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Abstract

Aggregate formation is a key process of soil development, which promotes carbon (C) stabilization by hindering decomposition of particulate organic matter (POM) and its interactions with mineral particles.
C stabilization processes lead to 13C fractionation and consequently to various d13C values of soil organic
matter (SOM) fractions. Differences in d13C within the aggregates and fractions may have two reasons: 1)
preferential stabilization of organic compounds with light or heavy d13C and/or 2) stabilization of organic
materials after passing one or more microbial utilization cycles, leading to heavier d13C in remaining C.
We hypothesized that: 1) 13C enrichment between the SOM fractions corresponds to successive steps of
SOM formation; 2) 13C fractionation (but not the d13C signature) depends mainly on the transformation
steps and not on the C precursors. Consequently, minimal differences between D13C of SOM fractions
between various ecosystems correspond to maximal probability of the SOM formation pathways.
We tested these hypotheses on three soils formed from cover loam during 45 years of growth of
coniferous or deciduous forests or arable crops. Organic C pools in large macroaggregates, small macroaggregates,
and microaggregates were fractionated sequentially for four density fractions to obtain free
POM with r < 1.6 g cm3, occluded POM with two densities (r < 1.6 and 1.6e2.0 g cm3), and mineral
fraction (r > 2.0 g cm3).
The density fractions were 13C enriched in the order: free POM < light occluded POM < heavy
occluded POM < mineral fraction. This, as well as their C/N ratios confirmed that free POM was close to
initial plant material, whereas the mineral fraction was the most microbially processed. To evaluate the
successive steps of SOM formation, the D13C values between d13C of SOM fractions and d13C of bulk SOM
were calculated. The D13C indicated that, parallel with biochemical transformations, the physical disintegration
strongly contributed to the formation of free and occluded light POM. In contrast, biochemical
transformations were more important than physical disintegration for formation of heavy occluded POM
from light occluded POM. This was confirmed by review of 70 D13C values from other studies analyzed
D13C depending on the density of SOM fractions. Accordingly, the successive steps of SOM formation
were: fLF<1.6 ¼ oLF<1.6/oDF1.6e2.0 ¼ MF>2.0. The obtained steps of C stabilization were independent on
the initial precursors (litter of coniferous forest, deciduous forest or grasses).
To test the second hypothesis, we proposed an extended scheme of C flows between the 3 aggregate
size classes and 4 SOM fractions. D13C enrichment of the SOM fractions showed that the main direction of
C flows within the aggregates and SOM fractions was from the macroaggregate-free POM to the mineral
microaggregate fraction. This confirmed the earlier concept of SOM turnover in aggregates, but for the
first time quantified the C flows within the aggregates and SOM density fractions based on d13C values.
So, the new 13C natural abundance approach is suitable for analysis of C pathways by SOM formation
under steady state without 13C or 14C labeling.