Response of the chemical structure of soil organic carbon to modes of maize straw return.

Elucidating the chemical structure of soil organic matter (SOM) is important for accurately evaluating the stability and function of SOM. Aboveground vegetation directly affects the quantity and quality of exogenous organic matter input into the soil through plant residues and root exudates, which in turn affects soil microbial species, community structure, and activity, and ultimately impacts the chemical structure of SOM. In this study, a 13C nuclear magnetic resonance technique was used to analyze the chemical structure characteristics of soil organic carbon (SOC) under various rates of straw returning combined with rotary tillage and under full straw mulching. The results showed that full straw returning with rotary tillage and full straw mulching more effectively increased the SOC content than reduced rate of straw returning (1/2 and 1/3 of full straw) with rotary tillage. The contents of alkyl C and alkoxy C in the functional groups of SOC under various straw returning treatments were increased compared with those under the treatment of maize stubble remaining in soil (CK). Furthermore, the contents of aromatic C and carboxyl C were decreased, which were consistent with the chemical shift changes of SOC. Compared with CK treatment, straw returning decreased the content of aromatic C in the functional groups of SOC, but increased the content of alkoxy C, which could be associated with the change in integral areas of absorption peaks of alkyl C and alkoxy C moving toward left and right, respectively. The content of total SOC was significantly positively (P < 0.05) correlated with that of alkoxy C and significantly negatively (P < 0.01) correlated with that of aromatic C. The molecular structure of SOC tends to be simplified due to the decreasing in refractory C and the increasing in easily decomposed C after straw returning to the field.

Investigations of the effect of the amount of biochar on soil porosity and aggregation and crop yields on fertilized black soil in northern China.

The combination of chemical fertilizer and biochar is regarded as a useful soil supplement for improving the properties of soil and crop yields, and this study describes how the biochar of maize straw can be used to improve the quality of the degraded black soil. This has been achieved by examining the effects of combining different amounts of biochar with chemical fertilizer on the porosities and aggregate formation of soil and exploring how these changes positively impact on crop yields. A field trial design combining different amounts of maize straw biochar [0 (NPK), 15.75 (BC1), 31.5 (BC2), and 47.25 t ha-1 (BC3)] with a chemical fertilizer (NPK) has been used to investigate changes in the formation of soil aggregate, clay content, soil organic carbon (SOC), and crop yields in Chinese black soil over a three year period from 2013 to 2015. The results of this study show that the addition of fertilizer and biochar in 2013 to black soil results in an increased soybean and maize yields from 2013 to 2015 for all the treatments, with BC1/BC2 affording improved crop yields in 2015, while BC3 gave a lower soybean yield in 2015. Total porosities and pore volumes were increased for BC1 and BC2 treatments but relatively decreased for BC3, which could be attributed to increased soil capillary caused by the presence of higher numbers of fine soil particles. The addition of biochar had a positive influence on the numbers and mean weight diameters (MWD) of soil macroaggregates (>0.25 mm) that were present, with the ratio of SOC to TN in soil macroaggregates found to be greater than in the microaggregates. The most significant amount of carbon present in macroaggregates (>2 mm and 0.25-2 mm) was observed when BC2 was applied as a soil additive. Increasing the levels of maze straw biochar to 47.25 t ha-1 led to an increase in the total organic carbon of soil, however, the overall amount of macroaggregates and MWD were decreased, which is possibly due to localized changes in microbial habitat. The supplementation of biochar increased in the amount of aromatic C present (most significant effect observed for BC2), with the ratio of aliphatic C to aromatic C found to be enhanced due to a relative reduction in the aliphatic C content with >2 mm particle fraction. These changes in organic carbon content and soil stability were analyzed using univariate quadratic equations to explain the relationship between the type of functional groups (polysaccharide C, aliphatic C, aromatic C, aliphatic C/aromatic C) present in the soil aggregates and their MWDs, which were found to vary significantly. Overall, the results of this study indicate that the use of controlled amounts of maize-straw biochar in black soil is beneficial for improving crop yields and levels of soil aggregation, however, the use of excessive amounts of biochar results in unfavorable aggregate formation which negatively impacts the yields of crop growth. The data produced suggest that aromatic C content can be used as a single independent variable to characterize the stability of soil aggregate when biochar/fertilizer mixtures are used as soil additives to boost growth yields. Analysis of soil and crop performance in black soil revealed that the application of maize-straw biochar at a rate of 15.75 and 31.5 t ha-1 had positive effects on crop yields, soil aggregation and accumulation of aromatic C in the aggregate fractions when a soybean-maize rotation system was followed over three years.

Variations in the profile distribution and protection mechanisms of organic carbon under long-term fertilization in a Chinese Mollisol.

Long term fertilization may have a significant effect on soil organic carbon (SOC) fractions and profile distribution. However, previous research mostly explored the SOC in the topsoil and provided little or no information about its distribution in deeper layers and various protection mechanisms particularly under long-term fertilization. The present study investigated the contents and profile distribution (0-100 cm) of distinct SOC protection mechanisms in the Mollisol (black soil) of Northeast China after 35 years of mineral and manure application. The initial Organic Matter content of the topsoil (0-20 cm) ranged from 26.4 to 27.0 g kg-1 soil, and ploughing depth was up to 20 cm. A combination of physical-chemical fractionation methods was employed to study various SOC fractions. There were significant variations throughout the profile among the various fractions and protection mechanisms. In topsoil (to 40 cm), mineral plus manure fertilization (MNPK) increased the total SOC content and accounted for 16.15% in the 0-20 cm and 12.34% in the 20-40 cm layer, while the manure alone (M) increased the total SOC by 56.14%, 48.73% and 27.73% in the subsoil (40-60, 60-80 and 80-100 cm, respectively). Moreover, MNPK and M in the topsoil and subsoil, respectively increased the unprotected coarse particulate organic carbon (cPOC) (48% and 26%, respectively), physically protected micro-aggregate (µagg) (20% and 18%, respectively) and occluded particulate organic carbon (iPOC) contents (279% and 93%, respectively) compared with the control (CK). A positive linear correlation was observed between total SOC and the cPOC, iPOC, physico-biochemically protected NH-µSilt and physico-chemically protected H-µSilt (p < 0.01) across the whole profile. Overall, physical, physico-biochemical and physico-chemical protection were the predominant mechanisms to sequester carbon in the whole profile, whereas the biochemical protection mechanisms were only relevant in the topsoil, thus demonstrating the differential mechanistic sensitivity of fractions for organic carbon cycling across the profile.

Profiles of antibiotic resistome with animal manure application in black soils of northeast China.

Black soils (Mollisols) are important soil resources for crop production and maintain food safety in China. For keeping soil fertility, the application of animal manure is commonly practiced in black soils. However, the impact of this application on abundance and diversity of antibiotic resistance genes (ARGs) in black soils of China remains unclear. Here, we surveyed the profiles of ARGs in 72 soil samples collected from four long-term experimental stations with different fertilization regimes and from open farmlands in two sites across northeast China using high-throughput quantitative PCR. Results showed that a total of 178 ARGs including mobile genetic elements (MGEs) were detected, and the diversity and abundance of ARGs were significantly increased with manure application. Additionally, the finding of a significant positive correlation between relative abundance of ARGs and MGEs (P < 0.0001), suggesting that horizontal gene transfer may potentially impact the transmission of ARGs. Furthermore, two genes aadA-1-01 and mexF, encoding resistance to aminoglycoside and multidrug, respectively, were recognized as indicators to estimate the abundance of other co-occurring ARGs. These findings provided insights into the soil resistome in black soils of northeast China and also highlighted the environmental risks caused by manure application should not be ignored.

Impacts of Long-term Fertilization on the Molecular Structure of Humic Acid and Organic Carbon Content in Soil Aggregates in Black Soil.

Soil aggregates are the basic units of soil structure, and their composition and carbon (C) stability directly affect soil fertility. As cementing agents, humic substances play an important role in the formation and stability of soil aggregates. Long-term fertilization not only changes the structure of humic acid (HA), but also affects the content and stability of organic C in soil aggregates. In this study, based on a long-term fertilization experiment, the relationship between the molecular structure of HA and the stability of organic C in the aggregates was examined. Compared with the non-fertilization control (CK), both the application of organic manure alone (M) and organic manure combined with inorganic fertilizer application (MNPK) increased organic C content in the bulk soil and in HA. In addition, the application of organic manure (M, MNPK) favored the formation of macroaggregates (>0.25 mm) and showed a higher organic C contents of soil aggregates with different sizes than the CK. Moreover, the content of aliphatic C, the ratios of aliphatic C/aromatic C and alkyl C/O-alkyl C was increased with the application of organic fertilizer. A significant negative correlation was observed between aromatic C and organic C content of the aggregates with sizes of >2 mm, 2-0.25 mm, and 0.25-0.053 mm (P < 0.05). The findings indicated that organic fertilization treatments (M and MNPK) increased the aliphatic C content of HA, which favored the increase in the organic C content and stability of the aggregates.

Long-term application of nitrogen, not phosphate or potassium, significantly alters the diazotrophic community compositions and structures in a Mollisol in northeast China.

This study investigated the effects of 35 years of application of inorganic fertilizers containing nitrogen (N), phosphate (P), or potassium (K) alone or in combination on the abundance and composition of diazotrophic community in a black soil (Mollisol) in northeast China. The abundance and composition of diazotrophic community were analyzed using qPCR and Illumina MiSeq sequencing targeting nifH genes. Nitrogen fertilization decreased the abundance and Shannon diversity of nifH genes. The diazotrophic community was dominated by Alphaproteobacteria (Bradyrhizobium at the genus level), with relatively higher abundance in the N fertilization treatments than in the non-N fertilization treatments. All diazotrophic communities were clustered into two groups with and without N fertilization history, and the soil pH, total C, total N, and NO3--N significantly influenced the structure of the whole diazotrophic community. Moreover, random matrix theory analysis elucidated a clear difference in network structures between the N and non-N fertilization treatments, with N fertilization causing a less stable network structure. These results highlighted that it was N fertilizer, but not P and K fertilizers, contributed to great changes in the diazotrophic community in this black agricultural soil.

Effects of long-term fertilization on soil humic acid composition and structure in Black Soil.

The composition and structure of humic acid (HA) can be affected by fertilization, but the short-term effects are difficult to detect using traditional analysis methods. Using a 35-year long-term experiment in Black Soil, the molecular structure of HA was analyzed with Fourier transform infrared spectroscopy (FTIR), 13C nuclear magnetic resonance spectroscopy (NMR), and fluorescence spectroscopy. Variation in HA was analyzed after long-term fertilization, including fertilization with manure (M), inorganic N, P and K fertilizer (NPK), manure combined with inorganic N, P, and K fertilizer (MNPK), and a no-fertilizer control (CK). The application of each fertilizer treatment increased crop yields compared with the CK treatment, and the MNPK treatment increased crop yield the most. The ratio of main IR absorption peak of HA at 2,920 cm-1 compared with the peak at 2,850 cm-1 (2920/2850) was higher in the NPK and MNPK treatments compared with the CK treatment. The application of manure (MNPK and M treatments) increased the ratio of hydrogen to carbon (H/C) in HA, and raised the ratio of the main IR absorption peak of HA at 2920 cm-1 to that at 1720 cm-1 (2920/1720). Manure treatments also raised the ratio of aliphatic carbon (C) to aromatic C, alkyl C to alkoxy C and hydrophobic C to hydrophilic C and the fluorescence index (f 450/500), but decreased the degree of aromatization of HA, when compared with the CK treatment. The ratio between each type of C in HA was similar among all the fertilizer treatments, but NPK had a lower ratio of H/C and a lower content of aliphatic C compared with the CK treatment. These results indicated that the molecular structure of HA in Black Soil tends to be aliphatic, simpler, and younger after the application of manure. While the application of inorganic fertilizers increased in the degree of condensation of HA and made HA structure complicated. The application of manure alone or combined with inorganic fertilizers may be an effective way to increase crop yield and improve the structure of soil organic matter.

[Study on Biochar Properties Analysis with Scanning Electron Microscope-EnergyDispersive X-Ray Spectroscopy (SEM-EDX)].

Scanning Electron Microscope-Energy Dispersive X-ray spectroscopy (SEM-EDX) was applied to analyze the chemical and structural properties of biochars produced under different temperatures. Results showed that average carbon content (SEM C aver.) and maximum carbon content (SEM C max.) of miscanthus (MS) biochar increased as temperature increasing. There were significant and positive relationships between SEM C max., SEM C aver. and highest treatment temperature (HTT) (r were 0.76 and 0.86). SEM C max., SEM C aver. and dry combustion total carbon content had significant and positive relationships (r were 0.83 and 0.91), SEM C max. which was better than SEM C aver. So the carbon content of MS biochar which had good correlationship with temperature analyzed by SEM-EDX, SEM C max. could be used for composition analysis of biochar, scanning results could analyze structural properties of biochar effectively. This method is rapid, simple and stable. It also could analyze structure and composition of biochar simultaneously. It is a promising method that would be useful to study the structure and composition of biochar and other materials.

[Distribution and enrichment characteristics of organic carbon and total nitrogen in mollisols under long-term fertilization].

The characteristics and changes of soil organic carbon (SOC) and total nitrogen (TN) in different size particles of soil under different agricultural practices are the basis for better understanding soil carbon sequestration of mollisols. Based on a 31-year long-term field experiment located at the Heilongjiang Academy of Agricultural Sciences (Harbin) , soil samples under six treatments were separated by size-fractionation method to explore changes and distribution of SOC and TN in coarse sand, fine sand, silt and clay from the top layer (0-20 cm) and subsurface layer (20-40 cm). Results showed that long-term application of manure (M) increased the percentages of SOC and TN in coarse sand and clay size fractions. In the top layer, application of nitrogen, phosphorus and potassium fertilizers combined with manure (NPKM) increased the percentages of SOC and TN in coarse sand by 191.3% and 179.3% compared with the control (CK), whereas M application increased the percentages of SOC and TN in clay by 45% and 47% respectively. For subsurface layers, the increase rates of SOC and TN in corresponding parts were lower than that in top layer. In the surface and subsurface layers, the percentages of SOC storage in silt size fraction accounted for 42%-63% and 48%-54%, TN storage accounted for 34%-59% and 41%-47%, respectively. The enrichment factors of SOC and TN in coarse sand and clay fractions of surface layers increased significantly under the treatments with manure. The SOC and TN enrichment factors were highest in the NPKM, being 2.30 and 1.88, respectively, while that in the clay fraction changed little in the subsurface layer.

Changes in Olsen Phosphorus Concentration and Its Response to Phosphorus Balance in Black Soils under Different Long-Term Fertilization Patterns.

The Olsen phosphorus (P) concentration of a soil is a key index that can be used to evaluate the P supply capacity of the soil and to estimate the optimal P fertilization rate. A study of the relationship between the soil Olsen P concentration and the P balance (P input minus P output) and their variations among different fertilization patterns will help to provide useful information for proper management of P fertilization. In this paper, the two investigated long-term experiments were established on black soils in the northeast region of China. Six fertilization treatments were selected: (1) unfertilized (CK); (2) nitrogen only (N); (3) nitrogen and potassium (NK); (4) nitrogen and phosphorus (NP); (5) nitrogen, phosphorus, and potassium (NPK); and (6) nitrogen, phosphorus, potassium and manure (NPKM). The results showed that the average Olsen P concentrations in the black soils at Gongzhuling and Harbin (16- and 31-year study periods, respectively), decreased by 0.49 and 0.56 mg kg-1 a-1, respectively, without P addition and increased by 3.17 and 1.78 mg kg-1 a-1, respectively, with P fertilization. The changes in soil Olsen P concentrations were significantly (P<0.05) correlated with the P balances at both sites except for the NP and NPK treatments at Gongzhuling. Under an average deficit of 100 kg ha-1 P, the soil Olsen P concentration at both sites decreased by 1.36~3.35 mg kg-1 in the treatments without P addition and increased by 4.80~16.04 mg kg-1 in the treatments with 100 kg ha-1 of P accumulation. In addition, the changes in Olsen P concentrations in the soil with 100 kg ha-1of P balance were significantly correlated with the P activation coefficient (PAC, percentage of Olsen P to total P, r=0.99, P<0.01) and soil organic matter content (r=0.91, P<0.01). A low pH was related to large changes of Olsen P by 1 kg ha-1 of P balance. These results indicated that soil organic matter and pH have important effects on the change in soil Olsen P by 1 kg ha-1 of P balance.

[Effect of long-term fertilization on microbial community functional diversity in black soil].

In order to study the effects of long-term different fertilization on microbial community functional diversity in arable black. soil, we examined microbial metabolic activities in two soil la- yers (0-20 cm, 20-40 cm) under four treatments (CK, NPK, M, MNPK) from a 35-year continuous fertilization field at the Ministry of Agriculture Key Field Observation Station of Harbin Black Soil Ecology Environment using Biolog-ECO method. The results showed that: in the 0-20 cm soil layer, combined application of organic and inorganic fertilizer(MNPK) increased the rate of soil microbial carbon source utilization and community metabolism richness, diversity and dominance; In the 20-40 cm layer, these indices of the MNPK treatment was lower than that of the NPK treat- ment; while NPK treatment decreased soil microbial community metabolism evenness in both layers. Six groups of carbon sources used by soil microbes of all the treatments were different between the two soil layers, and the difference was significant among all treatments in each soil layer (P < 0.05) , while the variations among treatments were different in the two soil layers. Canonical correspondence analysis (CCA) showed that soil microbial community metabolic function of all the treatments was different between the two soil layers, and there was difference among all treatments in each soil layer, while the influences of soil nutrients on soil microbial community metabolic function of all treatments were similar in each soil layer. It was concluded that long-term different fertilization affected soil microbial community functional diversity in both tillage soil layer and down soil layers, and chemical fertilization alone had a larger influence on the microbial community functional diversity in the 20-40 cm layer.

Bacillus daqingensis sp. nov., a halophilic, alkaliphilic bacterium isolated from saline-sodic soil in Daqing, China.

An alkaliphilic, moderately halophilic, bacterium, designated strain X10-1(T), was isolated from saline-alkaline soil in Daqing, Heilongjiang Province, China. Strain X10-1(T) was determined to be a Gram-positive aerobe with rod-shaped cells. The isolate was catalase-positive, oxidase-negative, non-motile, and capable of growth at salinities of 0-16% (w/v) NaCl (optimum, 3%). The pH range for growth was 7.5-11.0 (optimum, pH 10.0). The genomic DNA G+C content was 47.7 mol%. Its major isoprenoid quinone was MK-7 and its cellular fatty acid profile mainly consisted of anteiso-C15:0, anteiso-C17:0, iso-C15:0, C16:0, and iso-C16:0. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol. Phylogenetic analysis based on 16S rRNA gene sequences showed that X10-1(T) is a member of the genus Bacillus, being most closely related to B. saliphilus DSM15402(T) (97.8% similarity) and B. agaradhaerens DSM 8721(T) (96.2%). DNA-DNA relatedness to the type strains of these species was less than 40%. On the basis of the phylogenetic, physiological, and biochemical data, strain X10-1(T) represents a novel species of the genus Bacillus, for which the name Bacillus daqingensis sp. nov. is proposed. The type strain is X10-1(T) (=NBRC 109404(T) = CGMCC 1.12295(T)).

[Response of black soil organic carbon, nitrogen and its availability to longterm fertilization].

Based on the long-term fertilization experiments, effects of various fertilization practices on the soil organic carbon (SOC) and total nitrogen (TN) in the surface (0-20 cm) and subsurface (20-40 cm) black soil in northeast China were studied. Results showed that, compared with the CK, long-term application of organic manure, especially the combination of mineral fertilizers and organic manure significantly increased the organic SOC and TN in the surface soil. Application of mineral fertilizers plus organic manure with conventional (NPM) and high application (N2P2M2) rate increased SOC significantly by 24. 6% and 25.1% , and TN by 29.5% and 32.8%, respectively. However, there was no significant difference among the treatments for SOC and TN at the subsurface. Compared with the CK (CKh), mineral fertilizer plus organic manure (NPM and N2P2M2) did not only increase the soil microbial biomass carbon (SMBC) and nitrogen (SMBN) , dissolved organic carbon (DOC) and nitrogen (DN), but also significantly increased the ratio of SMBC and DOC to SOC, SMBN and TN to TN. Application of the NPM and N2P2M2 increased the value of SMBC/SOC by 0.36 to 0.59 and SMBN/TN by 1.21 to 1.95 percentage points, respectively. The value of DOC/SOC and DN/TN ranged from 0.53% to 0.72% and 1.41% to 1.78%, respectively. This result indicated that SMBC, SMBN, DOC, DN and SMBC/ SOC, SMBN/TN, DOC/SOC, DN/TN were more sensitive than SOC and TN to long-term fertilization in the soil profile, and were better indicators for the impact of long-term fertilization soil fertility. The concluded that the application of manure especially manure plus mineral fertilizers can increase soil nutrients activity in the surface and subsurface black soil, acting as a helpful practice to improve soil fertility and the ability of nutrient supply, while it may cause potential environment pollution on carbon and nitrogen loss in the agroecosystem.

[Fluorescence spectroscopic characteristics of fulvic acid from the long-term located fertilization in black soil].

In order to investigate the effect of long-term located fertilization on soil fulvic acid (FA), in this study, four soil samples were taken from black soil with long-term located fertilization (about 30 year) in Harbin, Heilongjiang province. The fertilization treatments included control (CK), N, P and K fertilization (NPK), horse manure (OM), combination of organic manure and chemical fertilizations (MNPK). Soil FA was extracted from the samples and purified. The excitation, emission, synchronous, and three-dimensional-excitation emission matrix fluorescence spectroscopy (3DEEM) characteristics of the FA were determined. The excitation, emission and synchronous scan spectra all indicated that the main peaks of FA in the NPK treatment exhibited a significantly blue shift compared with CK, while those of MNPK, OM treatment caused a red shift to some extent. 3DEEM spectra of FA in all treatments exhibited four peaks (peak a, peak b, peak c, and peak d), compared with FA in CK, the wavelengths shift tendency of peak a, peak b, and peak c of FA 3DEEM in NPK, MNPK and OM treatments were similar to that of traditional spectra in FA. In order to provide quantitative information of FA humification degree in different treatments, we investigated the fluorescence index f450/500 (FI), area integration (A370-600 nm, A1 370-412 nm, A4 538-600 nm). Compared with CK, the f450/500, ratio of A1/A in NPK and A4/A in MNPK treatment increased by 4.62%, 6.12%, 7.22%, respectively. However, the f450/500, the ratio of A1/A in MNPK and A4/A in NPK treatment decreased by 3.86%, 15.31%, and 7.22% respectively. This indicated that NPK application gave a lower degree of FA humification, and combination of organic manure and chemical fertilizations would lead to a greater degree of FA aromatization in black soil with long-term located fertilization than CK.