Festuca arundinacea is a model to work on the mechanisms of drought resistance in grasses. The crucial components of that resistance still remain not fully recognized. It was suggested that deep root system could be a crucial trait for drought avoidance strategy but the other components of root performance under water deficit have not paid much attention of scientists. In this study, two genotypes of F. arundinacea with a different ability to withstand soil water deficit were selected to perform comprehensive research, including analysis of root architecture, phytohormones, proteome, primary metabolome and lipidome under progressive stress conditions, followed by a rewatering period. The experiments were performed in tubes, thus enabling undisturbed development of root systems. We demonstrated that long roots are not sufficient to perfectly avoid drought damage in F. arundinacea and to withstand adverse environmental conditions without a disturbed cellular metabolism (with respect to leaf relative water potential and cellular membrane integrity). Furthermore, we proved that metabolic performance of roots is as crucial as its architecture under water deficit, to cope with drought stress via avoidance, tolerance and regeneration strategies. We believe that the presented studies could be a good reference for the other, more applied experiments, in closely related species.

Nitric oxide (NO) has been proven to be involved in the regulation of many physiological processes in plants. Though the contribution of NO in plant response to drought has been demonstrated in numerous studies, this phenomenon remains still not fully recognized. The research presented here was performed to decipher the role of NO metabolism in drought tolerance and the ability to recover after stress cessation in two closely related species of forage grasses, important for agriculture in European temperate regions: Festuca arundinacea and F. glaucescens. In both species, two genotypes with distinct levels of drought tolerance were selected to compare their physiological reactions to simulated water deficit and further re-watering, combined with a simultaneous application of NO scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO). The results clearly indicated a strong relationship between scavenging of NO in leaves and physiological response of both analyzed grass species to water deficit and re-watering. It was revealed that NO generated under drought was mainly located in mesophyll cells. In plants with reduced NO level a higher photosynthetic capacity and delay in stomatal closure under drought, were observed. Moreover, NO scavenging resulted also in the increased membrane permeability and higher accumulation of ROS in cells of analyzed plants both under drought and re-watering. This phenomena indicate that lower NO level might reduce drought tolerance and the ability of F. arundinacea and F. glaucescens to recover after stress cessation.

Impact of photosynthetic and antioxidant capacities on drought tolerance of two closely related forage grasses, Festuca arundinacea and Festuca glaucescens, was deciphered. Within each species, two genotypes distinct in drought tolerance were subjected to a short-term drought, followed by a subsequent re-watering. The studies were focused on: (i) analysis of plant physiological performance, including: water uptake, abscisic acid (ABA) content, membrane integrity, gas exchange, and relative water content in leaf tissue; (ii) analysis of plant photosynthetic capacity (chlorophyll fluorescence; gene expression, protein accumulation, and activity of selected enzymes of the Calvin cycle); and (iii) analysis of plant antioxidant capacity (reactive oxygen species (ROS) generation; gene expression, protein accumulation and activity of selected enzymes). Though, F. arundinacea and F. glaucescens revealed different strategies in water uptake, and partially also in ABA signaling, their physiological reactions to drought and further re-watering, were similar. On the other hand, performance of the Calvin cycle and antioxidant system differed between the analyzed species under drought and re-watering periods. A stable efficiency of the Calvin cycle in F. arundinacea was crucial to maintain a balanced network of ROS/redox signaling, and consequently drought tolerance. The antioxidant capacity influenced mostly tolerance to stress in F. glaucescens.

Triacylglycerol is a key lipid compound involved in the maintaining homeostasis of both membrane lipids and free fatty acids in plant cells under adverse environmental conditions. However, its role in the process of lipid remodeling has not been fully recognized, especially in monocots, including grass species. For our study, two closely related introgression forms of Lolium multiflorum (Italian ryegrass) and Festuca arundinacea (tall fescue), distinct in their level of drought tolerance, were selected as plant models to study rearrangements in plant lipidome under water deficit and further re-watering. The low drought tolerant (LDT) form revealed an elevated level of cellular membrane damage accompanied by an increased content of polyunsaturated free fatty acids and triacylglycerol under water deficit, compared with the high drought tolerant (HDT) form. However, the LDT introgression form demonstrated also the ability to regenerate its membranes after stress cessation. The obtained results clearly indicated that accumulation of triacylglycerol under advanced drought in the LDT form could serve as a cellular protective mechanism against over-accumulation of toxic polyunsaturated free fatty acids and other lipid intermediates. Furthermore, accumulation of triacylglycerol under drought conditions could serve also as a storage of substrates required for further regeneration of membranes after stress cessation. The rearrangements in triacylglycerol metabolism were supported by the up-regulation of several genes, involved in a biosynthesis of triacylglycerol. With respect to this process, diacylglicerol O-acyltransferase DGAT2 seems to play the most important role in the analyzed grasses.