The drought sensitivity of the pedunculate oak (Quercus robur L.) poses a threat to its survival in light of climate change. Mycorrhizal fungi, which orchestrate biogeochemical cycles and particularly have an impact on the plant’s defense mechanisms and metabolism of carbon, nitrogen, and phosphorus, are among the microbes that play a significant role in the mitigation of the effects of climate change on trees. The study’s main objectives were to determine whether ectomycorrhizal (ECM) fungi alleviate the effects of drought stress in pedunculate oak and to investigate their priming properties. The effects of two levels of drought (mild and severe, corresponding to 60% and 30% of field capacity, respectively) on the biochemical response of pedunculate oak were examined in the presence and absence of ectomycorrhizal fungi. To examine whether the ectomycorrhizal fungi modulate the drought tolerance of pedunculate oak, levels of plant hormones and polyamines were quantified using UPLC-TQS and HPLC-FD techniques in addition to gas exchange measurements and the main osmolyte amounts (glycine betaine-GB and proline-PRO) which were determined spectrophotometrically. Droughts increased the accumulation of osmolytes, such as proline and glycine betaine, as well as higher polyamines (spermidine and spermine) levels and decreased putrescine levels in both, mycorrhized and non-mycorrhized oak seedlings. In addition to amplifying the response of oak to severe drought in terms of inducible proline and abscisic acid (ABA) levels, inoculation with ECM fungi significantly increased the constitutive levels of glycine betaine, spermine, and spermidine regardless of drought stress. This study found that compared to non-mycorrhized oak seedlings, unstressed ECM-inoculated oak seedlings had higher levels of salicylic (SA) and abscisic acid (ABA) but not jasmonic acid (JA), indicating a priming mechanism of ECM is conveyed via these plant hormones. According to a PCA analysis, the effect of drought was linked to the variability of parameters along the PC1 axe, such as osmolytes PRO, GB, polyamines, and plant hormones such as JA, JA-Ile, SAG, and SGE, whereas mycorrhization was more closely associated with the parameters gathered around the PC2 axe (SA, ODPA, ABA, and E). These findings highlight the beneficial function of the ectomycorrhizal fungi, in particular Scleroderma citrinum, in reducing the effects of drought stress in pedunculate oak.

Understanding intraspecific genetic variation is one of the principal requirements for the evaluation of tree species capacity to cope with intensive climatic changes, as well as designing long-term conservation programs. Herein, we evaluated the genetic diversity and genetic structure of seven pedunculate oak (Quercus robur L.) populations, located at the southern margin of its distribution range on the Balkan Peninsula (Serbia). The objective of the study was to propose future in situ conservation measures aimed at protection of pedunculate oak adaptive and neutral genetic diversity at the species rear-edge. Genetic diversity and structure were estimated using twelve highly polymorphic simple sequence repeat (SSR) markers. The mean expected heterozygosity (H_e) was 0.769, allelic richness (A_R) 9.63, and private allelic richness (p_AR) 0.79, indicating high genetic diversity in the studied populations. Genetic differentiation among the populations was low (F_st = 0.032). Structure analysis, the unweighted pair group method with arithmetic mean (UPGMA) showed the existence of two gene pools unrelated to the populations’ area of occurrence. Taking into consideration the results of the current study and previous conservation activities on the pedunculate oak in Serbia, as well as the importance of rear-edge populations in the long-term conservation of the species genetic diversity, we suggested establishing three additional gene conservation units for securing long-term sustainability of the species.

Quercus robur L. is one of the most valued tree species of deciduous temperate forests. However, in the last decade, serious oak declines and loss of adaptation plasticity have been reported throughout Europe as a consequence of drought. Therefore, the aim of the present study was to define the adaptation potential of five Q. robur half-sib lines from the UNESCO Biosphere Reserve Mura-Drava-Danube to drought, using physiological, anatomical and biometrical traits. Half-sib lines that exhibited drought tolerance had particular suites of trait expression regarding biometrical traits (seedling height, root length, root to shoot ratio of dry mass and specific leaf area), leaf stomatal traits (stomatal density per mm², stomata guard cell length and width, stomatal aperture length and width) and leaf structural traits (adaxial epidermis thickness, palisade parenchyma thickness, spongy parenchyma thickness, lamina thickness). All of the observed parameters of chlorophyll a fluorescence were shown to be good indicators of short-term and severe drought. For the selection of drought-tolerant half-sib lines, all studied chlorophyll a fluorescence parameters associated with the heat dissipation of light energy (coefficient of non-photochemical quenching, quantum yield of regulated energy dissipation, Stern-Volmer type non-photochemical fluorescence quenching) and one parameter related to photochemical dissipation of light energy (effective quantum yield (efficiency) of PS II photochemistry) were proven to be suitable. On the other hand, the coefficient of photochemical quenching, coefficient of photochemical fluorescence quenching assuming interconnected photosystem II antennae and electron transport rate were not suitable for distinguishing the different responses of the studied half-sib lines under drought. The importance of results of the present study is in the selection of drought-tolerant Q. robur half-sib lines for future reforestation programs, particularly in protected areas with sensitive forest management and restricted activities for mitigation of the adverse effects of climate changes.