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The Conservation Reserve Program (CRP) in the USA plays a major role in carbon (C) sequestration to help mitigate rising CO2 levels and climate change. The Southern High Plains (SHP) region contains >900.000 ha enrolled in CRP, but a regionally specific C sequestration rate has not been studied, and identification of the C pools and processes important in controlling C sequestration rates remain unresolved. We aimed to address these gaps by utilizing a CRP chronosequence with historical rangeland as a reference ecosystem. Soil samples (0-10 and 10-30 cm) were collected in 2012 and 2014 from a total of 26 fields across seven counties within the SHP and included seven croplands (0 y in CRP), 16 CRP fields that ranged from 6 to 26 y (as of 2012), plus three rangelands. Multiple regression analysis was conducted to gauge the rate of C sequestration under CRP within C pools: soil organic C (SOC), particulate organic matter C (POM-C), and microbial biomass C (MBC), with two additional predictors (soil clay + silt content and precipitation). Despite attempts to control for soil texture by targeting a dominant soil series (Amarillo fine sandy loam), the percent of clay + silt (15.2-48.7%) significantly influenced C accrual. The C sources (C-3 from previous cropping systems or C-4 from CRP grasses) in SOC and POMC were assessed using stable C isotope signatures. Additionally, the role of soil microbes in C sequestration was evaluated by investigating the relationship between MBC and CO2 flux and C sequestration. SOC increased at a rate of 69.82 and 132.87 kg C ha(-1) y(-1) and would take approximately 74 and 77 y to reach the rangeland C stocks at 0-10 and 0-30 cm, respectively. The C-4-C primarily from the introduced grasses was the main source of C sequestration. SOC gains were essentially due to increases in POM-C and MBC, accounting for 50.04 and 15.64% of SOC sequestration at 0-30 cm, respectively. The highest semi-partial correlation coefficients between the increasing years under CRP restoration and MBC indicated CRP had the strongest effect on MBC compared to other C pools. In addition, increasing soil CO2 flux and MBC:SOC ratio with years of CRP restoration indicated MBC played a critical role in the C sequestration process. Conservation of CRP lands and efforts to sustain perennial systems in this highly erodible landscape should be a high priority of conservation programs. In doing so, significant offsets to increasing atmospheric CO2 levels may be achieved in addition to erosion control and improved wildlife habitat. (C) 2017 Elsevier B.V. All rights reserved.