Losing ground: projections of climate-driven bloom shifts and their implications for the future of California's almond orchards.

Climate change is expected to impact the spring phenology of perennial trees, potentially altering the suitability of land for their cultivation. In this study, we investigate the effects of climate change on the bloom timing of almond orchards, focusing on California, the world's leading region for almond production. By analyzing historical climatic data, employing a model that considers hourly temperatures and fall non-structural carbohydrates to predict bloom dates, and examining various Coupled Model Intercomparison Project Phase 6 (CMIP6) scenarios, we assess the potential impacts of climate shifts on plant phenology and, consequently, on land suitability for almond farming. Our findings reveal that, within the next 30 years, the land suitable for almond production will not undergo significant changes. However, under unchanged emission scenarios, the available land to support almond orchard farming could decline between 48 to 73% by the end of the century. This reduction corresponds with an early shift in bloom time from the average Day of Year (DOY) 64 observed over the past 40 years to a projected earlier bloom between DOY 28-33 by 2100. These results emphasize the critical role climate shifts have in shaping future land use strategies for almond production in Central Valley, California. Consequently, understanding and addressing these factors is essential for the sustainable management and preservation of agricultural land, ensuring long-term food security and economic stability in the face of a rapidly changing climate.

Interactive effect of branch source-sink ratio and leaf aging on photosynthesis in pistachio.

Tree source-sink ratio has a predominant and complex impact on tree performance and can affect multiple physiological processes including vegetative and reproductive growth, water and nutrient use, photosynthesis, and productivity. In this study, we manipulated the branch level source-sink ratio by reduction of photosynthetic activity (partial branch defoliation) or thinning branch fruit load early in the growing season (after fruit set) in pistachio (Pistacia vera) trees. We then characterized the leaf photosynthetic light response curves through leaf aging. In addition, we determined changes in leaf non-structural carbohydrates (NSC) and nitrogen (N) concentrations. In leaves with high source-sink ratios, there was a gradual decrease in maximum net photosynthetic rate (ANmax) over the growing season, while in branches with low source-sink ratios, there was a sharp decline in ANmax in the first two weeks of August. Branches with high-sink showed an up-regulation (increase) in photosynthesis toward the end of July (at 1,500 growing degree days) during the period of rapid kernel growth rate and increased sink strength, with ANmax being about 7 µmol m-1 s-1 higher than in branches with low-sink. In August, low source-sink ratios precipitated leaf senescence, resulting in a drastic ANmax decline, from 25 to 8 µmol m-1 s-1 (70% drop in two weeks). This reduction was associated with the accumulation of NSC in the leaves from 20 to 30 mg g-1. The mechanisms of ANmax reduction differ between the two treatments. Lower photosynthetic rates of 8-10 µmol m-1 s-1 late in the season were associated with lower N levels in high-sink branches, suggesting N remobilization to the kernels. Lower photosynthesis late in the season was associated with lower respiration rates in low-source branches, indicating prioritization of assimilates to storage. These results can facilitate the adaptation of management practices to tree crop load changes in alternate bearing species.

Acclimation of interacting leaf surface traits affects foliar water uptake.

Absorption of water across the surfaces of leaves is an ecologically important aspect of tree physiology. Variation in foliar water uptake capacity depends on environmental conditions when traits associated with the uptake pathway respond to climatic signals. Using a series of experiments, we verify that water enters Sequoia sempervirens (D. Don) Endl. leaves by crossing the cuticle, and show that surface-trait acclimation alters the kinetic parameters of foliar water uptake. Under our experimental conditions, the cuticle was the primary pathway for water entry into the leaf. Exposure to climatic variation may induce surface acclimations, such as increased waxiness, that reduce water-film formation over stomata at the expense of dry-season foliar uptake rates. We found that water uptake is negatively associated with the interaction of leaf-surface wax coverage and stomatal density, and provide an accessible protocol to measure these key traits in Sequoia. Linking absorptive pathways and trait acclimation to physiological performance can provide a foundation for range-wide or genomic investigations of forest interactions with water and a mechanism-centered means to monitor canopy hydraulic parameters over time.

Tracheid buckling buys time, foliar water uptake pays it back: Coordination of leaf structure and function in tall redwood trees.

Tracheid buckling may protect leaves in the dynamic environments of forest canopies, where rapid intensifications of evaporative demand, such as those brought on by changes in light availability, can result in sudden increases in transpiration rate. While treetop leaves function in reliably direct light, leaves below the upper crown must tolerate rapid, thermally driven increases in evaporative demand. Using synchrotron-based X-ray microtomography, we visualized impacts of experimentally induced water stress and subsequent fogging on living cells in redwood leaves, adding ecological and functional context through crown-wide explorations of variation in leaf physiology and microclimate. Under drought, leaf transfusion tracheids buckle, releasing water that supplies sufficient temporal reserves for leaves to reduce stomatal conductance safely while stopping the further rise of tension. Tracheid buckling fraction decreases with height and is closely coordinated with transfusion tissue capacity and stomatal conductance to provide temporal reserves optimized for local variation in microclimate. Foliar water uptake fully restores collapsed and air-filled transfusion tracheids in leaves on excised shoots, suggesting that trees may use aerial water sources for recovery. In the intensely variable deep-crown environment, foliar water uptake can allow for repetitive cycles of tracheid buckling and unbuckling, protecting the tree from damaging levels of hydraulic tension and supporting leaf survival.

Shoot dimorphism enables Sequoia sempervirens to separate requirements for foliar water uptake and photosynthesis.

PREMISE: Trees in wet forests often have features that prevent water films from covering stomata and inhibiting gas exchange, while many trees in drier environments use foliar water uptake to reduce water stress. In forests with both wet and dry seasons, evergreen trees would benefit from producing leaves capable of balancing rainy-season photosynthesis with summertime water absorption. METHODS: Using samples collected from across the vertical gradient in tall redwood (Sequoia sempervirens) crowns, we estimated tree-level foliar water uptake and employed physics-based causative modeling to identify key functional traits that determine uptake potential by setting hydraulic resistance. RESULTS: We showed that Sequoia has two functionally distinct shoot morphotypes. While most shoots specialize in photosynthesis, the axial shoot type is capable of much greater foliar water uptake, and its within-crown distribution varies with latitude. A suite of leaf surface traits cause hydraulic resistance, leading to variation in uptake capacity among samples. CONCLUSIONS: Shoot dimorphism gives tall Sequoia trees the capacity to absorb up to 48 kg H2 O h-1 during the first hour of leaf wetting, ameliorating water stress while presumably maintaining high photosynthetic capacity year round. Geographic variation in shoot dimorphism suggests that plasticity in shoot-type distribution and leaf surface traits helps Sequoia maintain a dominate presence in both wet and dry forests.

The impact of non-structural carbohydrates (NSC) concentration on yield in Prunus dulcis, Pistacia vera, and Juglans regia.

Successful yield in orchards is the culmination of a series of events that start with plants entering dormancy with adequate energy reserves (non-structural carbohydrates; NSC). These NSC are responsible for the maintenance of activities during dormancy and extending onto the period of activeness. Using multi-year yield information and monthly NSC content in twigs, we show that high levels of carbohydrate in Prunus dulcis, Pistachio vera, and Juglans regia during the winter months are indeed associated with high yield, while high levels of the NSC in late summer often correlate with low yield. An evaluation of monthly NSC level importance on yield revealed that for P. dulcis high levels in February were a good predictor of yield and that low levels throughout summer were associated with high yield. In P. vera, high levels of NSC in December were best predictors of yield. J. regia exhibited peculiar patterns; while high pre-budbreak reserves were associated with high yields they only played a minor role in explaining crop, the most important months for predicting yields were June and July. Results suggest that NSC levels can serve as good predictors of orchard yield potential and should be monitored to inform orchard management.

Spring phenology is affected by fall non-structural carbohydrate concentration and winter sugar redistribution in three Mediterranean nut tree species.

Deciduous trees mostly rely on non-structural carbohydrates (NSC-soluble carbohydrates and starch) stored prior to dormancy to sustain both spring bloom and the initial phase of spring growth prior to the transition of leaves from sink to source. Winter management of NSC, their loss due to respiration, reallocation patterns and remobilization during spring, seems to be key to a timely and synchronous bloom. To assess tree dependence on NSC during dormancy, we tested whether the interruption of local branch NSC accumulation prior to dormancy by defoliation and the interruption of NSC translocation by phloem girdling influence spring phenology in three major deciduous Mediterranean nut crop species: Prunus dulcis (Mill.) D.A Webb, a hybrid between Pistacia integerrima (J. L. Stewart ex Brandis) and P. atlantica Desf. (referred to as P. integerrima), and Juglans regia L. Defoliation treatments had different effects on NSC concentration in different species depending on the time of application. However, despite the significant initial impact (increase or decrease of NSC concentration), with time this impact diminished resulting in overall similar concentrations between control and defoliated branches suggesting the presence of NSC reallocation during dormancy. Phloem girdling in P. dulcis and P. integerrima resulted in reduced export activity and greater NSC concentrations, while in J. regia girdling resulted in lower NSC concentrations, indicating that this species requires a net import of NSC during dormancy. Bud break was distinctly delayed by both defoliation and phloem girdling in all the three species, providing evidence of the significant roles that fall NSC accumulation and winter NSC management play in priming trees for spring growth resumption.

Winding up the bloom clock-do sugar levels at senescence determine how trees respond to winter temperature?

Variable winter temperatures cause a year-to-year discrepancy in the phenology of deciduous trees. This implies that an intrinsic 'winter clock' synchronizes bloom with the progression of winter to spring. The carbohydrate-temperature (C-T) model established a mechanistic association between carbohydrate metabolism in dormant trees and hourly winter temperatures. Using historical winter temperature and bloom times of Prunus dulcis (Mill.) D. A. Webb (almond), Malus domestica L. (apple), Pistachia vera L. (pistachio) and Juglans regia L. (walnut) in California and Washington states, we parametrized species-specific metabolic parameters to the C-T model. There was a sound fit between actual and projected bloom dates with a deviation (root mean square error) of 4-7 days in all species. The parameterized model enabled us to study how the observed variability in soluble carbohydrate concentrations at senescence (SC0) could affect bloom time. The C-T model projected that low SC0 could advance, while high SC0 possibly delays, the bloom of the early blooming almond trees. In contrast, high SC0 would advance the bloom of apple, pistachio and walnut trees. These novel projections suggest that after experimental validation, SC0 could guide post-harvest farming applications that affect fall carbohydrate accumulation to mediate the effects of climate shifts.

Unravelling foliar water uptake pathways: The contribution of stomata and the cuticle.

Plants can absorb water through their leaf surfaces, a phenomenon commonly referred to as foliar water uptake (FWU). Despite the physiological importance of FWU, the pathways and mechanisms underlying the process are not well known. Using a novel experimental approach, we parsed out the contribution of the stomata and the cuticle to FWU in two species with Mediterranean (Prunus dulcis) and temperate (Pyrus communis) origin. The hydraulic parameters of FWU were derived by analysing mass and water potential changes of leaves placed in a fog chamber. Leaves were previously treated with abscisic acid to force stomata to remain closed, with fusicoccin to remain open, and with water (control). Leaves with open stomata rehydrated two times faster than leaves with closed stomata and attained approximately three times higher maximum fluxes and hydraulic conductance. Based on FWU rates, we propose that rehydration through stomata occurs primarily via diffusion of water vapour rather than in liquid form even when leaf surfaces are covered with a water film. We discuss the potential mechanisms of FWU and the significance of both stomatal and cuticular pathways for plant productivity and survival.

Maximum CO2 diffusion inside leaves is limited by the scaling of cell size and genome size.

Maintaining high rates of photosynthesis in leaves requires efficient movement of CO2 from the atmosphere to the mesophyll cells inside the leaf where CO2 is converted into sugar. CO2 diffusion inside the leaf depends directly on the structure of the mesophyll cells and their surrounding airspace, which have been difficult to characterize because of their inherently three-dimensional organization. Yet faster CO2 diffusion inside the leaf was probably critical in elevating rates of photosynthesis that occurred among angiosperm lineages. Here we characterize the three-dimensional surface area of the leaf mesophyll across vascular plants. We show that genome size determines the sizes and packing densities of cells in all leaf tissues and that smaller cells enable more mesophyll surface area to be packed into the leaf volume, facilitating higher CO2 diffusion. Measurements and modelling revealed that the spongy mesophyll layer better facilitates gaseous phase diffusion while the palisade mesophyll layer better facilitates liquid-phase diffusion. Our results demonstrate that genome downsizing among the angiosperms was critical to restructuring the entire pathway of CO2 diffusion into and through the leaf, maintaining high rates of CO2 supply to the leaf mesophyll despite declining atmospheric CO2 levels during the Cretaceous.

Chemical inhibition of xylem cellular activity impedes the removal of drought-induced embolisms in poplar stems - new insights from micro-CT analysis.

In drought-stressed plants a coordinated cascade of chemical and transcriptional adjustments occurs at the same time as embolism formation. While these processes do not affect embolism formation during stress, they may prime stems for recovery during rehydration by modifying apoplast pH and increasing sugar concentration in the xylem sap. Here we show that in vivo treatments modifying apoplastic pH (stem infiltration with a pH buffer) or reducing stem metabolic activity (infiltration with sodium vanadate and sodium cyanide; plant exposure to carbon monoxide) can reduce sugar accumulation, thus disrupting or delaying the recovery process. Application of the vanadate treatment (NaVO3, an inhibitor of many ATPases) completely halted recovery from drought-induced embolism for up to 24 h after re-irrigation, while partial recovery was observed in vivo in control plants using X-ray microcomputed tomography. Our results suggest that stem hydraulic recovery in poplar is a biological, energy-dependent process that coincides with accumulation of sugars in the apoplast during stress. Recovery and damage are spatially coordinated, with embolism formation occurring from the inside out and refilling from the outside in. The outside-in pattern highlights the importance of xylem proximity to the sugars within the phloem to the embolism recovery process.

Comparison of phenological traits, growth patterns, and seasonal dynamics of non-structural carbohydrate in Mediterranean tree crop species.

Despite non-structural carbohydrate (NSC) importance for tree productivity and resilience, little is known about their seasonal regulations and trade-off with growth and reproduction. We characterize the seasonal dynamics of NSC in relation to the aboveground phenology and temporal growth patterns of three deciduous Mediterranean species: almond (Prunus dulcis (Mill.) D. A. Webb), walnut (Juglans regia L.) and pistachio (Pistacia vera L.). Seasonal dynamics of NSC were synchronous between wood tissues from trunk, branches and twigs. Almond had almost identical levels and patterns of NSC variation in twigs, branches and trunks whereas pistachio and walnut exhibited clear concentration differences among plant parts whereby twigs had the highest and most variable NSC concentration, followed by branches and then trunk. While phenology had a significant influence on NSC seasonal trends, there was no clear trade-off between NSC storage and growth suggesting that both were similarly strong sinks for NSC. A temporal trade-off observed at the seasonal scale was influenced by the phenology of the species. We propose that late senescing species experience C allocation trade-off at the end of the growing season because of C-limiting thermal conditions and priority allocation to storage in order to survive winter.

Ray fractions and carbohydrate dynamics of tree species along a 2750 m elevation gradient indicate climate response, not spatial storage limitation.

Parenchyma cells in the xylem store nonstructural carbohydrates (NSC), providing reserves of energy that fuel woody perennials through periods of stress and/or limitations to photosynthesis. If the capacity for storage is subject to selection, then the fraction of wood occupied by living parenchyma should increase towards stressful environments. Ray parenchyma fraction (RPF) and seasonal NSC dynamics were quantified for 12 conifers and three oaks along a transect spanning warm dry foothills (500 m above sea level) to cold wet treeline (3250 m asl) in California's central Sierra Nevada. Mean RPF was lower for both conifer and oak species with warmer dryer ranges. RPF variability increased with elevation or in relation to associated climatic variables in conifers - treeline-dominant Pinus albicaulis had the lowest mean RPF measured (c. 3.7%), but the highest environmentally standardized variability index. Conifer RPF variability was explained by environment, increasing predominantly towards cooler wetter range edges. In oaks, NSC was explained by environment - values increasing for evergreen and decreasing for deciduous oaks with elevation. Lastly, all species surveyed appear to prioritize filling available RPF with sugar to achieve molarities that balance reasonable tensions over starch to maximize stored carbon. RPF responds to environment but is unlikely to spatially constrain NSC storage.

Measuring Phloem Transport Velocity on a Tissue Level Using a Phloem-Mobile Dye.

Here we describe an in vivo dye-tracking method for measuring phloem transport velocity in seedlings, leaves and petioles and potentially other translucent plant tissues. The method requires measurement of the fluorescent signal of a phloem-mobile fluorescent dye using sensitive photo-sensors placed external to the plant. Following dye application, velocity is determined using laser fluorescence bleaching and measuring the time it takes for the bleach front to reach a light sensor. This method can be used to measure phloem transport velocity on intact plants with minimal disturbance and has a potential to be used under a variety of growth conditions and in the field. Because there are large differences among species in their anatomy, this method should be optimized to individual plants and tissue types.

Sodium interception by xylem parenchyma and chloride recirculation in phloem may augment exclusion in the salt tolerant Pistacia genus: context for salinity studies on tree crops.

Working in tandem with root exclusion, stems may provide salt-tolerant woody perennials with some additional capacity to restrict sodium (Na) and chloride (Cl) accumulation in leaves. The Pistacia genus, falling at the nexus of salt tolerance and human intervention, provided an ideal set of organisms for studying the influences of both variable root exclusion and potentially variable discontinuities at the bud union on stem processes. In three experiments covering a wide range of salt concentrations (0 to 150 mM NaCl) and tree ages (1, 2 and 10 years) as well as nine rootstock-scion combinations we show that proportional exclusion of both Na and Cl reached up to ~85% efficacy, but efficacy varied by both rootstock and budding treatment. Effective Na exclusion was augmented by significant retrieval of Na from the xylem sap, as evidenced by declines in the Na concentrations of both sap and wood tissue along the transpiration stream. However, while we observed little to no differences between the concentrations of the two ions in leaves, analogous declines in sap concentrations of Cl were not observed. We conclude that some parallel but separate mechanism must be acting on Cl to provide leaf protection from toxicity specific to this ion and suggest that this mechanism is recirculation of Cl in the phloem. The presented findings underline the importance of holistic assessments of salt tolerance in woody perennials. In particular, greater emphasis might be placed on the dynamics of salt sequestration in the significant storage volumes offered by the stems of woody perennials and on the potential for phloem discontinuity introduced with a bud/graft union.

Priming xylem for stress recovery depends on coordinated activity of sugar metabolic pathways and changes in xylem sap pH.

Some plant species are capable of significant reduction of xylem embolism during recovery from drought despite stem water potential remains negative. However, the functional biology underlying this process is elusive. We subjected poplar trees to drought stress followed by a period of recovery. Water potential, hydraulic conductivity, gas exchange, xylem sap pH, and carbohydrate content in sap and woody stems were monitored in combination with an analysis of carbohydrate metabolism, enzyme activity, and expression of genes involved in sugar metabolic and transport pathways. Drought resulted in an alteration of differential partitioning between starch and soluble sugars. Upon stress, an increase in the starch degradation rate and the overexpression of sugar symporter genes promoted the efflux of disaccharides (mostly maltose and sucrose) to the apoplast. In turn, the efflux activity of the sugar-proton cotransporters caused a drop in xylem pH. The newly acidic environment induced the activity of apoplastic invertases leading to the accumulation of monosaccharides in the apoplast, thus providing the main osmoticum necessary for recovery. During drought and recovery, a complex network of coordinated molecular and biochemical signals was activated at the interface between xylem and parenchyma cells that appeared to prime the xylem for hydraulic recovery.

The prospects for constraining productivity through time with the whole-plant physiology of fossils.

Anatomically preserved fossils allow estimation of hydraulic parameters, potentially providing constraints on interpreting whole-plant physiology. However, different organ systems have typically been considered in isolation - a problem given common mismatches of high and low conductance components coupled in the hydraulic path of the same plant. A recent paper addressed the issue of how to handle resistance mismatches in fossil plant hydraulics, focusing on Carboniferous medullosan seed plants and arborescent lycopsids. Among other problems, however, a fundamental error was made: the transpiration stream consists of resistances in series (where resistances are additive and the component with the largest resistance can dominate the behavior of the system), but emphasis was instead placed on the lowest resistance, effectively treating the system as resistances in parallel (where the component with the smallest resistance will dominate the behavior). Instead of possessing high assimilation capacities to match high specific stem conductances, it is argued here that individual high conductance components in these Paleozoic plants are nonetheless associated with low whole-plant productivity, just as can be commonly seen in living plants. Resolution of how to handle these issues may have broad implications for the Earth system including geobiological feedbacks to rock weathering, atmospheric composition, and climate.

Diurnal Variation in Nonstructural Carbohydrate Storage in Trees: Remobilization and Vertical Mixing.

Nonstructural carbohydrate (NSC) storage plays a critical role in tree function and survival, but understanding and predicting local NSC storage dynamics is challenging because NSC storage pools are dispersed throughout the complex architecture of trees and continuously exchange carbon between source and sink organs at different time scales. To address these knowledge gaps, characterization and understanding of NSC diel variation are necessary. Here, we analyzed diurnal NSC dynamics in the overall architecture of almond (Prunus dulcis) trees. We also analyzed the allocation of newly assimilated carbon using isotopic labeling. We show that both components of NSC (i.e. soluble carbohydrates and starch) are highly dynamic at the diurnal time scale and that these trends are influenced by tissue type, age, and/or position within the canopy. In leaves, starch reserves can be depleted completely during the night, while woody tissue starch levels may vary by more than 50% over a daily cycle. Recently assimilated carbon showed a dispersed downward allocation across the entire tree. NSC diurnal fluctuations within the tree's structure in combination with dispersed carbon allocation patterns provide evidence for the presence of vertical mixing and suggest that the xylem acts as a secondary NSC redistribution pathway.

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.