Research highlights: Interaction effects of coniferous on deciduous species have been investigated before the background of climate change. Background and objectives: The cultivation of European beech (Fagus sylvatica L.) in mixed stands has currently received attention, since the future performance of beech in mid-European forest monocultures in a changing climate is under debate. We investigated water relations and nitrogen (N) nutrition of beech in monocultures and mixed with silver-fir (Abies alba Mill.) in the Black Forest at different environmental conditions, and in the Croatian Velebit at the southern distribution limit of beech, over a seasonal course at sufficient water availability. Material and methods: Water relations were analyzed via δ13C signatures, as integrative measures of water supply assuming that photosynthesis processes were not impaired. N nutrition was characterized by N partitioning between soluble N fractions and structural N. Results: In the relatively wet year 2016, water relations of beech leaves, fir needles and roots differed by season, but generally not between beech monocultures and mixed cultivation. At all sites, previous and current year fir needles revealed significantly lower total N contents over the entire season than beech leaves. Fir fine roots exhibited higher or similar amounts of total N compared to needles. Correlation analysis revealed a strong relationship of leaf and root δ13C signatures with soil parameters at the mixed beech stands, but not at pure beech stands. While glutamine (Gln) uptake capacity of beech roots was strongly related to soil N in the monoculture beech stands, arginine (Arg) uptake capacities of beech roots were strongly related to soil N in mixed stands. Conclusions: Leaf N contents indicated a facilitative effect of silver-fir on beech on sites where soil total N concentrations where low, but an indication of competition effect where it was high. This improvement could be partially attributed to protein contents, but not to differences in uptake capacity of an individual N source. From these results it is concluded that despite similar performance of beech trees at the three field sites investigated, the association with silver-fir mediated interactive effects between species association, climate and soil parameters even at sufficient water supply.

Hydraulic redistribution (HR) of water from wet- to dry-soil zones is suggested as an important process in the resilience of forest ecosystems to drought stress in semiarid and tropical climates. Scenarios of future climate change predict an increase of severe drought conditions in temperate climate regions. This implies the need for adaptations of locally managed forest systems, such as European beech (Fagus sylvatica L.) monocultures, for instance, through the admixing of deep-rooting silver fir (Abies alba Mill.). We designed a stable-isotope-based split-root experiment under controlled conditions to test whether silver fir seedlings could perform HR and therefore reduce drought stress in neighboring beech seedlings. Our results showed that HR by silver fir does occur, but with a delayed onset of three weeks after isotopic labelling with 2H2O (δ2H ≈ +6000‰), and at low rates. On average, 0.2% of added ²H excess could be recovered via HR. Fir roots released water under dry-soil conditions that caused some European beech seedlings to permanently wilt. On the basis of these results, we concluded that HR by silver fir does occur, but the potential for mitigating drought stress in beech is limited. Admixing silver fir into beech stands as a climate change adaptation strategy needs to be assessed in field studies with sufficient monitoring time.

Beech (Fagus sylvatica) and silver fir (Abies alba) are often cultivated in mixed stands and, hence, compete for water and nutrients. Besides nitrogen (N), also phosphorus (P) is an important nutrient for growth and development. Beech trees in Central Europe grow on both P-poor and P-rich soils, thereby showing similar growth and low variation in foliar P. The central aim of the present study was to test the hypothesis that variations in foliar P contents of beech are driven by seasonal changes rather than by the competition with silver fir. It was further hypothesized that P contents in silver fir needles depend on needle age and forest site. To test these hypotheses, P contents and P fractions, i.e. organic-bound P (Porg) and inorganic phosphate P (Pi), were measured in the foliage of beech trees from pure beech and mixed beech/silver fir plots as well as in needles of silver fir of the mixed plots. The forest sites investigated are located in Central Europe in the Black Forest, Germany, and in Croatia near the south-eastern distribution limit of beech and are all poor in plant-available soil P. The analyses showed that the main driver of P contents and P fractions in beech leaves at all forest sites is the season and that competition with silver fir had no effect. Hence, the present results demonstrate the high plasticity of beech trees to adapt to both poor plant-available soil P and competition with silver fir. Total P contents of silver fir needles were higher at the Croatian site compared to the Black Forest sites and originated from higher foliar Pi contents. One third of the P present in current-year needles in late summer was remobilized and exported until the needles reached the age of 1 year. The difference in P contents between current-year and 1-year-old needles can be seen as the amount of P resorbed from 1-year-old needles in summer during the generation of new needles to support the P demand of current-year needles for growth and development.

Very limited information is available on the drought tolerance of European beech and silver fir in mixed cultivation, both for mature forests and natural regeneration. Particularly, little information is available regarding the significance on memory effects of drought exposure. Therefore, drought memory was analyzed in seedlings of these species grown in mixed cultivation in the present study. The results showed that previous-year drought hardening mediated enhanced biomass accumulation of beech leaves and root in the subsequent year, but did not impact fir growth. Total carbon (C) content was decreased by drought hardening in both the leaves and roots of beech and previous-year needles and roots of fir, in beech probably as a consequence of increased growth. Previous-year drought hardening had no significant effect on relative water contents, total nitrogen (N), or soluble protein contents in leaves and roots of beech and fir, but resulted in decreased total amino acid contents of beech leaves and fir needles. It further reduced structural N in current-year fir needles and decreased C/N ratios in roots of both beech and silver fir seedlings. Generally, the number of interspecific neighbors had no considerable effect on biomass or total C or N contents, as well as N partitioning in leaves and roots of beech and fir seedlings. The present study highlights that drought hardening induces memory effects in European beech and silver fir seedlings in their mixture in the subsequent year of growth, but these memory effects are stronger in beech than in fir.

To alleviate the enhanced frequency, duration, and intensity of drought as a consequence of global warming, admixing drought-sensitive European beech (Fagus sylvatica L.) with deep rooting silver fir (Abies alba Mill.) has been proposed. However, information on the performance of the admixtures of seedlings of these tree species at limited water availability has so far not been reported. In the present study, we investigated the significance of water deprivation in mixtures of beech and fir seedlings on the foliar relative water content (RWC), δ13C signature, total C and N contents, and C:N ratios of both species in a drought-rewetting cycle. Surprisingly, moderate drought triggered increased RWC in beech leaves and current year fir needles indicating drought hardening. The enhanced foliar RWC was preserved after rewatering in beech leaves, but not in current year fir needles. Drought did not significantly affect δ13C abundance in beech leaves, but enhanced the δ13C abundance (less negative values) in current and one-year old fir needles, indicating stomatal control in fir needles but not in beech leaves upon moderate drought. Total C contents of beech leaves were significantly increased upon drought and rewatering, but remained constant in fir needles. Foliar total N increased in both species upon drought and decreased upon rewatering. Accordingly, C:N ratios decreased in response to drought and recovered after rewatering. These results suggest that drought hardening may be achieved at least partially via osmotic adjustment by different compatible solutes in beech leaves and fir needles. No apparent effects of the number of neighbours were observed, although more fir neighbours tended to increase the RWC and total C contents of beech leaves. These results indicate that drought hardening in mixtures of beech and fir seedlings is largely independent of the number of interspecies neighbours.