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Soil microbial diversity is extremely vulnerable to fertilization, which is one of the main anthropogenic activities associated with global changes. Yet we know little about how and why soil microbial diversity responds to fertilization across contrasting local ecological contexts. This knowledge is fundamental for predicting changes in soil microbial diversity in response to ongoing global changes. We analyzed soils from ten 20-year field fertilization (organic and/or inorganic) experiments across China and found that the national-scale responses of soil bacterial diversity to fertilization are dependent on ecological context. In acidic soils from regions with high precipitation and soil fertility, inorganic fertilization can result in further acidification, resulting in negative impacts on soil bacterial diversity. In comparison, organic fertilization causes a smaller disturbance to soil bacterial diversity. Despite the overall role of environmental contexts in driving soil microbial diversity, a small group of bacterial taxa were found to respond to fertilization in a consistent way across contrasting regions throughout China. Taxa such as Nitrosospira and Nitrososphaera, which benefit from nitrogen fertilizer addition, as well as Chitinophagaceae, Bacilli, and phototrophic bacteria, which respond positively to organic fertilization, could be used as bioindicators for soil fertility in response to fertilization at the national scale. Overall, our work provides new insights into the importance of local environmental context in determining the responses of soil microbial diversity to fertilization, and identifies regions with acidic soils wherein soil microbial diversity is more vulnerable to fertilization at the national scale.

Amphotericin-like glycosylated polyene macrolides (GPMs) are a clinically and industrially important family of natural products, but the mechanisms by which they exert their extraordinary biological activities have remained unclear for more than half a century. Amphotericin B exerts fungicidal action primarily via self-assembly into an extramembranous sponge that rapidly extracts ergosterol from fungal membranes, but it has remained unclear whether this mechanism is applicable to other GPMs. Using a highly conserved polyene–hemiketal region of GPMs that we hypothesized to represent a conserved ergosterol-binding domain, we bioinformatically mapped the entirety of the GPM sequence-function space and expanded the number of GPM biosynthetic gene clusters (BGCs) by 10-fold. We further leveraged bioinformatic predictions and tetrazine-based reactivity screening targeting the electron-rich polyene region of GPMs to discover a first-in-class methyltetraene- and diepoxide-containing GPM, kineosporicin, and to assign BGCs to many new producers of previously reported members. Leveraging a range of structurally diverse known and newly discovered GPMs, we found that the sterol sponge mechanism of fungicidal action is conserved., A combination of genome-mining and chemical reactivity identified new antifungal natural products. Such glycosylated polyene macrolides share a common mode of action via ergosterol extraction.

Wheat (Triticum aestivum L.) cultivars vary in their resistance to Fusarium head blight (FHB), while it is poorly understood how different cultivars influence FHB-causing Fusarium graminearum abundance in rhizosphere soil. A field experiment was conducted to investigate the influence of three wheat cultivars on FHB index, rhizosphere soil F. graminearum abundance, chemical properties, and microbial community composition and metabolic profiles. The cultivars were Sumai3 (SM), Yangmai13 (YM), and Annong8455 (AN), representing high, intermediate, and low resistance to FHB, respectively. Both wheat FHB index and rhizosphere F. graminearum abundance were ranked as SM

Ammonia oxidizers are key players in the global nitrogen cycle, yet little is known about their ecological performances and adaptation strategies for growth in saline terrestrial ecosystems. This study combined (13)C-DNA stable-isotope probing (SIP) microcosms with amplicon and shotgun sequencing to reveal the composition and genomic adaptations of active ammonia oxidizers in a saline-sodic (solonetz) soil with high salinity and pH (20.9 cmol(c) exchangeable Na(+) kg(−1) soil and pH 9.64). Both ammonia-oxidizing archaea (AOA) and bacteria (AOB) exhibited strong nitrification activities, although AOB performed most of the ammonia oxidation observed in the solonetz soil and in the farmland soil converted from solonetz soil. Members of the Nitrosococcus, which are more often associated with aquatic habitats, were identified as the dominant ammonia oxidizers in the solonetz soil with the first direct labeling evidence, while members of the Nitrosospira were the dominant ammonia oxidizers in the farmland soil, which had much lower salinity and pH. Metagenomic analysis of “Candidatus Nitrosococcus sp. Sol14”, a new species within the Nitrosococcus lineage, revealed multiple genomic adaptations predicted to facilitate osmotic and pH homeostasis in this extreme habitat, including direct Na(+) extrusion/H(+) import and the ability to increase intracellular osmotic pressure by accumulating compatible solutes. Comparative genomic analysis revealed that variation in salt-tolerance mechanisms was the primary driver for the niche differentiation of ammonia oxidizers in saline-sodic soils. These results demonstrate how ammonia oxidizers can adapt to saline-sodic soil with excessive Na(+) content and provide new insights on the nitrogen cycle in extreme terrestrial ecosystems.

Chemotherapy against the neglected tropical disease visceral leishmaniasis (VL) is suboptimal with only four licensed drugs. Amphotericin B (AmB), despite its toxicity, remained a second line drug for a long time. However, the demonstration that liposomal AmB is highly effective against VL propelled it, despite its cost, to a first line drug in many countries. While several ongoing efforts are aiming at finding cheaper and stable AmB-formulations, an alternative strategy is the development of less-toxic AmB derivatives. We show here that two less-toxic AmB derivatives with the carboxylate at position 16 of AmB derivatized to a methyl urea (AmB-MU) or amino urea (AmB-AU) are active in vitro against Leishmania donovani, both as free-living parasites as well as their intracellular form. Both less-toxic derivatives, similarly to AmB, target the ergosterol pathway of L. donovani. While the AmB-AU derivative showed female-specific liver toxicity in vivo, the AmB-MU derivative was well-tolerated and more effective than AmB against experimental VL. These studies are an important step for improving AmB-based therapy against a prevalent parasitic disease.

In this paper, we proposed a dual-signal fluorometric and colorimetric system based on silicon quantum dots (SiQDs) and 4-nitrophenol (4-NP) for pH and urease sensing. SiQDs with fluorescence emission of 460 nm were prepared via aqueous-phase synthesis. As the pH of the system gradually increased, the absorption band of 4-NP at 400 nm increased and a color reaction from colorless to yellow occurred. The absorption of 4-NP overlapped quiet well with the fluorescence excitation spectrum of SiQDs, which can effectively quench the fluorescence of SiQDs. Therefore, the change of fluorescence and absorption intensities could be used to quantify pH value. The fluorometric and colorimetric pH-sensing systems both exhibited a linear respond to pH ranging from 6.0 to 7.8 with an interval of 0.2 pH unit. Urease could specifically hydrolyze urea to generate carbon dioxide and ammonia, causing an obvious increase of the pH value. Thus, urease could also be detected quantitatively by the above dual-signal pH sensing system. The linear ranges of the fluorometric and colorimetric methods for urease detection were both 2–40 U L-1. The limits of detection were 1.67 and 1.07 U L-1, respectively. More importantly, this established dual-signal system has been successfully exploited in the detection of urease in real samples with satisfactory recoveries. Compared with other traditional single-signal assay strategies, the results obtained by dual-signal methods are more accurate and reliable.

The leaves of Aster glehni Fr. Schm. (Asteraceae) have been used to treat insomnia in Korea. Insomnia is a common adverse effect of therapeutic agents for Alzheimer's disease (AD), and the control of sleep disturbance may prevent dementia. We hypothesized that the leaves of A. glehni can attenuate cognitive dysfunctions observed in AD. We observed the ameliorating effects of the ethanolic extract of leaves of A. glehni (AG-D) on memory dysfunction through the Morris water maze test, the passive avoidance test, and the Y-maze test. We performed acetylcholinesterase (AChE) activity assay and Western blotting to determine the mechanism of action of AG-D. AG-D significantly attenuated memory dysfunction observed in the above behavior studies and inhibited the activity of AChE. AG-D also increased the levels of phosphorylation extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), and glycogen synthase kinase 3β (GSK-3β) and the expression levels of brain-derived neurotrophic factor (BDNF) in the hippocampi. These results suggest that AG-D ameliorates memory impairments by AChE inhibition and activation of ERK-CREB-BDNF and PI3K-Akt-GSK-3β signaling pathways. Taken together, this study suggests that AG-D could be used as a potential treatment for cognitive dysfunction.

The ability of soil microorganisms to uphold a variety of ecosystem services is of increasing interest, but our understanding of the linkage between cropland ecosystem services and the soil microbiome under long-term different fertilizer regimes is still limited. Here, we measured 15 response variables commonly used as indicators of cropland ecosystem services, and employed a shotgun metagenomic sequencing technology to determine microbiome taxonomic composition and functional composition in a sandy loam soil amended with organic and inorganic fertilizers for 28 years. Long-term application of fertilizers had overall positive effects on the variables that indicate soil C and nutrient status, organic material decomposition, nutrient transformation, and plant productivity, but the strength of these effects differed between organic and inorganic fertilization. In long term, organic fertilizers significantly increased the abundance of Sphingomonas and Acidobacteria as well as genes involved in C and N metabolism and degradation of aromatic organic compounds. The observed differences in the indicators of overall biological activity, litter decomposition, nutrient transformation, cropland fertility and productivity were associated with changes in taxonomic composition and functional composition of the soil microbiome. Network analysis revealed that soil organic C, total N, L-leucine aminopeptidase, phosphatase, microbial biomass C and N, and the multifunctionality index showed strong positive associations with Acidobacterium, Lysobacter and Nitrososphaera. These microbes are implicated in the decomposition of organic material, soil C and N transformation, and plant growth promotion. The present study deepens our understanding of the relationship between the soil microbiome and agro-ecosystem functioning, and provides support for the use of organic fertilization strategy to enhance multiple ecosystem services in croplands.

The present study determined the dynamic changes of bacterial community structures and its metabolism functions in rice straw composting process with different types of biochar addition. Results showed that both wheat and maize straw biochar addition greatly increased the temperatures and germination index. Biochar addition influenced microbial community structure and metabolic characteristics of the compost. Firmicutes, Actinobacteria and Proteobacteria were the dominant phyla throughout the process, and both biochar addition significantly increased the relative abundance of Actinobacteria phylum, especially genus Saccharomonospora. The abundance of bacterial genes related to amino acid metabolism, carbohydrate metabolism and energy metabolism were also increased with biochar addition. Moreover, total nitrogen was strongly affected by biochar addition and had the greatest influence on the bacterial community structure. These results indicated that biochar addition improved the maturity and fertility of the compost product as well as significantly regulated the microbial community structure and functions during composting process.

Many works have been performed to understand the mechanisms of the formation and persistence of memory. However, it is not fully understood whether the decay of long-term memory can be modulated by the activation of dopamine D1 receptor. A Barnes maze task was employed to measure long-term spatial memory. We observed that the spatial memory acquired through 3 trials per session for 4 days had begun to fade out by the 14th day and had completely disappeared by 21 days after the first probe test. The intraperitoneal administration of SKF 38393 (a dopamine D1 receptor agonist) for 7 days beginning on the 14th day after the first probe test prevented natural memory forgetting, and the intraperitoneal administration of SCH 23390 (a dopamine D1 receptor antagonist) prevented this memory persistence. In the Western blotting, the administration of SKF 38393 increased the phosphorylation levels of PKA, ERK1/2, CaMKII, and CREB in the hippocampus. In addition, such increased levels were decreased by the corresponding antagonist (SCH 23390). Moreover, the inhibition of PKA could completely reverse the preservation of spatial memory induced by dopamine D1 receptor activation. These results suggest that the activation of the dopamine D1 receptor plays a critical role in the persistence of long-term spatial memory through the PKA signaling pathway.

Long-term nitrogen field fertilization often results in significant changes in nitrifying communities that catalyze a key step in the global N cycle. However, whether microcosm studies are able to inform the dynamic changes in communities of ammonia-oxidizing bacteria (AOB) and archaea (AOA) under field conditions remains poorly understood. This study aimed to evaluate the transcriptional activities of nitrifying communities under in situ conditions, and we found that they were largely similar to those of 13C-labeled nitrifying communities in the urea-amended microcosms of soils that had received different N fertilization regimens for 22 years. High-throughput sequencing of 16S rRNA genes and transcripts suggested that Nitrosospira cluster 3-like AOB and Nitrososphaera viennensis-like AOA were significantly stimulated in N-fertilized fresh soils. Real-time quantitative PCR demonstrated that the significant increase of AOA and AOB in fresh soils upon nitrogen fertilization could be preserved in the air-dried soils. DNA-based stable-isotope probing (SIP) further revealed the greatest labeling of Nitrosospira cluster 3-like AOB and Nitrosospira viennensis-like AOA, despite the strong advantage of AOB over AOA in the N-fertilized soils. Nitrobacter-like nitrite-oxidizing bacteria (NOB) played more important roles than Nitrospira-like NOB in urea-amended SIP microcosms, while the situation was the opposite under field conditions. Our results suggest that long-term fertilization selected for physiologically versatile AOB and AOA that could have been adapted to a wide range of substrate ammonium concentrations. It also provides compelling evidence that the dominant communities of transcriptionally active nitrifiers under field conditions were largely similar to those revealed in 13C-labeled microcosms. IMPORTANCE The role of manipulated microcosms in microbial ecology has been much debated, because they cannot entirely represent the in situ situation. We collected soil samples from 20 field plots, including 5 different treatments with and without nitrogen fertilizers for 22 years, in order to assess active nitrifying communities by in situ transcriptomics and microcosm-based stable-isotope probing. The results showed that chronic N enrichment led to competitive advantages of Nitrosospira cluster 3-like AOB over N. viennensis-like AOA in soils under field conditions. Microcosm labeling revealed similar results for active AOA and AOB, although an apparent discrepancy was observed for nitrite-oxidizing bacteria. This study suggests that the soil microbiome represents a relatively stable community resulting from complex evolutionary processes over a large time scale, and microcosms can serve as powerful tools to test the theory of environmental filtering on the key functional microbial guilds.

The effects of land use and fertilization regimes on soil denitrification potential (SDP) are often investigated, but less is known about the responses of SDP to different types of land use with different fertilization regimes in soils derived from the same parent material. We conducted a study using soil samples from 2 long-term (over 30 years) fertilization experiments to determine the difference in SDP between paddy soil and upland soil derived from the same quaternary red clay parent material. The results showed that the SDP in paddy soil was 6.82 times higher than the upland soil, which was due to the higher abundances of narG, nirS, and nirK genes and nirS-denitrifying bacteria (Bradyrhizobium, Cupriavidus, and Herbaspirillum) in paddy soil. Inorganic fertilization regimes did not significantly affect the SDP of upland soil over the control group, whereas SDP in the NPK and 2NPK treatments in paddy soil were reduced relative the control group by 26.48% and 75.65%, respectively. Compared with the control, the NPKOM treatment consistently yielded the highest SDP in both soils, with 2.47 times and 2.86 times higher for paddy soil and upland soil, respectively. The SDP of paddy soil was significantly correlated with narG and nirS genes mainly regulated by amorphous aluminum oxide, whereas the treatment effect on SDP for upland soil largely depended on differences in nirS-denitrifying bacteria at the genus level (Herbaspirillum, Sulfuritalea, and Cupriavidus) and species level, which were mainly controlled by soil pH. Partial least squares path models further demonstrated that the direct effects of functional genes on SDP were greatest for paddy soil, whereas nirS-denitrifying bacterial communities played a larger role in upland soil. The results presented herein represent a key step toward understanding the mechanisms that govern SDP under land use and long-term fertilization.

The formation of aggregates plays a key role in shaping soil microenvironment, which in turn influences microbial community structure and organic carbon (C) dynamics in soil. The objective of this study was to identify the linkages between soil macroaggregation and microbial community as well as organic C accumulation using field samples following 9 years of conservation tillage. A wet-sieving method was used to fractionate soil aggregates and soil microbial abundance and community composition were determined using phospholipid fatty acid (PLFA) analysis. Compared to continuous tillage, reduced/no-tillage and straw returning significantly promoted soil macroaggregation and consequently increased organic C, C/N ratio and soil moisture but decreased the porosity and computed effective oxygen diffusion coefficients. The changes in these soil characteristics were closely related to the abundance and composition of microbial community. The abundance of microbial PLFAs in reduced/no-tillage soils was 30.0% higher in Gram-positive bacteria, 11.6% higher in Gram-negative bacteria, 71.7% higher in fungi and 45.4% lower in actinobacteria, whereas straw returning significantly increased all microbial PLFA abundances relative to straw removing. The ratios of bacteria/fungi (B/F) and monounsaturated/branched (M/B) PLFAs were significantly correlated with the volumetric soil water content, porosity, or computed effective oxygen diffusion coefficients. Soil macroaggregation and microbial community composition collectively explained 82.4% variation in organic C accumulation and their interaction made the largest contribution. Overall, soil macroaggregation under conservation tillage might cause a shift in microbial community to more fungi and anaerobes through primarily influencing soil moisture and aeration, which could effectively promote organic C accumulation in soil.

Ethnopharmacological relevance The seeds of Ziziphus jujuba var. spinosa (Bunge) Hu ex H.F Chow (Rhamnaceae) and the roots of Codonopsis lanceolata (Siedbold & Zucc.) Benth. & Hook. f ex Trautv. (Campanulaceae), contained in the DHP1402, have long been used for treating dementia or hypomnesia as folk medicine. Aim of the study It has been reported that Z. jujuba var. spinosa and C. lanceolata are effective in improving cognitive function, but via different mechanisms. Therefore, in the present study, we evaluated the synergistic effects of Z. jujuba var. spinosa and C. lanceolata on scopolamine-induced memory impairment. Materials and methods Scopolamine, a cholinergic muscarinic receptor antagonist, was used to induce cognitive dysfunction. We employed several behavioral tasks to estimate the synergistic effect of the seeds of Z. jujuba var. spinosa and the roots of C. lanceolata . In addition, we introduced the Western blotting, the antagonism passive avoidance task to investigate a synergistic effect of an herbal formulation. Results Synergistic effects of a combination of Z. jujuba var. spinosa and C. lanceolata at a 5:1 ratio [(w/w), DHP1402] were observed against cognitive dysfunction in the passive avoidance and Y-maze tasks. DHP1402 also ameliorated memory deficits in a dose-dependent manner in these behavioral tasks, as well as in the Morris water maze task. According to the Western blot results, the phosphorylation levels of protein kinase A (PKA), extracellular signal-regulated kinase (ERK) and cAMP response element-binding protein (CREB) in the hippocampus were also increased in a synergistic manner after the administration of DHP1402. In addition, we found that the effects of DHP1402 on cognitive function were mediated by N -methyl- D -aspartate (NMDA) receptor signalling, based on the antagonism studies. Furthermore, we found that DHP1402 has inhibitory activity against acetylcholinesterase (AChE). Conclusion DHP1402 attenuates cholinergic blockade-induced cognitive dysfunction through NMDA receptor modulation, PKA-ERK-CREB pathway activation, and AChE inhibition. Therefore, DHP1402 could be a candidate for alleviating cognitive dysfunction.

In the present study, we investigated whether 1-palmitoyl-2-linoleoyl-3-acetylglycerol (PLAG), a component of antlers of Cervus nippon Temminck, would have memory-ameliorating properties against cholinergic blockade-induced memory impairment in mice. In the passive avoidance task to investigate the effects of PLAG on long-term memory, PLAG (10 mg/kg, p.o.) administration ameliorated scopolamine-induced memory impairment. PLAG also reversed the impairments of working memory in the Y-maze task and spatial memory as shown in the Morris water maze. To identify the mechanism of the memory-ameliorating effect of PLAG, acetylcholinesterase (AChE) inhibition assay and the Western blot analysis were conducted. In the AChE inhibition assay, PLAG inhibited the AChE activity in mice and PLAG increased the expression levels of phosphorylated CaMKII, ERK, and CREB in the hippocampus. Additionally, long-term potentiation (LTP) of synaptic strength occurred by PLAG treatment in the hippocampal cultures. Overall, the present study suggests that PLAG reversed memory deficits in an animal model and that it affects biochemical pathways related to learning and memory.

Acanthopanax koreanum Nakai (Araliaceae) is one of the most widely cultivated medicinal plants in Jeju Island, Korea, and the roots and stem bark of A. koreanum have been traditionally used as a tonic agent for general weakness. However, the use of A. koreanum for general weakness observed in the elderly, including those with declined cognitive function, has not been intensively investigated. This study was performed to investigate the effect of the ethanol extract of A. koreanum (EEAK) on cholinergic blockade-induced memory impairment in mice. To evaluate the ameliorating effects of EEAK against scopolamine-induced memory impairment, mice were orally administered EEAK (25, 50, 100, or 200 mg/kg), and several behavioral tasks, including a passive avoidance task, the Y-maze, and a novel object recognition task, were employed. Besides, western blot analysis was conducted to examine whether EEAK affected memory-associated signaling molecules, such as protein kinase B (Akt), Ca2+/calmodulin-dependent protein kinase II (CaMKII), and cAMP response element-binding protein (CREB). The administration of EEAK (100 or 200 mg/kg, p.o.) significantly ameliorated the scopolamine-induced cognitive impairment in the passive avoidance task, the Y-maze, and the novel object recognition task. The phosphorylation levels of both Akt and CaMKII were significantly increased by approximately two-fold compared with the control group because of the administration of EEAK (100 or 200 mg/kg) (p

Oleanolic acid is a naturally occurring triterpenoid and is widely present in food and medicinal plants. To examine the effect of oleanolic acid on memory deficits, we employed a cholinergic blockade-induced cognitive deficit mouse model. A single administration of oleanolic acid significantly increased the latency on the passive avoidance task and affected the alternation behavior on the Y-maze task and the exploration time on the novel object recognition task, indicating that oleanolic acid reverses the cognitive impairment induced by scopolamine. In accordance with previous reports, oleanolic acid enhanced extracellular-signal-regulated kinase 1/2 (ERK1/2) and cAMP response element-binding protein (CREB) phosphorylation and brain-derived neurotrophic factor (BDNF) expression in the hippocampus. Interestingly, ameliorating effect of oleanolic acid on scopolamine-induced memory impairment was abolished by N2-(2-{[(2-oxoazepan-3-yl)amino]carbonyl}phenyl)benzo[b]thiophene-2-carboxamide (ANA-12), a potent and specific inhibitor of tropomyosin receptor kinase B (TrkB), in the passive avoidance task. Similarly, oleanolic acid significantly evoked long-term potentiation in a dose-dependent manner, which was diminished by ANA-12 treatment as shown in the electrophysiology study. Together, these results imply that oleanolic acid ameliorates scopolamine-induced memory impairment by modulating the BDNF-ERK1/2-CREB pathway through TrkB activation in mice, suggesting that oleanolic acid would be a potential therapeutic agent for the treatment of cognitive deficits.

Despite our current understanding of soil C and N interactions, less is known about the response and feedback of C mineralization to soil P availability driven by microorganisms. To better understand these interactions, soils with long-term P-sufficient NPK fertilization (available P: 13.4 mg kg −1 ) and P-deficient NK fertilization (available P: 0.96 mg kg −1 ) were incubated with and without glucose. CO 2 emissions were monitored to characterize C mineralization during the incubation. The soil bacterial community structure, quantity and metabolic activity were evaluated using high-throughput sequencing, qPCR and microcalorimetric dynamics. Compared with P-sufficient soils, P-deficient soils had significantly lower basal respiration, but a significantly higher net mineralization of added glucose, probably due to higher energy cost of soil microorganisms. Glucose addition promoted microbial biomass and activity, particularly in P-deficient soils, and this improvement was maintained for at least 70 days. Shifts in bacterial community composition were induced by a predominance of several specific taxonomic groups, all of which were capable of solubilizing P in soils. P-deficiency decreases the retention of exogenous labile C into soil. Adding labile C to P-deficient soils may shift P from relatively unavailable soil-bound pools into microbial biomass pools through pool cycling. Our results indicated negative effects of P-deficiency on soil C retention, as well as positive effects of labile C on soil P availability in arable soils.

Zinc(II) complexes of curcumin display moderate cytotoxicity towards cancer cells at low micromolar concentrations. However, the clinical use of zinc(II) complexes is hampered by hydrolytic insolubility and poor bioavailability and their anticancer mechanisms remain unclear. Here, we investigated the efficacy and mechanism of action of a polyvinylpyrrolidone (PVP-k30)-based solid dispersion of Zn(II)-curcumin (ZnCM-SD) against hepatocellular carcinoma (HCC) in vitro and in vivo. In vitro assays revealed ZnCM-SD not only reduced the viability of HepG2 cells and SK-HEP1 cells in a dose-dependent manner, but also potently and synergistically enhanced cell growth inhibition and cell death in response to doxorubicin by regulating cellular zinc homeostasis. ZnCM-SD was internalized into the cells via non-specific endocytosis and degraded to release curcumin and Zn2+ ions within cells. The anticancer effects also occur in vivo in animals following the oral administration of ZnCM-SD, without significantly affecting the weight of the animals. Interestingly, ZnCM-SD did not reduce tumor growth or affect zinc homeostasis in HepG2-bearing mice after gut microbiome depletion. Moreover, administration of ZnCM-SD alone or in combination with doxorubicin significantly attenuated gut dysbiosis and zinc dyshomeostasis in a rat HCC model. Notably, fecal microbiota transplantation revealed the ability of ZnCM-SD to regulate zinc homeostasis and act as a chemosensitizer for doxorubicin were dependent on the gut microbiota. The crucial role of the gut microbiota in the chemosensitizing ability of ZnCM-SD was confirmed by broad-spectrum antibiotic treatment. Collectively, ZnCM-SD could represent a simple, well-tolerated, safe, effective therapy and function as a novel chemosensitizing agent for cancer.

The objective of this study was to investigate microbial responses when seaweed fertilizer was added to maize straw compost. The metabolic characteristics and bacterial communities were assessed using 16S rRNA sequencing and Biolog analysis. Results showed that the urease, dehydrogenase, and metabolic activities improved on day 21 after the addition of seaweed fertilizer. Firmicutes, Proteobacteria, and Actinobacteria were the dominant phyla in the compost. The relative abundances of Proteobacteria and Actinobacteria were increased by addition of seaweed fertilizer during the early composting stage. Temperature, NH4+-N, NO3--N concentrations, and carbon source metabolism had close relationships with bacterial community variation during composting. A network analysis revealed that NO3--N content had a positive association with Aeromicrobium, and seaweed fertilizer application improved the growth of keystone species related to N cycling. These results suggested that seaweed fertilizer influenced bacterial community succession through its effects on N concentrations during the composting process.

Bee pollen consists of floral pollen mixed with bee secretions and nectar. It has been considered as a functional food and has different kinds of biologically active ingredients, such as flavonoids, polyphenols, phytosterols and minerals. However, its function in cognition has yet been investigated. In the present study, we investigated the ameliorating effect of bee pollen against scopolamine-caused cognitive impairment through the passive avoidance test, the Y-maze test and the Morris water maze test. In addition, Western blotting was employed to verify the effects of bee pollen on memory-related signaling molecules in the hippocampus. Bee pollen extract (100 or 300 mg/kg, per os (p.o.)) obviously reversed scopolamine-caused cognitive impairment in the passive avoidance test, ameliorated spontaneous alternation versus the scopolamine-treated group in the Y-maze test and prolonged swimming time in the target zone in the Morris water maze test. In addition, the phosphorylation levels of extracellular signal-regulated kinase (ERK), cAMP response element-binding protein (CREB), protein kinase B (Akt) and glycogen synthase kinase-3β (GSK-3β), and the expression levels of brain-derived neurotrophic factor (BDNF) and tissue plasminogen activator (tPA) in the hippocampi, were increased in response to the treatment with bee pollen extract (100 or 300 mg/kg, p.o.). These results indicated that bee pollen ameliorates cognitive impairment induced by cholinergic blockade through the enhancing conversion of proBDNF to mature BDNF by tPA, probably, through the ERK-CREB pathway or Akt-GSK-3β signaling pathway and would be a useful agent for the treatment of cognitive dysfunction.

Succession and metabolism functions of bacterial communities were determined in maize straw composting with earthworm casts and zeolite addition by using high-throughput sequencing, Biolog and PICRUSt. Results showed that earthworm casts and zeolite addition increased the temperature, decreased NH4+ contents and affected bacterial community structure. The relative abundances of Firmicutes and Betaproteobacteria increased with earthworm casts and zeolite addition in the late stage. The abundances of genes related to carbohydrate metabolism, amino acid metabolism, and energy metabolism were increased by these two additives in the early stage, but decreased in the late stage. Network analysis demonstrated that members of Bacillaceae were identified as the keystone taxa. Temperature showed negative relationship with Georgenia, while NH4+ exhibited positive associations with Georgenia, Devosia, Ruania and Mycobacterium. These results indicated that earthworm casts and zeolite addition benefitted the keystone species and enhanced the metabolism capacity of bacterial community, thereby improved the quality of compost.

Background China exhibits a great diversity of ecosystems and abundant cattle resources, with nearly 30 million cattle from 53 indigenous breeds reared in specific geographic regions. To explore the genetic diversity and population structure of Chinese indigenous cattle, a population genetic analysis at both the individual and population levels was conducted and the admixture analysis was performed. We genotyped 572 samples from 20 Chinese indigenous cattle breeds using GeneSeek Genomic Profiler Bovine LD (GGP-LD, 30 K) and downloaded the published data of 77 samples from 4 worldwide commercial breeds genotyped with Illumina BovineSNP50 Beadchip (SNP50, 50 K). Results In principal component analysis (PCA) and neighbour-joining (NJ) tree analysis, samples of the same breeds were grouped together, leading to clear separation from other breeds. And Chinese indigenous cattle were clustered into two groups of southern and northern breeds, originated from Asian Bos indicus lineage and Eurasian Bos taurus lineage, respectively. In STRUCTURE K = 2, a clear transition occurred from the northern breeds to the southern breeds. Additionally, the northern breeds contained a smaller Eurasian taurine (62.5%) descent proportion than that reported previously (more than 90%). In STRUCTURE K = 3, a distinct descent was detected in the southern Chinese breeds, which could reflect a long-term selection history of Chinese indigenous cattle. The results from TreeMix and f3 statistic provided the evidence of an admixture history between southern breeds and northern breeds. Conclusions Consistent with the observed geographical distributions, Chinese indigenous cattle were divided into two genetic clusters, northern indigenous cattle and southern indigenous cattle. Three improved breeds in the northern area also exhibited northern indigenous ancestry. We found that the breeds distributed in the northern China showed more southern lineage introgression than previously reported. Central-located populations appeared to the admixture between southern and northern lineages, and introgression events from European cattle were observed in Luxi Cattle, Qinchuan Cattle and Jinnan Cattle. The study revealed the population structures and levels of admixture pattern among Chinese indigenous cattle, shedding light on the origin and evolutionary history of these breeds. Electronic supplementary material The online version of this article (10.1186/s12863-018-0705-9) contains supplementary material, which is available to authorized users.

Manure application is widely recognized as a method of improving soil structure and soil fertility due to additional organic matter and nutrient inputs. However, the salinity of animal manure may have a detrimental effect on soil aggregation. The objective of this study was to determine the effects of long-term animal manure application on soil aggregation, binding agents (soil organic carbon, SOC and glomalin-related soil protein, GRSP), and dispersing agents (e.g., Na+) and their relationships based on nine long-term fertilization experiments (12 to 39 yr) across China. The two red soil experiments (Qiyang, QY and Jinxian, JX) and one paddy soil experiment in Jinxian (JX-P) were conducted in southern China (precipitation above 1200 mm yr−1), whereas the other six experiments were established in semi-humid or arid regions in China with precipitation in the range of 500–900 mm yr−1. Each experiment included three treatments as follows: no fertilization (Control), inorganic fertilizer (NP or NPK), and a combination of inorganic fertilizer and animal manure (NPM or NPKM). Long-term animal manure application not only significantly increased the biological binding agents (i.e., SOC and GRSP) in the nine experiments but also considerably increased the dispersing agents (i.e., exchangeable Na+) (P

High-throughput sequencing and network analysis were used to investigate the dynamic changes and the effects on bacterial community structures in green waste compost after biochar addition. Biochar addition led to higher thermophilic temperatures and total nitrogen (TN) concentrations and a longer thermophilic period compared with no biochar addition. Biochar also greatly influenced the composition of the bacterial community. Nitriliruptoraceae and Bacillaceae abundances increased in the poultry manure after biochar treatment, and Alcaligenaceae, Rhodispirillaceae and Xanthomonadaceae were more abundant in the cattle manure. Dissolved organic carbon (DOC) and TN emerged as the key determinants of bacterial community composition. Network analysis revealed that DOC had strong positive associations with some taxa (e.g. Comamonas, Leucobacter and Acidimicrobiales), whereas TN had negative associations with other taxa (e.g. Microbacteriaceae and Aeromicrobium). This study has revealed the key taxa related to the carbon and nitrogen cycle during composting with biochar.

The importance of soil saprotrophic fungi in agroecosystems is receiving increasing attention. However, the linkages between saprotrophic fungal communities and soil fertility and stoichiometry in agroecosystems are poorly understood. Here, four long-term field fertilization experiments with three treatments (i.e., no fertilizer (control), chemical nitrogen (N), phosphorus (P), and potassium (K) fertilizer (NPK), chemical combined with organic fertilizer (NPKM)) were performed to evaluate the associations of saprotrophic fungal communities with the soil fertility index (SFI) and stoichiometric carbon (C):N:P ratios. Long-term NPKM application increased the SFI by 57.66%–129.50% at the four sites, and the SFI was strongly correlated with the saprotrophic fungal community composition, which indicated that exogenous organic matter improved soil fertility to a large extent and stimulated the growth of saprotrophic fungi in arable soils. The soil C:P and N:P ratios were notably decreased in the fertilization treatment, indicated that P was sufficient. Further, the composition of saprotrophic fungi was significantly related to the C:N and N:P ratios, suggested that N status is an important factor for soil saprotrophic fungal communities. Our study provides empirical evidence that the saprotrophic fungal communities in arable soils are strongly associated with soil fertility and stoichiometry and offers guidance for improving soil fertility by regulating the saprophytic fungal community.

The supposition that cropping system might affect fertilization-induced changes in chemical composition of soil organic carbon (SOC) and microbial community composition has mostly derived from field experiments in different sites, which may have been disturbed by different climate conditions and parent materials. Here, two adjacent long-term field experiments were used, which contained similar four fertilization regimes of NPK, 2NPK, NPKOM, and an unamended control (Control), while contrasting annual cropping system of rice–rice and maize–maize, to investigate how fertilization-induced changes in SOC chemistry differ in paddy soil from upland soil, and whether they correlate with different bacterial taxa. The bacterial community composition of the paddy soil was mainly determined by NO3–-N and total N, and that of the upland was mostly explained by SOC. The NPK, 2NPK, and NPKOM treatments from the paddy soil increased CCH3, decreased OCH, and reduced CH/CH2, while those from the upland soil decreased CH/CH2, increased OCH, and raised CH/CH2, respectively, indicating opposite shifts of OCH and alkyl C (i.e., CCH3 and CH/CH2) in each paired fertilizer treatment of the two soils. In the paddy soil, the CCH3 from the NPK treatment showed no association with any bacterial taxa, the OCH from the 2NPK showed a negative association with NO3–-related species, and the CH/CH2 from the NPKOM showed a negative association with Proteobacteria and Bacteroidetes. In the upland soil, the CH/CH2 from the NPK negatively associated with Thermogemmatisporales and Acidobacteriaceae subgroup 1, the OCH from the 2NPK positively associated with Thermogemmatisporales, and the CH/CH2 from the NPKOM positively associated with copiotrophic bacterial taxa. Our results provided direct evidence that cropping system mediated the shift direction of specific functional group under specific fertilization regimes, and implied that completely opposite shifts of OCH and alkyl C in each paired fertilizer treatment of the paddy soil and upland soil can likely be attributed to different fertilization-induced shifts in the microbial community composition resulting from soil N change in the paddy soil and soil C change in the upland soil.

Soil microbes play a key role in terrestrial ecosystems. In the saline-sodic environment, the microbial communities and their community assembly processes, especially throughout soil profiles, however, are not well understood. Here, we characterized the soil microbial communities across three sodicity/salinity gradients to 1.0 m depth in the western Songnen Plain, Northeast China. Soil sodicity (exchangeable sodium percentage) and salinity (electrical conductivity) exerted a strong influence on bacterial community compositions compared with other soil properties. These filtering effects of sodicity/salinity were strong on the microbial communities in the layer of 0–60 cm across the three sites, with more contributions of deterministic processes over stochastic processes. As sodicity/salinity decreased, some potential biomarkers for a high community tolerance to salt or soda, such as KSA1, Halomonas, Euzebya and Bacillus, were depleted; meanwhile, some taxa associated with decomposition of organic matter and nutrient transformations, such as Arthrobacter, Mesorhizobium, Pseudomonas and Nitrospira, were enriched. Within each soil profile, the turnover in community composition was lower than between sites, and the stochastic processes (homogenizing dispersal) dominated the microbial assembly. Yet, the surface and deepest depth (80–100 cm) of saline-sodic soils still harbored some distinct microbial communities. The near-surface (20–40 cm) may serve as a gating role for limiting surface microbial penetrance into the deeper depth. Taken together, our results show that sodicity/salinity can strongly structure soil microbial communities, and emphasize the importance of soil depth as an environmental gradient on the assembly processes in saline-sodic soils.

The microbiomes inhabiting plant roots and their adjacent soil environment play significant roles in plant growth, health, and ecological services. Recent studies have provided detailed insights into many phytomicrobiomes, but our understanding of the root-associated microbiomes of perennial ryegrass is still very limited. Here we carried out a stratified rhizobox experiment involving a fluvo-aquic soil and red soil under elevated (750 ppm) and ambient (350–400 ppm) CO2, and used deep amplicon sequencing to investigate the ryegrass root-associated bacterial microbiomes across compartments (the bulk soil, 2-mm outer rhizosphere, rhizosphere, rhizoplane, and endosphere). We observed increased relative abundances of Proteobacteria (mainly Rhizobiaceae and Enterobacteriaceae) in the rhizosphere, rhizoplane, and endosphere compared with the bulk soil and outer rhizosphere. The dominant bacterial genera (mainly Methylobacterium, Rhizobium, Pseudomonas, Stenotrophomonas, and Enterobacter) which facilitate plant growth and soil nutrient cycling were enriched in the rhizosphere, rhizoplane, and endosphere compared with outer rhizosphere. PICRUSt predicted moderate functional differentiation of bacterial microbiomes across compartments. The bacterial α-diversity showed a decreased trend along a spatial gradient from the bulk soil to the endosphere. The β-diversity and network analyses indicated significant differences in the root-associated bacterial microbiomes between the fluvo-aquic soil and red soil. In conclusion, our study suggests that the filtration and acquisition of the ryegrass root-associated bacterial microbiomes are largely controlled by indigenous soil microbial communities, rather than the changes in root rhizodeposition caused by elevated CO2.

Variations in temperature and moisture play an important role in soil organic matter (SOM) decomposition. However, relationships between changes in microbial community composition induced by increasing temperature and SOM decomposition are still unclear. The present study was conducted to investigate the effects of temperature and moisture levels on soil respiration and microbial communities involved in straw decomposition and elucidate the impact of microbial communities on straw mass loss. A 120-d litterbag experiment was conducted using wheat and maize straw at three levels of soil moisture (40%, 70%, and 90% of water-holding capacity) and temperature (15, 25, and 35 °C). The microbial communities were then assessed by phospholipid fatty acid (PLFA) analysis. With the exception of fungal PLFAs in maize straw at day 120, the PLFAs indicative of Gram-negative bacteria and fungi decreased with increasing temperatures. Temperature and straw C/N ratio significantly affected the microbial PLFA composition at the early stage, while soil microbial biomass carbon (C) had a stronger effect than straw C/N ratio at the later stage. Soil moisture levels exhibited no significant effect on microbial PLFA composition. Total PLFAs significantly influenced straw mass loss at the early stage of decomposition, but not at the later stage. In addition, the ratio of Gram-negative and Gram-positive bacterial PLFAs was negatively correlated with the straw mass loss. These results indicated that shifts in microbial PLFA composition induced by temperature, straw quality, and microbial C sources could lead to changes in straw decomposition.

Heat stress dramatically decreases bull sperm quality and has recently received more attention due to the warmer global climate and more intensive production. However, no data exist regarding sperm quality or the related molecular mechanisms under heat stress. Recent studies showed that inducible heat shock proteins (HSPs) play an important role in the dairy heat stress regulation. In this article, to investigate the impacts of heat stress on sperm quality and the associated molecular mechanisms, sperm quality and enzyme activities concerning acrosome reaction were assessed in Simmental, Limousin and Yanbian bulls under heat stress. Subsequently, changes in heat shock protein expression profiles of Simmental bulls were observed, because we observed that sperm quality of these bulls was most sensitive to heat stress. Finally, the relationship between sperm quality and heat shock protein expression under heat stress was analyzed. The results show that summer heat stress decreased the sperm quality of the three bull breeds significantly. Moreover, different levels of heat stimulation induced various enzyme activity changes, among which the activity change in acrosomal enzyme was the most remarkable. Furthermore, the expression of heat shock proteins in the sperm was influenced by the imposed heat stress, among which the expression levels of HSP60 and HSP70 were increased while HSP90 decreased. In summary, our data show that heat stress seriously affects sperm quality and that HSP90 was most sensitive, although it should be noted that seasonal effects may confound these results. This change in heat shock protein expression may be the major factor that affected the sperm quality of the bulls. The findings may provide a new hypothesis for how heat stress impacts reproduction mechanistically.

Solonetz soils are typically characterized with excessive saline-alkali content, and physiochemical methods are often employed for solonetz reclamation. However, increasing lines of evidences have suggested that microbiomes play a pivotal role in sustaining soil fertility than previously appreciated, and it remains poorly understood about the taxonomic identity of microbiomes in solonetz soils and the underlying mechanisms that govern the community assembly of solonetz restoration. In this study, we show that significant increase of rare phylotypes occurred after solonetz restoration by natural grassland and reclaimed farmland, and prokaryotic community assembly was dominated by deterministic rather than stochastic processes. The exchangeable sodium decreased significantly from 27.04% in solonetz, to 4.35% in grassland and 0.52% in farmland. Soil organic carbon (SOC) increased significantly with 4.17- and 2.18- fold in grassland and farmland soils relative to solonetz, respectively. Real-time quantitative PCR indicated drastic increases in the abundance of functional genes responsible for nitrogen fixation and nitrification. High-throughput sequencing of 16S rRNA genes further revealed that 180 and 215 phylotypes could be considered as key taxa showing significant increase after solonetz restoration by grassland and farmland. Intriguingly, more than 96% of these key phylotypes were originally rare phylotypes with

Soil bacteria and fungi are essential constituents of arable ecosystems. Long-term phosphorus (P) deficiency has been proven to significantly affect microbial community diversity and composition. However, how P deficiency impacts the interplay between bacteria and fungi remains unclear. The objective was to explore the bacterial-fungal co-occurrence patterns in soil as influenced by long-term P deficiency, using network analysis. Bacterial and fungal community datasets were obtained from soils which have consistently and continuously been fertilized with P (+P) or without P (−P) for >20 years in three long-term field experiments in China. Our results showed that the −P network was less complex and less efficient as indicated by fewer edges, lower average degree and higher average path length. Chronic P deficiency simultaneously decreased positive and negative bacterial-fungal edges, implying that both synergistic and antagonistic interactions were enhanced by P application. The keystone species differed between +P and −P network, and those from the +P network showed significant correlations with less soil properties, indicating the −P network was more sensitive to environmental disturbances. Four major modules were observed in both +P and −P networks, and they related with different soil properties, suggesting distinct habitat heterogeneity or niche partitioning. All the modules had similar ecological roles, while the module composition was altered by long-term P deficiency, indicating a possible ecological redundancy. Most of the fungi from the −P treatment were distributed in a special module negatively correlated with available P, indicating that fungi may prefer niches with low P concentration. Taken together, our results illustrated the crucial roles of P availability in microbial community assembly and investigated the responses of co-occurrence patterns to long-term P deficiency by regulating the topological properties, keystone species and module composition of the bacterial-fungal network.

The stem of Opuntia ficus-indica var. saboten is edible and has been used as a medicinal herb on Jeju Island in Korea. We previously reported that the butanolic extract of O. ficus-indica var. saboten exerts the enhancement of long-term memory in mice. However, the antiamnesic effects of O. ficus-indica var. saboten and its mode of action has not been clearly elucidated. In the present study, we explored the effects of the ethanolic extract of stems of O. ficus-indica var. saboten (EOFS) on cognitive performance in mouse and attempted to delineate its mechanism of action. We used the passive avoidance, Y-maze, and novel object recognition tests to assess its effects on cognitive functions in scopolamine-induced memory-impaired mice. We observed that EOFS (100, 200, and 400 mg/kg) ameliorated scopolamine-induced cognitive dysfunction. We also explored its mechanism of action by conducting an acetylcholinesterase (AChE) activity assay using the mouse whole brain and Western blot using the mouse hippocampal tissue. Western blot analysis and the ex vivo study revealed that EOFS increased the levels of phosphorylated extracellular signal-regulated kinase and cAMP response element-binding protein (CREB) and the levels of brain-derived neurotrophic factor (BDNF) expression in the hippocampus. It also inhibited AChE activity in the brain. Our findings suggest that EOFS would be useful for the treatment of cholinergic blockade-induced cognitive dysfunction.

Membrane association of estrogen receptors (ER) depends on cysteine palmitoylation and two leucines in the ligand binding domain (LBD), conserved in most steroid receptors. The role of this region, corresponding to helix 8 of the glucocorticoid receptor (GR) LBD, on membrane association of GR was studied in 4B cells, expressing endogenous GR, and Cos-7 cells transfected EGFP-GR constructs. 4B cells preloaded with radiolabeled palmitic acid showed no radioactivity incorporation into immunoprecipitated GR. Moreover, mutation C683A (corresponding to ER palmitoylation site) did not affect corticosterone-induced membrane association of GR. Mutations L687-690A, L682A, E680G and K685G, prevented membrane and also nuclear localization through reduced ligand binding. L687-690A mutation decreased association of GR with heat shock protein 90 and transcriptional activity, without overt effects on receptor protein stability. The data demonstrate that palmitoylation does not mediate membrane association of GR, but that the region 680-690 (helix 8), is critical for ligand binding and receptor function.

The quick assessment approaches for accurately measuring water-use efficiency (WUE) in maize under water stress are important to water-saving agriculture. We investigated how and whether carbon isotope composition (δ 13C) and specific leaf area (SLA) could be used to assess WUE for maize as influenced by water stress. A pot experiment was conducted twice during six typical maize (Zea mays) growth stages of seedling, jointing, booting, tasseling, filling and maturity, respectively. The ratio between the activities of ribulose 1,5-bisphosphate carboxylase (Rubisco) and phosphoenolpyruvate carboxylase decreased bundle sheath leakiness (φ) under water deficiency, caused more 13C to be assimilated, and resulted in increased δ 13C in leaves. Water stress increased the fractionation of 13C when assimilates were transported from leaf to stem, indicating that water stress affected leaf expansion and translocation of assimilates from leaf to stem, and resulted in thicker leaves and lower SLA. WUE showed significant positive correlations with leaf δ 13C and SLA, implying that leaf δ 13C and SLA could effectively reflect the drought adaptation and high WUE under different water conditions.

It is of ecological significance to investigate microbial communities in response to straw amendment and moisture in arable soils. However, in Chinese fluvo-aquic soils, these responses are still poorly understood. We designed an incubation experiment involving two soils with and without the addition of maize stubble at two moisture levels, and bacterial community structure at days 20, 80, and 200 after the start of incubation was assessed via bar-coded pyrosequencing of the 16S rDNA amplicons. In the presence of stubble with identical moisture level, we observed higher bacterial diversity and richness in long-term organic manure-fertilized soil compared with the unfertilized soil at days 20 and 80, which we attributed to the different quality and quantity of organic matter between the two soils. However, there was no significant difference in bacterial diversity and richness between the two soils at day 200, indicating that long-term straw amendment probably lessens the difference in bacterial community structure between the two soils. In the amended soils bacterial diversity, richness, and community composition at 25% of the water-holding capacity distinctly differed from those at 55% of the water-holding capacity, indicating that moisture strongly affects bacterial distribution in the amended soils. As stubble-C availability declined over time, the dominance of copiotrophic population weakened, and oligotrophic population was moderately abundant. Finally, our study suggests that dissolved organic carbon, which drives redistribution in copiotrophic and oligotrophic categories in response to the varying water and stubble-C availability, is a determinant of bacterial community composition in the amended soils. (C) 2014 Elsevier B.V. All rights reserved.

Organic manure (OM) fertilization has a profound impact on agroecosystems. However, little is known about temporal responses and roles of the specific soil microbial guilds involved in the increases of soil fertility and crop yield triggered by OM fertilization. To unravel these interactions, a series of fresh and archived soil samples from a fertilization experiment started in 1989 in North China Plain (NCP) was systematically investigated. Molecular assays of contemporary fresh samples unravel that Bacillus asahii responded most distinctly to OM fertilization, while no shifts in microbial community structure were observed between chemical fertilizations and the control without fertilization; a series of archived soil samples from 1989 to 2009 reveal that the indigenous B. asahii took 2e4 years to become specifically dominant and its ratio fluctuated between 40% and 72% during 20 years. Culture-dependent assessments of isolated B. asahii strain further indicate that its rise subsequently played a key role in the increases of both crop yield and soil fertility, especially via accelerating carbon and phosphorus cycling. This insight deepens our understanding of how OM impacts agroecosystems through soil microbial processes, and highlights the possibility of using archived microbial information as a reference to develop an efficient and sustainable agricultural strategy.

Although increasing evidences have emerged for responses of soil microorganisms to fertilizations, the knowledge regarding community assemblages that cause variations in composition is still lacking, as well as the possible feedback to soil fertility. Phylogenetic conservatism of species indicates their similar environmental preferences and/or function traits and phylogenetic signals further can infer community assemblages and influenced ecological processes. Here, we calculated the mean pairwise phylogenetic distance (MPD) and nearest relative index (NRI), characterizing phylogenetic signal and the undergone ecological process to evaluate the community assembly of soil bacterial phylotypes in 20-year fertilized soils. The bacterial community assembly is structured by environmental filtering, regardless of fertilization regime. Soil phosphorous (P) availability imposes selection on community assemblage and influences their community turnover among fertilizations. When P nutrient lacks, the effect of environmental filtering becomes stronger, hence bacterial functional traits become more coherent; this process results into increased intraspecific interactions characterized by co-occurrence network analysis. In contrast, when P nutrient becomes abundant, the environmental selection is mitigated; function traits are evened. This process reduces intraspecific interactions and increases carbon sequestration efficiency, which is finally of great favor to the increases in soil fertility. This study has made the first attempt, at the bacterial level, to understand how fertilization affects agroecosystems. When more phylogenetic information on how nutrient cycling-related microbes respond to fertilization becomes available, the systematic knowledge will eventually provide guidance to optimal fertilization strategies.

Fertilization has a large impact on the soil microbial communities, which play pivotal roles in soil biogeochemical cycling and ecological processes. While the effects of changes in nutrient availability due to fertilization on the soil microbial communities have received considerable attention, specific microbial taxa strongly influenced by long-term organic and inorganic fertilization, their potential effects and associations with soil nutrients remain unclear. Here, we use deep 16S amplicon sequencing to investigate bacterial community characteristics in a fluvo-aquic soil treated for 24 years with inorganic fertilizers and organics (manure and straw)-inorganic fertilizers, and uncover potential links between soil nutrient parameters and specific bacterial taxa. Our results showed that combined organic-inorganic fertilization increased soil organic carbon (SOC) and total nitrogen (TN) contents and altered bacterial community composition, while inorganic fertilization had little impact on soil nutrients and bacterial community composition. SOC and TN emerged as the major determinants of community composition. The abundances of specific taxa, especially Arenimonas, Gemmatimonas, and an unclassified member of Xanthomonadaceae, were substantially increased by organic-inorganic amendments rather than inorganic amendments only. A co-occurrence based network analysis demonstrated that SOC and TN had strong positive associations with some taxa (Gemmatimonas and the members of Acidobacteria subgroup 6, Myxococcales, Betaproteobacteria, and Bacteroidetes), and Gemmatimonas, Flavobacterium, and an unclassified member of Verrucomicrobia were identified as the keystone taxa. These specific taxa identified above are implicated in the decomposition of complex organic matters and soil carbon, nitrogen, and phosphorus transformations. The present work strengthens our current understanding of the soil microbial community structure and functions under long-term fertilization management and provides certain theoretical support for selection of rational fertilization strategies.

Returning crop straw into fields is a typical agricultural practice to resolve an oversupply of straw and improve soil fertility. Soil microorganisms, especially eukaryotic microorganisms, play a critical role in straw decomposition. To date, microbial communities in response to straw amendment at different moisture levels in Chinese fluvo-aquic soil are poorly understood. The aim of this study was to explore the effects of straw amendment and moisture on microbial communities in Chinese fluvo-aquic soil. Two soils (one was applied with organic manure, and the other was not applied with any fertilizer) from a long-term field experiment in the North China Plain were collected. Soils with and without straw amendment at 25 and 55 % of the average water-holding capacities of the two soils were incubated at 25 °C for 80 days. All treatments were sampled 20 and 80 days after the start of incubation. Microbial biomass and community structure were analyzed by phospholipid fatty acids (PLFA) assay, and the eukaryotic diversity and community composition were assessed via barcoded pyrosequencing of the 18S ribosomal RNA (rRNA) gene amplicons. PLFA analysis showed that straw amendment increased the biomass of Gram-positive bacteria, Gram-negative bacteria, actinobacteria, and fungi and shifted microbial community structure. The varied straw availability resulted in a large variation in microbial community structure. In the presence of straw, actinobacterial and fungal biomass both decreased under high moisture content. 18S rRNA gene pyrosequencing indicated that straw amendment decreased eukaryotic diversity and richness and probably restructured the eukaryotic community. Under identical moisture content, long-term organic manure-fertilized soil had higher eukaryotic diversity and richness than the unfertilized soil. In the amended soils under high moisture content, the relative abundance of dominant fungal taxa (Dikarya subkingdom, Ascomycota phylum, and Pezizomycotina subphylum) decreased. Straw amendment increases microbial biomass, shifts microbial community structure, and decreases eukaryotic diversity and richness. High moisture content probably has a negative effect on fungal growth in the amended soils. In conclusion, microbial communities in Chinese fluvo-aquic soil are significantly affected by straw amendment at different moisture levels.

The objective of this study was to examine the effects of addition of inoculant to high-nitrogen (N) fertilized timothy on fermentation, nutritive value, and feed intake of silage. The silage of timothy cultivated with a N fertilizer rate at two levels (high level (H), standard level (S)) was prepared with (SI, HI) or without (SC, HC) an inoculants. The CP content of H increased by 38 g&middotkg-1 DM compared with that of S, and the WSC and ADF contents decreased compared with those of S. Regarding the fermentation of silage, the pH and NH3-N ratio was significantly lower in the silage with inoculant (SI and HI), showing improvement of the fermentation compared with those of the silage without the addition (SC and HC). The CP content was significantly higher in HC and HI than in SC and SI, and the NDF content was the lowest in HI among the four treatments. The CP digestibility of HC and HI was significantly higher than those of SC and SI. The EE digestibility of SI was significantly higher than that of SC, and that of HI was significantly higher than that of HC. The DCP contents were significantly higher in HC and HI than in SC and SI. The TDN content was the lowest in SC and highest in HI. The DCP intakes of HC and HI were significantly higher than those of SC and SI. When the N fertilizer rate was increased, the DCP content and DCP intake of the silage increased, and the addition of inoculant improved the fermentation and increased the TDN content.

Nitrate-nitrogen (NO − 3 -N) dynamics and nitrogen (N) budgets in rice ( Oryza sativa L.)-wheat ( Triticum aestivum L.) rotations in the Taihu Lake region of China were studied to compare the effects of N fertilizer management over a two-year period. The experiment included four N rates for rice and wheat, respectively: N1 (125 and 94 kg N ha −1 ), N2 (225 and 169 kg N ha −1 ), N3 (325 and 244 kg N ha −1 ), and N0 (0 kg N ha −1 ). The results showed that an overlying water layer during the rice growing seasons contributed to moderate concentrations of NO − 3 -N in sampled waters and the concentrations of NO − 3 -N only showed a rising trend during the field drying stage. The NO − 3 -N concentrations in leachates during the wheat seasons were much higher than those during the rice seasons, particularly in the wheat seedling stage. In the wheat seedling stage, the NO − 3 -N concentrations of leachates were significantly higher in N treatments than in N0 treatment and increased with increasing N rates. As the NO − 3 -N content (below 2 mg N L −1 ) at a depth of 80 cm during the rice-wheat rotations did not respond to the applied N rates, the high levels of NO − 3 -N in the groundwater of paddy fields might not be directly related to NO − 3 -N leaching. Crop growth trends were closely related to variations of NO − 3 -N in leachates. A reduction in N application rate, especially in the earlier stages of crop growth, and synchronization of the peak of N uptake by the crop with N fertilizer application are key measures to reduce N loss. Above-ground biomass for rice and wheat increased significantly with increasing N rate, but there was no significant difference between N2 and N3. Increasing N rates to the levels greater than N2 not only decreased N use efficiency, but also significantly increased N loss. After two cycles of rice-wheat rotations, the apparent N losses of N1, N2 and N3 amounted to 234, 366 and 579 kg N ha −1 , respectively. With an increase of N rate from N0 to N3, the percentage of N uptake in total N inputs decreased from 63.9% to 46.9%. The apparent N losses during the rice seasons were higher than those during the wheat seasons and were related to precipitation; therefore, the application of fertilizer should take into account climate conditions and avoid application before heavy rainfall.

Inorganic phosphate solubilizing bacteria (iPSB) are essential to facilitate phosphorus (P) mobilization in alkaline soil, however, the phylogenetic structure of iPSB communities remains poorly characterized. Thus, we use a reference iPSB database to analyze the distribution of iPSB communities based on 16S rRNA gene illumina sequencing. Additionally, a noval pqqC primer was developed to quantify iPSB abundance. In our study, an alkaline soil with 27-year fertilization treatment was selected. The percentage of iPSB was 1.10~2.87% per sample, and the dominant iPSB genera were closely related to Arthrobacter, Bacillus, Brevibacterium and Streptomyces. Long-term P fertilization had no significant effect on the abundance of iPSB communities. Rather than P and potassium (K) additions, long-term nitrogen (N) fertilization decreased the iPSB abundance, which was validated by reduced relative abundance of pqqC gene (pqqC/16S). The decreased iPSB abundance was strongly related to pH decline and total N increase, revealing that the long-term N additions may cause pH decline and subsequent P releases relatively decreasing the demands of the iPSB community. The methodology and understanding obtained here provides insights into the ecology of inorganic P solubilizers and how to manipulate for better P use efficiency.

Conjugates (A1–A5) of the Pt(IV) derivative (A6) with amino groups from peracetyl glucose, rhamnose and mannose with a propyl amino or ethyl amino linker at the reducing end were synthesized and exhibited significant therapeutic efficacy in tumour cells, especially for prostate cancer (PCa). The antitumor activities are greatly affected by glycosyl groups. Cytotoxic experiments in vitro indicated that the antitumor activities were increased by 5-fold when its Pt(IV) derivative was conjugated to S18 (IC50 = 4.82 ± 0.45 μM) and by 12-fold when conjugated to S21 (IC50 = 1.9 ± 0.67 μM). The mannose substituted Pt(IV) complexes A4 and A5 were also over an order of magnitude more potent towards HeLa, A549, MCF-7 and PC3 than cisplatin and oxaliplatin. Importantly, the glycosylated Pt(IV) derivatives A4 and A5 displayed potential safety for clinical therapeutic exposure with IC50 of 84 μM and 169 μM compared with cisplatin (IC50 = 8 μM) to 3T3. Cellular uptake and DNA platination are higher than cisplatin and oxaliplatin. ESI-MS analysis of A5 binding to 5′-dGMP revealed that bifunctional DNA lesions were formed. The antitumor activities in vivo showed that the MTD and LD50 for A4 and A5 are nearly 4-fold higher than that of oxaliplatin indicating the potential safety for the glycosylated Pt(IV) complexes.

Differences in the composition of a bacterial community engaged in decomposing wheat straw in a fluvo-aquic soil at 15 °C, 25 °C and 35 °C were identified using barcode pyrosequencing. Functional carbon groups in the decomposing wheat straw were evaluated by 13C-NMR (nuclear magnetic resonance). Actinobacteria and Firmicutes were more abundant, whereas Alphaproteobacteria and Bacteroidetes were less abundant, at higher temperatures during the later stages of decomposition. Differences in the chemical properties of straw accounted for 19.3% of the variation in the community composition, whereas soil properties accounted for more (24.0%) and temperature, for less (7.4%). Carbon content of the soil microbial biomass and nitrogen content of straw were significantly correlated with the abundance of Alphaproteobacteria, Actinobacteria and Bacteroidetes. The chemical properties of straw, especially the NCH/OCH3, alkyl O-C-O and O-alkyl functional groups, exercised a significant effect on the composition of the bacterial community at different temperatures during decomposition—results that extend our understanding of bacterial communities associated with the decomposition of straw in agro-ecosystems and of the effects of temperature and chemical properties of the decomposing straw and soil on such communities.

Arbuscular mycorrhizal (AM) fungi are crucial for ecosystem functioning and can contribute to the formation and maintenance of soil aggregates through the exudation of glomalin by extraradical hyphae. Monitoring fertilization effects on AM fungi may help us to develop sound management strategies. The objectives of this study were to investigate the impacts of long-term fertilization on AM fungal parameters and to find out the key factor that affects the diversity and function of AM fungi. A long-term fertilization experiment established in a sandy loam soil at northern China has received continuous fertilization treatments for 21 years, including control; mineral fertilizers of NK, PK, NP, and NPK; organic manure (OM); and half organic manure N plus half mineral fertilizer N (1/2 OMN). Top soil samples (0–15 cm) from three individual plots per treatment were collected for the analysis of chemical properties and fungal parameters. The population size of soil AM fungi was determined by real-time PCR, and the community composition was analyzed using PCR-denature gradient gel electrophoresis (DGGE), cloning, and sequencing techniques. The external mycelium of AM fungi was assessed using the grid-line intersect method, and the glomalin-related soil protein (GRSP) was extracted with citrate solution using bovine serum albumin as a standard. Long-term fertilization significantly increased (P

Soil nematode abundance, community composition and biomass were determined in the Fengqiu State Key Agro-Ecological Experimental Station, North China, in order to evaluate the effects of tillage system (conventional tillage and no-tillage) and residue management (0, 50% and 100% wheat residue incorporation/coverage) on the nematode communities. Two kinds of indicators (descriptive and evaluative) were categorized. Of the descriptive indicators, residue management had a significant effect on the total nematode abundance, biomass and trophic groups except for bacterivores. Of the evaluative indicators, Shannon diversity (H′), generic richness (GR), nematode channel ratio (NCR) and enrichment index (EI) significantly increased with increasing residue quantity, whereas dominance (λ), basal index (BI) and channel index (CI) exhibited an opposite trend. Significant tillage effects were observed on the trophic diversity (TD), EI, CI and carbon production (P). The responses of nematodes to tillage and residue were genus-dependent. Canonical correspondence analysis indicated that tillage explained 4.9% and 15.4%, and residue management explained 5.2% and 13.1% of the variations in soil nematode abundance and biomass, respectively. Different metabolic footprint characteristics of the food web were demonstrated graphically by enrichment and structure footprints. The evaluative indicators, such as EI and CI, were sensitive to both tillage and residue management. The descriptive indicators could be used to obtain an intuitive answer to the effect of residue management and the evaluative indicators were more comprehensive for interpreting the structure and function of the soil food web under different tillage and residue management regimes.