On the mechanism for winter stem pressure build-up in walnut trees.

Xylem embolism is a significant factor in tree mortality. Restoration of hydraulic conductivity after massive embolization of the vascular system requires the application of positive pressure to the vessels and/or the creation of new conductive elements. Some species generate positive pressure from the root system to propagate pressure in distal, aboveground organs in spring, whereas other species generate positive pressure locally at the stem level during winter. We provide a mechanistic explanation for winter stem pressure build-up in the walnut tree. We have developed a physical model that accounts for temperature fluctuations and phase transitions. This model is based on the exchange of water and sugars between living cells and vessels. Our computations demonstrate that vessel pressurization can be attributed to the transfer of water between vessels across the parenchyma rays, which is facilitated by a radial imbalance in sugar concentration. The ability to dispose of soluble sugars in living cells, and to transport them between living cells and up to the vessels, is identified as the main drivers of stem pressure build-up in the walnut tree.

Unpacking the point of no return under drought in poplar: insight from stem diameter variation.

A specific, robust threshold for drought-induced tree mortality is needed to improve the prediction of forest dieback. Here, we tested the relevance of continuous measurements of stem diameter variations for identifying such a threshold, their relationship with hydraulic and cellular damage mechanisms, and the influence of growth conditions on these relationships. Poplar saplings were grown under well-watered, water-limited, or light-limited conditions and then submitted to a drought followed by rewatering. Stem diameter was continuously measured to investigate two parameters: the percentage loss of diameter (PLD) and the percentage of diameter recovery (DR) following rewatering. Water potentials, stomatal conductance, embolism, and electrolyte leakage were also measured, and light microscopy allowed investigating cell collapse induced by drought. The water release observed through loss of diameter occurred throughout the drought, regardless of growth conditions. Poplars did not recover from drought when PLD reached a threshold and this differed according to growth conditions but remained linked to cell resistance to damage and collapse. Our findings shed new light on the mechanisms of drought-induced tree mortality and indicate that PLD could be a relevant indicator of drought-induced tree mortality, regardless of the growth conditions.

Freeze dehydration vs supercooling in tree stems: physical and physiological modelling.

Frost resistance is the major factor affecting the distribution of plant species at high latitude and elevation. The main effects of freeze-thaw cycles are damage to living cells and formation of gas embolism in tree xylem vessels. Lethal intracellular freezing can be prevented in living cells by two mechanisms, such as dehydration and deep supercooling. We developed a multiphysics numerical model coupling water flow, heat transfer and phase change, considering different cell types in plant tissues, to study the dynamics and extent of cell dehydration, xylem pressure changes and stem diameter changes in response to freezing and thawing. Results were validated using experimental data for stem diameter changes of walnut trees (Juglans regia). The effect of cell mechanical properties was found to be negligible as long as the intracellular tension developed during dehydration was sufficiently low compared with the ice-induced cryostatic suction. The model was finally used to explore the coupled effects of relevant physiological parameters (initial water and sugar content) and environmental conditions (air temperature variations) on the dynamics and extent of dehydration. It revealed configurations where cell dehydration could be sufficient to protect cells from intracellular freezing, and situations where supercooling was necessary. This model, freely available with this paper, could easily be extended to explore different anatomical structures, different species and more complex physical processes.

Dynamic modeling of stem water content during the dormant period in walnut trees.

Water content (WC) is a key variable in plant physiology even during the winter period. To simulate stem WC during the dormant season, a series of experiments were carried out on walnut trees under controlled conditions. In the field, WC was significantly correlated with soil temperature at 50 cm depth (R2 = 0.526). In the greenhouse, WC remained low as long as soil temperature was kept cold (<+5 °C) and increased after the soil temperature was warmed to +15 °C regardless of the date. Stem dehydration rate was significantly influenced by the WC and evaporative demand. A parsimonious model with functions describing the main experimental results was calibrated and validated with field data from 13 independent winter dynamics in Juglans regia L. orchards. Three functions of water uptake were tested, and these gave equivalent accuracies (root-mean-square error (RMSE) = 0.127-8; predictive root-mean-square error = 0.116). However, only a sigmoid function describing the relationship between the root water uptake and soil temperature gave values in agreement with the experimental results. Finally, the simulated WC provided a similar accuracy in predicting frost hardiness compared with the measured WC (RMSE ca 3 °C) and was excellent in spring (RMSE ca 2 °C). This model may be a relevant tool for predicting the risk of spring frost in walnut trees. Its genericity should be tested in other fruit and forest tree species.

Xylem embolism and bubble formation during freezing suggest complex dynamics of pressure in Betula pendula stems.

Freeze-thaw-induced embolism, a key limiting factor for perennial plants results from the formation of gas bubbles during freezing and their expansion during thawing. However, the ice volumetric increase generates local pressures, which can affect the formation of bubbles. To characterize local dynamics of pressure tension and the physical state of the sap during freeze-thaw cycles, we simultaneously used ultrasonic acoustic emission analysis and synchrotron-based high-resolution computed tomography on the diffuse-porous species Betula pendula. Visualization of individual air-filled vessels and the distribution of gas bubbles in frozen xylem were performed.. Ultrasonic emissions occurred after ice formation, together with bubble formation, whereas the development of embolism took place after thawing. The pictures of frozen tissues indicated that the positive pressure induced by the volumetric increase of ice can provoke inward flow from the cell wall toward the lumen of the vessels. We found no evidence that wider vessels within a tissue were more prone to embolism, although the occurrence of gas bubbles in larger conduits would make them prone to earlier embolism. These results highlight the need to monitor local pressure as well as ice and air distribution during xylem freezing to understand the mechanism leading to frost-induced embolism.

ConnecSenS, a Versatile IoT Platform for Environment Monitoring: Bring Water to Cloud.

Climate change is having an increasingly rapid impact on ecosystems and particularly on the issue of water resources. The Internet of Things and communication technologies have now reached a level of maturity that allows sensors to be deployed more easily on sites to monitor them. The communicating node based on LoRaWAN technology presented in this article is open and allows the interfacing of numerous sensors for designing long-term environmental monitoring systems of isolated sites. The data integration in the cloud is ensured by a workflow driving the storage and indexing of data, allowing a simple and efficient use of the data for different users (scientists, administration, citizens) through specific dashboards and extractions. This article presents this infrastructure through environmental monitoring use cases related to water resources.

Molinia caerulea alters forest Quercus petraea seedling growth through reduced mycorrhization.

Oak regeneration is jeopardized by purple moor grass, a well-known competitive perennial grass in the temperate forests of Western Europe. Below-ground interactions regarding resource acquisition and interference have been demonstrated and have led to new questions about the negative impact of purple moor grass on ectomycorrhizal colonization. The objective was to examine the effects of moor grass on root system size and ectomycorrhization rate of oak seedlings as well as consequences on nitrogen (N) content in oak and soil. Oak seedlings and moor grass tufts were planted together or separately in pots under semi-controlled conditions (irrigated and natural light) and harvested 1 year after planting. Biomass, N content in shoot and root in oak and moor grass as well as number of lateral roots and ectomycorrhizal rate in oak were measured. Biomass in both oak shoot and root was reduced when planting with moor grass. Concurrently, oak lateral roots number and ectomycorrhization rate decreased, along with a reduction in N content in mixed-grown oak. An interference mechanism of moor grass is affecting oak seedlings performance through reduction in oak lateral roots number and its ectomycorrhization, observed in conjunction with a lower growth and N content in oak. By altering both oak roots and mycorrhizas, moor grass appears to be a species with a high allelopathic potential. More broadly, these results show the complexity of interspecific interactions that involve various ecological processes involving the soil microbial community and need to be explored in situ.

Detection of acoustic events in lavender for measuring xylem vulnerability to embolism and cellular damage.

Acoustic emission analysis is promising to investigate the physiological events leading to drought-induced injury and mortality. However, their nature and source are not fully understood, making this technique difficult to use as a direct measure of the loss of xylem hydraulic conductance. Acoustic emissions were recorded during severe dehydration in lavender plants (Lavandula angustifolia) and compared with the dynamics of embolism development and cell damage. The timing and characteristics of acoustic signals from two independent recording systems were compared by principal component analysis (PCA). Changes in water potential, branch diameter, loss of hydraulic conductance, and cellular damage were also measured to quantify drought-induced damages. Two distinct phases of acoustic emissions were observed during dehydration: the first one associated with a rapid loss of diameter and a significant increase in loss of xylem conductance (90%), and the second with slower changes in diameter and a significant increase in cellular damage. Based on PCA, a developed algorithm discriminated hydraulic-related acoustic signals from other sources, proposing a reconstruction of hydraulic vulnerability curves. Cellular damage preceded by hydraulic failure seems to lead to a lack of recovery. The second acoustic phase would allow detection of plant mortality.

Seasonal changes in carbohydrates and water content predict dynamics of frost hardiness in various temperate tree species.

Predicting tree frost tolerance is critical to select adapted species according to both the current and predicted future climate. The relative change in water to carbohydrate ratio is a relevant trait to predict frost acclimation in branches from many tree species. The objective of this study is to demonstrate the interspecific genericity of this approach across nine tree species. In the studied angiosperm species, frost hardiness dynamics were best correlated to a decrease in water content at the early stage of acclimation (summer and early autumn). Subsequently, frost hardiness dynamics were more tightly correlated to soluble carbohydrate contents until spring growth resumption. Based on different model formalisms, we predicted frost hardiness at different clade levels (angiosperms, family, genus and species) with high to moderate accuracy (1.5-6.0 °C root mean squared error (RMSE)) and robustness (2.8-6.1 °C prediction RMSE). The TOT model, taking all soluble carbohydrate and polyols into account, was more effective and adapted for large scale studies aiming to explore frost hardiness across a wide range of species. The ISC model taking the individual contribution of each soluble carbohydrate molecule into account was more efficient at finer scale such as family or species. The ISC model performance also suggests that the role of solutes cannot be reduced to a 'bulk' osmotic effect as could be computed if all of them were located in a single, common, compartment. This study provides sets of parameters to predict frost hardiness in a wide range of species, and clues for targeting specific carbohydrate molecules to improve frost hardiness.

Drought-Induced Mortality: Branch Diameter Variation Reveals a Point of No Recovery in Lavender Species.

In the context of climate change, determining the physiological mechanisms of drought-induced mortality in woody plants and identifying thresholds of drought survivorship will improve forecasts of forest and agroecosystem die-off. Here, we tested whether continuous measurements of branch diameter variation can be used to identify thresholds of hydraulic failure and physiological recoverability in lavender (Lavandula angustifolia and Lavandula × intermedia) plants exposed to severe drought. Two parameters of branch diameter variation were tested: the percentage loss of diameter and the percentage loss of rehydration capacity. In two greenhouse experiments with different growth conditions, we monitored variation in branch diameter in the two lavender species exposed to a series of drought/rewatering cycles that varied in drought-stress intensity. Water potential, stomatal conductance, loss of xylem hydraulic conductance, and electrolyte leakage were also measured. We observed that plants were not able to recover when percentage loss of diameter reached maximum values of 21.3% ± 0.6% during drought, regardless of species and growth conditions. A percentage loss of rehydration capacity of 100% was defined as the point of no recovery, and was observed with high levels of cellular damage as estimated by electrolyte leakage measured at 75.4% ± 9.3% and occurred beyond 88% loss of xylem hydraulic conductance. Our study demonstrates that lavender plants are not able to recover from severe drought when they have used up their elastic water storage. Additionally, drought-induced mortality in these species was not linked to xylem hydraulic failure but rather to high levels of cell damage.

Soil organic matter rather than ectomycorrhizal diversity is related to urban tree health.

Urban trees provide many ecosystem services, including carbon sequestration, air quality improvement, storm water attenuation and energy conservation, to people living in cities. Provisioning of ecosystem services by urban trees, however, may be jeopardized by the typically poor quality of the soils in urban areas. Given their well-known multifunctional role in forest ecosystems, ectomycorrhizal fungi (EcM) may also contribute to urban tree health and thus ecosystem service provisioning. Yet, no studies so far have directly related in situ EcM community composition to urban tree health indicators. Here, two previously collected datasets were combined: i) tree health data of 175 Tilia tomentosa trees from three European cities (Leuven, Strasbourg and Porto) estimated using a range of reflectance, chlorophyll fluorescence and physical leaf indicators, and ii) ectomycorrhizal diversity of these trees as characterized by next-generation sequencing. Tree health indicators were related to soil characteristics and EcM diversity using canonical redundancy analysis. Soil organic matter significantly explained variation in tree health indicators whereas no significant relation between mycorrhizal diversity variables and the tree health indicators was found. We conclude that mainly soil organic matter, through promoting soil aggregate formation and porosity, and thus indirectly tree water availability, positively affects the health of trees in urban areas. Our results suggest that urban planners should not overlook the importance of soil quality and its water holding capacity for the health of urban trees and potentially also for the ecosystem services they deliver. Further research should also study other soil microbiota which may independently, or in interaction with ectomycorrhiza, mediate tree performance in urban settings.

Dynamic Modeling of Carbon Metabolism During the Dormant Period Accurately Predicts the Changes in Frost Hardiness in Walnut Trees Juglans regia L.

The leafless period is often considered as inactive, although trees have to actively modulate their metabolism through the cold acclimation/deacclimation processes, to cope with frost exposure during winter and to restore growth ability in spring. Carbon metabolism is a key component of these processes through the osmotic control of extracellular ice formation and the trophic control of bud growth. The influence of temperature on the inter-conversion between starch and soluble carbohydrate has been evidenced for years, but we are currently missing an operational tool to predict starch vs. soluble carbohydrate contents during this period, which should allow to better predict frost hardiness. For this purpose, we exposed 1-year-old branches of Juglans regia to constant temperature for one to 3 weeks and measured the changes in carbohydrate composition at three periods (autumn, winter, and spring). As expected, the temperature significantly affected the changes in carbohydrate composition, but the water content and the sampling period were also relevant. Higher starch hydrolysis was observed at low temperature (<5°C) for all sampling periods. Starch hydrolysis was also observed at warm temperature, but in autumn only. These data were used to compare three modeling approaches simulating the changes in carbohydrate composition through enzymatic analogy. The most empirical and the most mechanistic approach did not succeed to simulate external observations (Root Mean Standard Error of Prediction (RMSEP) > 30 mg.g DM-1, Efficiency (Eff) <0), whereas the intermediate model was more efficient (RMSEP = 15.19 mg.g DM-1, Eff = 0.205 and 16.61 mg.g DM-1, Eff = 0.366, for GFS (Glucose + Fructose + Sucrose) and starch, respectively). The accuracy of the model was further improved when using field data for calibration (RMSEP = 5.86 mg.g DM-1, Eff = 0.962; RMSEP = 10.56 mg.g DM-1, Eff = 0.752, for GFS and starch, respectively). This study provided an operative tool to simulate carbohydrate dynamics over leafless period that could predict frost hardiness with approx. 3.4°C accuracy with temperature, water content and initial starch and soluble carbohydrate measurements. It should now be tested under various meteorological conditions and biological systems.

Vegetation reflectance spectroscopy for biomonitoring of heavy metal pollution in urban soils.

Heavy metals in urban soils may impose a threat to public health and may negatively affect urban tree viability. Vegetation spectroscopy techniques applied to bio-indicators bring new opportunities to characterize heavy metal contamination, without being constrained by laborious soil sampling and lab-based sample processing. Here we used Tilia tomentosa trees, sampled across three European cities, as bio-indicators i) to investigate the impacts of elevated concentrations of cadmium (Cd) and lead (Pb) on leaf mass per area (LMA), total chlorophyll content (Chl), chlorophyll a to b ratio (Chla:Chlb) and the maximal PSII photochemical efficiency (Fv/Fm); and ii) to evaluate the feasibility of detecting Cd and Pb contamination using leaf reflectance spectra. For the latter, we used a partial-least-squares discriminant analysis (PLS-DA) to train spectral-based models for the classification of Cd and/or Pb contamination. We show that elevated soil Pb concentrations induced a significant decrease in the LMA and Chla:Chlb, with no decrease in Chl. We did not observe pronounced reductions of Fv/Fm due to Cd and Pb contamination. Elevated Cd and Pb concentrations induced contrasting spectral changes in the red-edge (690-740 nm) region, which might be associated with the proportional changes in leaf pigments. PLS-DA models allowed for the classifications of Cd and Pb contamination, with a classification accuracy of 86% (Kappa = 0.48) and 83% (Kappa = 0.66), respectively. PLS-DA models also allowed for the detection of a collective elevation of soil Cd and Pb, with an accuracy of 66% (Kappa = 0.49). This study demonstrates the potential of using reflectance spectroscopy for biomonitoring of heavy metal contamination in urban soils.

Variation in ectomycorrhizal fungal communities associated with Silver linden (Tilia tomentosa) within and across urban areas.

Trees in urban areas face harsh environmental conditions. Ectomycorrhizal fungi (EcM) form a symbiosis with many tree species and provide a range of benefits to their host through their extraradical hyphal network. Although our understanding of the environmental drivers and large scale geographical variation of EcM communities in natural ecosystems is growing, our knowledge of EcM communities within and across urban areas is still limited. Here, we characterized EcM communities using Illumina miseq sequencing on 175 root samples of the urban tree Tilia tomentosa from three European cities, namely Leuven (Belgium), Strasbourg (France) and Porto (Portugal). We found strong differences in EcM richness and community composition between cities. Soil acidity, organic matter and moisture content were significantly associated with EcM community composition. In agreement, the explained variability in EcM communities was mostly attributed to general soil characteristics, whereas very little variation was explained by city and heavy metal pollution. Overall, our results suggest that EcM communities in urban areas are significantly associated with soil characteristics, while heavy metal pollution and biogeography had little or no impact. These findings deliver new insights into EcM distribution patterns in urban areas and contribute to specific inoculation strategies to improve urban tree vitality.

Monitoring of Freezing Dynamics in Trees: A Simple Phase Shift Causes Complexity.

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.

Can phenological models predict tree phenology accurately in the future? The unrevealed hurdle of endodormancy break.

The onset of the growing season of trees has been earlier by 2.3 days per decade during the last 40 years in temperate Europe because of global warming. The effect of temperature on plant phenology is, however, not linear because temperature has a dual effect on bud development. On one hand, low temperatures are necessary to break bud endodormancy, and, on the other hand, higher temperatures are necessary to promote bud cell growth afterward. Different process-based models have been developed in the last decades to predict the date of budbreak of woody species. They predict that global warming should delay or compromise endodormancy break at the species equatorward range limits leading to a delay or even impossibility to flower or set new leaves. These models are classically parameterized with flowering or budbreak dates only, with no information on the endodormancy break date because this information is very scarce. Here, we evaluated the efficiency of a set of phenological models to accurately predict the endodormancy break dates of three fruit trees. Our results show that models calibrated solely with budbreak dates usually do not accurately predict the endodormancy break date. Providing endodormancy break date for the model parameterization results in much more accurate prediction of this latter, with, however, a higher error than that on budbreak dates. Most importantly, we show that models not calibrated with endodormancy break dates can generate large discrepancies in forecasted budbreak dates when using climate scenarios as compared to models calibrated with endodormancy break dates. This discrepancy increases with mean annual temperature and is therefore the strongest after 2050 in the southernmost regions. Our results claim for the urgent need of massive measurements of endodormancy break dates in forest and fruit trees to yield more robust projections of phenological changes in a near future.

Cavitation and water fluxes driven by ice water potential in Juglans regia during freeze-thaw cycles.

Freeze-thaw cycles induce major hydraulic changes due to liquid-to-ice transition within tree stems. The very low water potential at the ice-liquid interface is crucial as it may cause lysis of living cells as well as water fluxes and embolism in sap conduits, which impacts whole tree-water relations. We investigated water fluxes induced by ice formation during freeze-thaw cycles in Juglans regia L. stems using four non-invasive and complementary approaches: a microdendrometer, magnetic resonance imaging, X-ray microtomography, and ultrasonic acoustic emissions analysis. When the temperature dropped, ice nucleation occurred, probably in the cambium or pith areas, inducing high water potential gradients within the stem. The water was therefore redistributed within the stem toward the ice front. We could thus observe dehydration of the bark's living cells leading to drastic shrinkage of this tissue, as well as high tension within wood conduits reaching the cavitation threshold in sap vessels. Ultrasonic emissions, which were strictly emitted only during freezing, indicated cavitation events (i.e. bubble formation) following ice formation in the xylem sap. However, embolism formation (i.e. bubble expansion) in stems was observed only on thawing via X-ray microtomography for the first time on the same sample. Ultrasonic emissions were detected during freezing and were not directly related to embolism formation. These results provide new insights into the complex process and dynamics of water movements and ice formation during freeze-thaw cycles in tree stems.

Effects of environmental factors and management practices on microclimate, winter physiology, and frost resistance in trees.

Freezing stress is one of the most important limiting factors determining the ecological distribution and production of tree species. Assessment of frost risk is, therefore, critical for forestry, fruit production, and horticulture. Frost risk is substantial when hazard (i.e., exposure to damaging freezing temperatures) intersects with vulnerability (i.e., frost sensitivity). Based on a large number of studies on frost resistance and frost occurrence, we highlight the complex interactive roles of environmental conditions, carbohydrates, and water status in frost risk development. To supersede the classical empirical relations used to model frost hardiness, we propose an integrated ecophysiologically-based framework of frost risk assessment. This framework details the individual or interactive roles of these factors, and how they are distributed in time and space at the individual-tree level (within-crown and across organs). Based on this general framework, we are able to highlight factors by which different environmental conditions (e.g., temperature, light, flood, and drought), and management practices (pruning, thinning, girdling, sheltering, water aspersion, irrigation, and fertilization) influence frost sensitivity and frost exposure of trees.

Ultrasonic emissions during ice nucleation and propagation in plant xylem.

Ultrasonic acoustic emission analysis enables nondestructive monitoring of damage in dehydrating or freezing plant xylem. We studied acoustic emissions (AE) in freezing stems during ice nucleation and propagation, by combining acoustic and infrared thermography techniques and controlling the ice nucleation point. Ultrasonic activity in freezing samples of Picea abies showed two distinct phases: the first on ice nucleation and propagation (up to 50 AE s(-1) ; reversely proportional to the distance to ice nucleation point), and the second (up to 2.5 AE s(-1) ) after dissipation of the exothermal heat. Identical patterns were observed in other conifer and angiosperm species. The complex AE patterns are explained by the low water potential of ice at the ice-liquid interface, which induced numerous and strong signals. Ice propagation velocities were estimated via AE (during the first phase) and infrared thermography. Acoustic activity ceased before the second phase probably because the exothermal heating and the volume expansion of ice caused decreasing tensions. Results indicate cavitation events at the ice front leading to AE. Ultrasonic emission analysis enabled new insights into the complex process of xylem freezing and might be used to monitor ice propagation in natura.

Characteristics of ultrasonic acoustic emissions from walnut branches during freeze-thaw-induced embolism formation.

Ultrasonic acoustic emission (UAE) methods have been applied for the detection of freeze-thaw-induced embolism formation in water conduits of tree species. Until now, however, the exact source(s) of UAE has not been identified especially in angiosperm species, in which xylem tissues are composed of diverse types of cells. In this study, UAE was recorded from excised branches of walnut (Juglans regia cv. Franquette) during freeze-thaw cycles, and attempts were made to characterize UAEs generated by cavitation events leading to embolism formation according to their properties. During freeze-thaw cycles, a large number of UAEs were generated from the sample segments. However, the cumulative numbers of total UAE during freeze-thawing were not correlated with the percentage loss of hydraulic conductivity after thawing, suggesting that the sources of UAE were not only cavitation leading to embolism formation in vessels. Among the UAEs, cumulative numbers of UAEs with absolute energy >10.0 fJ strongly correlated with the increase in percentage loss of hydraulic conductivity. The high absolute energy of the UAEs might reflect the formation of large bubbles in the large lumen of vessels. Therefore, UAEs generated by cavitation events in vessels during freeze-thawing might be distinguished from other signals according to their magnitudes of absolute energy. On the other hand, the freezing of xylem parenchyma cells was followed by a certain number of UAEs. These results indicate the possibility that UAE methods can be applied to the detection of both freeze-thaw-induced embolism and supercooling breakdown in parenchyma cells in xylem.