Xylem embolism induced by freeze-thaw and drought are influenced by different anatomical traits in subtropical montane evergreen angiosperm trees.

Subtropical evergreen broadleaved forests distributed in montane zones of southern China experience seasonal droughts and winter frost. Previously, studies have recognized that xylem anatomy is a determinant of its vulnerability to embolism caused by drought and freezing events. We hypothesized that there is a coordination of xylem resistance to freeze-thaw and drought-induced embolism for the subtropical montane evergreen broadleaved tree species because they are influenced by common xylem structural traits (e.g., vessel diameter). We examined the branch xylem anatomy, resistance to drought-induced embolism (P50), and the percent loss of branch hydraulic conductivity after a severe winter frost (PLCwinter) for 15 evergreen broadleaved tree species in a montane forest in South China. Our results showed that P50 of the studied species ranged from -2.81 to -5.13 MPa, which was not associated with most xylem anatomical properties except for the axial parenchyma-to-vessel connectivity. These tree species differed substantially in PLCwinter, ranging from 0% to 76.41%. PLCwinter was positively related to vessel diameter and negatively related to vessel density, vessel group index, and vessel-to-vessel connectivity, but no coordination with P50. This study suggests that hydraulic adaptation to frost is important to determine the distributional limit of subtropical montane evergreen woody angiosperms.

An appreciation of apex-to-base variation in xylem traits will lead to more precise understanding of xylem phenotypic plasticity.

Xylem air embolism is the primary cause of drought-related tree mortality. Phenotypic plasticity of xylem traits is key for species acclimation to environmental variability and evolution. It is widely believed that plants increase xylem embolism resistance in response to drought. However, I argue that this hypothesis, based on extensive literature, relies on sampling methods that overlook predictable anatomical patterns, potentially biasing our understanding of acclimation and adaptation strategies.

Weak link or strong foundation? Vulnerability of fine root networks and stems to xylem embolism.

Resolving the position of roots in the whole-plant hierarchy of drought-induced xylem embolism resistance is fundamental for predicting when species become isolated from soil water resources. Published research generally suggests that roots are the most vulnerable organ of the plant vascular system, although estimates vary significantly. However, our knowledge of root embolism excludes the fine roots (< 2 mm diameter) that form the bulk of total absorptive surface area of the root network for water and nutrient uptake. We measured fine root and stem xylem vulnerability in 10 vascular plant species from the major land plant clades (five angiosperms, three conifers, a fern and lycophyte), using standardised in situ methods (Optical Methods and MicroCT). Mean fine root embolism resistance across the network matched or exceeded stems in all study species. In six of these species (one fern, one lycophyte, three conifers and one angiosperm), fine roots were significantly more embolism resistant than stems. No clear relationship was found between root xylem conduit diameter and vulnerability. These results provide insight into the resistance of the plant hydraulic pathway at the site of water and nutrient uptake, and challenge the long-standing assumption that fine roots are more vulnerable than stems.

Growing at the arid edge: Anatomical variations in leaves are more extensive than in stems of five Mediterranean species across contrasting moisture regimes.

PREMISE: Increasing aridity in the Mediterranean region affects ecosystems and plant life. Various anatomical changes in plants help them cope with dry conditions. This study focused on anatomical differences in leaves and xylem of five co-occurring Mediterranean plant species namely Quercus calliprinos, Pistacia palaestina, Pistacia lentiscus, Rhamnus lycioides, and Phillyrea latifolia in wet and dry sites. METHODS: Stomatal density, stomatal length, leaf mass area, lamina composition, percentage of intercellular air spaces, and mesophyll cell area in leaves of plants in wet and dry sites were analyzed. Xylem anatomy was assessed through vessel length and area in branches. RESULTS: In the dry site, three species had increased stomatal density and decreased stomatal length. Four species had increased palisade mesophyll and reduced air space volume. In contrast, phenotypic changes in the xylem were less pronounced; vessel length was unaffected by site conditions, but vessel diameter decreased in two species. Intercellular air spaces proved to be the most dynamic anatomical feature. Quercus calliprinos had the most extensive anatomical changes; Rhamnus lycioides had only minor changes. All these changes were observed in comparison to the species in the wet site. CONCLUSIONS: This study elucidated variations in anatomical responses in leaves among co-occurring Mediterranean plant species and identified the most dynamic traits. Understanding these adaptations provides valuable insights into the ability of plants to thrive under changing climate conditions.

Born suckers: the cibarial pump of Philaenus spumarius scales across ontogeny to ensure functional equivalence.

A select group of hemipterans within the suborder Auchenorrhyncha are the only animals that feed exclusively on xylem sap - a nutritionally poor liquid that exists under negative pressure within a plant's xylem vessels. To consume it, xylem-feeding bugs have evolved enlarged cibarial pumps capable of generating enormous negative pressures. A previous study examining the allometry of this feeding model suggested that small xylem feeders pay relatively higher energetic costs while feeding, favouring the evolution of larger-bodied species. However, this interspecific analysis only considered adult xylem-feeding insects and neglected the considerable intraspecific change in size that occurs across the insect's development. Here, we examine the changes in cibarial pump morphology and function that occur during the development of Philaenus spumarius, the common meadow spittlebug. We show that the cibarial pump scales largely as expected from isometry and that the maximum negative pressure is mass independent, indicating that size has no effect on the xylem-feeding capacity of juvenile spittlebugs. We conclude that a first instar nymph with a body mass 2% of the adult can still feed at the >1 MPa tension present in a plant's xylem vessels without a substantial energetic disadvantage.

Ecological stress memory in wood architecture of two Neotropical hickory species from central-eastern Mexico.

BACKGROUND: Drought periods are major evolutionary triggers of wood anatomical adaptive variation in Lower Tropical Montane Cloud Forests tree species. We tested the influence of historical drought events on the effects of ecological stress memory on latewood width and xylem vessel traits in two relict hickory species (Carya palmeri and Carya myristiciformis) from central-eastern Mexico. We hypothesized that latewood width would decrease during historical drought years, establishing correlations between growth and water stress conditions, and that moisture deficit during past tree growth between successive drought events, would impact on wood anatomical features. We analyzed latewood anatomical traits that developed during historical drought and pre- and post-drought years in both species. RESULTS: We found that repeated periods of hydric stress left climatic signatures for annual latewood growth and xylem vessel traits that are essential for hydric adaptation in tropical montane hickory species. CONCLUSIONS: Our results demonstrate the existence of cause‒effect relationships in wood anatomical architecture and highlight the ecological stress memory linked with historical drought events. Thus, combined time-series analysis of latewood width and xylem vessel traits is a powerful tool for understanding the ecological behavior of hickory species.

Tip-to-base conduit widening remains consistent across cambial age and climates in Fagus sylvatica L.

Water transport, mechanical support and storage are the vital functions provided by the xylem. These functions are carried out by different cells, exhibiting significant anatomical variation not only within species but also within individual trees. In this study, we used a comprehensive dataset to investigate the consistency of predicted hydraulic vessel diameter widening values in relation to the distance from the tree apex, represented by the relationship Dh ∝ Lß (where Dh is the hydraulic vessel diameter, L the distance from the stem apex and ß the scaling exponent). Our analysis involved 10 Fagus sylvatica L. trees sampled at two distinct sites in the Italian Apennines. Our results strongly emphasize that vessel diameter follows a predictable pattern with the distance from the stem apex and ß ~ 0.20 remains consistent across cambial age and climates. This finding supports the hypothesis that trees do not alter their axial configuration represented by scaling of vessel diameter to compensate for hydraulic limitations imposed by tree height during growth. The study further indicates that within-tree variability significantly contributes to the overall variance of the vessel diameter-stem length exponent. Understanding the factors that contribute to the intraindividual variability in the widening exponent is essential, particularly in relation to interspecific responses and adaptations to drought stress.

Limited effects of xylem anatomy on embolism resistance in cycad leaves.

Drought-induced xylem embolism is a primary cause of plant mortality. Although c. 70% of cycads are threatened by extinction and extant cycads diversified during a period of increasing aridification, the vulnerability of cycads to embolism spread has been overlooked. We quantified the vulnerability to drought-induced embolism, pressure-volume curves, in situ water potentials, and a suite of xylem anatomical traits of leaf pinnae and rachises for 20 cycad species. We tested whether anatomical traits were linked to hydraulic safety in cycads. Compared with other major vascular plant clades, cycads exhibited similar embolism resistance to angiosperms and pteridophytes but were more vulnerable to embolism than noncycad gymnosperms. All 20 cycads had both tracheids and vessels, the proportions of which were unrelated to embolism resistance. Only vessel pit membrane fraction was positively correlated to embolism resistance, contrary to angiosperms. Water potential at turgor loss was significantly correlated to embolism resistance among cycads. Our results show that cycads exhibit low resistance to xylem embolism and that xylem anatomical traits - particularly vessels - may influence embolism resistance together with tracheids. This study highlights the importance of understanding the mechanisms of drought resistance in evolutionarily unique and threatened lineages like the cycads.

Comparative venation costs of monocotyledon and dicotyledon species in the eastern Colorado steppe.

MAIN CONCLUSION: Leaf vein network cost (total vein surface area per leaf volume) for major veins and vascular bundles did not differ between monocot and dicot species in 21 species from the eastern Colorado steppe. Dicots possessed significantly larger minor vein networks than monocots. Across the tree of life, there is evidence that dendritic vascular transport networks are optimized, balancing maximum speed and integrity of resource delivery with minimal resource investment in transport and infrastructure. Monocot venation, however, is not dendritic, and remains parallel down to the smallest vein orders with no space-filling capillary networks. Given this departure from the "optimized" dendritic network, one would assume that monocots are operating at a significant energetic disadvantage. In this study, we investigate whether monocot venation networks bear significantly greater carbon/construction costs per leaf volume than co-occurring dicots in the same ecosystem, and if so, what physiological or ecological advantage the monocot life form possesses to compensate for this deficit. Given that venation networks could also be optimized for leaf mechanical support or provide herbivory defense, we measured the vascular system of both monocot and dicots at three scales to distinguish between leaf investment in mechanical support (macroscopic vein), total transport and capacitance (vascular bundle), or exclusively water transport (xylem) for both parallel and dendritic venation networks. We observed that vein network cost (total vein surface area per leaf volume) for major veins and vascular bundles was not significantly different between monocot species and dicot species. Dicots, however, possess significantly larger minor vein networks than monocots. The 19 species subjected to gas-exchange measurement in the field displayed a broad range of Amax and but demonstrated no significant relationships with any metric of vascular network size in major or minor vein classes. Given that monocots do not seem to display any leaf hydraulic disadvantage relative to dicots, it remains an important research question why parallel venation (truly parallel, down to the smallest vessels) has not arisen more than once in the history of plant evolution.

Parametrization of biological assumptions to simulate growth of tree branching architectures.

Modeling and simulating the growth of the branching of tree species remains a challenge. With existing approaches, we can reconstruct or rebuild the branching architectures of real tree species, but the simulation of the growth process remains unresolved. First, we present a tree growth model to generate branching architectures that resemble real tree species. Secondly, we use a quantitative morphometric approach to infer the shape similarity of the generated simulations and real tree species. Within a functional-structural plant model, we implement a set of biological parameters that affect the branching architecture of trees. By modifying the parameter values, we aim to generate basic shapes of spruce, pine, oak and poplar. Tree shapes are compared using geometric morphometrics of landmarks that capture crown and stem outline shapes. Five biological parameters, namely xylem flow, shedding rate, proprioception, gravitysense and lightsense, most influenced the generated tree branching patterns. Adjusting these five parameters resulted in the different tree shapes of spruce, pine, oak, and poplar. The largest effect was attributed to gravity, as phenotypic responses to this effect resulted in different growth directions of gymnosperm and angiosperm branching architectures. Since we were able to obtain branching architectures that resemble real tree species by adjusting only a few biological parameters, our model is extendable to other tree species. Furthermore, the model will also allow the simulation of structural tree-environment interactions. Our simplifying approach to shape comparison between tree species, landmark geometric morphometrics, showed that even the crown-trunk outlines capture species differences based on their contrasting branching architectures.

Climate of origin shapes variations in wood anatomical properties of 17 Picea species.

BACKGROUND: Variations in hydraulic conductivity may arise from species-specific differences in the anatomical structure and function of the xylem, reflecting a spectrum of plant strategies along a slow-fast resource economy continuum. Spruce (Picea spp.), a widely distributed and highly adaptable tree species, is crucial in preventing soil erosion and enabling climate regulation. However, a comprehensive understanding of the variability in anatomical traits of stems and their underlying drivers in the Picea genus is currently lacking especially in a common garden. RESULTS: We assessed 19 stem economic properties and hydraulic characteristics of 17 Picea species grown in a common garden in Tianshui, Gansu Province, China. Significant interspecific differences in growth and anatomical characteristics were observed among the species. Specifically, xylem hydraulic conductivity (Ks) and hydraulic diameter exhibited a significant negative correlation with the thickness to span ratio (TSR), cell wall ratio, and tracheid density and a significant positive correlation with fiber length, and size of the radial tracheid. PCA revealed that the first two axes accounted for 64.40% of the variance, with PC1 reflecting the trade-off between hydraulic efficiency and mechanical support and PC2 representing the trade-off between high embolism resistance and strong pit flexibility. Regression analysis and structural equation modelling further confirmed that tracheid size positively influenced Ks, whereas the traits DWT, D_r, and TSR have influenced Ks indirectly. All traits failed to show significant phylogenetic associations. Pearson's correlation analysis demonstrated strong correlations between most traits and longitude, with the notable influence of the mean temperature during the driest quarter, annual precipitation, precipitation during the wettest quarter, and aridity index. CONCLUSIONS: Our results showed that xylem anatomical traits demonstrated considerable variability across phylogenies, consistent with the pattern of parallel sympatric radiation evolution and global diversity in spruce. By integrating the anatomical structure of the stem xylem as well as environmental factors of origin and evolutionary relationships, our findings provide novel insights into the ecological adaptations of the Picea genus.

Negative allometry of leaf xylem conduit diameter and double-wall thickness: implications for implosion safety.

Xylem conduits have lignified walls to resist crushing pressures. The thicker the double-wall (T) relative to its diameter (D), the greater the implosion safety. Having safer conduits may incur higher costs and reduced flow, while having less resistant xylem may lead to catastrophic collapse under drought. Although recent studies have shown that conduit implosion commonly occurs in leaves, little is known about how leaf xylem scales T vs D to trade off safety, flow efficiency, mechanical support, and cost. We measured T and D in > 7000 conduits of 122 species to investigate how T vs D scaling varies across clades, habitats, growth forms, leaf, and vein sizes. As conduits become wider, their double-cell walls become proportionally thinner, resulting in a negative allometry between T and D. That is, narrower conduits, which are usually subjected to more negative pressures, are proportionally safer than wider ones. Higher implosion safety (i.e. higher T/D ratios) was found in asterids, arid habitats, shrubs, small leaves, and minor veins. Despite the strong allometry, implosion safety does not clearly trade off with other measured leaf functions, suggesting that implosion safety at whole-leaf level cannot be easily predicted solely by individual conduits' anatomy.

Root topological order drives variation of fine root vessel traits and hydraulic strategies in tropical trees.

Vessel traits contribute to plant water transport from roots to leaves and thereby influence how plants respond to soil water availability, but the sources of variation in fine root anatomical traits remain poorly understood. Here, we explore the variations of fine root vessel traits along topological orders within and across tropical tree species. Anatomical traits were measured along five root topological orders in 80 individual trees of 20 species from a tropical forest in southwestern China. We found large variations for most root anatomical traits across topological orders, and strong co-variations between vessel traits. Within species, theoretical specific xylem hydraulic conductivity (Kth) increased with topological order due to increased mean vessel diameter, size heterogeneity, and decreased vessel density. Across species, Kth was associated with vessel fraction in low-order roots and correlated with mean vessel diameter and vessel density in high-order roots, suggesting a shift in relative anatomical contributors to Kth from the second- to fifth-order roots. We found no clear relationship between Kth and stele: root diameter ratios. Our study shows strong variations in root vessel traits across topological orders and species, and highlights shifts in the anatomical underpinnings by varying vessel-related anatomical structures for an optimized water supply.

Phenotyping xylem connections in grafted plants using X-ray micro-computed tomography.

Plants are able to naturally graft or inosculate their trunks, branches and roots together, this mechanism is used by humans to graft together different genotypes for a range of purposes. Grafts are considered successful if functional vascular connections between the two genotypes occur. Various techniques can evaluate xylem connections across the graft interface. However, these methods are generally unable to assess the heterogeneity and three-dimensional (3D) structure of xylem vessel connections. Here we present the use of X-ray micro-computed tomography to characterize the 3D morphology of grafts of grapevine. We show that xylem vessels form between the two plants of natural root and human-made stem grafts. The main novelty of this methodology is that we were able to visualize the 3D network of functional xylem vessels connecting the scion and rootstock in human-made stem grafts thanks to the addition of a contrast agent to the roots and improved image analysis pipelines. In addition, we reveal the presence of extensive diagonal xylem connections between the main axial xylem vessels in 2-year old grapevine stems. In conclusion, we present a method that has the potential to provide new insights into the structure and function of xylem vessels in large tissue samples.

Variations in wood anatomy in Afrotropical trees with a particular emphasis on radial and axial parenchyma.

BACKGROUND AND AIMS: Understanding anatomical variations across plant phylogenies and environmental gradients is vital for comprehending plant evolution and adaptation. Previous studies on tropical woody plants have paid limited attention to quantitative differences in major xylem tissues, which serve specific roles in mechanical support (fibres), carbohydrate storage and radial conduction (radial parenchyma, rays), wood capacitance (axial parenchyma) and water transport (vessels). To address this gap, we investigate xylem fractions in 173 tropical tree species spanning 134 genera and 53 families along a 2200-m elevational gradient on Mount Cameroon, West Africa. METHODS: We determined how elevation, stem height and wood density affect interspecific differences in vessel, fibre, and specific axial (AP) and radial (RP) parenchyma fractions. We focus on quantifying distinct subcategories of homogeneous or heterogeneous rays and apotracheal, paratracheal and banded axial parenchyma. KEY RESULTS: Elevation-related cooling correlated with reduced AP fractions and vessel diameters, while fibre fractions increased. Lower elevations exhibited elevated AP fractions due to abundant paratracheal and wide-banded parenchyma in tall trees from coastal and lowland forests. Vasicentric and aliform AP were predominantly associated with greater tree height and wider vessels, which might help cope with high evaporative demands via elastic wood capacitance. In contrast, montane trees featured a higher fibre proportion, scarce axial parenchyma, smaller vessel diameters and higher vessel densities. The lack of AP in montane trees was often compensated for by extended uniseriate ray sections with upright or squared ray cells or the presence of living fibres. CONCLUSIONS: Elevation gradient influenced specific xylem fractions, with lower elevations showing elevated AP due to abundant paratracheal and wide-banded parenchyma, securing greater vessel-to-parenchyma connectivity and lower embolism risk. Montane trees featured a higher fibre proportion and smaller vessel diameters, which may aid survival under greater environmental seasonality and fire risk.

Modeling Xylem Functionality Aspects.

Depending on the questions to be answered, water flow in the xylem can be modelled following different approaches with varying spatial and temporal resolution. When focussing on the influence of hydraulic architecture upon flow dynamics, distribution of water potentials in a tree crown or questions of vulnerability of the hydraulic system, functional-structural plant models, which link representations of morphological structure with simulated processes and with a virtual environment, can be a promising tool. Such a model will then include a network of idealized xylem segments, each representing the conducting part of a stem or branch segment, and a numerical machinery suitable for solving a system of differential equations on it reflecting the hydrodynamic laws, which are the basis of the broadly accepted cohesion-tension theory of water flow in plants. We will discuss functional-structural plant models, the simplifications that are useful for hydraulic simulations within this framework, the deduction of the used differential equations from basic physical conservation laws, and their numerical solution, as well as additional necessary models of radiation, photosynthesis, and stomatal conductance. In some supplementary notes, we are shortly addressing some related questions, for example, about root systems or about the relation between macro-scale hydraulic parameters and fine-grained (anatomical) xylem structure.

Do root secondary xylem functional traits differ between growth forms in Fabaceae species in a seasonally dry Neotropical environment?

BACKGROUND AND AIMS: Whole-plant performance in water-stressed and disturbance-prone environments depends on a suitable supply of water from the roots to the leaves, storage of reserves during periods of shortage, and a morphological arrangement that guarantees the maintenance of the plants anchored to the soil. All these functions are performed by the secondary xylem of roots. Here, we investigate whether different growth forms of Fabaceae species from the seasonally dry Neotropical environment have distinct strategies for water transport, mechanical support and non-structural carbon and water storage in the root secondary xylem. METHODS: We evaluated cross-sections of root secondary xylem from species of trees, shrubs and subshrubs. We applied linear models to verify the variability in secondary xylem anatomical traits among growth forms. KEY RESULTS: Secondary xylem with larger vessels and lower vessel density was observed in tree species. Vessel wall thickness, vessel grouping index, potential hydraulic conductivity and cell fractions (vessels, fibres, rays and axial parenchyma) were not statistically different between growth forms, owing to the high interspecific variation within the groups studied. CONCLUSION: Our results showed that the variability in anatomical traits of the secondary xylem of the root is species specific. In summary, the cellular complexity of the secondary xylem ensures multiple functional strategies in species with distinct growth forms, a key trait for resource use in an environment with strong water seasonality.

Toward a better understanding of angiosperm xylogenesis: a new method for a cellular approach.

The kinetics of wood formation in angiosperms are largely unknown because their complex xylem anatomy precludes using the radial position of vessels and fibers to infer their time of differentiation. We analyzed xylogenesis in ring-porous ash (Fraxinus angustifolia) and diffuse-porous beech (Fagus sylvatica) over 1 yr and proposed a novel procedure to assess the period of vessel and fiber enlargement using a referential radial file (RRF). Our approach captured the dynamics of wood formation and provided a robust estimation of the kinetics of vessel and fiber enlargement. In beech, fibers and vessels had a similar duration of enlargement, decreasing from 14 to 5 d between April and July. In ash, wide vessels formed in April enlarged at a rate of 27 × 103 µm2 d-1 , requiring half the time of contemporary fibers (6 vs 12 d), and less time than the narrower vessels (14 d) formed in May. These findings reveal distinct cell-type-dependent mechanisms for differentiation in diffuse-porous and ring-porous trees, enhancing our understanding of angiosperm wood cell kinetics. Our approach presents an effective method for investigating angiosperm wood formation and provides a more accurate representation of vessel and fiber morphogenesis in wood formation models.

Xylem resistance to cavitation increases during summer in Pinus halepensis.

Cavitation resistance has often been viewed as a relatively static trait, especially for stems of forest trees. Meanwhile, other hydraulic traits, such as turgor loss point (Ψtlp ) and xylem anatomy, change during the season. In this study, we hypothesized that cavitation resistance is also dynamic, changing in coordination with Ψtlp . We began with a comparison of optical vulnerability (OV), microcomputed tomography (µCT) and cavitron methods. All three methods significantly differed in the slope of the curve,Ψ12 and Ψ88 , but not in Ψ50 (xylem pressures that cause 12%, 88%, 50% cavitation, respectively). Thus, we followed the seasonal dynamics (across 2 years) of Ψ50 in Pinus halepensis under Mediterranean climate using the OV method. We found that Ψ50 is a plastic trait with a reduction of approximately 1 MPa from the end of the wet season to the end of the dry season, in coordination with the dynamics of the midday xylem water potential (Ψmidday ) and the Ψtlp . The observed plasticity enabled the trees to maintain a stable positive hydraulic safety margin and avoid cavitation during the long dry season. Seasonal plasticity is vital for understanding the actual risk of cavitation to plants and for modeling species' ability to tolerate harsh environments.

Addressing controversies in the xylem embolism resistance-vessel diameter relationship.

Although xylem embolism is a key process during drought-induced tree mortality, its relationship to wood anatomy remains debated. While the functional link between bordered pits and embolism resistance is known, there is no direct, mechanistic explanation for the traditional assumption that wider vessels are more vulnerable than narrow ones. We used data from 20 temperate broad-leaved tree species to study the inter- and intraspecific relationship of water potential at 50% loss of conductivity (P50 ) with hydraulically weighted vessel diameter (Dh ) and tested its link to pit membrane thickness (TPM ) and specific conductivity (Ks ) on species level. Embolism-resistant species had thick pit membranes and narrow vessels. While Dh was weakly associated with TPM , the P50 -Dh relationship remained highly significant after accounting for TPM . The interspecific pattern between P50 and Dh was mirrored by a link between P50 and Ks , but there was no evidence for an intraspecific relationship. Our results provide robust evidence for an interspecific P50 -Dh relationship across our species. As a potential cause for the inconsistencies in published P50 -Dh relationships, our analysis suggests differences in the range of trait values covered, and the level of data aggregation (species, tree or sample level) studied.