BAHD acetyltransferase contributes to wound-induced biosynthesis of oleo-gum resin triterpenes in Boswellia.

Triterpenes (30-carbon isoprene compounds) represent a large and highly diverse class of natural products that play various physiological functions in plants. The triterpene biosynthetic enzymes, particularly those catalyzing the late-stage regio-selective modifications are not well characterized. The bark of select Boswellia trees, e.g., B. serrata exudes specialized oleo-gum resin in response to wounding, which is enriched with boswellic acids (BAs), a unique class of C3α-epimeric pentacyclic triterpenes with medicinal properties. The bark possesses a network of resin secretory structures comprised of vertical and horizontal resin canals, and amount of BAs in bark increases considerably in response to wounding. To investigate BA biosynthetic enzymes, we conducted tissue-specific transcriptome profiling and identified a wound-responsive BAHD acetyltransferase (BsAT1) of B. serrata catalyzing the late-stage C3α-O-acetylation reactions in the BA biosynthetic pathway. BsAT1 catalyzed C3α-O-acetylation of αBA, ßBA, and 11-keto-ßBA in vitro and in planta assays to produce all the major C3α-O-acetyl-BAs (3-acetyl-αBA, 3-acetyl-ßBA, and 3-acetyl-11-keto-ßBA) found in B. serrata bark and oleo-gum resin. BsAT1 showed strict specificity for BA scaffold, whereas it did not acetylate the more common C3ß-epimeric pentacyclic triterpenes. The analysis of steady-state kinetics using various BAs revealed distinct substrate affinity and catalytic efficiency. BsAT1 transcript expression coincides with increased levels of C3α-O-acetyl-BAs in bark in response to wounding, suggesting a role of BsAT1 in wound-induced biosynthesis of C3α-O-acetyl-BAs. Overall, the results provide new insights into the biosynthesis of principal chemical constituents of Boswellia oleo-gum resin.

Contribution of plant anatomy to forensic investigation: Tree bark morphology.

Plant science has been more and more utilized in forensic investigation, although its full potential is still to be reached. Plant macroremains are a powerful tool to link a body or other evidence back to a primary crime scene as they can provide detailed information about its previous ecological and geographic location. However, plant macroremains are often poorly preserved and difficult to identify, as diagnostic elements are seldom present within the assemblage occurring on the scene. Plant fragments most likely to be found are those exposed to the environment and resistant to degradation. The bark of woody plants meets these requirements but the possibility of its identification at species level from small fragments is not known. Starting from a real homicide case, where bark splinters were found on the victim, we aimed to assess the forensic potential of bark identification from small fragments like those likely to occur on a crime scene. Two identification keys were prepared for 16 common lowland tree species from Northern Italy; one key used all the available anatomical traits, the second only those from the outer bark. The second key was not able to discriminate some couples of species unambiguously, but could identify the bark fragments of the homicide as Robinia pseudoacacia, as confirmed from direct comparison with a reference sample. Bark fragments deserve to be included into the macroremains to be analyzed during an investigation, but small samples could easily lack diagnostic traits, and the building of a reference collection should be encouraged.

Inner bark as a crucial tissue for non-structural carbohydrate storage across three tropical woody plant communities.

Non-structural carbohydrates (NSC) are crucial for forest resilience, but little is known regarding the role of bark in NSC storage. However, bark's abundance in woody stems and its large living fraction make it potentially key for NSC storage. We quantified total NSC, soluble sugar (SS) and starch concentrations in the most living region of bark (inner bark, IB), and sapwood of twigs, trunks and roots of 45 woody species from three contrasting tropical climates spanning global extremes of bark diversity and wide phylogenetic diversity. NSC concentrations were similar (total NSC, starch) or higher (SS) in IB than wood, with concentrations co-varying strongly. NSC concentrations varied widely across organs and species within communities and were not significantly affected by climate, leaf habit or the presence of photosynthetic bark. Starch concentration tended to increase with density, but only in wood. IB contributed substantially to NSC storage, accounting for 17-36% of total NSC, 23-47% of SS and 15-33% of starch pools. Further examination of the drivers of variation in IB NSC concentration, and taking into account the substantial contribution of IB to NSC pools, will be crucial to understand the role of storage in plant environmental adaptation.

The role of bud protection and bark density in frost resistance of savanna trees.

Frost events occur with a significant frequency in savannas of the Southern Hemisphere, especially in the Cerrados of Brazil. One of the main strategies to deal with such events is to invest in thick and dense bark, which can insulate internal branch tissues and protect buds, essential to ensure resprouting if frost damage causes plant canopy die-back. Such strategies may be fundamental to determine the persistence of savanna species in regions where low temperatures and frost events are recurrent. Here we describe bud protection and bark strategies of 53 woody species growing in typical savanna vegetation of central Brazil. In addition, we used an experimental approach exposing branches to 0 °C to measure temperature variation in internal branch tissue and test its relationship to bud protection and bark properties. We found that the majority of species (69%) showed medium to high bud protection against extreme temperatures; however, the degree of bud protection was not clearly related to bark properties, such as bark thickness and density. Bark density is a fundamental trait in determining protection against low temperatures (0 °C), since species with low bark density showed lower temperature variation in their internal branch tissues, independently of the bud protection degree. Bark properties and bud protection are two different (albeit related) strategies for the protection and persistence of savanna trees under extreme environmental temperatures and can explain ecological observations related to savanna tree responses after frost events.

Patterns and dynamics of canopy-root coupling in tropical tree saplings vary with light intensity but not with root depth.

Carbon (C) dynamics in canopy and roots influence whole-tree carbon fluxes, but little is known about canopy regulation of tree-root activity. Here, the patterns and dynamics of canopy-root C coupling are assessed in tropical trees. Large aeroponics facility was used to study the root systems of Ceiba pentandra and Khaya anthotheca saplings directly at different light intensities. In Ceiba, root respiration (Rr ) co-varied with photosynthesis (An ) in large saplings (3-to-7-m canopy-root axis) at high-light, but showed no consistent pattern at low-light. At medium-light and in small saplings (c. 1-m axis), Rr tended to decrease transiently towards midday. Proximal roots had higher Rr and nonstructural carbohydrate concentrations than distal roots, but canopy-root coupling was unaffected by root location. In medium-sized Khaya, no Rr pattern was observed, and in both species, Rr was unrelated to temperature. The early-afternoon increase in Rr suggests that canopy-root coupling is based on mass flow of newly fixed C in the phloem, whereas the early-morning rise in Rr with An indicates an additional coupling signal that travels faster than the phloem sap. In large saplings and potentially also in higher trees, light and possibly additional environmental factors control the diurnal patterns of canopy-root coupling, irrespective of root location.

Bark in Woody Plants: Understanding the Diversity of a Multifunctional Structure.

Most biological structures carry out multiple functions. Focusing on only one function to make adaptive inferences overlooks that manifold selection pressures and tradeoffs shape the characteristics of a multifunctional structure. Focusing on single functions can only lead to a partial picture of the causes underlying diversity and the evolutionary origin of the structure in question. I illustrate this discussion using bark as a study case. Bark comprises all the tissues surrounding the xylem in woody plants. Broadly, bark includes an inner and mostly living region and an outer, dead one. Of all plant structures, bark has the most complex anatomical structure and ontogenetic origin involving two (and often three) different meristems. Traditionally, the wide diversity in bark traits, mainly bark thickness, has been interpreted as the result of the selective pressures imposed by fire regime. However, recent research has shown that explanations based on fire regime cannot account for salient patterns of bark variation globally including the very strong inner bark thickness-stem diameter scaling, which is likely due to metabolic needs, and the very high intracommunity variation in total, inner, and outer bark thickness, and in inner:outer proportions. Moreover, explanations based on fire disregard that in addition to fire protection, bark carries out several other crucial functions for plants including translocation of photosynthates; storage of starch, soluble sugars, water, and other compounds; protection from herbivores, pathogens, and high temperatures; wound closure, as well as mechanical support, photosynthesis, and likely being involved in xylem embolism repair. All these functions are crucial for plant performance and are involved in synergistic (e.g., storage of water and insulation) and trade-off relationships (e.g., protection from fire vs photosynthetic activity). Focusing on only one of these functions, protection from fire has provided an incomplete picture of the selective forces shaping bark diversity and has severely hindered our incipient understanding of the functional ecology of this crucial region of woody stems. Applying a multifunctional perspective to the study of bark will allow us to address why we observe such high intracommunity variation in bark traits, why some bark trait combinations are ontogenetically impossible or penalized by selection, how bark is coordinated functionally with other plant parts, and as a result, to understand how bark contributes to the vast diversity of plant ecological strategies across the globe.

Is stemflow a vector for the transport of small metazoans from tree surfaces down to soil?

BACKGROUND: Stemflow is an essential hydrologic process shaping the soil of forests by providing a concentrated input of rainwater and solutions. However, the transport of metazoans by stemflow has yet to be investigated. This 8-week study documented the organisms (< 2 mm) present in the stemflow of different tree species. Because the texture of the tree bark is a crucial determination of stemflow, trees with smooth bark (Carpinus betulus and Fagus sylvatica) and rough bark (Quercus robur) were examined. RESULTS: Up to 1170 individuals per liter of stemflow were collected. For rotifers and nematodes, a highly positive correlation between abundance and stemflow yield was determined. Both taxa were predominant (rotifers: up to 70%, nematodes: up to 13.5%) in the stemflow of smooth-barked trees whereas in that of the oak trees collembolans were the most abundant organisms (77.3%). The mean number of organisms collected per liter of stemflow from the two species of smooth-barked trees was very similar. A higher number of nematode species was found in the stemflow of these trees than in the stemflow of rough-barked oak and all were typical colonizers of soil- and bark-associated habitats. CONCLUSION: This pilot study showed for the first time that stemflow is a transport vector for numerous small metazoans. By connecting tree habitats (e.g., bark, moss, lichens or water-filled tree holes) with soil, stemflow may influence the composition of soil fauna by mediating intensive organismal dispersal.

Quantitative trait locus mapping of Populus bark features and stem diameter.

BACKGROUND: Bark plays important roles in photosynthate transport and storage, along with physical and chemical protection. Bark texture varies extensively among species, from smooth to fissured to deeply furrowed, but its genetic control is unknown. This study sought to determine the main genomic regions associated with natural variation in bark features and stem diameter. Quantitative trait loci (QTL) were mapped using an interspecific pseudo-backcross pedigree (Populus trichocarpa x P. deltoides and P. deltoides) for bark texture, bark thickness and diameter collected across three years, two sites and three biological replicates per site. RESULTS: QTL specific to bark texture were highly reproducible in shared intervals across sites, years and replicates. Significant positive correlations and co-localization between trait QTL suggest pleiotropic regulators or closely linked genes. A list of candidate genes with related putative function, location close to QTL maxima and with the highest expression level in the phloem, xylem and cambium was identified. CONCLUSION: Candidate genes for bark texture included an ortholog of Arabidopsis ANAC104 (PopNAC128), which plays a role in lignified fiber cell and ray development, as well as Pinin and Fasciclin (PopFLA) genes with a role in cell adhesion, cell shape and migration. The results presented in this study provide a basis for future genomic characterization of genes found within the QTL for bark texture, bark thickness and diameter in order to better understand stem and bark development in Populus and other woody perennial plants. The QTL mapping approach identified a list of prime candidate genes for further validation using functional genomics or forward genetics approaches.

Cell-type- and tissue-specific transcriptomes of the white spruce (Picea glauca) bark unmask fine-scale spatial patterns of constitutive and induced conifer defense.

Plant defenses often involve specialized cells and tissues. In conifers, specialized cells of the bark are important for defense against insects and pathogens. Using laser microdissection, we characterized the transcriptomes of cortical resin duct cells, phenolic cells and phloem of white spruce (Picea glauca) bark under constitutive and methyl jasmonate (MeJa)-induced conditions, and we compared these transcriptomes with the transcriptome of the bark tissue complex. Overall, ~3700 bark transcripts were differentially expressed in response to MeJa. Approximately 25% of transcripts were expressed in only one cell type, revealing cell specialization at the transcriptome level. MeJa caused cell-type-specific transcriptome responses and changed the overall patterns of cell-type-specific transcript accumulation. Comparison of transcriptomes of the conifer bark tissue complex and specialized cells resolved a masking effect inherent to transcriptome analysis of complex tissues, and showed the actual cell-type-specific transcriptome signatures. Characterization of cell-type-specific transcriptomes is critical to reveal the dynamic patterns of spatial and temporal display of constitutive and induced defense systems in a complex plant tissue or organ. This was demonstrated with the improved resolution of spatially restricted expression of sets of genes of secondary metabolism in the specialized cell types.

Exploring the bark thickness-stem diameter relationship: clues from lianas, successive cambia, monocots and gymnosperms.

Bark thickness is ecologically crucial, affecting functions from fire protection to photosynthesis. Bark thickness scales predictably with stem diameter, but there is little consensus on whether this scaling is a passive consequence of growth or an important adaptive phenomenon requiring explanation. With a comparative study across 913 species, we test the expectation that, if bark thickness-stem diameter scaling is adaptive, it should be possible to find ecological situations in which scaling is predictably altered, in this case between species with different types and deployments of phloem. 'Dicots' with successive cambia and monocots, which have phloem-free bark, had predictably thinner inner (mostly living) bark than plants with single cambia. Lianas, which supply large leaf areas with limited stem area, had much thicker inner bark than self-supporting plants. Gymnosperms had thicker outer bark than angiosperms. Inner bark probably scales with plant metabolic demands, for example with leaf area. Outer bark scales with stem diameter less predictably, probably reflecting diverse adaptive factors; for example, it tends to be thicker in fire-prone species and very thin when bark photosynthesis is favored. Predictable bark thickness-stem diameter scaling across plants with different photosynthate translocation demands and modes strongly supports the idea that this relationship is functionally important and adaptively significant.

Convergence of bark investment according to fire and climate structures ecosystem vulnerability to future change.

Fire regimes in savannas and forests are changing over much of the world. Anticipating the impact of these changes requires understanding how plants are adapted to fire. In this study, we test whether fire imposes a broad selective force on a key fire-tolerance trait, bark thickness, across 572 tree species distributed worldwide. We show that investment in thick bark is a pervasive adaptation in frequently burned areas across savannas and forests in both temperate and tropical regions where surface fires occur. Geographic variability in bark thickness is largely explained by annual burned area and precipitation seasonality. Combining environmental and species distribution data allowed us to assess vulnerability to future climate and fire conditions: tropical rainforests are especially vulnerable, whereas seasonal forests and savannas are more robust. The strong link between fire and bark thickness provides an avenue for assessing the vulnerability of tree communities to fire and demands inclusion in global models.

Height-related scaling of phloem anatomy and the evolution of sieve element end wall types in woody plants.

In the sieve elements (SEs) of the phloem, carbohydrates are transported throughout the whole plant from their site of production to sites of consumption or storage. SE structure, especially of the pore-rich end walls, has a direct effect on translocation efficiency. Differences in pore size and other features were interpreted as an evolutionary trend towards reduced hydraulic resistance. However, this has never been confirmed. Anatomical data of 447 species of woody angiosperms and gymnosperms were used for a phylogenetic analysis of end wall types, calculation of hydraulic resistance and correlation analysis with morphological and physiological variables. end wall types were defined according to pore arrangement: either grouped into a single area (simple) or into multiple areas along the end wall (compound). Convergent evolution of end wall types was demonstrated in woody angiosperms. In addition, an optimization of end wall resistance with plant height was discovered, but found to be independent of end wall type. While physiological factors also showed no correlation with end wall types, the number of sieve areas per end wall was found to scale with SE length. The results exclude the minimization of hydraulic resistance as evolutionary driver of different end wall types, contradicting this long-standing assumption. Instead, end wall type might depend on SE length.

Bark thickness across the angiosperms: more than just fire.

Global variation in total bark thickness (TBT) is traditionally attributed to fire. However, bark is multifunctional, as reflected by its inner living and outer dead regions, meaning that, in addition to fire protection, other factors probably contribute to TBT variation. To address how fire, climate, and plant size contribute to variation in TBT, inner bark thickness (IBT) and outer bark thickness (OBT), I sampled 640 species spanning all major angiosperm clades and 18 sites with contrasting precipitation, temperature, and fire regime. Stem size was by far the main driver of variation in thickness, with environment being less important. IBT was closely correlated with stem diameter, probably for metabolic reasons, and, controlling for size, was thicker in drier and hotter environments, even fire-free ones, probably reflecting its water and photosynthate storage role. OBT was less closely correlated with size, and was thicker in drier, seasonal sites experiencing frequent fires. IBT and OBT covaried loosely and both contributed to overall TBT variation. Thickness variation was higher within than across sites and was evolutionarily labile. Given high within-site diversity and the multiple selective factors acting on TBT, continued study of the different drivers of variation in bark thickness is crucial to understand bark ecology.

Functional and environmental determinants of bark thickness in fire-free temperate rain forest communities.

PREMISE OF THE STUDY: In fire-prone ecosystems, variation in bark thickness among species and communities has been explained by fire frequency; thick bark is necessary to protect cambium from lethal temperatures. Elsewhere this investment is deemed unnecessary, and thin bark is thought to prevail. However, in rain forest ecosystems where fire is rare, bark thickness varies widely among species and communities, and the causes of this variation remain enigmatic. We tested for functional explanations of bark thickness variation in temperate rain forest species and communities. METHODS: We measured bark thickness in 82 tree species throughout New Zealand temperate rain forests that historically have experienced little fire and applied two complementary analyses. First, we examined correlations between bark traits and leaf habit, and leaf and stem traits. Second, we calculated community-weighted mean (CWM) bark thickness for 272 plots distributed throughout New Zealand to identify the environments in which thicker-barked communities occur. KEY RESULTS: Conifers had higher size-independent bark thickness than evergreen angiosperms. Species with thicker bark or higher bark allocation coefficients were not associated with "slow economic" plant traits. Across 272 forest plots, communities with thicker bark occurred on infertile soils, and communities with thicker bark and higher bark allocation coefficients occurred in cooler, drier climates. CONCLUSIONS: In non-fire-prone temperate rain forest ecosystems, investment in bark is driven by soil resources, cool minimum temperatures, and seasonal moisture stress. The role of these factors in fire-prone ecosystems warrants testing.

Contribution of Light and Electron Microscopy to Identification of Bark from Frangula azorica, an Azorean Medicinal Plant.

Frangula azorica V. Grubow is a Macaronesian flora medicinal plant, endemic from Azores islands and inscribed on the International Union for the Conservation of Nature (IUCN) Red List of Threatened Species. This species, known as "sanguinho," belongs to the family Rhamnaceae, the same as Frangula alnus Mill. and Frangula purshiana (DC.) J. G. Cooper, two widely used official laxative herbal medicines of the western Pharmacopoeias constituted by the dried barks of each species. Morphological and chemical studies on F. azorica dried bark are scarce although it is potentially recognized as a Portuguese laxative herbal medicine. Macroscopically, the bark occurs in quills or nearly flat pieces. A channeled external surface with transversely elongated lenticels is characteristic. When the outer phellem layer is removed, a bright purple inner phellem layer is disclosed. Light and electron microscopy observations revealed flattened phellem cells with slightly thickened walls, cortical parenchyma with secretory ducts and groups of sclereids, phloem with groups of fibers and sheaths of parenchymatous cells containing druses or more frequently prismatic crystals of calcium oxalate, and parenchymatous medullary rays one to three cells wide with spherical starch grains. Observation of these botanical characteristics must be included in quality monographs of F. azorica bark herbal medicine.

Bark ecology of twigs vs. main stems: functional traits across eighty-five species of angiosperms.

Although produced by meristems that are continuous along the stem length, marked differences in bark morphology and in microenvironment would suggest that main stem and twig bark might differ ecologically. Here, we examined: (1) how closely associated main stem and twig bark traits were, (2) how these associations varied across sites, and (3) used these associations to infer functional and ecological differences between twig and main stem bark. We measured density, water content, photosynthesis presence/absence, total, outer, inner, and relative thicknesses of main stem and twig bark from 85 species of angiosperms from six sites of contrasting precipitation, temperature, and fire regimes. Density and water content did not differ between main stems and twigs across species and sites. Species with thicker twig bark had disproportionately thicker main stem bark in most sites, but the slope and degree of association varied. Disproportionately thicker main stem bark for a given twig bark thickness in most fire-prone sites suggested stem protection near the ground. The savanna had the opposite trend, suggesting that selection also favors twig protection in these fire-prone habitats. A weak main stem-twig bark thickness association was observed in non fire-prone sites. The near-ubiquity of photosynthesis in twigs highlighted its likely ecological importance; variation in this activity was predicted by outer bark thickness in main stems. It seems that the ecology of twig bark can be generalized to main stem bark, but not for functions depending on the amount of bark, such as protection, storage, or photosynthesis.

Outer bark thickness decreases more with height on stems of fire-resistant than fire-sensitive Floridian oaks (Quercus spp.; Fagaceae).

UNLABELLED: • PREMISE OF THE STUDY: In ecosystems maintained by low-intensity surface fires, tree bark thickness is a determinant of fire-survival because it protects underlying tissues from heat damage. However, it has been unclear whether relatively thick bark i S: maintained at all heights or only near the ground where damage is most likely.• METHODS: We studied six Quercus species from the red and white clades, with three species characteristic of fire-maintained savannas and three species characteristic of forests with infrequent fire. Inner and outer bark (secondary phloem and rhytidome, respectively) thicknesses were measured at intervals from 10 to 300 cm above the ground. We used linear mixed-effects models to test for relationships among height, habitat, and clade on relative thickness (stem proportion) of total, inner, and outer bark. Bark moisture and tissue density were measured for each species at 10 cm.• KEY RESULTS: Absolute and relative total bark thickness declined with height, with no difference in height-related changes between habitat groups. Relative outer bark thickness showed a height-by-habitat interaction. There was a clade effect on relative thickness, but no interaction with height. Moisture contents were higher in inner than outer bark, and red oaks had denser bark than white oaks, but neither trait differed by habitat.• CONCLUSIONS: Quercus species characteristic of fire-prone habitats invest more in outer bark near the ground where heat damage to outer tissues is most likely. Future investigations of bark should consider the height at which measurements are made and distinguish between inner and outer bark.

Modelling biomechanics of bark patterning in grasstrees.

BACKGROUND AND AIMS: Bark patterns are a visually important characteristic of trees, typically attributed to fractures occurring during secondary growth of the trunk and branches. An understanding of bark pattern formation has been hampered by insufficient information regarding the biomechanical properties of bark and the corresponding difficulties in faithfully modelling bark fractures using continuum mechanics. This study focuses on the genus Xanthorrhoea (grasstrees), which have an unusual bark-like structure composed of distinct leaf bases connected by sticky resin. Due to its discrete character, this structure is well suited for computational studies. METHODS: A dynamic computational model of grasstree development was created. The model captures both the phyllotactic pattern of leaf bases during primary growth and the changes in the trunk's width during secondary growth. A biomechanical representation based on a system of masses connected by springs is used for the surface of the trunk, permitting the emergence of fractures during secondary growth to be simulated. The resulting fracture patterns were analysed statistically and compared with images of real trees. KEY RESULTS: The model reproduces key features of grasstree bark patterns, including their variability, spanning elongated and reticulate forms. The patterns produced by the model have the same statistical character as those seen in real trees. CONCLUSIONS: The model was able to support the general hypothesis that the patterns observed in the grasstree bark-like layer may be explained in terms of mechanical fractures driven by secondary growth. Although the generality of the results is limited by the unusual structure of grasstree bark, it supports the hypothesis that bark pattern formation is primarily a biomechanical phenomenon.

Effects of Phyllanthus sellowianus Müll Arg. extracts on the rheological properties of human erythrocytes.

Phyllanthus sellowianus extracts have been used in Argentina since colonial times in the treatment of diabetes. The in vitro biorheological and hemoagglutinant action of different extracts of P. sellowianus bark on human erythrocytes (RBC) were studied. RBCs were incubated in vitro with four aqueous extracts: Maceration; Controlled Digestion (PD); Decoction; and Infusion. Biorheological parameters (deformability, membrane surface viscosity, elastic modulus, and dynamic viscolelasticity) were determined with an Erythrodeformeter, and erythrocyte adhesion was characterized by image digital analysis. Immunohematological assays in RBC incubated with all the extracts showed large globular aggregates and agglutination in human ABO blood group system. Isolated cell coefficient showed the increase of cell adhesion. Aggregated shape parameters were significantly higher than normal and they changed with the concentration, particularly of PD extracts. Rheological results showed that the extract biorheological action varies with the temperature used in the extract preparations. The results obtained are useful to study the action mechanism of extracts from P. sellowianus bark in order to evaluate its use as therapeutic agent in diabetes. Immunohematological Tests using ABO system showed its agglutinant power, which is of special interest in Immunohematology to be used as hemoclassifier.

Bark functional ecology: evidence for tradeoffs, functional coordination, and environment producing bark diversity.

The causes underlying bark diversity are unclear. Variation has been frequently attributed to environmental differences across sites. However, variation may also result from tradeoffs and coordination between bark's multiple functions. Bark traits may also covary with wood and leaf traits as part of major dimensions of plant variation. To assess hypotheses regarding tradeoffs and functional coordination, we measured bark traits reflecting protection, storage, mechanics, and photosynthesis in branches of 90 species spanning a wide phylogenetic and environmental range. We also tested associations between bark, wood, and leaf traits. We partitioned trait variation within species, and within and across communities to quantify variation associated with across-site differences. We observed associations between bark mechanics and storage, density and thickness, and thickness and photosynthetic activity. Increasing bark thickness contributed significantly to stiffer stems and greater water storage. Bark density, water content, and mechanics covaried strongly with the equivalent wood traits, and to a lesser degree with leaf size, xylem conductivity, and vessel diameter. Most variation was observed within sites and had low phylogenetic signal. Compared with relatively minor across-site differences, tradeoffs and coordination among functions of bark, leaves, and wood are likely to be major and overlooked factors shaping bark ecology and evolution.