During soil formation, the interaction of different biota (plants, soil fauna, microbes) with weathered mineral material shapes unique structures depending on the parental material and the site specific climatic conditions. While many of these interactions are known, the relative importance of the different biota is difficult to unravel and therefore difficult to quantify. Biological soil structure formation is often superimposed by soil management and swell-shrink dynamics, making it even more difficult to derive mechanistic understanding.

We here explore soil structure formation within a “space-for-time” chronosequence in the Rhenish lignite mining area. Loess material from a depth of 4–10 m has been used for reclamation in a standardized procedure for 24 years.

Changes in soil pore system are characterized by properties such as connectivity (Euler number) and pore size distribution using undisturbed soil columns with a diameter of 10 cm. They were taken from two different depths (0–20 cm and 40–60 cm) at different sites ranging in age from 0 to 24 years. X-ray CT is used for scanning the original columns as well as undisturbed subsamples of 3 and 0.7 cm diameter. This hierarchical sampling scheme was developed to overcome the trade-off between sample size and resolution.

For the first time also information on the development of biopores could be measured by separating them from other structural pores based on their unique shape. The data were complemented by destructive sampling and determination of root length with WinRHIZO to give an estimate of how many biopores are filled with roots. Furthermore HYPROP measurements of water retention curves were conducted and showed a general agreement with the image-derived pore size distribution merged across three scales.

An increase in biopore density throughout year zero to year 12, in particular in 40–60 cm soil depth, was observed. The biopore length densities of approximately 17 cm/cm³ obtained in year 12 was similar to the one measured in year 24, suggesting that equilibrium was reached. Only about 10% of these biopores were filled with roots. In the topsoil (0–20 cm) the equilibrium value in biopore density is already reached after six years due to a higher root length density. Ploughing lead to higher mean pore size and to lower connectivity compared to the well-connected, very stable pore network in 40–60 cm depth.

This study shows how fast plant roots create a stable and connected biopore system and how this is disrupted by soil tillage, which produces completely contrasting pore characteristics.