BackgroundTo counteract soil degradation, it is important to convert conventional agricultural practices to environmentally sustainable management practices. To this end, the application of biostimulants could be considered a good strategy. Compost, produced by the composting of biodegradable organic compounds, is a source of natural biostimulants, such as humic acids, which are naturally occurring organic compounds that arise from the decomposition and transformation of organic residues, and compost tea, a compost-derived liquid formulated produced by compost water-phase extraction. This study aimed to determine the molecular responses of the roots of tomato plants (cv. Crovarese) grown under hydroponic conditions and subjected to biostimulation with humic substances (HSs) and filtered sterile compost tea (SCT).ResultsThe 13C CPMAS NMR of humic acids (HA) and SCT revealed strong O-alkyl-C signals, indicating a high content of polysaccharides.Thermochemolysis identified over 100 molecules, predominantly from lignin, fatty acids, and biopolymers. RNA-Seq analysis of tomato roots treated with HA or SCT revealed differentially expressed genes (DEGs) with distinct patterns of transcriptional reprogramming.Notably, HA treatment affected carbohydrate metabolism and secondary metabolism, particularly phenylpropanoids and flavonoids, while SCT had a broader impact on hormone and redox metabolism. Both biostimulants induced significant gene expression changes within 24 h, including a reduction in cell wall degradation activity and an increase in the expression of hemicellulose synthesis genes, suggesting that the treatments prompted proactive cell wall development.ConclusionsThe results demonstrate that HS and SCT can mitigate stress by activating specific molecular mechanisms and modifying root metabolic pathways, particularly those involved in cell wall synthesis. However, gene regulation in response to these treatments is complex and influenced by various factors. These findings highlight the biostimulatory effects of HS and SCT, suggesting their potential application in crop biofertilization and the development of innovative breeding strategies to maximize the benefits of humic substances for crops. Further research is needed to fully elucidate these mechanisms across various contexts and plant species.