlling plant lignin and MNC accumulation. Structural equation modeling revealed that biochar application increased lignin phenols by stimulating root biomass, whereas enhanced MNC accumulation was primarily from increased microbial biomass and lignin phenols. Overall, our findings suggest that biochar application increases the accumulation of the two SOC sources but decreases their contributions to SOC in paddy soils.

Edible insects as a novel source of lecithin: Extraction and lipid characterization of black soldier fly larvae and yellow mealworm

Edible insects with high fat and phosphorus content are a potential novel source of lecithin, however, studies on their minor lipids are limited. In this study, lecithin was extracted from black soldier fly larvae and yellow mealworm. Herein, the effects of lecithin extraction method, matrix and ultrasound pretreatment were explored based on the fatty acid composition and phospholipid profile with soy lecithin as a reference. The use of a wet matrix and ultrasound pretreatment increased the extraction efficiency of total PLs from both insects. Insect lecithin contained a considerable amount of sphingomyelin compared to soy lecithin. In insect lecithin, a total of 47 glycerophospholipid and sphingomyelin molecular species, as well as four molecular species of fatty acyl esters of hydroxy fatty acid, were detected. This study is the first comprehensive investigation of insects as a new source of lecithin with applications in food, cosmetics and in the pharmaceutical industry.

Input of high-quality litter reduces soil carbon losses due to priming in a subtropical pine forest

To date, it is unclear how differences in litter quality affect soil organic matter (SOM) decomposition through a phenomenon called 'priming effects' (PEs), especially for low-fertility forest soils under field conditions. Here, the effects of low- and high-quality leaf litter on PE and microbial metabolism of litter-derived carbon (C) were explored in a low-fertility pine (Pinus massoniana) plantation. A 185-day in situ incubation experiment was carried out by adding two 13C-labeled leaf litters to the pine soil—a low-quality (high lignin: nitrogen ratio) litter sourced from pine and a broadleaved species Schima superba produced high-quality (low lignin: nitrogen ratio) litter. To determine the key microbial groups contributing to PEs, the abundance of 13C-labeled litter enrichment in soil phospholipid fatty acids (13C-PLFAs) was quantified. We found that high-quality litter decomposed more rapidly than low-quality litter, with both litter-derived CO2 efflux reaching a plateaued level during the experimental period. Low-quality litter induced net positive PEs, while high-quality litter induced net negative PEs during the litter decomposition processes. The 13C-PLFAs results showed that bacterial groups governed the negative PEs induced by high-quality litter, whereas fungal communities targeted the positive PEs induced by low-quality litter. Random forest model and variation partitioning analysis demonstrated that the direction and magnitude of PEs were driven by litter-induced changes in key microbial groups rather than the structure of the microbial community. Our results demonstrate that microorganisms preferentially utilized litter-derived C in high-quality litter treatment and SOM in low-quality litter treatment, respectively. In contrast to low-quality litter, adding high-quality litter promoted the microbial metabolism of litter-derived C, reducing SOM decomposition (strong negative PEs). Taken together, this study provides isotope-based suggestions for the improvement of degraded pine forests—introducing tree species that produce high-quality litter may benefit soil C sequestration by reducing soil C losses due to PE in nutrient-poor pine forests.

Effects of microbial groups on soil organic carbon accrual and mineralization during high- and low-quality litter decomposition

Litter input regulates the mineralization and accrual of soil organic carbon (SOC), depending on microbial groups and litter quality. Stable isotope probing was used after the addition of 13C-labeled litter of different recalcitrance, including Populus davidiana (characterized by low quality, high C:N, and lignin content) and Quercus wutaishanica (characterized by high quality, low C:N, and lignin content). The flow of litter C to microbial biomass and necromass was followed by quantification of 13C incorporation into phospholipid fatty acids (13C-PLFAs) and amino sugars (13C-AS), and their respiration. The CO2 efflux and positive priming effects (PE) were higher, but the 13C-necromass was lower (8.5% vs. 10%) in the soil with P. davidiana than with Q. wutaishanica. The 13C-necromass continuously increased with P. davidiana litter decomposition, but reached a peak at day 14 after Q. wutaishanica litter addition. These results indicated that the decomposition of resistance compounds mainly resulted in necromass formation after low-quality litter addition. The P. davidiana-derived 13C in fungal PLFAs increased with decomposition, suggesting the importance of these microorganisms in decomposing resistance compounds in low-quality litter despite the bacterial-dominated decomposition process of both litters. Bacteria frequently exhibit versatility in utilizing C from different sources, whereas a larger proportion of fungi display specialization. SOC mineralization increased with the remaining litter but decreased with 13C-G- bacterial PLFAs and 13C-actinobacterial PLFAs after both types of litter addition because starving microbes need to invest more soil C in maintenance respiration vs. biomass formation. 13C-necromass increased with 13C-PLFA because microbial biomass is the primary precursor of necromass. Altogether, low-quality compounds are more readily respired, whereas C from high-quality biopolymers is relatively more assimilated into the necromass. This study further explains the effect of soil microbial groups on SOC under different qualities of litter decomposition and deepens our understanding of the mechanism of SOC sequestration.

Adaptive multi-paddock grazing increases mineral associated soil carbon in Northern grasslands

Grassland soils play an important role in sequestering carbon (C) and are primarily used for livestock grazing. Grazing management can increase the amount of C stored in soils and the distribution of C in different soil fractions by altering soil microbial community structure, thereby influencing the persistence of soil C over time. Adaptive multi-paddock grazing (AMP), grazing cattle at high stock densities with long periods between grazing events, is considered to enhance grassland sustainability, productivity, and soil C compared to more conventional grazing practices (continuous to slow rotational). However, whether and how AMP grazing might affect the amount of C stored in different soil fractions has received little attention. Soil samples (0–15 cm depth) were collected from 24 ranches in a paired design, where 12 ranches practiced AMP grazing and 12 neighboring ranches practiced conventional grazing. Soil organic C (SOC) and total nitrogen were measured in different soil particle size [fine (<53 µm), medium (53–250 µm) and coarse (>250 µm)], and density [light (>1.6 g cm−3), and heavy (<1.6 g cm−3)] fractions. Soil bacterial and fungal biomass and abundances were obtained from phospholipid fatty acid analysis. Mean weights of soil microaggregates and heavy fractions were higher in soil exposed to AMP grazing, whereas mean weights of macroaggregates and light fractions were higher in conventionally grazed soils. In warmer and drier climatic conditions, SOC in both bulk soils and within various size and density fractions tended to decrease. Soil organic C (both concentration and stock) were significantly higher in the fine soil fractions under AMP management than conventionally grazed grasslands. Given that SOC in fine fractions, is more mineral associated and recalcitrant, our results indicate that AMP grazing increased the size of the stable SOC pool. The increased carbon pool in the fine fraction was associated with greater fungal to bacterial ratios, suggesting a possible biological mechanism for the increase. Overall, the results indicate that AMP grazing may be beneficial for sequestering more stable carbon that helps mitigate the effects of climate change.

Aggregate composition directly affects the characteristics of soil microbial and nematode communities in a chronosequence of Chinese fir plantations

AbstractMaintaining soil biological diversity is an effective approach in facilitating Chinese fir plantations' sustainable development. However, dynamic characteristics of soil microbial and nematode communities during Chinese fir planting remain unclear, particularly on the aggregate scales, which are the main sites of soil biological activity. Therefore, the research aims to explore how stand age (3, 9, 17, and 26 years) of Chinese fir plantations affects soil microbial biomass (including bacteria, fungi, and actinomycetes, based on the phospholipid fatty acid method), nematode abundances (including bacterivores, fungivores, plant parasites, and omnivores‐predators, based on the modified Baermann funnel method), and their diversity within aggregates of differing sizes (&gt;2, 2–1, 1–0.25, and &lt;0.25 mm). According to the findings, both aggregate size and stand age were major factors affecting soil microbes and nematodes in Chinese fir plantations. Specifically, soil microbial biomass (mainly bacteria), nematode abundances (except fungivores), and their diversity peaked within large macro‐aggregates (&gt;2 mm), and the ranking order for stand ages was 17 &gt; 9 ≈ 3 &gt; 26 years. As the main fractions of soil aggregates in Chinese fir plantations, large macro‐aggregates had suitable internal environments for soil microbial propagation and nematode growth and mainly accumulated at 17 years, which caused the prosperity of soil microbes and nematodes at this stand age. Moreover, the partial least squares path model demonstrated that the influence of stand age on soil microbes and nematodes was indirect; but rather, the aggregate composition played a direct role in shaping their communities. Therefore, in order to maintain soil biological diversity, large macro‐aggregates should be protected during the growth of Chinese fir, especially at the late stage (from 17 to 26 years). Overall, these results could provide valuable insights into Chinese fir plantations' sustainable development.

Revisiting soil fungal biomarkers and conversion factors: Interspecific variability in phospholipid fatty acids, ergosterol and rDNA copy numbers

Refined conversion factors for soil fungal biomarkers are proposed.High interspecific variability is present in all fungal biomarkers.A modeling approach supports the validity of biomarker estimates in diverse soils.ITS1 copies vary strongly, but are fungal-specific with least phylogenetic bias.A combination of fungal biomarkers will reveal soil fungal physiology and activity.The abundances of fungi and bacteria in soil are used as simple predictors for carbon dynamics, and represent widely available microbial traits. Soil biomarkers serve as quantitative estimates of these microbial groups, though not quantifying microbial biomass per se. The accurate conversion to microbial carbon pools, and an understanding of its comparability among soils is therefore needed. We refined conversion factors for classical fungal biomarkers, and evaluated the application of quantitative PCR (qPCR, rDNA copies) as a biomarker for soil fungi. Based on biomarker contents in pure fungal cultures of 30 isolates tested here, combined with comparable published datasets, we propose average conversion factors of 95.3 g fungal C g−1 ergosterol, 32.0 mg fungal C µmol−1 PLFA 18:2ω6,9 and 0.264 pg fungal C ITS1 DNA copy−1. As expected, interspecific variability was most pronounced in rDNA copies, though qPCR results showed the least phylogenetic bias. A modeling approach based on exemplary agricultural soils further supported the hypothesis that high diversity in soil buffers against biomarker variability, whereas also phylogenetic biases impact the accuracy of comparisons in biomarker estimates. Our analyses suggest that qPCR results cover the fungal community in soil best, though with a variability only partly offset in highly diverse soils. PLFA 18:2ω6,9 and ergosterol represent accurate biomarkers to quantify Ascomycota and Basidiomycota. To conclude, the ecological interpretation and coverage of biomarker data prior to their application in global models is important, where the combination of different biomarkers may be most insightful.

Effects of triglyceride and ethyl ester forms of EPA on hepatic lipid metabolism in mice with non-alcoholic fatty liver disease

In this study, the beneficial effects of EPA in ethyl esters (EE) and triacylglycerides (TAG) forms on high fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) were studied and compared. Results indicated that both EPA-TAG and EPA-EE can alleviate pathological features of NAFLD, such as hepatic lipid accumulation and lobular inflammation, with a more pronounced protective effects observed in EPA-TAG treated mice. The integrated hepatic transcriptomic, metabolic and gut microbiota profiles suggest that EPA-TAG and EPA-EE improved hepatic steatosis by suppression of food intake and lipogenesis, promotion of lipid and phospholipid PUFA remodeling, ketogenesis and fatty acids oxidation, possibly via activating the AMPK/adipocytokine signaling pathway. The protective effect of EPA was also associated with the regulation of intestinal flora structure. In general, EPA-TAG was found to be more effective than EPA-EE in preventing HFD-induced NAFLD by reducing appetite, improving hepatic lipid metabolism, increasing energy expenditure, and enhancing gut-liver cross-talk.

The Diversity and Composition of Soil Microbial Communities Differ in Three Land Use Types of the Sanjiang Plain, Northeastern China.

In recent years, the Sanjiang Plain has experienced drastic human activities, which have dramatically changed its ecological environment. Soil microorganisms can sensitively respond to changes in soil quality as well as ecosystem function. In this study, we investigated the changes in soil microbial community diversity and composition of three typical land use types (forest, wetland and cropland) in the Sanjiang Plain using phospholipid fatty acid analysis (PLFA) technology, and 114 different PLFA compounds were identified. The results showed that the soil physicochemical properties changed significantly (p < 0.05) among the different land use types; the microbial diversity and abundance in cropland soil were lower than those of the other two land use types. Soil pH, soil water content, total organic carbon and available nitrogen were the main soil physico-chemical properties driving the composition of the soil microbial community. Our results indicate that the soil microbial community response to the three different habitats is complex, and provide ideas for the mechanism by which land use changes in the Sanjiang Plain affect the structure of soil microbial communities, as well as a theoretical basis for the future management and sustainable use of the Sanjiang plain, in the northeast of China.

Mass Production of Arbuscular Mycorrhizal Fungi on the Sorghum Plants Inoculated with Burkholderia arboris Using Soybean Mill Waste and Vermicompost-Amended Soil-Sand Substrate.

Arbuscular mycorrhizal (AM) fungi are being used as a new generation of biofertilizers to increase plant growth by improving plant nutrition and bio-protection. However, because of the obligatory nature of the plant host, large-scale multiplication of AM propagules is challenging, which limits its applicability. This study evaluates the ability of Burkholderia arboris to increase AM production in soybean mill waste and vermicompost amended by soil-sand mixture planted with sorghum as a host plant. The experiment was conducted in a nursery using a completely randomized design with four inoculation treatments (B. arboris, AM fungi, B. arboris + AM fungi, and control) under sterilized and unsterilized conditions. AM production was investigated microscopically (spore density and root colonization), and biochemically (AM-specific lipid biomarker, 16:1ω5cis derived from neutral lipid fatty acid (NLFA), and phospholipid fatty acid (PLFA) fractions from both soil and roots). Integrating B. arboris with AM fungi in organically amended pots was found to increase AM fungal production by 62.16 spores g-1 soil and root colonization by 80.85%. Biochemical parameters also increased with B. arboris inoculation: 5.49nmol PLFA g-1 soil and 692.68nmol PLFA g-1 root and 36.72nmol NLFA g-1 soil and 3147.57nmol NLFA g-1 root. Co-inoculation also increased glomalin-related soil protein and root biomass. Principal component analysis (PCA) further supported the higher contribution of B. arboris to AM fungi production under unsterilized conditions. In conclusion, inoculation of AM plant host seeds with B. arboris prior to sowing into organic potting mix could be a promising and cost-effective approach for increasing AM inoculum density for commercial production. Furthermore, efforts need to be made for up-scaling the AM production with different plant hosts and soil-substrate types.

Arachidonic acid and docosahexaenoic acid levels correlate with the inflammation proteome in extremely preterm infants

Clinical trials supplementing the long-chain polyunsaturated fatty acids (LCPUFAs) docosahexaenoic acid (DHA) and arachidonic acid (AA) to preterm infants have shown positive effects on inflammation-related morbidities, but the molecular mechanisms underlying these effects are not fully elucidated. This study aimed to determine associations between DHA, AA, and inflammation-related proteins during the neonatal period in extremely preterm infants. A retrospective exploratory study of infants (n=183) born below 28 weeks gestation from the Mega Donna Mega trial, a randomized multicenter trial designed to study the effect of DHA and AA on retinopathy of prematurity. Serial serum samples were collected after birth until postnatal day 100 (median 7 samples per infant) and analyzed for phospholipid fatty acids and proteins using targeted proteomics covering 538 proteins. Associations over time between LCPUFAs and proteins were explored using mixed effect modeling with splines, including an interaction term for time, and adjusted for gestational age, sex, and center. On postnatal day one, 55 proteins correlated with DHA levels and 10 proteins with AA levels. Five proteins were related to both fatty acids, all with a positive correlation. Over the first 100 days after birth, we identified 57 proteins to be associated with DHA and/or AA. Of these proteins, 41 (72%) related to inflammation. Thirty-eight proteins were associated with both fatty acids and the overall direction of association did not differ between DHA and AA, indicating that both LCPUFAs similarly contribute to up- and down-regulation of the preterm neonate inflammatory proteome. Primary examples of this were the inflammation-modulating cytokines IL-6 and CCL7, both being negatively related to levels of DHA and AA in the postnatal period. This study supports postnatal non-antagonistic and potentially synergistic effects of DHA and AA on the inflammation proteome in preterm infants, indicating that supplementation with both fatty acids may contribute to limiting the disease burden in this vulnerable population. ClinicalTrials.gov (NCT03201588).

A Cross-Sectional Pilot Study on Association of Ready-to-Eat and Processed Food Intakes with Metabolic Factors, Serum Trans Fat and Phospholipid Fatty Acid Compositions in Healthy Japanese Adults.

Frequently consuming processed and ready-to-eat (RTE) foods is regarded as unhealthy, but evidence on the relationships with circulating metabolic parameters is lacking. Japanese residents of a metropolitan area, 20 to 50 years of age, were studied in terms of anthropometric and biochemical parameters, including circulating trans fat and serum phospholipid fatty acid levels. Processed foods, except drinks and dairy items, were categorized according to requirements for additional ingredients and cooking before eating. Processed and RTE foods were divided according to fat and/or oil content into non-fatty or fatty foods. The participants were grouped into tertiles based on the energy percent (En%) derived from fatty-RTE foods. Fatty-RTE En% showed negative associations with fish, soybean and soybean products, dairy, eggs, vegetables, seaweed/mushrooms/konjac, fruit and non-oily seasonings reflecting lower dietary fiber, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and mineral and vitamin intakes, while the associations with fat/oil, confectionaries, and sweet beverages were positive. Fatty-RTE En% consumption was positively associated with alkaline phosphatase, leucine aminopeptidase, direct bilirubin, elaidic acid, and C18:2 but inversely associated with HDL cholesterol, C15:0, C17:0, EPA, and DHA. A higher fatty-RTE food intake was suggested to contribute to unbalanced nutrient intakes, as reflected in lipid metabolic parameters. Further large-scale studies are needed to evaluate the quality and impacts of RTE foods.

Organic carbon source controlled microbial olivine dissolution in small-scale flow-through bioreactors, for CO2 removal

The development of carbon dioxide removal methods, coupled with decreased CO2 emissions, is fundamental to achieving the targets outlined in the Paris Agreement limiting global warming to 1.5 °C. Here we are investigating the importance of the organic carbon feedstock to support silicate mineral weathering in small-scale flow through bioreactors and subsequent CO2 sequestration. Here, we combine two bacteria and two fungi, widely reported for their weathering potential, in simple flow through bioreactors (columns) consisting of forsterite and widely available, cheap organic carbon sources (wheat straw, bio-waste digestate of pig manure and biowaste, and manure compost), over six weeks. Compared to their corresponding abiotic controls, the inoculated straw and digestate columns release more total alkalinity (~2 times more) and produce greater dissolved and solid inorganic carbon (29% for straw and 13% for digestate), suggesting an increase in CO2 sequestration because of bio-enhanced silicate weathering. Microbial biomass is higher in the straw columns compared to the digestate and manure compost columns, with a phospholipid fatty acid derived total microbial biomass 10 x greater than the other biotic columns. Scanning Electron Microscopy imaging shows the most extensive colonisation and biofilm formation on the mineral surfaces in the straw columns. The biotic straw and digestate columns sequester 50 and 14 mg C more than their abiotic controls respectively, while there is no difference in the manure columns. The selection of organic carbon sources to support microbial communities in the flow through bioreactors controlls the silicate weathering rates and CO2 sequestration.

Comparison of comprehensive fatty acid profile from different regions in Chinese human milk project (CHMP) study

In this study, we aimed to detect the comprehensive fatty acid (FA) profile in human milk, including total fatty acids (total FAs), Sn-2 fatty acids (Sn-2 FAs), and phospholipid fatty acids (PLFAs), and further analyzed the influence of the sampling regions on human milk FA profile. The results showed that oleic acid (C18:1n9c), palmitic acid (C16:0), and linoleic acid (C18:2n6c) were the main FA species in total FAs, and Sn-2 C16:0 was the dominant Sn-2 FA species. Moreover, the PLFAs were mainly comprised of PL C16:0, PL stearic acid (PL C18:0), PL C18:1n9c, and PL C18:2n6c. Furthermore, FAs showed geographical differences, such as the highest content of linolenic acid (C18:3n3) and docosahexaenoic acid (C22:6n3) of total FAs was observed in Lanzhou and Weihai, respectively. Therefore, these results provide more FA data for the Chinese human milk database, and further enrich the theoretical information for the development of infant formula.

Elevated CO2 enhanced the incorporation of 13C-residue into plant but depressed it in the microbe in the tomato (Solanum lycopersicum L.) rhizosphere soils

The translation of plant residue carbon in plant–soil–microbe systems under atmospheric CO2 (aCO2) and elevated CO2 (eCO2) is key to understanding the response of ecosystems to climatic change. Using stable isotope probing technology, we conducted an experiment on 3.5 g·kg−1 tomato 13C-residue (1050 ‰) added to tomato soils to investigate the decomposition and incorporation of 13C-residue under varied CO2 concentrations (aCO2 and eCO2: 400 and 800 μmol·mol−1, respectively). The results showed that the 13C-residue decomposition rate was stimulated by approximately 7.2 % and the 13C amount in plant stems was stimulated by 15.6 % (22.2 vs. 19.2 μg/pot), while the total 13C amount in soil microbial phospholipid fatty acids (PLFAs) was significantly decreased by 16.3 % (541 vs. 646 nmol·g−1), the DNA δ13C in soil microbes decreased by 5.1 % (−27.629 ‰ vs. −26.289 ‰), and soil δ13C decreased by 7.1 % (49.9 ‰ vs. 53.7 ‰) under eCO2. The microbial community structure involved in 13C-residue incorporation was significantly affected by the CO2 concentration. Firmicutes (Caldalkalibacillus 20.4 % and Bacillus 20.4 %) and Actinobacteria (Streptomyces 6.6 %) were the dominant phyla under aCO2 and eCO2, respectively. Proteobacteria (Dyella) were the most depleted microbes under eCO2, whereas almost no genera were found under aCO2. The 13C-residue decomposition rate was positively correlated with soil NO3−-N (r = 0.998, P < 0.05) at t1 (17 days of cultivation under varied CO2 concentrations), and DNA δ13C was positively related with soil pH (r = 0.978, P < 0.05) at t2 (35 days of cultivation under varied CO2 concentrations). Mantel tests showed that the microbial community composition involved in 13C incorporation was significantly correlated with the soil pH under various CO2 conditions (r = 0.879, P < 0.05). These results indicate that eCO2 stimulated the decomposition of 13C-residue and the transportation of 13C to the plant stem, but suppressed the incorporation of 13C into microbes (PLFA and DNA), which were adjusted by the rhizosphere N status and soil pH.

Linkages of soil and microbial stoichiometry to crop nitrogen use efficiency: Evidence from a long-term nitrogen addition experiment

Increasing crop nitrogen use efficiency (NUE) has important implications for food security and agricultural sustainability. Changes in nutrient availability due to stoichiometric imbalances in soil under long-term application of nitrogen (N) can limit crop NUE and yield. However, little is known about the linkages across stoichiometric balance, N fertilizer application, and N uptake. We investigated the changes in soil stoichiometry, microbial community, and crop NUE relative to N fertilizer application in a 16-year field experiment in which five levels (0, 70, 140, 210 and 280 kg N ha−1) of mineral N fertilizer treatment were applied to wheat and maize cropland. The results showed that the P storage decreased from 4.3 Mg P ha–1 under the fertilizer dose of 0 kg N ha−1 to 3.5 Mg P ha–1 under the fertilizer dose of 280 kg N ha−1. Thus, long-term N application increased soil C/P and N/P ratios, in a marked decrease in the content of soil available P. The microbial community based on phospholipid fatty acids (PLFAs) increased with increasing N addition rates, from 0 to 140 kg N ha−1, but significantly decreased at application rates above 210 kg N ha−1. Thus, applying N at a rate of 140 kg ha−1 resulted in the N threshold rate for microorganism survival being reached or exceeded. The crop NUE peak occurred at the urea application rate of 140 kg N ha−1 (60.4 %), after which NUE declined. The soil elementary and enzymatic stoichiometric ratios under long-term N addition directly affected crop NUE in the topsoil. Soil elementary stoichiometric ratios affected crop NUE by indirectly altering microbial biomass of the subsoil. Overall, soil stoichiometric imbalances under long-term N addition were the key factors driving NUE across N gradients.

Ancient prairies as a reference for soil organic carbon content and microbial community structure

The historic remnant prairies of Missouri display diversity above and below the soil surface and represent the few remaining acres of unconverted grassland habitat in the state. The soil microbiome is foundational to nutrient acquisition, resource partitioning, and ecosystem resilience; therefore, the intact plant-soil interactions of remnant prairies can serve as a reference for microbial community structure and ecosystem services lost due to land conversion. This research was conducted to show the importance of reference soil conditions and the variability that lies therein while contrasting degraded soil ecosystems to provide a standard to which comparable soils should be held. In this study, 35 sites across Missouri representing three prairie management conditions on similar soils and variable climatic conditions were compared to assess differences in the microbial community structure and biomass using phospholipid fatty acid (PLFA) analysis. Surface (0–7.5 cm) soil organic carbon (SOC) content provided a measure of substrate available to support microbial community health. Total microbial biomass, the biomass of individual microbial groups, selected biomarker ratios, and SOC, were significantly different across treatments and highest in remnant prairies apart from Saprophytic fungi and eukaryotes. This study demonstrated that the soil microbial community in reconstructed and restored prairie systems remained significantly diminished in biomass relative to remnant prairies, even after several decades. This study highlights the need to preserve remnant prairies, where soil is in its native state under native flora and abiotic conditions. These results also illustrate the importance of selecting remnant systems as the ecological reference standard for prairie lands.

A metabolomics approach reveals metabolic disturbance of human cholangiocarcinoma cells after parthenolide treatment

Ethnopharmacological relevanceTanacetum parthenium (L.) Schultz-Bip, commonly known as feverfew, has been traditionally used to treat fever, migraines, rheumatoid arthritis, and cancer. Parthenolide (PTL), the main bioactive ingredient isolated from the shoots of feverfew, is a sesquiterpene lactone with anti-inflammatory and antitumor properties. Previous studies showed that PTL exerts anticancer activity in various cancers, including hepatoma, cholangiocarcinoma, acute myeloid leukemia, breast, prostate, and colorectal cancer. However, the metabolic mechanism underlying the anticancer effect of PTL remains poorly understood. Aim of the studyTo explore the anticancer activity and underlying mechanism of PTL in human cholangiocarcinoma cells. Material and methodsIn this investigation, the effects and mechanisms of PTL on human cholangiocarcinoma cells were investigated via a liquid chromatography/mass spectrometry (LC/MS)-based metabolomics approach. First, cell proliferation and apoptosis were evaluated using cell counting kit-8 (CCK-8), flow cytometry analysis, and western blotting. Then, LC/MS-based metabolic profiling along with orthogonal partial least-squares discriminant analysis (OPLS-DA) has been constructed to distinguish the metabolic changes between the negative control group and the PTL-treated group in TFK1 cells. Next, enzyme-linked immunosorbent assay (ELISA) was applied to investigate the changes of metabolic enzymes associated with significantly alerted metabolites. Finally, the metabolic network related to key metabolic enzymes, metabolites, and metabolic pathways was established using MetaboAnalyst 5.0 and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Database. ResultsPTL treatment could induce the proliferation inhibition and apoptosis of TFK1 in a concentration-dependent manner. Forty-three potential biomarkers associated with the antitumor effect of PTL were identified, which primarily related to glutamine and glutamate metabolism, alanine, aspartate and glutamate metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, arginine biosynthesis, arginine and proline metabolism, glutathione metabolism, nicotinate and nicotinamide metabolism, pyrimidine metabolism, fatty acid metabolism, phospholipid catabolism, and sphingolipid metabolism. Pathway analysis of upstream and downstream metabolites, we found three key metabolic enzymes, including glutaminase (GLS), γ-glutamyl transpeptidase (GGT), and carnitine palmitoyltransferase 1 (CPT1), which mainly involved in glutamine and glutamate metabolism, glutathione metabolism, and fatty acid metabolism. The changes of metabolic enzymes associated with significantly alerted metabolites were consistent with the levels of metabolites, and the metabolic network related to key metabolic enzymes, metabolites, and metabolic pathways was established. PTL may exert its antitumor effect against cholangiocarcinoma by disturbing metabolic pathways. Furthermore, we selected two positive control agents that are considered as first-line chemotherapy standards in cholangiocarcinoma therapy to verify the reliability and accuracy of our metabolomic study on PTL. ConclusionThis research enhanced our comprehension of the metabolic profiling and mechanism of PTL treatment on cholangiocarcinoma cells, which provided some references for further research into the anti-cancer mechanisms of other drugs.

Effects of the Replacement of Chemical Fertilizers with Organic Fertilizers in Different Proportions on Microbial Biomass and Enzyme Activities of Soil Aggregates in Gravel-Mulched Field

Gravel-mulched fields are a unique form of drought-resistant agriculture in the northwest region of China. In recent years, continuous cropping obstacles caused by the perennial cultivation of a single crop have seriously constrained the sustainable development of sand fields. This study aimed to explore the distribution patterns of different particle sizes of aggregates (&gt;2, 1–2, 0.25–1, and &lt;0.25 mm) and the relationships between their microbial biomass and enzyme activities under different organic fertilization and to explore the effective measures for improving soil fertility in a gravel-mulched field with an 8-year positioning test. The results indicate that the mass percentage of soil aggregates of ≥1 mm and their mean weight diameter (MWD), microbial biomass (carbon and nitrogen, bacteria, fungi, actinomycetes, and total phospholipid fatty acids), and their related enzyme activities (leucine aminopeptidase, LAP; N-acetyl-β-d-glucosidase, NAG; β-glucosidase, BG; and polyphenol oxidase, PPO) in aggregates of different particle sizes increased with the increase in the proportion of organic fertilizers replacing the N fertilizer. Among them, the organic fertilizer replacing more than 50% of chemical nitrogen fertilizers exerted the most significant effect. With the decrease in agglomerate particle size, the contents of microbial carbon and nitrogen showed a decreasing trend, whereas LAP, NAG, and BG activities followed an increasing trend, and the change in microbial biomass was not obvious. The correlation analysis showed highly significant positive correlations between the MWD of soil aggregates, microbial biomass, and the activities of LAP, NAG, BG, and PPO. Therefore, the replacement of more than 50% of chemical fertilizer with organic fertilizer was observed to be conducive to promoting the formation of large aggregates in sandy soils and increasing the microbial biomass and enzyme activities in different sizes of aggregates.

Soil carbon dynamics in drained prairie pothole wetlands

Drainage leads to trade-offs between crop production efficiency and wetland conservation, with complex impacts on ecosystem services. In North America’s Prairie Pothole Region, wetland drainage is widespread, often to increase the available land for cultivation, prevent crop loss due to flooding, and manage soil salinity. Wetlands are known for providing key ecosystem services such as improved water quality, flood mitigation, and carbon storage. There is limited research on how changes to soil hydrology and soil redistribution through wetland drainage can impact soil carbon storage and persistence in this region. This research evaluates factors that contribute to soil carbon storage in drained prairie pothole wetland based on 33 drained wetlands in Saskatchewan, Canada. These analyses showed regional differences in the response of soil carbon storage to drainage that are driven by environmental factors such as annual precipitation, temperature, and wetland permanence. We observed increasing soil carbon storage from the Dark Brown to Black soil zones, as well as with longer wetland pond permanence. The sampling depth used for calculating soil carbon storage was especially important when comparing geographically across the soil zones as the Black soil zone had greater soil carbon stored at depth. Soil carbon was also intensively monitored over 2 years following installation of surface drainage across a wetland complex (8 drained wetlands) where water was partially directed to a consolidation wetland. We further assessed changes in soil carbon dynamics and protection from microbial decomposition based on three soil organic matter fractions, ATR-FTIR for organic matter functional groups, and phospholipid fatty acid analysis to understand the microbial community abundance and structure. After 2 years following drainage, ephemeral wetlands with short pond permanence were found to be most sensitive to drainage and the only wetland class with decreases in soil carbon. The temporary and seasonal wetland classes showed no significant differences in soil carbon content but there were changes in the organic matter with depth due to soil redistribution during drainage implementation. Jointly, this research provides region-specific estimates of soil carbon storage in drained prairie pothole wetlands that can be used to inform wetland soil carbon management in cultivated fields.