| Summary: | Adaptations in crop rotation with the inclusion of temporary grass-clover leys and organic amendments, have been promoted as effective ways to improve soil carbon (C) sequestration and mitigate climate change in agricultural systems. However, there are still a lot of uncertainties related to i) the combined effects of different crop rotations and different fertilisation sources, e.g., organic amendments, on soil C stocks; and ii) their potential effect on C stabilisation. The objective of this study was to evaluate the effect of different arable crop rotations with varying degrees of diversity in crop type and lengths of grass-clover ley periods and fertilisation sources on soil C stocks and C stabilisation down to 0.60 m soil depth. This was investigated in a long-term factorial field experiment-combining different crop rotation (cereal-intensive conventional vs. diversified legume-intensive organic) with different lengths of grass-clover ley periods (2 vs. 3 years), fertilisation sources (mineral vs. compost), and years (samples taken at the beginning and at the last year of one complete cycle of rotation; 8 years apart)-to explore their individual and interactive effect on soil C stock and C stabilisation at two soil depths (0–0.30 and 0.30–0.60 m). Soil C stabilisation was assessed using a unique combination of three different techniques: physical fractionation for separation of C associated to organic and mineral fractions, thermal analysis combined with differential scanning calorimetry and a quadrupole mass spectrometry (TG-DSC-QMS) for physical-chemical aspects, and pyrolysis coupled with gas chromatography-mass spectrometry (Py-GC/MS) for molecular structural information. The findings showed higher soil C stocks under the diversified organic rotation with 3 years of grass-clover ley period at both soil depths, regardless of the fertilisation source or sampling year. However, the organic rotation seemed to deliver stable soil C stocks only in the subsoil layer. Compost fertilisation, in turn, increased topsoil C stocks between the two sample dates under both rotations, and it appears to be stable. These results suggested that combining a diversified organic rotation with 3 years grass-clover ley with compost fertilisation could be one way for agricultural systems to deliver stable soil C sequestration.
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