Autotrophic and Mixotrophic Microbial Carbon Assimilation During Organic Residue Decomposition in Mollisols: Mechanisms and Controls

Mollisols represent foundational agricultural soils in which high organic carbon (C) and active microbiomes sustain fertility and mediate global C cycling. However, decades of intensive cultivation have depleted soil organic C (SOC) and degraded soil structure and function. Enhancing C sequestration in agricultural Mollisols through the incorporation of organic residue, such as crop residues, organic waste, and spent mushroom substrates has become an urgent scientific and management priority. This review integrates advances from the past decade, combining stable isotope probing, multi-omics analyses, and ultrahigh-resolution molecular characterization to elucidate how microorganisms mediate C sequestration during organic residue return and decomposition. We propose a four-dimensional conceptual framework, “substrate–microenvironment–metabolic pathway–residue stabilization,” that links microbial metabolism with long-term C persistence in Mollisols. We further highlight that organic residue inputs promote CO2 sequestration via fermentation–autotrophy coupling, nitrifying autotrophy, and microbial mixotrophy. Major C sequestration pathways operate synergistically across redox microenvironments, forming stratified metabolic networks that sustain continuous C cycling. The chemical composition and decomposition kinetics of organic residue governs substrate and energy fluxes for microbial C sequestration, while soil redox status, and nutrient coupling (Carbon–Nitrogen–Phosphorus–Sulfur) collectively direct C flow toward stabilization. Microbial necromass and extracellular polymers achieve long-term C storage through mineral adsorption and microaggregate formation. Finally, we summarize recent methodological advances for tracing microbial CO2 sequestration in agricultural Mollisols and identify key research needs on residue formation, C use efficiency, and aggregate-mineral protection mechanisms. This synthesis establishes a mechanistic foundation for biologically regulated C management and offers guidance for sustainable cropland restoration.