Summary Eastern Australia was subject to its hottest and driest year on record in 2019. This extreme drought resulted in massive canopy die‐back in eucalypt forests. The role of hydraulic failure and tree size on canopy die‐back in three eucalypt tree species during this drought was examined. We measured pre‐dawn and midday leaf water potential (Ψleaf), per cent loss of stem hydraulic conductivity and quantified hydraulic vulnerability to drought‐induced xylem embolism. Tree size and tree health was also surveyed. Trees with most, or all, of their foliage dead exhibited high rates of native embolism (78–100%). This is in contrast to trees with partial canopy die‐back (30–70% canopy die‐back: 72–78% native embolism), or relatively healthy trees (little evidence of canopy die‐back: 25–31% native embolism). Midday Ψleaf was significantly more negative in trees exhibiting partial canopy die‐back (−2.7 to −6.3 MPa), compared with relatively healthy trees (−2.1 to −4.5 MPa). In two of the species the majority of individuals showing complete canopy die‐back were in the small size classes. Our results indicate that hydraulic failure is strongly associated with canopy die‐back during drought in eucalypt forests. Our study provides valuable field data to help constrain models predicting mortality risk.

(S.M.).During winter, trees have to cope with harsh conditions, including extreme freeze-thaw stress. This study focused on ice nucleation and propagation, related water shifts and xylem cavitation, as well as cell damage and was based on in situ monitoring of xylem (thermocouples) and surface temperatures (infrared imaging), ultrasonic emissions, and dendrometer analysis. Field experiments during late winter on Picea abies growing at the alpine timberline revealed three distinct freezing patterns: (1) from the top of the tree toward the base, (2) from thin branches toward the main stem's top and base, and (3) from the base toward the top. Infrared imaging showed freezing within branches from their base toward distal parts. Such complex freezing causes dynamic and heterogenous patterns in water potential and probably in cavitation. This study highlights the interaction between environmental conditions upon freezing and thawing and demonstrates the enormous complexity of freezing processes in trees. Diameter shrinkage, which indicated water fluxes within the stem, and acoustic emission analysis, which indicated cavitation events near the ice front upon freezing, were both related to minimum temperature and, upon thawing, related to vapor pressure deficit and soil temperature. These complex patterns, emphasizing the common mechanisms between frost and drought stress, shed new light on winter tree physiology.

Summary 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 (Ψ 50 −1.13 MP a) than stems (−2.51 MP a) and roots (−1.78 MP a). The main leaf veins of F. sylvatica had similar Ψ 50 values (−2.26 MP a) to stems (−2.74 MP a) and roots (−2.75 MP a). 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.

Drought-induced xylem embolism is a serious threat to plant survival under future climate scenarios. Hence, accurate quantification of species-specific vulnerability to xylem embolism is a key to predict the impact of climate change on vegetation. Low-cost hydraulic measurements of embolism rate have been suggested to be prone to artefacts, thus requiring validation by direct visualization of the functional status of xylem conduits using nondestructive imaging techniques, such as X-ray microtomography (microCT). We measured the percentage loss of conductance (PLC) of excised stems of Laurus nobilis (laurel) dehydrated to different xylem pressures, and compared results with direct observation of gas-filled vs water-filled conduits at a synchrotron-based microCT facility using a phase contrast imaging modality. Theoretical PLC calculated on the basis of microCT observations in stems of laurel dehydrated to different xylem pressures overall were in agreement with hydraulic measurements, revealing that this species suffers a 50% loss of xylem hydraulic conductance at xylem pressures averaging -3.5 MPa. Our data support the validity of estimates of xylem vulnerability to embolism based on classical hydraulic techniques. We discuss possible causes of discrepancies between data gathered in this study and those of recent independent reports on laurel hydraulics.

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.