Salinity is a major abiotic stress factor that affects crops and has an adverse effect on plant growth. In recent years, there has been increasing evidence that microbial volatile organic compounds (mVOC) play a significant role in microorganism–plant interactions. In the present study, we evaluated the impact of microbial volatile organic compounds (mVOC) emitted by Bacillus amyloliquefaciens GB03 on the biosynthesis of secondary metabolites and the antioxidant status in Mentha piperita L. grown under 0, 75 and 100 mM NaCl. Seedlings were exposed to mVOCs, avoiding physical contact with the bacteria, and an increase in NaCl levels produced a reduction in essential oil (EO) yield. Nevertheless, these undesirable effects were mitigated in seedlings treated with mVOCs, resulting in an approximately a six-fold increase with respect to plants not exposed to mVOCs, regardless of the severity of the salt stress. The main components of the EOs, menthone, menthol, and pulegone, showed the same tendency. Total phenolic compound (TPC) levels increased in salt-stressed plants but were higher in those exposed to mVOCs than in stressed plants without mVOC exposure. To evaluate the effect of mVOCs on the antioxidant status from salt-stressed plants, the membrane lipid peroxidation was analyzed. Peppermint seedlings cultivated under salt stress and treated with mVOC showed a reduction in malondialdehyde (MDA) levels, which is considered to be an indicator of lipid peroxidation and membrane damage, and had an increased antioxidant capacity in terms of DPPH (2,2-diphenyl−1-picrylhydrazyl) radical scavenging activity in relation to plants cultivated under salt stress but not treated with mVOCs. These results are important as they demonstrate the potential of mVOCs to diminish the adverse effects of salt stress.

Plants are naturally subjected to various types of biotic stresses, including pathogenic microorganisms and herbivory by insects, which trigger different signaling pathways and related defense mechanisms. Inoculation with microorganisms, such as plant growth-promoting rhizobacteria (PGPR), can be seen as a form of stress because it triggers a systemic resistance response in plants similar to that caused by insect herbivory. However, these interactions have typically been studied independently, which has limited the understanding of their combined effects. This study examines the effects of Bacillus amyloliquefaciens GB03 inoculation and Spodoptera frugiperda herbivory on the total phenolic contents of Ocimum basilicum. We also analyze the levels of endogenous phytohormones and the activity of phenylalanine ammonia-lyase (PAL), a crucial enzyme involved in the biosynthesis of phenolic defense-related metabolites. The results indicate that the total phenolic content significantly increased only in plants that were both inoculated by GB03 and damaged by larvae. Additionally, PAL activity showed an increase in plants that were damaged by larvae and in those subjected to the combined treatment of larval damage and inoculation with GB03. Regarding phytohormones, in plants damaged by insects, the levels of salicylic acid (SA) increased, regardless of whether they were inoculated or not, while the levels of jasmonic acid–isoleucine (JA-ile) rose in all treatments compared to the control. This study highlights the intricate relationships among beneficial microbes, herbivores, and plant defense mechanisms, emphasizing their potential impact on improving plant resilience and the production of secondary metabolites. Furthermore, understanding the independent effects of PGPR inoculation, beyond its interaction with herbivory, could provide valuable insights into its role as a sustainable alternative for enhancing plant defense responses and promoting crop productivity.

Considering that inoculation with beneficial rhizobacteria promotes biomass and secondary metabolite biosynthesis and also the fact that drought stress can increase their production, we determined the effects when both of these situations were combined in EO yield. In addition, the levels of endogenous phytohormones and the gene expression of enzymes related to monoterpenes biosynthesis were measured. The experimental results showed that inoculation with PGPR in stressed plants produced the same effects on trichome density, the EO main components and total yield as in plants only inoculated or with moderate stress alone. In addition, the VOC emissions of inoculated stressed plants did not vary the release amount in relation to uninoculated stressed ones. The results observed have suggested a way of improving aromatic plant productivity, particularly that of peppermint, by managing drought stress through the inoculation of plants with PGPR. This inoculation under moderate drought stress is appropriate when the purpose of the crops is to obtain plants with a high secondary metabolites yield.

Inoculation with rhizobacteria and feeding by herbivores, two types of abiotic stress, have been shown to increase the production of secondary metabolites in plants as part of the defense response. This study explored the simultaneous effects of inoculation with Bacillus amyloliquefaciens GB03 (a PGPR species) and herbivory by third-instar Spodoptera frugiperda larvae on essential oil (EO) yield and volatile organic compound (VOC) emissions in Ocimum basilicum plants. The density of glandular trichomes was also examined, given that they are linked to EO production and VOC emission. Herbivory increased EO content, but inoculation on its own did not. When combined, however, the two treatments led to a 10-fold rise in EO content with respect to non-inoculated plants. VOC emissions did not significantly differ between inoculated and non-inoculated plants, but they doubled in plants chewed by the larvae with respect to their undamaged counterparts. Interestingly, no changes were observed in VOC emissions when the treatments were tested together. In short, the two biotic stressors elicited differing plant defense responses, mainly when EO was concerned. PGPR did not stimulate EO production, while herbivory significantly enhanced it and increased VOC emissions. The combined treatment acted synergistically, and in this case, PGPR inoculation may have had a priming effect that amplified plant response to herbivory. Peltate trichome density was higher in inoculated plants, those damaged by larvae, and those subjected to the combination of both treatments. The findings highlight the intricate nature of plant defense mechanisms against various stressors and hint at a potential strategy to produce essential oil through the combined application of the two stressors tested here.