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Background Soil carbon-rich organic amendments (biochar, humic substances) may improve the quality and fertility of arable soil. Their co-application can additively enhance the beneficial effect on soil. Hypothetically, the pre-treatment of biochar, by aging via soaking in a solution of commercially available humic substances, could result in synergism, which may exceed the benefit from simple co-application of both amendments to the soil. Therefore, the aim of this study was to investigate the impact of biochar, humic substances, the combination of both, and the impact of biochar aged by humic substances solution on soil microbial activities and plant growth in a short-term pot experiment with lettuce. Results The aging of biochar decreased the C:N ratio as compared to non-activated biochar. The co-application of biochar and humic substances into the soil resulted in the highest microbial biomass carbon and respiration activity. The majority of enzyme activities (β-glucosidase, arylsulfatase,  N -acetyl-β- d -glucosaminidase, phosphatase) were the highest in humic substances-amended soil. The application of humic substances and biochar with humic substances seemed to stimulate microbial growth and activity followed by the competition of microflora for nutrients with plants, whereas the aged biochar behaved differently. The plants treated by aged biochar achieved the highest values of dry aboveground and root biomass of all variants. However, the assumed rapid uptake of nutrients by plants resulted in lower nutrient availability for microflora, and a decline in microbial viability. Conclusions Based on this study, the positive effect of co-applied humic substances and biochar on soil fertility, quality, and health can be concluded. The usability of biochar aging by humic solution requires further study. Graphic abstract

Assessing the potential of biochar aged by humic substances to enhance plant growth and soil biological activity