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To understand the long-term effects of combined organic and chemical nitrogen fertilization on soil organic C (SOC) and total N (TN), we conducted a 30-year field experiment with a wheat–maize rotation system on the Huang-Huai-Hai Plain during 1990–2019. The experimental treatments consisted of five fertilizer regimes: no fertilizer (control), chemical fertilizer only (NPK), chemical fertilizer with straw (NPKS), chemical fertilizer with manure (NPKM), and 1.5 times the rate of NPKM (1.5NPKM). The NPK, NPKS, and NPKM treatments had equal N inputs. The crop yields were measured over the whole experimental duration. Soil samples were collected from the topsoil (0–10 and 10–20 cm) and subsoil (20–40 cm) layers for assessing soil aggregates and taking SOC and TN measurements. Compared with the NPK treatment, the SOC and TN contents increased significantly in both the topsoil (24.1–44.4% for SOC and 22.8–47.7% for TN) and subsoil layers (22.0–47.9% for SOC and 19.8–41.8% for TN) for the organically amended treatments (NPKS, NPKM and 1.5NPKM) after 30 years, while no significant differences were found for the average annual crop yields over the 30 years of the experiment. The 0–10 cm layer of the NPKS treatment and the 20–40 cm layer of the NPKM treatment had significantly higher macroaggregate fraction mass proportions (19.8 and 27.0%) than the NPK treatment. However, the 0–10 and 20–40 cm layers of the 1.5NPKM treatment had significantly lower macroaggregate fraction mass proportions (–19.2 and –29.1%) than the control. The analysis showed that the higher SOC and TN in the soil of organically amended treatments compared to the NPK treatment were related to the increases in SOC and TN protected in the stable fractions (i.e., free microaggregates and microaggregates within macroaggregates), in which the contributions of the stable fractions were 81.1–91.7% of the increase in SOC and 83.3–94.0% of the increase in TN, respectively. The relationships between average C inputs and both stable SOC and TN stocks were significantly positive with R2 values of 0.74 and 0.72 (P

Dissolved organic matter (DOM) plays important roles in soil biogeochemistry activity and nutrients transportation in soils, but studies regarding the long-term effects of green manures on the content and structure of DOM in red paddy soil have not been reported yet. A long-term green manure experiment established in 1982 was utilized to test the DOM contents in different treatments, and the spectral characteristics of DOM were investigated by using ultraviolet-visible (UV-Vis) spectrometry and Fourier transform infrared (FTIR) spectrometry. The experiment included four cropping systems: rice-rice-milk vetch (RRV), rice-rice-rape (RRP), rice-rice-ryegrass (RRG) and rice-rice-winter fallow (RRF), among them, milk vetch, rape, and ryegrass are popular winter green manure species in southern China. The results showed that the content of dissolved organic carbon (DOC), which is widely used to estimate the concentration of DOM, was significantly promoted after the incorporation of green manures compared with the other sampling stages. The contents of aromatic groups and the degree of humification of DOM increased in RRV and RRP, suggesting more complex compositions of the soil DOM after long-term application of milk vetch and rape. The contents of phenol, alcohol and carboxylic acid group at the mature stage of early rice were significantly higher than those at the stage of after green manures turned over, especially for the RRV treatment. The absorption ratio of FTIR indicated that winter plantation of rape increased the aromatic-C/aliphatic-C ratio, and ryegrass increased the aromatic-C/carboxyl-C ratio. In conclusion, long-term planting of milk vetch and rape as green manures increased the degree of aromaticity, humification and average molecular weight of DOM, and made the DOM more stable in red paddy soil.

Dissimilatory Fe(III) reduction is an important process in the geochemical cycle of iron in anoxic environment. As the main products of dissimilatory iron reduction, the Fe(II) species accumulation could indicate the reduction ability. The effects of different green manures on Fe(III) reduction in paddy soil were explored based on a 31-year rice-rice-winter green manure cropping experiment. Four treatments were involved, i.e., rice-rice-milk vetch (RRV), rice-rice-rape (RRP), rice-rice-rye-grass (RRG) and rice-rice-winter fallow (RRF). Soils were sampled at flowering stage of milk vetch and rape (S1), before transplantation (S2), at tillering (S3), jointing (S4), and mature (S5) stages of the early rice, and after the harvest of the late rice (S6). The contents of TFeHCl (HCl-extractable total Fe), Fe(II)HCl (HCl-extractable Fe(II) species) and Fe(III)HCl (HCl-extractable Fe(III) species) were measured. The correlations among those Fe species with selected soil environmental factors and the dynamic characteristics of Fe(II)HCl accumulation were investigated. The results showed that TFeHCl in RRF was significantly higher than those in the green manure treatments at most of the sampling stages. Fe(II)HCl increased rapidly after the incorporation of green manures in all treatments and kept rising with the growth of early rice. Fe(II)HCl in RRG was quite different from those in other treatments, i.e., it reached the highest at the S2 stage, then increased slowly and became the lowest one at the S4 and S5 stages. Fe(III)HCl showed oppositely, and Fe(II)HCl/Fe(III)HCl performed similarly to Fe(II)HCl. The maximum accumulation potential of Fe(II)HCl was significantly higher in RRF, while the highest maximum reaction rate of Fe(II)HCl accumulation appeared in RRG. Significant correlations were found between the indexes of Fe(II)HCl accumulation and soil pH, oxidation-reduction potential (Eh) and total organic acids, respectively. In together, we found that long-term application of green manures decreased the TFeHCl in red paddy soils, but promoted the ability of Fe(III) reduction, especially the ryegrass; Fe(II)HCl increased along with the growth of rice and was affected by soil conditions and environmental factors, especially the water and redox ability.

The development of more efficient management systems is crucial to achieving high grain yields with high nitrogen use efficiency (NUE). February Orchid-spring maize rotation system is a newly established planting system with the benefits of ground cover and potential wind erosion in northern China. A field experiment was conducted to evaluate the effects of integrated application of February Orchid as green manure with reduction of chemical fertilizers (INTEGRATED) on spring maize yield, N uptake, ammonium volatilization, and soil residual mineral N in northern China. Compared to farmers’ traditional fertilization (CON), integrated application of February Orchid as green manure with 30% reduction of nitrogen fertilizers (INTEGRATED) increased maize grain yield and biomass by 9.9 and 10.2%, respectively. The 0–100 cm soil residual Nmin at harvest was decreased by 58.5% and thus nitrogen use efficiency was increased significantly by 26.7%. The nitrogen balance calculation further demonstrated that the INTEGRATED approach performed better than CON with lower apparent nitrogen loss (decreased by 48.9%) which evidenced by the ammonium volatilization of top-dressing fertilizer was decreased by 31.1%, the Nmin movement to the deeper soil layers was reduced, and the apparent nitrogen leaching loss nearly equal to 0 under the INTEGRATED treatment. Therefore, in northern China, integrated application of green manure and chemical fertilizers is an efficient management approach for improving maize yields and NUE simultaneously.

On the basis of a long-term (30 years) field experiment that involved four rotation systems, rice-rice-winter fallow (RRF), rice-rice-ryegrass (RRG), rice-rice-rape (RRP), and rice-rice-milk vetch (RRV), this study described the effects of green manure on the microbial communities in the red paddy soils using 454 pyrosequencing for the 16S rRNA gene. The Chao1 richness and non-parametric Shannon's index increased in all soil samples that received green manure treatments. The communities’ structures with the green manure applications were significantly dissimilar from that under the winter fallow. Using Metastats tests, many genera in the RRG, RRP and RRV soils were significantly different from those in the RRF soil, including a number of genera that functioned in the nitrogen and sulfur cycles. Analyses of the genera with these functions revealed the shifts in microbial ecosystem functions after long-term green manuring. Changes in the microbial communities increased the ammonium supply and decreased the soil acidification in green-manure-amended soils. Together, these data suggested powerful effects of green manure on both the microbial communities and the biogeochemical cycle driven by the shifts in bacterial functional groups.