Green manure (GM) has been used to support rice production in southern China for thousands of years. However, the effects of GM on soil carbon sequestration (CS) and the carbon footprint (CF) at a regional scale remain unclear. Therefore, we combined the datasets from long-term multisite experiments with a meta-analysis approach to quantify the potential of GM to increase the CS and reduce the CF of paddy soils in southern China. Compared with the fallow– rice practice, the GM–rice practice increased the soil C stock at a rate of 1.62 Mg CO₂-eq ha^–1 yr^–1 and reduced chemical N application by 40% with no loss in the rice yield. The total CF varied from 7.51 to 13.66 Mg CO₂-eq ha^–1 yr^–1 and was dominated by CH₄ emissions (60.7–81.3%). GM decreased the indirect CF by 31.4% but increased the direct CH₄ emissions by 19.6%. In the low and high CH4 emission scenarios, the CH₄ emission factors of GM (EFgc) were 5.58 and 21.31%, respectively. The greater soil CS offset the increase in GM-derived CF in the low CH₄ scenario, but it could not offset the CF increase in the high CH₄ scenario. A trade-off analysis also showed that GM can simultaneously increase the CS and reduce the total CF of the rice production system when the EFgc was less than 9.20%. The variation in EFgc was mainly regulated by the GM application rates and water management patterns. Determining the appropriate GM application rate and drainage pattern warrant further investigation to optimize the potential of the GM–rice system to increase the CS and reduce the total CF in China.

The co-utilization of green manure (GM) and rice straw (RS) in paddy fields has been widely applied as an effective practice in southern China. However, its effects on soil aggregate and soil organic carbon (SOC) stability remain unclear. In the present study, the effect of GM, RS, and co-utilization of GM and RS on particle size distribution of soil aggregates and SOC density fractions were measured in a field experiment. The experiment included six treatments, i.e., winter fallow (WF) without RS return (Ctrl), WF with 50% RS return (1/2RS), WF with 100% RS return (RS), GM without RS return (GM), GM with 50% RS return (GM1/2RS) and GM with 100% RS return (GMRS). The results showed that the proportion of small macro-aggregates (0.25–2 mm) and the mean weight diameter (MWD) of aggregates in the GMRS treatment was greater (by 18.9 and 3.41%, respectively) than in the RS treatment, while the proportion of silt+clay particles (<0.053 mm) was lower (by 14.4%). The concentration of SOC in microaggregates (0.053–0.25 mm) and silt+clay particles was higher in the GMRS treatment than in GM and RS treatments individually. The concentration and proportion of free light organic carbon (fLOC) in aggregates of various particle sizes and bulk soil was greater in the GMRS treatment than the RS treatment, whereas the concentration and proportion of mineral-associated organic carbon in small macroaggregates, microaggregates, and bulk was lower in the GMRS treatment than in the RS treatment. The proportion of intra-aggregate particulate organic carbon (iPOC) was greater in the GMRS treatment than in GM treatment. The GMRS treatment had strong positive effects on iPOC in small macroaggregates, suggesting that SOC was transferred from fLOC to iPOC. In conclusion, co-utilizing green manure and rice straw cultivated the SOC pool by increasing the concentration of fLOC and improved soil carbon stability by promoting the sequestration of organic carbon in iPOC as a form of physical protection.

Green manure–crop rotation is a sustainable approach to protect crops against diseases and improve yield.  However, the mechanism by which green manuring manipulates the crop-associated microbial community remains to be elucidated.  In this study, we explored the horizontal processes of bacterial communities in different compartments of the soil–root interface (bulk soil, rhizosphere soil, rhizoplane and endosphere) of tobacco by performing a field experiment including four rotation practices, namely, tobacco rotated with smooth vetch, ryegrass, radish, and winter fallow (without green manure).  Results showed that the co-occurrence networks constructed by adjacent compartments of the soil–root interface with green manuring had more edges than without green manuring, indicating  dramatic microbial interactions.  Green manuring increased the dispersal-niche continuum index between bulk soil and other compartments, indicating that it facilitated the horizontal dispersal of microbes.  For the different green manuring practices, the neutral community model explained 24.6–27.6% of detection frequency for bacteria, and at least one compartment under each practice had a normalized stochasticity ratio higher than the 50% boundary point, suggesting that the deterministic and stochastic processes jointly shaped the tobacco microbiome.  In conclusion, green manuring generally facilitates bacterial community dispersal across different compartments and enhances potential interactions among adjacent compartments.  This study provides empirical evidence for understanding the microbiome assembly under green manure–crop rotation.

To ascertain the possibility of cultivating maize using biological nitrogen fixation (BNF) by leguminous green manure crops in maize/leguminous green manure intercropping systems, BNF and nitrogen (N) transfer were studied in Xining and Wuwei, two typical northwestern Chinese cities.  The experimental treatments included monocultured maize, monocultured green manures (hairy vetch and common vetch), and their intercropping systems.  The proportions of N derived from the atmosphere (%N_dfa) in intercropping systems were not significantly different from that in monocultured green manure systems at either experimental site, except for that in hairy vetch (HV) in Xining.  The amount of N derived from the atmosphere (N_dfa) of common vetch (CV) significantly decreased from 1.16 and 1.10 g/pot in monoculture to 0.77 and 0.55 g/pot when intercropped with maize, in Xining and Wuwei, respectively, and the N_dfa of HV when intercropped significantly decreased from 1.02 to 0.48 g/pot in Xining.  In the intercropping systems in Xining and Wuwei, the amounts of N transferred (N_transfer) from CV to maize were 21.54 and 26.81 mg/pot, accounting for 32.9 and 5.9% respectively of the N accumulation in maize, and the values of N_transfer from HV to maize were 39.61 and 46.22 mg/pot, accounting for 37.0 and 23.3%, respectively, of the N accumulation in maize.  Path analysis showed that soil nutrient and green manure biomass were mainly related to N_dfa, and that ^δ15N had a primary relationship with N_transfer.  We found that 5.9–37.0% of N accumulation in maize was transferred from green manures, and that the N transfer ability to maize of HV was higher than that of CV.  In conclusion, intercropping with leguminous green manures provided a feasible way for maize to effectively utilize biologically fixed N.

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-ryegrass (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 TFe_HCl (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 TFe_HCl 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 TFe_HCl 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.

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.

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.