Agricultural intensification accelerates the degradation of cropland, and restoration has been managed by changing its vegetation. However, the keystone microbiome that drives the decomposition of plant-associated organic matter in the restoration is poorly understood. In this study, we established a 14-year field restoration experiment on a degraded cropland with four treatments: (1) bare land soil without biomass input (BL), (2) maize cropland (CL) without fertilization and biomass input, (3) natural grassland (GL), and (4) alfalfa cropland (AL) with biomass left in the fields. The activity of total soil microbiome was assessed by community-level physiological profiling (CLPP) with Biolog EcoPlates analysis, and keystone microbiome was identified as phylotypes showing statistically significant increase in the restored soils (GL and AL) relative to the degraded BL soil. The results showed that GL and AL treatments improved soil fertility as indicated by significant increase in soil organic carbon, total nitrogen, and available phosphorus when compared to BL treatment. The significant difference was not observed between CL and BL treatments except for pH and available phosphorus, indicating that the input and microbial decomposition of plant-associated organic matter were the key for restoration of soil fertility. Similar results were obtained for soil microbial activities of carbon utilization efficiency via CLPP analysis, and real-time quantitative polymerase chain reaction of 16S rRNA genes further revealed significantly higher abundance of total soil microbial community in GL and AL soils than in BL and CL. High-throughput sequencing of total 16S rRNA genes revealed the Bacteroidetes as the only keystone taxa at phylum level, and 106 and 120 genera were keystone phylotypes. Compared with BL and CL, the genera that increase significantly in GL and AL are called keystone genera of ecological restoration. The dominant keystone genera includedReyranella,Mesorhizobium,Devosia,Haliangium,Nocardioides, andPseudonocardia. Significantly higher abundance ofBacillusgenus in BL soil implied it might serve as an indicator of agricultural land degradation. Statistical analysis showed that soil organic carbon and pH were significantly correlated with the input of plant-associated organic matters and dynamic changes of keystone taxa. These results suggest that the vegetation of natural grass (GL) and alfalfa plant (AL) and subsequent decomposition of plant-associated materials could serve as effective strategies for restoration of the degraded cropland by stimulating the keystone microbiomes and improving their physiological metabolisms of carbon utilization efficiency.

Agricultural intensification accelerates the degradation of cropland, and restoration has been managed by changing its vegetation. However, the keystone microbiome that drives the decomposition of plant-associated organic matter in the restoration is poorly understood. In this study, we established a 14-year field restoration experiment on a degraded cropland with four treatments: (1) bare land soil without biomass input (BL), (2) maize cropland (CL) without fertilization and biomass input, (3) natural grassland (GL), and (4) alfalfa cropland (AL) with biomass left in the fields. The activity of total soil microbiome was assessed by community-level physiological profiling (CLPP) with Biolog EcoPlates analysis, and keystone microbiome was identified as phylotypes showing statistically significant increase in the restored soils (GL and AL) relative to the degraded BL soil. The results showed that GL and AL treatments improved soil fertility as indicated by significant increase in soil organic carbon, total nitrogen, and available phosphorus when compared to BL treatment. The significant difference was not observed between CL and BL treatments except for pH and available phosphorus, indicating that the input and microbial decomposition of plant-associated organic matter were the key for restoration of soil fertility. Similar results were obtained for soil microbial activities of carbon utilization efficiency via CLPP analysis, and real-time quantitative polymerase chain reaction of 16S rRNA genes further revealed significantly higher abundance of total soil microbial community in GL and AL soils than in BL and CL. High-throughput sequencing of total 16S rRNA genes revealed the Bacteroidetes as the only keystone taxa at phylum level, and 106 and 120 genera were keystone phylotypes. Compared with BL and CL, the genera that increase significantly in GL and AL are called keystone genera of ecological restoration. The dominant keystone genera includedReyranella,Mesorhizobium,Devosia,Haliangium,Nocardioides, andPseudonocardia. Significantly higher abundance ofBacillusgenus in BL soil implied it might serve as an indicator of agricultural land degradation. Statistical analysis showed that soil organic carbon and pH were significantly correlated with the input of plant-associated organic matters and dynamic changes of keystone taxa. These results suggest that the vegetation of natural grass (GL) and alfalfa plant (AL) and subsequent decomposition of plant-associated materials could serve as effective strategies for restoration of the degraded cropland by stimulating the keystone microbiomes and improving their physiological metabolisms of carbon utilization efficiency.