Differential Responses of Arbuscular Mycorrhizal Fungal Communities to Long-Term Fertilization in the Wheat Rhizosphere and Root Endosphere.

Arbuscular mycorrhizal fungi (AMF) provide essential nutrients to crops and are critically impacted by fertilization in agricultural ecosystems. Understanding shifts in AMF communities in and around crop roots under different fertilization regimes can provide important lessons for improving agricultural production and sustainability. Here, we compared the responses of AMF communities in the rhizosphere (RS) and root endosphere (ES) of wheat (Triticum aestivum) to different fertilization treatments, nonfertilization (control), mineral fertilization only (NPK), mineral fertilization plus wheat straw (NPKS), and mineral fertilization plus cow manure (NPKM). We employed high-throughput amplicon sequencing and investigated the diversity, community composition, and network structure of AMF communities to assess their responses to fertilization. Our results elucidated that AMF communities in the RS and ES respond differently to fertilization schemes. Long-term NPK application decreased the RS AMF alpha diversity significantly, whereas additional organic amendments (straw or manure) had no effect. In contrast, NPK fertilization increased the ES AMF alpha diversity significantly, while additional organic amendments decreased it significantly. The effect of different fertilization schemes on AMF network complexity in the RS and ES were similar to their effects on alpha diversity. Changes to AMF communities in the RS and ES correlated mainly with the pH and phosphorus level of the rhizosphere soil under long-term inorganic and organic fertilization regimes. We suggest that the AMF community in the roots should be given more consideration when studying the effects of fertilization regimes on AMF in agroecosystems. IMPORTANCE Arbuscular mycorrhizal fungi are an integral component of rhizospheres, bridging the soil and plant systems and are highly sensitive to fertilization. However, surprisingly little is known about how the response differs between the roots and the surrounding soil. Decreasing arbuscular mycorrhizal fungal diversity under fertilization has been reported, implying a potential reduction in the mutualism between plants and arbuscular mycorrhizal fungi. However, we found opposing responses to long-term fertilization managements of arbuscular mycorrhizal fungi in the wheat roots and rhizosphere soil. These results suggested that changes in the arbuscular mycorrhizal fungal community in soils do not reflect those in the roots, highlighting that the root arbuscular mycorrhizal fungal community is pertinent to understand arbuscular mycorrhizal fungi and their crop hosts' responses to anthropogenic influences.

Enhanced yields and soil quality in a wheat-maize rotation using buried straw mulch.

BACKGROUND: Straw return may improve soil quality and crop yields. In a 2-year field study, a straw return method (ditch-buried straw return, DB-SR) was used to investigate the soil quality and crop productivity effects on a wheat-corn rotation system. This study consisted of three treatments, each with three replicates: (1) mineral fertilisation alone (CK0); (2) mineral fertilisation + 7500 kg ha-1 wheat straw incorporated at depth of 0-15 cm (NPKWS); and (3) mineral fertilisation + 7500 kg ha-1 wheat straw ditch buried at 15-30 cm (NPKDW). RESULTS: NPKWS and NPKDW enhanced crop yield and improved soil biotical properties compared to mineral fertilisation alone. NPKDW contributed to greater crop yields and soil nutrient availability at 15-30 cm depths, compared to NPKWS treatment. NPKDW enhanced soil microbial activity and bacteria species richness and diversity in the 0-15 cm layer. NPKWS increased soil microbial biomass, bacteria species richness and diversity at 15-30 cm. CONCLUSIONS: The comparison of the CK0 and NPKWS treatments indicates that a straw ditch buried by digging to the depth of 15-30 cm can improve crop yields and soil quality in a wheat-maize rotation system. © 2016 Society of Chemical Industry.

Screening hydroxyapatite for cadmium and lead immobilization in aqueous solution and contaminated soil: The role of surface area.

Hydroxyapatite (HAP) has been widely used to immobilize many cationic metals in water and soils. The specific reason why an increase in the surface area of HAP enhances cadmium (Cd) uptake, but has no effect on lead (Pb) uptake, is not clear. The aim of this study was to determine the factors causing the differences in sorption behavior between Cd and Pb by evaluating HAPs with different surface areas. We synthesized HAPs with two different surface areas, which were characterized by X-ray diffraction, N2 adsorption, and scanning electron microscopy, and then evaluated them as sorbents for Cd and Pb removal by testing in single and binary systems. The sorption capacity of large surface area HAP (1.85mmol/g) for Cd in the single-metal system was higher than that of small surface area HAP (0.64mmol/g), but there were no differences between single- and binary-metal solutions containing Pb. After the Cd experiments, the HAP retained a stable structure and intact morphology, which promotes the accessibility of reactive sites for Cd. However, a newly formed precipitate covered the surface and blocked the channels in the presence of Pb, which reduced the number of potential adsorption sites on HAP for Cd and Pb. Remediation experiments using Cd- and Pb-contaminated soil produced similar results to the solution tests. These results indicate that alterations of the structure and morphology during the reaction is an important factor influencing metal sorption to HAP.

Fungal community composition in soils subjected to long-term chemical fertilization is most influenced by the type of organic matter.

Organic matter application is a widely used practice to increase soil carbon content and maintain soil fertility. However, little is known about the effect of different types of organic matter, or the input of exogenous species from these materials, on soil fungal communities. In this study, fungal community composition was characterized from soils amended with three types of organic matter over a 30-year fertilization experiment. Chemical fertilization significantly changed soil fungal community composition and structure, which was exacerbated by the addition of organic matter, with the direction of change influenced by the type of organic matter used. The addition of organic matter significantly increased soil fungal richness, with the greatest richness achieved in soils amended with pig manure. Importantly, following addition of cow and pig manure, fungal taxa associated with these materials could be found in the soil, suggesting that these exogenous species can augment soil fungal composition. Moreover, the addition of organic matter decreased the relative abundance of potential pathogenic fungi. Overall, these results indicate that organic matter addition influences the composition and structure of soil fungal communities in predictable ways.

Phosphorus and Zinc Are Strongly Associated with Belowground Fungal Communities in Wheat Field under Long-Term Fertilization.

Belowground fungi are closely related to crop growth, and agricultural fertilization is widely known to affect soil fungal communities. Yet it remains unclear whether fungal communities in differing belowground habitats-root endosphere, rhizosphere soil, and bulk soil-respond differently to long-term fertilization. Here we investigated the variation in fungal communities of root endosphere, rhizosphere soil, and bulk soil under 35 years of fertilization in wheat fields. Specifically, the fertilization regimes were applied as five treatments: soils receiving NPK fertilizer, NPK and cow manure (NPK+CM), NPK and pig manure (NPK+PM), NPK and wheat straw (NPK+WS), and no fertilizer (Control). Long-term fertilization significantly impacted fungal community composition in all three habitats, and these effects were stronger in the rhizosphere and bulk soils than root endosphere. Mantel test results showed that fungal community composition was significantly correlated with phosphorus and zinc contents. Further, fungal alpha diversity was lowest in the NPK+PM treatment and was negatively correlated with both phosphorus and zinc contents. Moreover, NPK+PM treatment had the lowest complexity of fungal co-occurrence network, and in general network complexity was significantly negatively correlated with the zinc and phosphorus contents. Taken together, these results suggest that long-term fertilization can impact fungal communities not only in soils but in root endosphere, and this is strongly associated with the contents of phosphorus and zinc there, a finding important for guiding fertilization management practices and supporting sustainable agriculture. IMPORTANCE Fungi, an essential component in nutrient cycling and plant growth, are highly sensitive to fertilization. However, there are limited studies on fungi in root endosphere under long-term fertilization management. Our research extended the study on the endophytic fungal community of crop roots under agricultural management and found that its responses were similar to the communities in soil habitats. In addition, the type of organic materials was reported as the main driver affecting soil fungal community under long-term fertilization. Our research further revealed that the underlying mechanism of affecting the fungal communities in the soils and roots was the differences in phosphorus and zinc contents caused by the application of different organic materials. Therefore, our results highlight that except for phosphorus, zinc content of the organic materials should be considered in long-term organic fertilization systems.

Root stoichiometry explains wheat endophytes and their link with crop production after four decades of fertilization.

Fertilization can impact root endophytic microbiomes and food production. However, the impacts of decades of continued fertilization on root microbiomes, and their link with ongoing crop production, remain poorly understood. Here, we used a four decade-long fertilization experiment, including contrasting types of organic and inorganic fertilization, to investigate the effects of long-term fertilization on multi-kingdom root endophytic microbiomes, including keystone species (modules within microbial networks), and their indirect associations with the production of wheat, which is one of the most important crops worldwide. We found that long-term inorganic (nitrogen, phosphorus, potassium (NPK)) and organic (NPK with straw (NPKS) and NPK with cow manure (NPKM)) fertilization had significant impacts on the community composition of endophytic arbuscular mycorrhizal fungi (AMF), bacteria, and non-mycorrhizal fungi. In addition, compared with NPK fertilization, NPKS and NPKM amendments significantly decreased the microbial network complexity, which was associated with changes in the root iron content. Finally, we identified an important subset of keystone root endophyte species within the microbial network (Module #2), which was positively correlated with wheat yield, and affected by changes in root carbon to phosphorus ratio. This study provides evidence that long-term fertilization can affect keystone root endophytic species in the root microbiome, with implications for food security in an over-fertilized world.

Biodiversity of key-stone phylotypes determines crop production in a 4-decade fertilization experiment.

Cropping systems have fertilized soils for decades with undetermined consequences for the productivity and functioning of terrestrial ecosystems. One of the critical unknowns is the role of soil biodiversity in controlling crop production after decades of fertilization. This knowledge gap limits our capacity to assess how changes in soil biodiversity could alter crop production and soil health in changing environments. Here, we used multitrophic ecological networks to investigate the importance of soil biodiversity, in particular, the biodiversity of key-stone taxa in controlling soil functioning and wheat production in a 35-year field fertilization experiment. We found strong and positive associations between soil functional genes, crop production and the biodiversity of key-stone phylotypes; soils supporting a larger number of key-stone nematode, bacteria and fungi phylotypes yielded the highest wheat production. These key-stone phylotypes were also positively associated with plant growth (phototrophic bacteria, nitrogen fixers) and multiple functional genes related to nutrient cycling. The retrieved information on the genomes clustered with key-stone bacterial phylotypes indicated that the key-stone taxa had higher gene copies of oxidoreductases (participating most biogeochemical cycles of ecosystems and linking to microbial energetics) and 71 essential functional genes associated with carbon, nitrogen, phosphorus, and sulfur cycling. Altogether, our work highlights the fundamental role of the biodiversity of key-stone phylotypes in maintaining soil functioning and crop production after several decades of fertilization, and provides a list of key-stone phylotypes linking to crop production and soil nutrient cycling, which could give science-based guidance for sustainable food production.

Suppressed N fixation and diazotrophs after four decades of fertilization.

BACKGROUND: N fixation is one of the most important microbially driven ecosystem processes on Earth, allowing N to enter the soil from the atmosphere, and regulating plant productivity. A question that remains to be answered is whether such a fundamental process would still be that important in an over-fertilized world, as the long-term effects of fertilization on N fixation and associated diazotrophic communities remain to be tested. Here, we used a 35-year fertilization experiment, and investigated the changes in N fixation rates and the diazotrophic community in response to long-term inorganic and organic fertilization. RESULTS: It was found that N fixation was drastically reduced (dropped by 50%) after almost four decades of fertilization. Our results further indicated that functionality losses were associated with reductions in the relative abundance of keystone and phylogenetically clustered N fixers such as Geobacter spp. CONCLUSIONS: Our work suggests that long-term fertilization might have selected against N fixation and specific groups of N fixers. Our study provides solid evidence that N fixation and certain groups of diazotrophic taxa will be largely suppressed in a more and more fertilized world, with implications for soil biodiversity and ecosystem functions.

Prevalence of antibiotic resistance genes in soils after continually applied with different manure for 30 years.

A 30year field experiment with a wheat-soybean cropping system were performed to compare the long-term anthropogenic influence on soil ARGs. Compared with chemical fertilization, the occurrence of 38 ARGs and the abundance of seven ARGs (tetL, tetB(P), tetO, tetW, sul1, ermB, and ermF) were significantly increased by long term exposure of pig manure. However, application of wheat straw and cow manure not substantially affected the abundance of ARGs except cow manure increased the abundance of tetM and tetW to a detectable level. Relative abundance of Firmicutes, Gammaproteobacteria and Bacteroidetes and ARGs observed in the soil were significantly correlated. Integrase gene I1 (intI1) is commonly linked to genes conferring resistance to antibiotics, it was significantly increased in pig manure treated soils and showed a high positive correlation with the abundance of ARGs which were significantly affected by pig manure. Concentrations of Cu, Zn and Pb were also increased in manure treated soil and positive correlated with the relative abundance of intI1 and most of the ARGs. These results indicated that long-term animal manure application to soils has polluted the soil, especially for pig manure, and it should be scrutinized as part of future stewardship programs.

Effects of surface ligands on the uptake and transport of gold nanoparticles in rice and tomato.

Nanotechnology is advancing rapidly and substantial amounts of nanomaterials are released into the environment. Plants are an essential base component of the ecological environment and play a critical role in the fate and transport of nanomaterials in the environment through plant uptake and bioaccumulation. In this study, plant uptake of gold nanoparticles (GNPs) functionalized with three types of short ligands [cysteamine (CA), cysteine (CYS) and thioglycolic acid (TGA)] and of nearly identical hydrodynamic size (8-12nm) was investigated in the major crops rice (Oryza sativa L.) and tomato (Solanum lycopersicum). Uptake and translocation of GNPs not only depended on particle surface charge, but were also related to the species of ligand on the GNPs. The negatively charged GNPs capped with the CYS ligand (GNP-CYS) were more efficiently absorbed in roots and transferred to shoots (including stems and leaves) than that of GNPs capped with CA and TGA. The absorption process of GNPs involved a combination of both clathrin-dependent and -independent mechanisms. The endocytosis of GNPs was strongly inhibited by wortmannin, suggesting that clathrin-independent endocytosis was an important pathway of nanoparticle internalization in plants. Competition experiments with a free ligand (CYS) showed that the CYS ligand probably facilitated the endocytosis process of GNPs and increased the internalization of GNP-CYS in plants. The results will aid understanding of the mechanisms of nanoparticle uptake and translocation in plants.

The influence of biochar type on long-term stabilization for Cd and Cu in contaminated paddy soils.

Long-term effect of biochar on PTEs (potential toxic elements) immobilization depends upon biochar own property and its aging process in soil. To understand the role of biachar type on PTEs stabilization, two types of biochar, corn-straw-derived biochar (CB) and hardwood-derived biochar (HB), were compared for their efficacy in achieving a stable decrease in the bio-availability of Cd and Cu in soils. The 3-year pot-culture experiment showed that HB reduced the concentration of CaCl2-extractable Cd and Cu by 57.9 and 63.8% in soil, and Cd and Cu uptake by 63.6 and 56.3% in rice tissue respectively, in the first year, whereas these values increased in the next two years. On the other hand, CB decreased these values steadily year by year. At the end of the 3 years, CB at 5% level had lowered the levels of CaCl2-extractable Cd and Cu by 53.6 and 66.8%, respectively. These variations between CB and HB were due to the differences in the way the two types of biochar age in the soil. The aging process was simulated in the laboratory, and the XPS results showed that the oxidization of the biochars introduced more oxygen-containing groups (especially carboxyl) on the surface of CB than HB, leading to a correspondingly greater number of oxygenated binding sites for Cd and Cu in the case of CB. The content of lignin was the major factor resulting in the variation of oxidation degree in two biochars. These results suggest that it is important to select the right kind of biochar to stably decrease the bio-availability of potential toxic elements (Cd and Cu) in contaminated soils.

Productivity and sustainability of rainfed wheat-soybean system in the North China Plain: results from a long-term experiment and crop modelling.

A quantitative understanding of yield response to water and nutrients is key to improving the productivity and sustainability of rainfed cropping systems. Here, we quantified the effects of rainfall, fertilization (NPK) and soil organic amendments (with straw and manure) on yields of a rainfed wheat-soybean system in the North China Plain (NCP), using 30-years' field experimental data (1982-2012) and the simulation model-AquaCrop. On average, wheat and soybean yields were 5 and 2.5 times higher in the fertilized treatments than in the unfertilized control (CK), respectively. Yields of fertilized treatments increased and yields of CK decreased over time. NPK + manure increased yields more than NPK alone or NPK + straw. The additional effect of manure is likely due to increased availability of K and micronutrients. Wheat yields were limited by rainfall and can be increased through soil mulching (15%) or irrigation (35%). In conclusion, combined applications of fertilizer NPK and manure were more effective in sustaining high crop yields than recommended fertilizer NPK applications. Manure applications led to strong accumulation of NPK and relatively low NPK use efficiencies. Water deficiency in wheat increased over time due to the steady increase in yields, suggesting that the need for soil mulching increases.

Carbon sequestration efficiency of organic amendments in a long-term experiment on a vertisol in Huang-Huai-Hai Plain, China.

Soil organic carbon (SOC) sequestration is important for improving soil fertility of cropland and for the mitigation of greenhouse gas emissions to the atmosphere. The efficiency of SOC sequestration depends on the quantity and quality of the organic matter, soil type, and climate. Little is known about the SOC sequestration efficiency of organic amendments in Vertisols. Thus, we conducted the research based on 29 years (1982-2011) of long-term fertilization experiment with a no fertilizer control and five fertilization regimes: CK (control, no fertilizer), NPK (mineral NPK fertilizers alone), NPK+1/2W (mineral NPK fertilizers combined with half the amount of wheat straw), NPK+W (mineral NPK fertilizers combined with full the amount of wheat straw), NPK+PM (mineral NPK fertilizers combined with pig manure) and NPK+CM (mineral NPK fertilizers combined cattle manure). Total mean annual C inputs were 0.45, 1.55, 2.66, 3.71, 4.68 and 6.56 ton/ha/yr for CK, NPK, NPKW1/2, NPKW, NPKPM and NPKCM, respectively. Mean SOC sequestration rate was 0.20 ton/ha/yr in the NPK treatment, and 0.39, 0.50, 0.51 and 0.97 ton/ha/yr in the NPKW1/2, NPKW, NPKPM, and NPKCM treatments, respectively. A linear relationship was observed between annual C input and SOC sequestration rate (SOCsequestration rate  = 0.16 Cinput -0.10, R = 0.95, P<0.01), suggesting a C sequestration efficiency of 16%. The Vertisol required an annual C input of 0.63 ton/ha/yr to maintain the initial SOC level. Moreover, the C sequestration efficiencies of wheat straw, pig manure and cattle manure were 17%, 11% and 17%, respectively. The results indicate that the Vertisol has a large potential to sequester SOC with a high efficiency, and applying cattle manure or wheat straw is a recommendable SOC sequestration practice in Vertisols.

[Effects of application time and basal/topdressing ratio of nitrogen fertilizer on the spatiotemporal variation of soil NO3- -N and NH4+ -N contents and the grain yield and its quality of wheat].

Field trials were conducted to study the effects of different application time and basal/topdressing ratio of nitrogen fertilizer on the spatiotemporal variation of soil NO3- -N and NH4+ -N contents and the grain yield and its quality of wheat. The results showed that soil NO3- -N and NH4+ -N contents decreased with increasing soil depth. Both the application time and the basal/topdressing ratio of nitrogen fertilizer had significant effects on the NO3- -N and NH4+ -N contents in 0-20 cm soil layer. Compared with basal application, later fertilization and higher topdressing ratio could promote the nitrogen uptake by wheat plant and increase the plant nitrogen recovery significantly, decrease the soil nitrogen recovery during wheat growth, and improve the grain quality significantly, while had less effects on the grain yield. Topdressing too much nitrogen fertilizer at booting stage could result in a significant decrease of grain yield. Under the condition of this experiment, the optimal nitrogen fertilization mode for good wheat grain yield and its quality and good ecological benefits was 5:3:2 of basal application: topdressing at jointing stage: topdressing at booting stage.