Substrate pH mediates growth promotion and resilience to water stress of Tilia tomentosa seedlings after Ectomycorrhizal inoculation

Colonization by Ectomycorrhizal (EcM) fungi is key for the health and performance of plants under different stress scenarios, such as those faced by trees in urban environments. Because urban environments can be lacking EcM fungi, we here assessed the benefits of inoculating Tilia tomentosa seedlings in a pre-transplantation nursery context with the EcM fungi Lactarius deliciosus and Paxillus involutus, using substrates of different pH and facing water-stress. P. involutus had a more evident positive effect in T. tomentosa seedlings and had a good performance in both acidic and alkaline substrate. In acidic substrate the fungus increased the plant height by 0.91-fold, increased the mycorrhization rate by 3.23-fold, expansion rate by 5.03-fold and formation of secondary roots by 0.46-fold, compared to the non-inoculated control. This species also improved the phosphorus content of leaves, which revealed a promotion of nutrient uptake. In alkaline substrate P. involutus increased root dry weight by 3.92-fold and the mycorrhization parameters. In contrast, L. deliciosus only had a positive effect in the improvement of mycorrhization and expansion rates and phosphorus content in the root, effects visible only in alkaline substrate. When exposed to water-stress the increase of proline content was visible in acidic substrate for both fungi, L. deliciosus and P. involutus, and in alkaline substrate for the fungus P. involutus, a response indicative of the enhancement of defenses in stressing scenarios such as water scarcity. We conclude that fungal inoculation improves the vigour and resilience of Tilia seedlings and that it is of utmost importance to select a suitable EcM fungus and to consider the soil pH of the transplanting site. The inoculation approach can be a valuable tool to produce robust seedlings which may have a better performance when transplanted to the challenging urban environment.