Nitrogen (N) fertilization is a crucial agricultural practice for boosting production traits in vegetables. However, N synthetic fertilizers—commonly adopted by farmers—have several counterproductive effects on the environment and on humans. The research was performed to assess the combined influence of plant growth promoting bacteria (PGPB) (Azospirillum brasilense DSM 1690, A. brasilense DSM 2298 and Pseudomonas sp. DSM 25356) and various N fertilization doses (0, 30, 60 or 120 kg ha⁻¹) on growth, yield, quality and nitrogen indices of lettuce in protected cultivation. Plant height, root collar diameter, number of leaves and fresh weight were enhanced by A. brasilense DSM 2298 inoculation and N at 30 or 60 kg ha⁻¹. Overall, soluble solids content (SSC), ascorbic acid, total phenolics, carotenoids, total chlorophyll and total sugars were augmented by the combined effect of A. brasilense strains and 30, 60 or 120 kg N ha⁻¹. Furthermore, PGBP inoculation improved potassium (K) and magnesium (Mg) concentrations in leaf tissues. PGPB inoculation increased N leaf concentration; however, it hastened N indices. These results suggest that the PGPB tested can be considered an eco-friendly tool to improve lettuce yield, particularly when combined with N at 30 or 60 kg ha⁻¹.

The use of chemical fertilizers and pesticides has caused harmful impacts on the environment with the increase in economic burden. Biofertilizers are biological products containing living microorganisms capable of improving plant growth through eco-friendly mechanisms. In this work, three actinobacterial strains Streptomyces violaceoruber, Streptomyces coelicolor, and Kocuria rhizophila were characterized for multiple plant growth promoting (PGP) traits such as indole acetic acid production, phosphate solubilization, N₂-fixation, and drought and salt tolerance. Then, these strains were investigated for their secreted and cellular metabolome, revealing a rich arsenal of bioactive molecules, including antibiotics and siderophores, with S. violaceoruber being the most prolific strain. Furthermore, the in vivo assays, performed on tomato (Solanum lycopersicum L.), resulted in an improved germination index and the growth of seedlings from seeds treated with PGP actinobacteria, with a particular focus on S. violaceoruber cultures. In particular, this last strain, producing volatile organic compounds having antimicrobial activity, was able to modulate volatilome and exert control on the global DNA methylation of tomato seedlings. Thus, these results, confirming the efficacy of the selected actinobacteria strains in promoting plant growth and development by producing volatile and non-volatile bioactive molecules, can promote eco-friendly alternatives in sustainable agriculture.