Local carbon reserves are insufficient for phloem terpene induction during drought in Pinus edulis in response to bark beetle-associated fungi.

Stomatal closure during drought inhibits carbon uptake and may reduce a tree's defensive capacity. Limited carbon availability during drought may increase a tree's mortality risk, particularly if drought constrains trees' capacity to rapidly produce defenses during biotic attack. We parameterized a new model of conifer defense using physiological data on carbon reserves and chemical defenses before and after a simulated bark beetle attack in mature Pinus edulis under experimental drought. Attack was simulated using inoculations with a consistent bluestain fungus (Ophiostoma sp.) of Ips confusus, the main bark beetle colonizing this tree, to induce a defensive response. Trees with more carbon reserves produced more defenses but measured phloem carbon reserves only accounted for c. 23% of the induced defensive response. Our model predicted universal mortality if local reserves alone supported defense production, suggesting substantial remobilization and transport of stored resin or carbon reserves to the inoculation site. Our results show that de novo terpene synthesis represents only a fraction of the total measured phloem terpenes in P. edulis following fungal inoculation. Without direct attribution of phloem terpene concentrations to available carbon, many studies may be overestimating the scale and importance of de novo terpene synthesis in a tree's induced defense response.

Carbon starvation following a decade of experimental drought consumes old reserves in Pinus edulis.

Shifts in the age or turnover time of non-structural carbohydrates (NSC) may underlie changes in tree growth under long-term increases in drought stress associated with climate change. But NSC responses to drought are challenging to quantify, due in part to large NSC stores in trees and subsequently long response times of NSC to climate variation. We measured NSC age (Δ14 C) along with a suite of ecophysiological metrics in Pinus edulis trees experiencing either extreme short-term drought (-90% ambient precipitation plot, 2020-2021) or a decade of severe drought (-45% plot, 2010-2021). We tested the hypothesis that carbon starvation - consumption exceeding synthesis and storage - increases the age of sapwood NSC. One year of extreme drought had no impact on NSC pool size or age, despite significant reductions in predawn water potential, photosynthetic rates/capacity, and twig and needle growth. By contrast, long-term drought halved the age of the sapwood NSC pool, coupled with reductions in sapwood starch concentrations (-75%), basal area increment (-39%), and bole respiration rates (-28%). Our results suggest carbon starvation takes time, as tree carbon reserves appear resilient to extreme disturbance in the short term. However, after a decade of drought, trees apparently consumed old stored NSC to support metabolism.

Fire intensity impacts on physiological performance and mortality in Pinus monticola and Pseudotsuga menziesii saplings: a dose-response analysis.

Fire is a major cause of tree injury and mortality worldwide, yet our current understanding of fire effects is largely based on ocular estimates of stem charring and foliage discoloration, which are error prone and provide little information on underlying tree function. Accurate quantification of physiological performance is a research and forest management need, given that declining performance could help identify mechanisms of-and serve as an early warning sign for-mortality. Many previous efforts have been hampered by the inability to quantify the heat flux that a tree experiences during a fire, given its highly variable nature in space and time. In this study, we used a dose-response approach to elucidate fire impacts by subjecting Pinus monticola var. minima Lemmon and Pseudotsuga menziesii (Mirb.) Franco var. glauca (Beissn.) Franco saplings to surface fires of varying intensity doses and measuring short-term post-fire physiological performance in photosynthetic rate and chlorophyll fluorescence. We also evaluated the ability of spectral reflectance indices to quantify change in physiological performance at the individual tree crown and stand scales. Although physiological performance in both P. monticola and P. menziesii declined with increasing fire intensity, P. monticola maintained a greater photosynthetic rate and higher chlorophyll fluorescence at higher doses, for longer after the fire. Pinus monticola also had complete survival at lower fire intensity doses, whereas P. menziesii had some mortality at all doses, implying higher fire resistance for P. monticola at this life stage. Generally, individual-scale spectral indices were more accurate at quantifying physiological performance than those acquired at the stand-scale. The Photochemical Reflectance Index outperformed other indices at quantifying photosynthesis and chlorophyll fluorescence, highlighting its potential use to quantify crown scale physiological performance. Spectral indices that incorporated near-infrared and shortwave infrared reflectance, such as the Normalized Burn Ratio, were accurate at characterizing stand-scale mortality. The results from this study were included in a conifer cross-comparison using physiology and mortality data from other dose-response studies. The comparison highlights the close evolutionary relationship between fire and species within the Pinus genus, assessed to date, given the high survivorship of Pinus species at lower fire intensities versus other conifers.

No carbon storage in growth-limited trees in a semi-arid woodland.

Plant survival depends on a balance between carbon supply and demand. When carbon supply becomes limited, plants buffer demand by using stored carbohydrates (sugar and starch). During drought, NSCs (non-structural carbohydrates) may accumulate if growth stops before photosynthesis. This expectation is pervasive, yet few studies have combined simultaneous measurements of drought, photosynthesis, growth, and carbon storage to test this. Using a field experiment with mature trees in a semi-arid woodland, we show that growth and photosynthesis slow in parallel as [Formula: see text] declines, preventing carbon storage in two species of conifer (J. monosperma and P. edulis). During experimental drought, growth and photosynthesis were frequently co-limited. Our results point to an alternative perspective on how plants use carbon that views growth and photosynthesis as independent processes both regulated by water availability.

Death from hunger or thirst? Phloem death, rather than xylem hydraulic failure, as a driver of fire-induced conifer mortality.

Disruption of photosynthesis and carbon transport due to damage to the tree crown and stem cambial cells, respectively, can cause tree mortality. It has recently been proposed that fire-induced dysfunction of xylem plays an important role in tree mortality. Here, we simultaneously tested the impact of a lethal fire dose on nonstructural carbohydrates (NSCs) and xylem hydraulics in Pinus ponderosa saplings. Saplings were burned with a known lethal fire dose. Nonstructural carbohydrates were assessed in needles, main stems, roots and whole plants, and xylem hydraulic conductivity was measured in the main stems up to 29 d postfire. Photosynthesis and whole plant NSCs declined postfire. Additionally, all burned saplings showed 100% phloem/cambium necrosis, and roots of burned saplings had reduced NSCs compared to unburned and defoliated saplings. We further show that, contrary to patterns observed with NSCs, water transport was unchanged by fire and there was no evidence of xylem deformation in saplings that experienced a lethal dose of heat from fire. We conclude that phloem and cambium mortality, and not hydraulic failure, were probably the causes of death in these saplings. These findings advance our understanding of the physiological response to fire-induced injuries in conifer trees.

The osmotic balancing act: When sugars matter for more than metabolism in woody plants.

Sugars and other non-structural carbohydrates are known to serve as currency in plants, to either fuel metabolic activities or as storage for later use. They can also serve non-metabolic purposes of osmoregulation and cryoprotection, especially in perennial woody plants.

Global field observations of tree die-off reveal hotter-drought fingerprint for Earth's forests.

Earth's forests face grave challenges in the Anthropocene, including hotter droughts increasingly associated with widespread forest die-off events. But despite the vital importance of forests to global ecosystem services, their fates in a warming world remain highly uncertain. Lacking is quantitative determination of commonality in climate anomalies associated with pulses of tree mortality-from published, field-documented mortality events-required for understanding the role of extreme climate events in overall global tree die-off patterns. Here we established a geo-referenced global database documenting climate-induced mortality events spanning all tree-supporting biomes and continents, from 154 peer-reviewed studies since 1970. Our analysis quantifies a global "hotter-drought fingerprint" from these tree-mortality sites-effectively a hotter and drier climate signal for tree mortality-across 675 locations encompassing 1,303 plots. Frequency of these observed mortality-year climate conditions strongly increases nonlinearly under projected warming. Our database also provides initial footing for further community-developed, quantitative, ground-based monitoring of global tree mortality.

Hotter droughts alter resource allocation to chemical defenses in piñon pine.

Heat and drought affect plant chemical defenses and thereby plant susceptibility to pests and pathogens. Monoterpenes are of particular importance for conifers as they play critical roles in defense against bark beetles. To date, work seeking to understand the impacts of heat and drought on monoterpenes has primarily focused on young potted seedlings, leaving it unclear how older age classes that are more vulnerable to bark beetles might respond to stress. Furthermore, we lack a clear picture of what carbon resources might be prioritized to support monoterpene synthesis under drought stress. To address this, we measured needle and woody tissue monoterpene concentrations and physiological variables simultaneously from mature piñon pines (Pinus edulis) from a unique temperature and drought manipulation field experiment. While heat had no effect on total monoterpene concentrations, trees under combined heat and drought stress exhibited ~ 85% and 35% increases in needle and woody tissue, respectively, over multiple years. Plant physiological variables like maximum photosynthesis each explained less than 10% of the variation in total monoterpenes for both tissue types while starch and glucose + fructose measured 1-month prior explained ~ 45% and 60% of the variation in woody tissue total monoterpene concentrations. Although total monoterpenes increased under combined stress, some key monoterpenes with known roles in bark beetle ecology decreased. These shifts may make trees more favorable for bark beetle attack rather than well defended, which one might conclude if only considering total monoterpene concentrations. Our results point to cumulative and synergistic effects of heat and drought that may reprioritize carbon allocation of specific non-structural carbohydrates toward defense.

Seawater exposure causes hydraulic damage in dying Sitka-spruce trees.

Sea-level rise is one of the most critical challenges facing coastal ecosystems under climate change. Observations of elevated tree mortality in global coastal forests are increasing, but important knowledge gaps persist concerning the mechanism of salinity stress-induced nonhalophytic tree mortality. We monitored progressive mortality and associated gas exchange and hydraulic shifts in Sitka-spruce (Picea sitchensis) trees located within a salinity gradient under an ecosystem-scale change of seawater exposure in Washington State, USA. Percentage of live foliated crown (PLFC) decreased and tree mortality increased with increasing soil salinity during the study period. A strong reduction in gas exchange and xylem hydraulic conductivity (Ks) occurred during tree death, with an increase in the percentage loss of conductivity (PLC) and turgor loss point (πtlp). Hydraulic and osmotic shifts reflected that hydraulic function declined from seawater exposure, and dying trees were unable to support osmotic adjustment. Constrained gas exchange was strongly related to hydraulic damage at both stem and leaf levels. Significant correlations between foliar sodium (Na+) concentration and gas exchange and key hydraulic parameters (Ks, PLC, and πtlp) suggest that cellular injury related to the toxic effects of ion accumulation impacted the physiology of these dying trees. This study provides evidence of toxic effects on the cellular function that manifests in all aspects of plant functioning, leading to unfavourable osmotic and hydraulic conditions.

Foliar respiration is related to photosynthetic, growth and carbohydrate response to experimental drought and elevated temperature.

Short-term plant respiration (R) increases exponentially with rising temperature, but drought could reduce respiration by reducing growth and metabolism. Acclimation may alter these responses. We examined if species with different drought responses would differ in foliar R response to +4.8°C temperature and -45% precipitation in a field experiment with mature piñon and juniper trees, and if any differences between species were related to differences in photosynthesis rates, shoot growth and nonstructural carbohydrates (NSCs). Short-term foliar R had a Q10 of 1.6 for piñon and 2.6 for juniper. Piñon foliar R did not respond to the +4.8°C temperatures, but R increased 1.4× for juniper. Across treatments, piñon foliage had higher growth, lower NSC content, 29% lower photosynthesis rates, and 44% lower R than juniper. Removing 45% precipitation had little impact on R for either species. Species differences in the response of R under elevated temperature were related to substrate availability and stomatal response to leaf water potential. Despite not acclimating to the higher temperature and having higher R than piñon, greater substrate availability in juniper suggests it could supply respiratory demand for much longer than piñon. Species responses will be critical in ecosystem response to a warmer climate.

Short- and long-term effects of fire on stem hydraulics in Pinus ponderosa saplings.

Understanding tree physiological responses to fire is needed to accurately model post-fire carbon processes and inform management decisions. Given trees can die immediately or at extended time periods after fire, we combined two experiments to assess the short- (one-day) and long-term (21-months) fire effects on Pinus ponderosa sapling water transport. Native percentage loss of conductivity (nPLC), vulnerability to cavitation and xylem anatomy were assessed in unburned and burned saplings at lethal and non-lethal fire intensities. Fire did not cause any impact on nPLC and xylem cell wall structure in either experiment. However, surviving saplings evaluated 21-months post-fire were more vulnerable to cavitation. Our anatomical analysis in the long-term experiment showed that new xylem growth adjacent to fire scars had irregular-shaped tracheids and many parenchyma cells. Given conduit cell wall deformation was not observed in the long-term experiment, we suggest that the irregularity of newly grown xylem cells nearby fire wounds may be responsible for decreasing resistance to embolism in burned plants. Our findings suggest that hydraulic failure is not the main short-term physiological driver of mortality for Pinus ponderosa saplings. However, the decrease in embolism resistance in fire-wounded saplings could contribute to sapling mortality in the years following fire.

Drought Tolerance and Competition in Eastern Redcedar (Juniperus virginiana) Encroachment of the Oak-Dominated Cross Timbers.

On the dry, western edge of the eastern deciduous forest of the USA (Cross Timbers), the drought-tolerant, evergreen eastern redcedar (Juniperus virginiana) is encroaching into post oak- (Quercus stellata) dominated woodlands. The overall goal of this study was to examine whether the drought tolerance strategies of eastern redcedar provide it a competitive advantage over post oak and whether this is a key attribute facilitating its successful establishment in the Cross Timbers. Specifically, we assessed xylem water potential and leaf gas exchange of these two species growing in single-species stands and in a mixed-species stand. We found that both species exhibit a similar degree of isohydry and close their stomates to the same extent in response to declining xylem water potentials. Both species had similar relative reductions in gas exchange in response to drought, despite differences in xylem anatomy. However, post oak had leaf-level gas exchange rates approximately 5× greater than eastern redcedar during periods of high moisture availability. Therefore, it did not appear that eastern redcedar encroachment into an oak-dominated forest is facilitated by growing season differences in carbon gain, although evergreen eastern redcedar can conduct gas exchange year-round when conditions are favorable while post oak is deciduous. We found that volumetric soil water content (0-45 cm) was lower in the pure eastern redcedar stand than the mixed-species or pure post oak stand which may indicate that eastern redcedar may experience favorable soil moisture conditions when encroaching into open oak woodlands. Moreover, water potentials in eastern redcedar tended to be more negative in pure stands compared to the mixed stand. Our results suggest the two species may be using water from different depths, reducing competition. Overall, our findings indicate that eastern redcedar encroachment into formerly oak-dominated Cross Timbers forests likely will continue under moderate drought, in the absence of fire, with consequences for water budgets, carbon cycling, grazing forage, wildlife habitat, and wildfire risk.

Conifers depend on established roots during drought: results from a coupled model of carbon allocation and hydraulics.

Trees may survive prolonged droughts by shifting water uptake to reliable water sources, but it is unknown if the dominant mechanism involves activating existing roots or growing new roots during drought, or some combination of the two. To gain mechanistic insights on this unknown, a dynamic root-hydraulic modeling framework was developed that set up a feedback between hydraulic controls over carbon allocation and the role of root growth on soil-plant hydraulics. The new model was tested using a 5 yr drought/heat field experiment on an established piñon-juniper stand with root access to bedrock groundwater. Owing to the high carbon cost per unit root area, modeled trees initialized without adequate bedrock groundwater access experienced potentially lethal declines in water potential, while all of the experimental trees maintained nonlethal water potentials. Simulated trees were unable to grow roots rapidly enough to mediate the hydraulic stress, particularly during warm droughts. Alternatively, modeled trees initiated with root access to bedrock groundwater matched the hydraulics of the experimental trees by increasing their water uptake from bedrock groundwater when soil layers dried out. Therefore, the modeling framework identified a critical mechanism for drought response that required trees to shift water uptake among existing roots rather than growing new roots.

Tree defence and bark beetles in a drying world: carbon partitioning, functioning and modelling.

Drought has promoted large-scale, insect-induced tree mortality in recent years, with severe consequences for ecosystem function, atmospheric processes, sustainable resources and global biogeochemical cycles. However, the physiological linkages among drought, tree defences, and insect outbreaks are still uncertain, hindering our ability to accurately predict tree mortality under on-going climate change. Here we propose an interdisciplinary research agenda for addressing these crucial knowledge gaps. Our framework includes field manipulations, laboratory experiments, and modelling of insect and vegetation dynamics, and focuses on how drought affects interactions between conifer trees and bark beetles. We build upon existing theory and examine several key assumptions: (1) there is a trade-off in tree carbon investment between primary and secondary metabolites (e.g. growth vs defence); (2) secondary metabolites are one of the main component of tree defence against bark beetles and associated microbes; and (3) implementing conifer-bark beetle interactions in current models improves predictions of forest disturbance in a changing climate. Our framework provides guidance for addressing a major shortcoming in current implementations of large-scale vegetation models, the under-representation of insect-induced tree mortality.

Divergences in hydraulic conductance and anatomical traits of stems and leaves in three temperate tree species coping with drought, N addition and their interactions.

Drought and nitrogen (N) addition have been shown to affect tree hydraulic traits, but few studies have been made on their interactions across species with different wood types or leaf forms. We examined the responses of hydraulic conductance and xylem anatomical traits of Quercus mongolica (ring porous with simple leaves), Fraxinus mandshurica (ring porous with compound leaves) and Tilia amurensis (diffuse porous with simple leaves) to drought, N addition and their interactions. Drought stress decreased current-year xylem-specific conductivity in stems (Ksx) and leaf hydraulic conductance (Kleaf ), but N addition affected Ksx and Kleaf differently among species and watering regimes. These divergent effects were associated with different responses of anatomical traits and leaf forms. Higher mean vessel diameter in stems and lower vessel density in leaves were observed with N addition. The three-way interactive effects of drought, N addition and tree species were significant for most values of anatomical traits. These results were also reflected in large differences in vessel diameter and density among species with different wood types or leaf forms. The two-way interactive effects of drought and N addition were significant on Kleaf and predawn water potential, but not Ksx, indicating that leaves were more sensitive than stems to a combination of drought stress and N addition. Our results provide mechanistic insight into the variable responses of xylem water transport to the interactions of drought and N availability.

Embolism recovery strategies and nocturnal water loss across species influenced by biogeographic origin.

Drought-induced tree mortality is expected to increase in future climates with the potential for significant consequences to global carbon, water, and energy cycles. Xylem embolism can accumulate to lethal levels during drought, but species that can refill embolized xylem and recover hydraulic function may be able to avoid mortality. Yet the potential controls of embolism recovery, including cross-biome patterns and plant traits such as nonstructural carbohydrates (NSCs), hydraulic traits, and nocturnal stomatal conductance, are unknown. We exposed eight plant species, originating from mesic (tropical and temperate) and semi-arid environments, to drought under ambient and elevated CO2 levels, and assessed recovery from embolism following rewatering. We found a positive association between xylem recovery and NSCs, and, surprisingly, a positive relationship between xylem recovery and nocturnal stomatal conductance. Arid-zone species exhibited greater embolism recovery than mesic zone species. Our results indicate that nighttime stomatal conductance often assumed to be a wasteful use of water, may in fact be a key part of plant drought responses, and contribute to drought survival. Findings suggested distinct biome-specific responses that partially depended on species climate-of-origin precipitation or aridity index, which allowed some species to recover from xylem embolism. These findings provide improved understanding required to predict the response of diverse plant communities to drought. Our results provide a framework for predicting future vegetation shifts in response to climate change.

Dead or dying? Quantifying the point of no return from hydraulic failure in drought-induced tree mortality.

Determining physiological mechanisms and thresholds for climate-driven tree die-off could help improve global predictions of future terrestrial carbon sinks. We directly tested for the lethal threshold in hydraulic failure - an inability to move water due to drought-induced xylem embolism - in a pine sapling experiment. In a glasshouse experiment, we exposed loblolly pine (Pinus taeda) saplings (n = 83) to drought-induced water stress ranging from mild to lethal. Before rewatering to relieve drought stress, we measured native hydraulic conductivity and foliar color change. We monitored all measured individuals for survival or mortality. We found a lethal threshold at 80% loss of hydraulic conductivity - a point of hydraulic failure beyond which it is more likely trees will die, than survive, and describe mortality risk across all levels of water stress. Foliar color changes lagged behind hydraulic failure - best predicting when trees had been dead for some time, rather than when they were dying. Our direct measurement of native conductivity, while monitoring the same individuals for survival or mortality, quantifies a continuous probability of mortality risk from hydraulic failure. Predicting tree die-off events and understanding the mechanism involved requires knowledge not only of when trees are dead, but when they begin dying - having passed the point of no return.

Belowground annual ring growth coordinates with aboveground phenology and timing of carbon storage in two tallgrass prairie forb species.

PREMISE OF THE STUDY: Herb chronology, the study of belowground annual growth rings in perennial forbs, has much potential as a tool for monitoring plant growth as a function of environment. To harness this potential, understanding of the coordination between ring ontogeny, aboveground phenology, and the temporal allocation of carbon products belowground in herbaceous forbs must be improved. METHODS: We investigated these relationships in two southern United States tallgrass prairie perennial forb species, Asclepias viridis and Lespedeza stuevei, making monthly excavations for a year. KEY RESULTS: Belowground xylogenesis began when starch reserves were at their seasonal low in the spring as shoots reached maximum height. The highest relative radial growth of the ring occurred concurrently with replenishment of root starch reserves in early summer. Xylogenesis concluded with leaf senescence in late summer and belowground starch reserves near saturation. CONCLUSIONS: By demonstrating that ring ontogeny is tied to early summer starch replenishment, our results illustrate the mechanisms behind previous findings where ring width was highly correlated with summer climatic conditions for these two species. This study provides a new physiological link between how ring chronologies in herbs often accord with growing-season environment; further dissecting this phenomenon is vital in unlocking the potential of herb chronology.