Biochar aging, soil microbiota and chemistry of charcoal kilns in Mediterranean forests

Abstract Charcoal kilns, old structures used for charcoal production in the forest, preserve a charcoal-enriched topsoil representing a suitable proxy for studying the long-term effect of biochar addition to soil. Two kiln platforms located at Gelbison and Vesole mountain sites in Southern Italy were selected due to their comparable climates but distinct parent rocks. We conducted standard soil chemical analyses and used next-generation sequencing to explore bacterial and fungal microbiome. Anthracology identified charcoal species, while scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDS) characterized charcoal particles. Reflectance Fourier transform infrared spectroscopy (DRIFTS) assessed biochar surface oxidation. Additionally, a bioassay with soybean, maize, and Tomato investigated the impact of kiln soil on plant performance. Our results showed that kiln soils did not exhibit higher pH, cation exchange capacity, or greater richness in cations. EDS and FTIR analyses showed that charcoal buried in forest soil for decades undergoes significant oxidation, with increased O/C ratio and the presence of oxygenated functional groups. Charcoal surfaces were selectively enriched with Ca2+ on limestone substrate sites but with Al and Si over sedimentary (flysch) substrate. While differences in the kiln soil and its surroundings were noticeable, they were not drastic in terms of microbial diversity and composition. Surprisingly, the bioassay indicated that the kiln microbiota had a more positive impact on plant growth compared to external forest soil. In conclusion, this study highlights the unique nature of kiln microsites and begins to unveil the enduring effects of charcoal accumulation on soil chemistry and microbiota in forest soil. Graphical Abstract