Maize functional requirements drive the selection of rhizobacteria under long-term fertilization practices.

Rhizosphere microbiomes are pivotal for crop fitness, but the principles underlying microbial assembly during root-soil interactions across soils with different nutrient statuses remain elusive. We examined the microbiomes in the rhizosphere and bulk soils of maize plants grown under six long-term (≥ 29 yr) fertilization experiments in three soil types across middle temperate to subtropical zones. The assembly of rhizosphere microbial communities was primarily driven by deterministic processes. Plant selection interacted with soil types and fertilization regimes to shape the structure and function of rhizosphere microbiomes. Predictive functional profiling showed that, to adapt to nutrient-deficient conditions, maize recruited more rhizobacteria involved in nutrient availability from bulk soil, although these functions were performed by different species. Metagenomic analyses confirmed that the number of significantly enriched Kyoto Encyclopedia of Genes and Genomes Orthology functional categories in the rhizosphere microbial community was significantly higher without fertilization than with fertilization. Notably, some key genes involved in carbon, nitrogen, and phosphorus cycling and purine metabolism were dominantly enriched in the rhizosphere soil without fertilizer input. In conclusion, our results show that maize selects microbes at the root-soil interface based on microbial functional traits beneficial to its own performance, rather than selecting particular species.

Soil enzyme activities, soil physical properties, photosynthetic physical characteristics and water use of winter wheat after long-term straw mulch and organic fertilizer application.

Introduction: Inappropriate residue and nutrient management leads to soil degradation and the decline of soil quality and water storage capacity. Methods: An ongoing field experiment has been conducted since 2011 to investigate the effects of straw mulching (SM), and straw mulching combined with organic fertilizer (SM+O), on winter wheat yield, including a control treatment (CK, no straw). We studied the effects of these treatments on soil microbial biomass nitrogen and carbon, soil enzyme activity in 2019, photosynthetic parameters, evapotranspiration (ET), water use efficiency (WUE), and yields over five consecutive years (2015-2019). We also analyzed the soil organic carbon, soil structure, field capacity, and saturated hydraulic conductivity in 2015 and 2019. Results: Results indicate that compared with CK, SM and SM+O treatments increased the proportion of >0.25mm aggregates, soil organic carbon, field capacity, and saturated hydraulic conductivity, but decreased the soil bulk density. In addition, the SM and SM+O treatments also increased soil microbial biomass nitrogen and carbon, the activity of soil enzymes, and decreased the carbon-nitrogen ratio of microbial biomass. Therefore, SM and SM+O treatments both increased the leaf water use efficiency (LWUE) and photosynthetic rate (Pn), and improved the yields and water use efficiency (WUE) of winter wheat. The combination SM (4.5 t/ha)+O (0.75 t/ha) was more effective than SM alone, and both treatments were superior to the control. Conclusion: Based on the results of this study, SM+O is recommended as the most effective cultivation practice.

A highly conserved core bacterial microbiota with nitrogen-fixation capacity inhabits the xylem sap in maize plants.

Microbiomes are important for crop performance. However, a deeper knowledge of crop-associated microbial communities is needed to harness beneficial host-microbe interactions. Here, by assessing the assembly and functions of maize microbiomes across soil types, climate zones, and genotypes, we found that the stem xylem selectively recruits highly conserved microbes dominated by Gammaproteobacteria. We showed that the proportion of bacterial taxa carrying the nitrogenase gene (nifH) was larger in stem xylem than in other organs such as root and leaf endosphere. Of the 25 core bacterial taxa identified in xylem sap, several isolated strains were confirmed to be active nitrogen-fixers or to assist with biological nitrogen fixation. On this basis, we established synthetic communities (SynComs) consisting of two core diazotrophs and two helpers. GFP-tagged strains and 15N isotopic dilution method demonstrated that these SynComs do thrive and contribute, through biological nitrogen fixation, 11.8% of the total N accumulated in maize stems. These core taxa in xylem sap represent an untapped resource that can be exploited to increase crop productivity.

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Canopy nitrogen content in wheat is a key indicator of wheat grain yield and quality. When using remote sensing technology to predict wheat canopy nitrogen content, a hyperspectral mode with high adaptability and high accuracy is needed to improve the inversion efficiency. We developed a new three-band spectral vegetation index (NEW-NDRE) by combining a two-band spectral index NDRE and the spectral reflectance at 550 nm based on field data collected from different sites, years, with different varieties and nitrogen levels and at multiple growth stages. The NEW-NDRE was compared with 11 traditional spectral vegetation indices in terms of wheat canopy nitrogen content inversion. NEW-NDRE and three traditional indices (NDRE, NDDA and RI-1dB) all closely correlated with wheat canopy nitrogen content. NEW-NDRE displayed the highest correlation with wheat canopy nitrogen content at early grain filling stage, with a coefficient (R2) of 0.9 and a root mean squared error (RMSE) of 0.4. The inversion model developed with the NEW-NDRE was validated with an independent dataset. The relative error (RE) of the model was 9.3%, which was significantly lower than that of NDRE, NDDA and RI-1dB. Generally, NEW-NDRE is a more robust index for wheat canopy nitrogen content inversion than traditional indices through eliminating environmental limitation, and it could be used as a new tool for precise fertilizer application.

[Effects of Biochar on N2O Emission from Four Typical Soils in the North China Plain].

As a potential soil conditioner, biochar plays an important role in alleviating greenhouse gas (GHG) emissions. To clarify the influence of biochar on soil N2O emissions during the winter wheat seedling stage, four typical soils in the North China Plain (paddy soil, shajiang black soil, cinnamon soil, and fluvo-aquic soil) were adopted for field experiments, and four treatments were set:Control (CK), Fertilizer (NPK), Biochar (BC), and Fertilizer+Biochar (NPK+BC). The results showed that fertilization (NPK) significantly increased the N2O emissions of the four soils. Compared with that of the CK, the N2O emissions of four soils were increased by 314%, 116%, 240%, and 282%, respectively. The effect of biochar addition on N2O emissions of the four soils in the North China Plain was different. Compared with that of the CK treatment, the N2O emissions of paddy soil and cinnamon soil in the BC treatment significantly increased by 72.4% and 50.9%, respectively, whereas the shajiang black soil and fluvo-aquic soil exhibited no significant differences. The combined application of biochar and fertilizer significantly reduced the N2O emissions of the four soils, compared to that of NPK. The addition of biochar increased the pH of soil. In particular, paddy soil had the lowest initial pH and was most affected by biochar. Fertilization reduced the pH of the four soils. N2O flux under fertilizer treatment for the shajiang black soil, cinnamon soil, and fluvo-aquic soil was significantly positively correlated with ammonium nitrogen content, whereas N2O emission fluxes under single biochar treatment on paddy soil and shajiang black soil were significantly positively correlation with nitrate nitrogen content.

Linkages between straw decomposition rate and the change in microbial fractions and extracellular enzyme activities in soils under different long-term fertilization treatments.

In order to study the linkages between straw decomposition rate and the change in soil biological properties after straw addition to different fertilized soils, we collected soils from three long-term fertilization treatments (no-fertilizer, CK; nitrogen, phosphorus, and potassium fertilizers, NPK; NPK plus straw (S), NPKS), and incubated maize straw with these soils at 25°C for 75 days. The average straw carbon dioxide (CO2) emission rate in the CK+straw (S), NPK+S, and NPKS+S treatments was 0.58±0.51, 0.66±0.53, and 0.74±0.58 µg C g-1soil h-1, respectively. The average increase in the contents of fungi, bacteria, and Actinomycetes under straw addition treatments than the control soils (CK, NPK, and NPKS, respectively) changed in the order of CK+S≤NPK+S

Long-term manure amendments and chemical fertilizers enhanced soil organic carbon sequestration in a wheat (Triticum aestivum L.)-maize (Zea mays L.) rotation system.

BACKGROUND: The carbon sequestration potential is affected by cropping system and management practices, but soil organic carbon (SOC) sequestration potential under fertilizations remains unclear in north China. This study examined SOC change, total C input to soil and, via integration of these estimates over years, carbon sequestration efficiency (CSE, the ratio of SOC change over C input) under no fertilization (control), chemical nitrogen fertilizer alone (N) or combined with phosphorus and potassium fertilizers (NP, NK, PK and NPK), or chemical fertilizers combined with low or high (1.5×) manure input (NPKM and 1.5NPKM). RESULTS: Results showed that, as compared with the initial condition, SOC content increased by 0.03, 0.06, 0.05, 0.09, 0.16, 0.26, 0.47 and 0.68 Mg C ha-1 year-1 under control, N, NK, PK, NP, NPK, NPKM and 1.5NPKM treatments respectively. Correspondingly, the C inputs of wheat and maize were 1.24, 1.34, 1.55, 1.33, 2.72, 2.96, 2.97 and 3.15 Mg ha-1 year-1 respectively. The long-term fertilization-induced CSE showed that about 11% of the gross C input was transformed into SOC pool. CONCLUSION: Overall, this study demonstrated that decade-long manure input combined with chemical fertilizers can maintain high crop yield and lead to SOC sequestration in north China. © 2016 Society of Chemical Industry.

Effect of long-term combined application of organic and inorganic fertilizers on soil nematode communities within aggregates.

A long-term fertilization experiment was conducted to examine the effects of different fertilization practices on nematode community composition within aggregates in a wheat-maize rotation system. The study was a randomized complete block design with three replicates. The experiment involved the following four treatments: no fertilizer, inorganic N, P and K fertilizer (NPK), NPK plus manure (NPKM) and NPK plus maize straw (NPKS). Soil samples were taken at 0-20 cm depth during the wheat harvest stage. Based on our results, NPKS contributed to soil aggregation and moisture retention, with a positive effect on soil total nitrogen accumulation, particularly within small macroaggregates (0.25-1 mm) and microaggregates (<0.25 mm). The C/N ratio was correlated to the distribution of the soil nematode community. Both manure application and straw incorporation increased the nematode functional metabolic footprints within all aggregates. Additionally, the functional metabolic footprints decreased with a decline in aggregate size. The accumulation of total nitrogen within <1 mm aggregates under NPKS might play a key role in maintaining the survival of soil nematodes. In our study, both crop straw incorporation and inorganic fertilizer application effectively improved soil physicochemical properties and were also beneficial for nematode survival within small aggregate size fractions.

[Characteristics of soil organic carbon mineralization at different temperatures in paddy soils under long-term fertilization].

Dynamics of soil organic carbon mineralization affected by long-term fertilizations and temperature in relation to different soil carbon fractions were investigated in paddy soils. Soil samples were collected from the plough layer of 3 long-term national experimental sites in Xinhua, Ningxiang and Taojiang counties of Hunan Province. Mineralization of soil organic C was estimated by 33-day aerobic incubation at different temperatures of 10, 20 and 30 degrees C. The results showed that the rates of CO2 production were higher during the earlier phase (0-13 d) in all treatments, and then decreased according to a logarithm function. Higher incubation temperature strengthened C mineralization in the different treatments. The quantities of cumulative CO2 production in NPK with manure or straw treatments were greater than in inorganic fertilizers treatments. The Q10 values in the different soil treatments ranged from 1.01-1.53. There were significantly positive correlations between the Q10 values and soil total organic carbon (TOC), easy oxidation organic carbon (EOOC), humic acid carbon (C(HA)), fulvic acid carbon (CFA). The cumulative amount of mineralized C was significantly positively correlated with microbial biomass carbon (MBC) at 10 and 20 degrees C, but not significantly at 30 degrees C. Significant correlations were found between the cumulative amount of mineralized C and different soil carbon fractions and C(HA)/C(FA). The correlations of differ- ent soil carbon fractions with the ratio of cumulative mineralized C to TOC were negatively correlated at 10 degrees C, but not significantly at 20 and 30 degrees C. These results suggested that the application of NPK with manure or straw would be helpful to increase the sequestration of C in paddy soils and reduce its contribution of CO2 release in the atmosphere.

Immunization of aged pigs with attenuated pseudorabies virus vaccine combined with CpG oligodeoxynucleotide restores defective Th1 immune responses.

BACKGROUND AND AIMS: Attempts to immunize aged subjects often result in the failure to elicit a protective immune response. Murine model studies have shown that oligonucleotides containing CpG motifs (CpG ODN) can stimulate immune system in aged mice as effectively as in young mice. Since many physiological and pathophysiological data of pigs can be transferred to humans, research in pigs is important to confirm murine data. Here we investigated whether immunization of aged pig model with attenuated pseudorabies virus vaccine (PRV vaccine) formulated with CpG ODN could promote a successful development of immune responses that were comparable to those induced in young pigs in a similar manner. METHODOLOGY: Young and aged pigs were immunized IM with PRV vaccine alone, or in combination with CpG ODN respectively. At days 3, 7, 14 post immunization sera were assayed by ELISA for IgG titres, at day 7 for IgG1 and IgG2 subtypes titres. All blood samples collected in evacuated test tubes with K-EDTA at day 7 were analyzed for flow cytometer assay. Blood samples at day 7 collected in evacuated test tubes with heparin were analysed for antigen-specific cytokines production and peripheral blood mononuclear cells (PBMCs) proliferative responses. RESULTS: CpG ODN could enhance Th1 responses (PRV-specific IgG2/IgG1 ratio, proliferative responses, Th1 cytokines production) when used as an adjuvant for the vaccination of aged pigs, which were correlated with enhanced CD4+ T cells percentage, decreased CD4+CD8+CD45RO+ T cells percentage and improved PRV-specific CD4+ T cells activation. CONCLUSIONS: Our results demonstrate a utility for CpG ODN, as a safe vaccine adjuvant for promoting effective systemic immune responses in aged pig model. This agent could have important clinical uses in overcoming some of age-associated depressions in immune function that occur in response to vaccination.

Innate defense regulator peptide synergizes with CpG ODN for enhanced innate intestinal immune responses in neonate piglets.

The in vivo immunoadjuvant effects of the combination of CpG oligodeoxynucleotide (CpG ODN) and innate defense-regulator peptides (IDRs) have been studied in mice. However, little is known about the efficacy of these molecules in stimulating the innate intestinal immune system in neonatal piglets. In this study, we observed that intranasal (IN) administration of CpG-IDR (peptide HH2 (VQLRIRVAVIRA)) complex significantly increased intestinal mRNA expression of Th1 cytokines, CC chemokines and CXC chemokines when compared to HH2 and CpG ODN alone. Also an obvious cellular infiltration was observed in the intestine of CpG-HH2-treated neonatal piglets, which was associated with increased protection against Enterotoxigenic Escherichia coli (ETEC). Moreover, we showed that pro-inflammatory cytokine TNF-α was inhibited when CpG ODN combined with HH2. This was the first report that deciphered the role played by CpG-HH2 complex in the intestine of neonatal piglets. This work clearly demonstrated that efficiency of the IN route inducing intestinal responses in neonatal piglets might be taken into consideration for further vaccine development against neonatal intestinal diseases.

Characterization of human cytomegalovirus UL145 and UL136 genes in low-passage clinical isolates from infected Chinese infants.

BACKGROUND: Human cytomegalovirus (HCMV) is a leading cause of morbidity and mortality in immunocompromised individuals. The unique long b' (ULB') region of HCMV contains at least 19 open reading frames (ORFs); however, little is known about the function of UL145 and UL136. We characterized UL145 and UL136 in low-passage clinical isolates from Chinese infants. MATERIAL/METHODS: The clinical strains of HCMV were recovered from the urine from HCMV-infected infants. Human embryonic lung fibroblasts (HELFs) were infected with clinical isolates of HCMV, and the viral DNA and mRNA for UL145 and UL136 were analyzed by polymerase chain reaction (PCR) and sequencing techniques. We also predicted the structure and function of UL145 and UL136 proteins. RESULTS: Sixty-two Chinese infants infected with HCMV were recruited into this study and the clinical isolates were recovered from the urine. Two strains among the low-passage isolates, D2 and D3, were obtained. The UL145 and UL136 sequences were deposited with GenBank under accession numbers of DQ180367, DQ180381, DQ180377, and DQ180389. The mRNA expression of both UL145 and UL136 was confirmed by reverse transcription (RT-PCR) assays. UL145 was predicted to contain 1 protein kinase C phosphorylation site, 2 casein kinase II phosphorylation sites and a zinc finger structure. UL136 was predicted to contain a protein kinase C phosphorylation site, N-myristoylation site, cAMP- and cGMP-dependent protein kinase phosphorylation site and tyrosine kinase II phosphorylation site. Both UL145 and UL136 are highly conserved. CONCLUSIONS: UL145 may act as an intranuclear regulating factor by direct binding to DNA, while UL136 may be a membrane receptor involving signal transduction.

[Effects of long-term application of organic fertilizer and superphosphate on accumulation and leaching of Olsen-P in Fluvo-aquic soil].

Based on a 20-year experiment of fertilization with organic and chemical fertilizers on a Fluvo-aquic soil under wheat-corn cropping system, this paper studied the relationships between Olsen-P concentration in plough layer and crop yields as well as the accumulation and vertical translocation of Olsen-P in soil profile. The results showed that when the Olsen-P concentration in plough layer maintained at 10-40 mg x kg(-1), the grain yields of wheat and corn were higher, whereas when the concentration of Olsen-P in plough layer was higher than 40 mg x kg(-1), it started to leach, which meant that in light loam Fluvo-aquic soil, the threshold value for P leaching might be 40 mg x kg(-1). In the treatments of chemical fertilization (NPK) and corn straw returning (SNPK) with the P application rate of 77-90 kg x hm(-2), the Olsen-P concentration in plough layer was increased by 0.63-0.72 mg x kg(-1) per 100 kg x hm(-2) of applied P, with an annual increment of 0.49-0.65 mg x kg(-1) and needed 45-60 years for reaching the threshold value for P leaching. In the treatments of chemical fertilization combined with manure application (MNPK, MNPK2, and 1.5MNPK), the formula of Olsen-P accumulation in 0-20 cm soil layer were Y(MNPK) = 3.1097x + 6.9615 (R2 = 0.8562), Y(MNPK2) = 2.4765x + 13.563 (R2 = 0.9307), and Y1.5MNPK = 4.506x + 6.4464 (R2 = 0.8862). It might take 8 years to reach the threshold value for Olsen-P leaching when the P application rate in treatment 1.5MNPK was 210 kg x hm(-2), 11 years when the P application rate in treatments MNPK2 and MNPK was 125 and 140 kg x hm(-2). Organic fertilization combined with chemical fertilization increased the Olsen-P accumulation rate being 2.5 times higher than chemical fertilization. Excessive application of organic fertilizer could increase the accumulation and leaching of Olsen-P in soil profile.