Soil health relies on the actions and interactions of an abundant and diverse biological community. Current soil health assessments rely heavily on a suite of soil biological, chemical, and physical indicators, often excluding molecular information. Soil health is critical for sustainable agricultural production, and a comprehensive understanding of how microbial communities provide ecosystem services can help guide management practices. To explore the role of microbial function in soil health, 536 soil samples were collected from 26 U.S. states, representing 52 different crops and grazing lands, and analyzed for various soil health indicators. The bacterial functional profile was characterized using 16S ribosomal RNA gene sequencing paired with PICRUSt2 to predict metagenome functions. Functional data were used as predictors in eXtreme Gradient Boosting (XGBoost), a powerful machine learning algorithm, and enzymes important to soil health indicators were compiled into a Molecular Index of Soil Health (MISH). The overall MISH score significantly correlated with non-molecular measures of soil health and management practice adoption. Additionally, several new enzymes were identified as potential targets to better understand microbial mediation of soil health. This low-cost, DNA-based approach to measuring soil health is robust and generalizable across climates.

The goal of our study was to evaluate the long-term (>12 years) influence of stocking density and herd rotation frequency on plant and soil microbial community and carbon dynamics in three working ranches in Texas. One ranch utilized a high stocking density and high-frequency (HIGH) rotation where cattle were moved multiple times each day; the second ranch used a medium stocking density and rotation frequency (MED) where herds were moved every 2–3 weeks; and the third ranch used a low stocking density with continuous grazing (LOW). Neither plant nor microbial diversity measures differed between the ranches, but plant functional and microbial community compositions differentiated management strategies. The MED ranch was characterized by a plant community dominated by little bluestem (Schizachyrium scoparium) and had the greatest soil organic matter content (2.8%) and soil respiration rates compared to the LOW (SOM = 2.2%) and HIGH (SOM = 2.1%) ranches. The HIGH ranch had a relatively high abundance and diversity of forbs and introduced grasses, and the LOW ranch had an even mixture of tall, introduced, and cool-season grasses. All three ranches had relatively high levels of Gram-positive bacteria (>70%) with MED having a higher relative abundance of bacteria important for carbon cycling. Furthermore, network analyses suggest that soil microbial communities at all ranches were highly synergistic and exhibited well-defined ecological niches. Differences in soil properties between ranches tended to be minor and suggest that grazing strategies can differ without any substantial shifts in soil and microbial function.