Advances in understanding Norway spruce natural resistance to needle bladder rust infection: transcriptional and secondary metabolites profiling.

BACKGROUND: Needle rust caused by the fungus Chrysomyxa rhododendri causes significant growth decline and increased mortality of young Norway spruce trees in subalpine forests. Extremely rare trees with enhanced resistance represent promising candidates for practice-oriented reproduction approaches. They also enable the investigation of tree molecular defence and resistance mechanisms against this fungal disease. Here, we combined RNA-Seq, RT-qPCR and secondary metabolite analyses during a period of 38 days following natural infection to investigate differences in constitutive and infection-induced defence between the resistant genotype PRA-R and three susceptible genotypes. RESULTS: Gene expression and secondary metabolites significantly differed among genotypes from day 7 on and revealed already known, but also novel candidate genes involved in spruce molecular defence against this pathogen. Several key genes related to (here and previously identified) spruce defence pathways to needle rust were differentially expressed in PRA-R compared to susceptible genotypes, both constitutively (in non-symptomatic needles) and infection-induced (in symptomatic needles). These genes encoded both new and well-known antifungal proteins such as endochitinases and chitinases. Specific genetic characteristics concurred with varying phenolic, terpene, and hormone needle contents in the resistant genotype, among them higher accumulation of several flavonoids (mainly kaempferol and taxifolin), stilbenes, geranyl acetone, α-ionone, abscisic acid and salicylic acid. CONCLUSIONS: Combined transcriptional and metabolic profiling of the Norway spruce defence response to infection by C. rhododendri in adult trees under subalpine conditions confirmed the results previously gained on artificially infected young clones in the greenhouse, both regarding timing and development of infection, and providing new insights into genes and metabolic pathways involved. The comparison of genotypes with different degrees of susceptibility proved that several of the identified key genes are differently regulated in PRA-R, and that the resistant genotype combines a strong constitutive defence with an induced response in infected symptomatic needles following fungal invasion. Genetic and metabolic differences between the resistant and susceptible genotypes indicated a more effective hypersensitive response (HR) in needles of PRA-R that prevents penetration and spread of the rust fungus and leads to a lower proportion of symptomatic needles as well as reduced symptom development on the few affected needles.

Alpine dwarf shrubs show high proportions of nonfunctional xylem: Visualization and quantification of species-specific patterns.

Xylem conductive capacity is a key determinant of plant hydraulic function and intimately linked to photosynthesis and productivity, but can be impeded by temporary or permanent conduit dysfunctions. Here we show that persistent xylem dysfunctions in unstressed plants are frequent in Alpine dwarf shrubs and occur in various but species-specific cross-sectional patterns. Combined synchrotron micro-computed tomography (micro-CT) imaging, xylem staining, and flow measurements in saturated samples of six widespread Ericaceae species evidence a high proportion (19%-50%) of hydraulically nonfunctional xylem areas in the absence of drought stress, with regular distribution of dysfunctions between or within growth rings. Dysfunctions were only partly reversible and reduced the specific hydraulic conductivity to 1.38 to 3.57 ×10-4 m2 s-1 MPa-1 . Decommission of inner growth rings was clearly related to stem age and a higher vulnerability to cavitation of older rings, while the high proportion of nonfunctional conduits in each annual ring needs further investigations. The lower the xylem fraction contributing to the transport function, the higher was the hydraulic efficiency of conducting xylem areas. Improved understanding of the functional lifespan of xylem elements and the prevalence and nature of dysfunctions is critical to correctly assess structure-function relationships and whole-plant hydraulic strategies.

Hydraulic-stomatal coordination in tree seedlings: tight correlation across environments and ontogeny in Acer pseudoplatanus.

Hydraulic conductance is recognized as a major determinant of gas exchange and productivity. However, whether this also applies to seedlings, a critically important stage for vegetation regeneration, has been largely unknown. We analyzed the hydraulic and stomatal conductance of leaves and shoots for 6-wk-old Acer pseudoplatanus seedlings emerging in different lowland and treeline habitats and under glasshouse conditions, respectively, as well as on 9-, 15- and 18-wk-old plants, and related findings to leaf and xylem anatomical traits. Treeline seedlings had higher leaf area-specific shoot hydraulic conductance (Kshoot-L ), and stomatal conductance (gs ), associated with wider xylem conduits, lower leaf area and higher stomatal density than lowland and glasshouse-grown plants. Across the first 18 wk of development, seedlings increased four-fold in absolute shoot hydraulic conductance (Kshoot ) and declined by half in Kshoot-L , with correlated shifts in xylem and leaf anatomy. Distal leaves had higher leaf hydraulic conductance (Kleaf ) and gs compared to basal leaves. Seedlings show strong variation across growth environments and ontogenetic shifts in hydraulic and anatomical parameters. Across growth sites, ontogenetic stages and leaf orders, gs was tightly correlated with Kshoot-L and Kleaf , balancing hydraulic supply with demand for the earliest stages of seedling establishment.

Subalpine dwarf shrubs differ in vulnerability to xylem cavitation: An innovative staining approach enables new insights.

Dwarf shrubs are a key functional group of the high-elevation vegetation belt. Despite their ecological relevance and high sensitivity to environmental changes, the hydraulic adaptations and species-specific variations in drought tolerance within this growth form are mostly unknown. Here, we assess the xylem vulnerability to cavitation of important character species of the Alpine dwarf shrub heaths in Central Europe. Due to the high percentage of nonfunctional xylem areas in these species, vulnerability curves were gained by an innovative staining approach with safranin, validated by hydraulic and xylem anatomical measurements. The loss of 50% conducting xylem area appeared in the range of -1.78 to -2.91 MPa. Midday plant water potential during an intense summer drought remained above these critical thresholds but was correlated with cavitation resistance. No trade-off between hydraulic safety and efficiency was detected across analyzed species. We conclude that the characteristic clustered occurrence of species in the heterogeneous mountain terrain (mainly interpreted as a consequence of varying snow cover dynamics) is also reflected in species-specific adjustments in xylem hydraulics. The interspecific variety in vulnerability thresholds and plant water potentials during summer drought indicates different hydraulic strategies and adjustments in water relations between these co-occurring shrubs.

RNA-Seq and secondary metabolite analyses reveal a putative defence-transcriptome in Norway spruce (Picea abies) against needle bladder rust (Chrysomyxa rhododendri) infection.

BACKGROUND: Norway spruce trees in subalpine forests frequently face infections by the needle rust fungus Chrysomyxa rhododendri, which causes significant growth decline and increased mortality of young trees. Yet, it is unknown whether trees actively respond to fungal attack by activating molecular defence responses and/or respective gene expression. RESULTS: Here, we report results from an infection experiment, in which the transcriptomes (via RNA-Seq analysis) and phenolic profiles (via UHPLC-MS) of control and infected trees were compared over a period of 39 days. Gene expression between infected and uninfected ramets significantly differed after 21 days of infection and revealed already known, but also novel candidate genes involved in spruce molecular defence against pathogens. CONCLUSIONS: Combined RNA-Seq and biochemical data suggest that Norway spruce response to infection by C. rhododendri is restricted locally and primarily activated between 9 and 21 days after infestation, involving a potential isolation of the fungus by a hypersensitive response (HR) associated with an activation of phenolic pathways. Identified key regulatory genes represent a solid basis for further specific analyses in spruce varieties with varying susceptibility, to better characterise resistant clones and to elucidate the resistance mechanism.

Insights into trunks of Pinus cembra L.: analyses of hydraulics via electrical resistivity tomography.

KEY MESSAGE: The lack of elevational changes in electrical resistivity in Pinus cembra trunks indicated consistent growth and hydraulics across elevations. Though, electrical resistivity tomograms exhibited pronounced temperature-driven seasonal changes. ABSTRACT: Alpine conifers growing at high elevation are exposed to low temperatures, which may limit xylogenesis and cause pronounced seasonal changes in tree hydraulics. Electrical resistivity (ER) tomography enables minimal invasive monitoring of stems in situ. We used this technique to analyze Pinus cembra trunks along a 400 m elevational gradient up to the timberline and over seasons. Furthermore, ER data of earlywood across tree rings were compared with the respective specific hydraulic conductivity (K S), measured on extracted wood cores. ER tomograms revealed pronounced changes over the year and a strong correlation between average ER (ERmean) and air and xylem temperatures. Surprisingly, no elevational changes in ERmean, earlywood ER or K S were observed. ER data corresponded to variation in earlywood K S, which decreased from the youngest (ca. 4-5 cm2s-1 MPa-1) to the oldest tree rings (0.63 ± 0.22 cm2s-1 MPa-1). The lack of changes in ER data and earlywood K S along the study transect indicated consistent growth patterns and no major changes in structural and functional hydraulic traits across elevation. The constant decrease in earlywood K S with tree ring age throughout all elevations highlights the hydraulic relevance of the outermost tree rings in P. cembra. Seasonal measurements demonstrated pronounced temperature effects on ER, and we thus recommend a detailed monitoring of trunk temperatures for ER tomography.

Insights from in vivo micro-CT analysis: testing the hydraulic vulnerability segmentation in Acer pseudoplatanus and Fagus sylvatica seedlings.

The seedling stage is the most susceptible one during a tree's life. Water relations may be crucial for seedlings due to their small roots, limited water buffers and the effects of drought on water transport. Despite obvious relevance, studies on seedling xylem hydraulics are scarce as respective methodical approaches are limited. Micro-CT scans of intact Acer pseudoplatanus and Fagus sylvatica seedlings dehydrated to different water potentials (Ψ) allowed the simultaneous observation of gas-filled versus water-filled conduits and the calculation of percentage loss of conductivity (PLC) in stems, roots and leaves (petioles or main veins). Additionally, anatomical analyses were performed and stem PLC measured with hydraulic techniques. In A. pseudoplatanus, petioles showed a higher Ψ at 50% PLC (Ψ50 -1.13MPa) than stems (-2.51 MPa) and roots (-1.78 MPa). The main leaf veins of F. sylvatica had similar Ψ50 values (-2.26 MPa) to stems (-2.74 MPa) and roots (-2.75 MPa). In both species, no difference between root and stems was observed. Hydraulic measurements on stems closely matched the micro-CT based PLC calculations. Micro-CT analyses indicated a species-specific hydraulic architecture. Vulnerability segmentation, enabling a disconnection of the hydraulic pathway upon drought, was observed in A. pseudoplatanus but not in the especially shade-tolerant F. sylvatica. Hydraulic patterns could partly be related to xylem anatomical traits.

Are hydraulic patterns of lianas different from trees? New insights from Hedera helix.

Lianas form long and flexible but disproportionately narrow stems, and thus require particular strategies to maintain the integrity of xylem water transport and ensure supply to large crown areas. The hydraulic architecture of lianas and the respective within-plant coordination of transport efficiency and safety, and the underlying anatomical variations in xylem, are largely unexplored. We analysed Hedera helix, a liana widespread in European temperate forests, with respect to hydraulic and xylem anatomical variations between the main stem and branches, between juvenile and adult life phases, and along the vertical axis. Main stems were significantly less embolism resistant but exhibited a higher hydraulic conductivity than branches. In branches, the cell turgor loss point of leaves decreased, while the embolism resistance and conductivity of xylem, as well as conduit diameters, increased with height. High water-transport capacities allow ivy to compensate for the small cross-section of stems, while the limited resistance to drought-induced xylem dysfunction of the main stem is probably linked to conservative stomatal regulation. Pronounced differences in xylem anatomy, hydraulic efficiency, and safety between the main stem and branches and along the vertical axis are surprisingly similar to those of self-supporting plants, and indicate that the coordination of carbon and water economies requires similar internal adjustments in tall plants.

The pitfalls of in vivo imaging techniques: evidence for cellular damage caused by synchrotron X-ray computed micro-tomography.

Synchrotron X-ray computed micro-tomography (microCT) has emerged as a promising noninvasive technique for in vivo monitoring of xylem function, including embolism build-up under drought and hydraulic recovery following re-irrigation. Yet, the possible harmful effects of ionizing radiation on plant tissues have never been quantified. We specifically investigated the eventual damage suffered by stem living cells of three different species exposed to repeated microCT scans. Stem samples exposed to one, two or three scans were used to measure cell membrane and RNA integrity, and compared to controls never exposed to X-rays. Samples exposed to microCT scans suffered serious alterations to cell membranes, as revealed by marked increase in relative electrolyte leakage, and also underwent severe damage to RNA integrity. The negative effects of X-rays were apparent in all species tested, but the magnitude of damage and the minimum number of scans inducing negative effects were species-specific. Our data show that multiple microCT scans lead to disruption of fundamental cellular functions and processes. Hence, microCT investigation of phenomena that depend on physiological activity of living cells may produce erroneous results and lead to incorrect conclusions.

Association genetics of phenolic needle compounds in Norway spruce with variable susceptibility to needle bladder rust.

KEY MESSAGE: Accumulation of phenolic needle metabolites in Norway spruce is regulated by many genes with small and additive effects and is correlated with the susceptibility against fungal attack. Norway spruce accumulates high foliar concentrations of secondary phenolic metabolites, with important functions for pathogen defence responses. However, the molecular genetic basis underlying the quantitative variation of phenolic compounds and their role in enhanced resistance of spruce to infection by needle bladder rust are unknown. To address these questions, a set of 1035 genome-wide single nucleotide polymorphisms (SNPs) was associated to the quantitative variation of four simple phenylpropanoids, eight stilbenes, nine flavonoids, six related arithmetic parameters and the susceptibility to infection by Chrysomyxa rhododendri in an unstructured natural population of Norway spruce. Thirty-one significant genetic associations for the flavonoids gallocatechin, kaempferol 3-glucoside and quercetin 3-glucoside and the stilbenes resveratrol, piceatannol, astringin and isorhapontin were discovered, explaining 22-59% of phenotypic variation, and indicating a regulation of phenolic accumulation by many genes with small and additive effects. The phenolics profile differed between trees with high and low susceptibility to the fungus, underlining the importance of phenolic compounds in the defence mechanisms of Norway spruce to C. rhododendri. Results highlight the utility of association studies in non-model tree species and may enable marker-assisted selection of Norway spruce adapted to severe pathogen attack.

Herb Hydraulics: Inter- and Intraspecific Variation in Three Ranunculus Species.

The requirements of the water transport system of small herbaceous species differ considerably from those of woody species. Despite their ecological importance for many biomes, knowledge regarding herb hydraulics remains very limited. We compared key hydraulic features (vulnerability to drought-induced hydraulic decline, pressure-volume relations, onset of cellular damage, in situ variation of water potential, and stomatal conductance) of three Ranunculus species differing in their soil humidity preferences and ecological amplitude. All species were very vulnerable to water stress (50% reduction in whole-leaf hydraulic conductance [kleaf] at -0.2 to -0.8 MPa). In species with narrow ecological amplitude, the drought-exposed Ranunculus bulbosus was less vulnerable to desiccation (analyzed via loss of kleaf and turgor loss point) than the humid-habitat Ranunculus lanuginosus Accordingly, water stress-exposed plants from the broad-amplitude Ranunculus acris revealed tendencies toward lower vulnerability to water stress (e.g. osmotic potential at full turgor, cell damage, and stomatal closure) than conspecific plants from the humid site. We show that small herbs can adjust to their habitat conditions on interspecific and intraspecific levels in various hydraulic parameters. The coordination of hydraulic thresholds (50% and 88% loss of kleaf, turgor loss point, and minimum in situ water potential) enabled the study species to avoid hydraulic failure and damage to living cells. Reversible recovery of hydraulic conductance, desiccation-tolerant seeds, or rhizomes may allow them to prioritize toward a more efficient but vulnerable water transport system while avoiding the severe effects that water stress poses on woody species.

Dwarf shrub hydraulics: two Vaccinium species (Vaccinium myrtillus, Vaccinium vitis-idaea) of the European Alps compared.

Vaccinium myrtillus and Vaccinium vitis-idaea are two dwarf shrubs widespread in the European Alps. We studied the hydraulics of these species hypothesizing that (1) the hydraulic architecture of dwarf shrubs differs from trees, (2) hydraulic properties reflect the species' ecological amplitude and (3) hydraulic properties vary spatially and seasonally. Key hydraulic parameters (osmotic potential, turgor loss point, xylem hydraulic conductivity, vulnerability to drought-induced embolism, stomata closure, drought-induced cell damage and embolism repair) and related wood anatomical traits (conduit diameter and conduit wall reinforcement) were analyzed at four sites in Tyrol, Austria. Both species exhibited low hydraulic safety as well as low hydraulic efficiency. Fifty percentage embolism accumulated at -2.08 (V. myrtillus) and -1.97 MPa (V. vitis-idaea), 88% stomata closure was at -2.19 and -2.35 MPa, respectively. After drought, both species showed embolism repair on re-watering. Site-specific variation within species was low, while seasonal changes in embolism resistance and turgor loss point were observed. Results indicate that studied Vaccinium species have a high risk for embolism formation. This is balanced by refilling capacities, which are probably based on the small growth height of dwarf shrubs. V. vitis-idaea, which occurs on drier sites, showed more efficient repair and a lower turgor loss point than V. myrtillus.

Elevated nutrient supply can exert worse effects on Norway spruce than drought, viewed through chemical defence against needle rust.

Abiotic factors such as water and nutrient availability can exert a dominant influence on the susceptibility of plants to various pathogens. Effects of abiotic environmental factors on phenolic compound concentrations in the plant tissue may represent one of the major underlying mechanisms, as these compounds are known to play a substantial role in plant resistance to pests. In particular, this applies to conifer trees, in which a large range of phenolic compounds are produced constitutively and/or induced by pathogen attack. We subjected Norway spruce saplings to water limitation and elevated nutrient supply over 2 years and subsequently controlled infection with the needle rust Chrysomyxa rhododendri (DC.) de Bary and analysed both constitutive and inducible phenolic compound concentrations in the needles as well as the degree of infection. Compared with the control group, both drought and fertilization profoundly modified the constitutive and pathogen-induced profiles of phenolic compounds, but had little impact on the total phenolic content. Fertilization predominantly affected the inducible phenolic response and led to higher infection rates by C. rhododendri. Drought stress, in contrast, mainly shaped the phenolic profiles in healthy plant parts and had no consequences on the plant susceptibility. The results show that specific abiotic effects on individual compounds seem to be decisive for the infection success of C. rhododendri, whereby the impaired induced response in saplings subjected to nutrient supplementation was most critical. Although drought effects were minor, they varied depending on the time and length of water limitation. The results indicate that prolonged drought periods in the future may not significantly alter the foliar defence of Norway spruce against C. rhododendri, but fertilization, often propagated to increase tree growth and forest productivity, can be counterproductive in areas with high pathogen pressure.

The 2018 European heatwave led to stem dehydration but not to consistent growth reductions in forests.

Heatwaves exert disproportionately strong and sometimes irreversible impacts on forest ecosystems. These impacts remain poorly understood at the tree and species level and across large spatial scales. Here, we investigate the effects of the record-breaking 2018 European heatwave on tree growth and tree water status using a collection of high-temporal resolution dendrometer data from 21 species across 53 sites. Relative to the two preceding years, annual stem growth was not consistently reduced by the 2018 heatwave but stems experienced twice the temporary shrinkage due to depletion of water reserves. Conifer species were less capable of rehydrating overnight than broadleaves across gradients of soil and atmospheric drought, suggesting less resilience toward transient stress. In particular, Norway spruce and Scots pine experienced extensive stem dehydration. Our high-resolution dendrometer network was suitable to disentangle the effects of a severe heatwave on tree growth and desiccation at large-spatial scales in situ, and provided insights on which species may be more vulnerable to climate extremes.

Tracheid and Pit Dimensions Hardly Vary in the Xylem of Pinus sylvestris Under Contrasting Growing Conditions.

Maintaining sufficient water transport via the xylem is crucial for tree survival under variable environmental conditions. Both efficiency and safety of the water transport are based on the anatomical structure of conduits and their connections, the pits. Yet, the plasticity of the xylem anatomy, particularly that of the pit structures, remains unclear. Also, trees adjust conduit dimensions to the water transport distance (i.e., tree size), but knowledge on respective adjustments in pit dimensions is scarce. We compared tracheid traits [mean tracheid diameter d, mean hydraulic diameter d h , cell wall reinforcement (t/b)2], pit dimensions (diameters of pit aperture D a , torus D t , margo D m , and pit border D p ), and pit functional properties (margo flexibility F, absolute overlap O a , torus overlap O, and valve effect V ef ) of two Scots pine (Pinus sylvestris L.) stands of similar tree heights but contrasting growth rates. Furthermore, we analyzed the trends of these xylem anatomical parameters across tree rings. Tracheid traits and pit dimensions were similar on both sites, whereas O a , O, and F were higher at the site with a lower growth rate. On the lower growth rate site, d h and pit dimensions increased across tree rings from pith to bark, and in trees from both sites, d h scaled with pit dimensions. Adjusted pit functional properties indicate slightly higher hydraulic safety in trees with a lower growth rate, although a lack of major differences in measured traits indicated overall low plasticity of the tracheid and pit architecture. Mean hydraulic diameter and pit dimension are well coordinated to increase the hydraulic efficiency toward the outer tree rings and thus with increasing tree height. Our results contribute to a better understanding of tree hydraulics under variable environmental conditions.

From the comfort zone to crown dieback: Sequence of physiological stress thresholds in mature European beech trees across progressive drought.

Drought responses of mature trees are still poorly understood making it difficult to predict species distributions under a warmer climate. Using mature European beech (Fagus sylvatica L.), a widespread and economically important tree species in Europe, we aimed at developing an empirical stress-level scheme to describe its physiological response to drought. We analysed effects of decreasing soil and leaf water potential on soil water uptake, stem radius, native embolism, early defoliation and crown dieback with comprehensive measurements from overall nine hydrologically distinct beech stands across Switzerland, including records from the exceptional 2018 drought and the 2019/2020 post-drought period. Based on the observed responses to decreasing water potential we derived the following five stress levels: I (predawn leaf water potential >-0.4 MPa): no detectable hydraulic limitations; II (-0.4 to -1.3): persistent stem shrinkage begins and growth ceases; III (-1.3 to -2.1): onset of native embolism and defoliation; IV (-2.1 to -2.8): onset of crown dieback; V (<-2.8): transpiration ceases and crown dieback is >20%. Our scheme provides, for the first time, quantitative thresholds regarding the physiological downregulation of mature European beech trees under drought and therefore synthesises relevant and fundamental information for process-based species distribution models. Moreover, our study revealed that European beech is drought vulnerable, because it still transpires considerably at high levels of embolism and because defoliation occurs rather as a result of embolism than preventing embolism. During the 2018 drought, an exposure to the stress levels III-V of only one month was long enough to trigger substantial crown dieback in beech trees on shallow soils. On deep soils with a high water holding capacity, in contrast, water reserves in deep soil layers prevented drought stress in beech trees. This emphasises the importance to include local data on soil water availability when predicting the future distribution of European beech.

Noninvasive Analysis of Tree Stems by Electrical Resistivity Tomography: Unraveling the Effects of Temperature, Water Status, and Electrode Installation.

The increasing demand for tree and forest health monitoring due to ongoing climate change requires new future-oriented and nondestructive measurement techniques. Electrical resistivity (ER) tomography represents a promising and innovative approach, as it allows insights into living trees based on ER levels and ER cross-sectional distribution patterns of stems. However, it is poorly understood how external factors, such as temperature, tree water status, and electrode installation affect ER tomograms. In this study, ER measurements were carried out on three angiosperms (Betula pendula, Fagus sylvatica, Populus nigra) and three conifers (Larix decidua, Picea abies, Pinus cembra) exposed to temperatures between -10 and 30°C and to continuous dehydration down to -6.3 MPa in a laboratory experiment. Additionally, effects of removal of peripheral tissues (periderm, phloem, cambium) and electrode installation were tested. Temperature changes above the freezing point did not affect ER distribution patterns but average ER levels, which increased exponentially and about 2.5-fold from 30 to 0°C in all species. In contrast, freezing of stems caused a pronounced raise of ER, especially in peripheral areas. With progressive tree dehydration, average ER increased in all species except in B. pendula, and measured resistivities in the peripheral stem areas of both angiosperms and conifers were clearly linearly related to the tree water status. Removal of the periderm resulted in a slight decrease of high ER peaks. Installation of electrodes for a short period of 32-72 h before conducting the tomography caused small distortions in tomograms. Distortions became serious after long-term installation for several months, while mean ER was only slightly affected. The present study confirms that ER tomography of tree stems is sensitive to temperature and water status. Results help to improve ER tomogram interpretation and suggest that ER analyses may be suitable to nondestructively determinate the hydraulic status of trees. They thus provide a solid basis for further technological developments to enable presymptomatic detection of physiological stress in standing trees.

Electrical resistivity tomography: patterns in Betula pendula, Fagus sylvatica, Picea abies and Pinus sylvestris.

Electrical resistivity (ER) tomography is a promising technique to minimally invasively study stems of living trees. It allows insights into xylem properties based on the cross-sectional distribution of ER that is governed by the wood's electrical conductance. In this study, ER measurements were carried out on four forest tree species, Betula pendula, Fagus sylvatica, Picea abies and Pinus sylvestris, to demonstrate interspecific, intraspecific and within-tree variation of ER tomograms. Further, ER patterns were linked to xylem moisture content (MC), electrolyte content and density obtained from wood core analyses. The ER patterns of both coniferous species, P. abies and P. sylvestris, were found to be more homogenous and concentric compared with the complex tomograms of angiosperms. However, the ER range of coniferous trees showed considerable intraspecific variation. Measurements near ground level showed pronounced effects on ER tomograms, highlighting the importance of the chosen measurement height. A strong relation between ER and wood density was found in F. sylvatica while ER patterns of conifers were mainly influenced by MC. Results demonstrate a high species specificity of ER tomograms and of respective influencing xylem traits. They underline the importance of reference measurements for a correct interpretation of ER studies.

Robustness of xylem properties in conifers: analyses of tracheid and pit dimensions along elevational transects.

In alpine regions, tree hydraulics are limited by low temperatures that restrict xylem growth and induce winter frost drought and freezing stress. While several studies have dealt with functional limitations, data on elevational changes in functionally relevant xylem anatomical parameters are still scarce. In wood cores of Pinus cembra L. and Picea abies (L.) Karst. trunks, harvested along five elevational transects, xylem anatomical parameters (tracheid hydraulic diameter dh, wall reinforcement (t/b)2), pit dimensions (pit aperture Da, pit membrane Dm and torus Dt diameters) and respective functional indices (torus overlap O, margo flexibility) were measured. In both species, tracheid diameters decreased and (t/b)2 increased with increasing elevation, while pit dimensions and functional indices remained rather constant (P. cembra: Dt 10.3 ± 0.2 µm, O 0.477 ± 0.005; P. abies: Dt 9.30 ± 0.18 µm, O 0.492 ± 0.005). However, dh increased with tree height following a power trajectory with an exponent of 0.21, and also pit dimensions increased with tree height (exponents: Dm 0.18; Dt 0.14; Da 0.11). Observed elevational trends in xylem structures were predominantly determined by changes in tree size. Tree height-related changes in anatomical traits showed a remarkable robustness, regardless of the distributional ranges of study species. Despite increasing stress intensities towards the timberline, no adjustment in hydraulic safety at the pit level was observed.

Foliar Phenolic Compounds in Norway Spruce with Varying Susceptibility to Chrysomyxa rhododendri: Analyses of Seasonal and Infection-Induced Accumulation Patterns.

Secondary phenolic metabolites are involved in plant responses to various biotic stress factors, and are apparently important for the defense against fungal pathogens. In this study, we investigated their role in defense against the rust Chrysomyxa rhododendri in Norway spruce. The fungal pathogen undergoes a seasonal lifecycle with host shift; after overwintering in rhododendron shrubs, it attacks the sprouting current-year spruce needles and causes needle fall in autumn. Repeated infections lead to reduced timber yield and severe problems with rejuvenation in subalpine Norway spruce forests. Trees with varying susceptibility to infection by C. rhododendri were selected and foliar phenolic composition was assessed using UHPLC-MS. We report on seasonal accumulation patterns and infection-related changes in the concentrations of 16 metabolites, including flavonoids, stilbenes, simple phenylpropanoids and the precursor shikimic acid, and their correlation with the infection degree of the tree. We found significant variation in the phenolic profiles during needle development: flavonoids were predominant in the first weeks after sprouting, whereas stilbenes, picein and shikimic acid increased during the first year. Following infection, several flavonoids and resveratrol increased up to 1.8 fold in concentration, whereas picein and shikimic acid were reduced by about 70 and 60%, respectively. The constitutive and early stage infection-induced concentrations of kaempferol, quercetin and taxifolin as well as the late stage infection-induced concentrations of stilbenes and picein were negatively correlated with infection degree. We conclude that a combination of constitutive and inducible accumulation of phenolic compounds is associated with the lower susceptibility of individual trees to C. rhododendri. The potentially fungicidal flavonoid aglycones may limit hyphal growth and prevent development of infection symptoms, and high levels of stilbenes may impede the infection of older needles. The presented results underline a highly compound-specific seasonal accumulation and defense response of Norway spruce and may facilitate the selection of promising trees for breeding programs.