Bark structure is coordinated with xylem hydraulic properties in branches of five Cupressaceae species.

The properties of bark and xylem contribute to tree growth and survival under drought and other types of stress conditions. However, little is known about the functional coordination of the xylem and bark despite the influence of selection on both structures in response to drought. To this end, we examined relationships between proportions of bark components (i.e. thicknesses of tissues outside the vascular cambium) and xylem transport properties in juvenile branches of five Cupressaceae species, focusing on transport efficiency and safety from hydraulic failure via drought-induced embolism. Both xylem efficiency and safety were correlated with multiple bark traits, suggesting that xylem transport and bark properties are coordinated. Specifically, xylem transport efficiency was greater in species with thicker secondary phloem, greater phloem-to-xylem thickness ratio and phloem-to-xylem cell number ratio. In contrast, species with thicker bark, living cortex and dead bark tissues were more resistant to embolism. Thicker phellem layers were associated with lower embolism resistance. Results of this study point to an important connection between xylem transport efficiency and phloem characteristics, which are shaped by the activity of vascular cambium. The link between bark and embolism resistance affirms the importance of both tissues to drought tolerance.

Bark wounding triggers gradual embolism spreading in two diffuse-porous tree species.

Xylem transport is essential for the growth, development and survival of vascular plants. Bark wounding may increase the risk of xylem transport failure by tension-driven embolism. However, the consequences of bark wounding for xylem transport are poorly understood. Here, we examined the impacts of the bark wounding on embolism formation, leaf water potential and gas exchange in the terminal branches of two diffuse-porous tree species (Acer platanoides L. and Prunus avium L.). The effects of bark removal were examined on field-grown mature trees exposed to increased evaporative demands on a short-term and longer-term basis (6 h vs 6 days after bark wounding). Bark removal of 30% of branch circumference had a limited effect on the xylem hydraulic conductivity when embolized vessels were typically restricted to the last annual ring near the bark wound. Over the 6-day exposure, the non-conductive xylem area had significantly increased in the xylem tissue underneath the bark wound (from 22-29% to 51-52% of the last annual ring area in the bark wound zone), pointing to gradual yet relatively limited embolism spreading to deeper xylem layers over time. In both species, the bark removal tended to result in a small but non-significant increase in the percent loss of hydraulic conductivity compared with control intact branches 6 days after bark wounding (from 6 to 8-10% in both species). The bark wounding had no significant effects on midday leaf water potential, CO2 assimilation rates, stomatal conductance and water-use efficiency of the leaves of the current-year shoot, possibly due to limited impacts on xylem transport. The results of this study demonstrate that bark wounding induces limited but gradual embolism spreading. However, the impacts of bark wounding may not significantly limit water delivery to distal organs and leaf gas exchange at the scale of several days.

Relationships between trunk radial growth and fruit yield in apple and pear trees on size-controlling rootstocks.

BACKGROUND AND AIMS: Understanding the mutual co-ordination of vegetative and reproductive growth is important in both agricultural and ecological settings. A competitive relationship between vegetative growth and fruiting is often highlighted, resulting in an apparent trade-off between structural growth and fruit production. However, our understanding of factors driving this relationship is limited. METHODS: We used four scions grafted onto a series of size-controlling rootstocks to evaluate the relationships between the annual fruit yield and radial growth of trunks, branches and roots. To assess tree radial growth, we measured ring widths on extracted tree cores, which is an approach not frequently used in a horticultural setting. KEY RESULTS: We found that the yield and radial growth were negatively related when plotted in absolute terms or as detrended and normalized indices. The relationship was stronger in low vigour trees, but only after the age-related trend was removed. In contrast, when trunk radial growth was expressed as basal area increment, the negative relationship disappeared, suggesting that the relationship between trunk radial growth and fruit yield might not be a true trade-off related to the competition between the two sinks. The effect of low yield was associated with increased secondary growth not only in trunks but also in branches and roots. In trunks, we observed that overcropping was associated with reduced secondary growth in a subsequent year, possibly due to the depletion of reserves. CONCLUSIONS: Our results show that variation in annual fruit yield due to tree ageing, weather cueing and inherent alternate bearing behaviour is reflected in the magnitude of secondary growth of fruit trees. We found little support for the competition/architecture theory of rootstock-induced growth vigour control. More broadly, our study aimed at bridging the gap between forest ecology and horticulture.

Trunk radial growth, water and carbon relations of mature apple trees on two size-controlling rootstocks during severe summer drought.

The use of size-controlling rootstocks is central to modern high-density fruit production systems. While biological mechanisms responsible for vigor control are not fully understood, differences in water relations and carbohydrate storage ability have been suggested as two potential factors. To better understand the processes that control growth vigor, we analyzed the trunk radial variation at seasonal and diurnal timescales and measured the midday leaf water potential (ΨMD), leaf gas exchange and concentrations of non-structural carbohydrates (NSC) in apple trees of variety 'Jonagold' grafted on two rootstocks of contrasting growth vigor (dwarfing J-TE-G vs invigorating J-TE-H). The measurements were conducted during an exceptionally hot and dry summer. We found that smaller annual trunk radial increments in dwarfed trees were primarily due to an earlier cessation of trunk secondary growth. The interdiurnal trunk circumference changes (ΔC) were slightly lower in dwarfed trees, and these trees also had fewer days with positive ΔC values, particularly during the driest summer months. The trunks of dwarfed trees shrank gradually during the drought, showed less pronounced diurnal variation of trunk circumference and the maximum trunk daily shrinkage was only weakly responsive to the vapor pressure deficit. These results indicated that lower turgidity in the cambial region may have limited the trunk radial expansion in dwarfed trees during the hot and dry days. Dwarfed trees also maintained lower ΨMD and leaf gas exchange rates during the summer drought. These parameters decreased in parallel for both rootstock combinations, suggesting their similar drought sensitivity. Similar concentrations and seasonal dynamics of NSC in both rootstock combinations, together with their similar spring growth rates, suggest that NSC reserves were not directly limiting for growth. Our results support the prominent role of water relations in rootstock-induced size-controlling mechanisms and highlight the complexity of this topic.

Do angiosperm tree species adjust intervessel lateral contact in response to soil drought?

During soil drought (i.e. limited soil water availability to plants), woody species may adjust the structure of their vessel network to improve their resistance against future soil drought stress. Impacts of soil drought on intervessel lateral contact remain poorly understood despite of its significance to xylem transport efficiency and safety. Here, we analysed drought-induced modifications in xylem structures of temperate angiosperm trees with a focus on intervessel lateral contact. Anatomical analyses were performed both in stems of seedlings cultivated under different substrate water availability and annual rings of mature individuals developed during years of low and high soil drought intensities. In response to limited water availability, a decrease in vessel diameter (up to -20%) and simultaneous increase in vessel density (up to +60%) were observed both in seedlings and mature trees. Conversely, there were only small and inconsistent drought-induced changes in intervessel contact frequency and intervessel contact fraction (typically up to ±15%) observed across species, indicating that intervessel lateral contact is a conservative trait. The small adjustments in intervessel lateral contacts were primarily driven by changes in the contact frequencies between neighbouring vessels (i.e. vessel grouping) rather than by changes in proportions of shared cell walls. Our results demonstrate that angiosperm tree species, despite remarkable adjustments in vessel dimensions and densities upon soil drought, exhibit surprisingly invariant intervessel lateral contact architecture.

Abscisic acid-mediated modifications in water transport continuum are involved in cadmium hyperaccumulation in Sedum alfredii.

Abscisic acid (ABA) play a crucial role in plant acclimation to heavy-metals stresses. Nevertheless, the effects of ABA on long-distance transport and its consequences for cadmium (Cd) accumulation are insufficiently understood. Here, we investigated the effects of ABA on the development of the whole-plant water transport pathway and implications for Cd uptake and transport to the shoot of Sedum alfredii. Exposure to Cd stimulated the production of endogenous ABA levels in the non-hyperaccumulating ecotype (NHE), but not in the hyperaccumulating ecotype (HE). Increased ABA levels in NHE significantly reduced aquaporin expressions in roots, the number of xylem vessel in stem, dimensions and densities of stomata in leaves, but induced leaf osmotic adjustment. Furthermore, the ABA-driven modifications in NHE plants showed typically higher sensitivity to ABA content in leaves compared to HE, illustrating ecotype-specific responses to ABA level. In NHE, the ABA-mediated modifications primarily affected the xylem transport of Cd ions and, at the cost of considerable water delivery limitations, significantly reduced delivery of Cd ions to shoots. In contrast, maintenance of low ABA levels in HE failed to t limit transpiration rates and maximized Cd accumulation in shoots. Our results demonstrated that ABA regulates Cd hyperaccumulation of S. alfredii through specific modifications in the water transport continuum.

Ethylene-mediated apoplastic barriers development involved in cadmium accumulation in root of hyperaccumulator Sedum alfredii.

Ethylene is an important phytohormone for plant adaptation to heavy metal stress. However, the effects of ethylene on radial apoplastic transport of Cd remain elusive. This study investigated the role of ethylene on apoplastic barriers development and consequences for Cd uptake in Sedum alfredii. In response to Cd, endogenous ethylene production in hyperaccumulating ecotype (HE) roots was decreased due to the down-regulated expressions of ethylene biosynthesis genes, while the opposite result was observed in non-hyperaccumulating ecotype (NHE). Interestingly, the ethylene emission in HE was always higher than that in NHE, regardless of Cd concentrations. Results of exogenous application of ethylene biosynthesis precursor/inhibitor indicate that ethylene with high level would delay the formation of apoplastic barriers in HE through restraining phenylalanine ammonia lyase activity and gene expressions related to lignin/suberin biosynthesis. Simultaneously, correlation analyses suggest that Cd-induced apoplastic barriers formation may be also regulated by ethylene signaling. By using an apoplastic bypass tracer and scanning ion-selected electrode, we observed that the delayed deposition of apoplastic barriers significantly promoted Cd influx in roots. Taken together, high endogenous ethylene in HE postponed the formation of apoplastic barriers and thus promoted the Cd accumulation in the apoplast of roots.

Simultaneous remediation of sediments contaminated with sulfamethoxazole and cadmium using magnesium-modified biochar derived from Thalia dealbata.

In situ remediation and assessment of sediments contaminated with both antibiotics and heavy metals remains a technological challenge. In this study, MgCl2-modified biochar (BCM) was obtained at 500 °C through slow pyrolysis of Thalia dealbata and used for remediation of sediments contaminated by sulfamethoxazole (SMX) and Cd. The BCM showed greater surface area (110.6 m2 g-1) than pristine biochar (BC, 7.1 m2 g-1). The SMX sorption data were well described by Freundlich model while Langmuir model was better for the Cd2+ sorption data. The addition of 5.0% BCM significantly increased the sorption of SMX (by 50.8-58.6%) and Cd (by 24.2-25.6%) on sediments in both single and binary systems as compared with 5.0% BC. SMX sorption in sediments was significantly improved by addition of Cd2+, whereas SMX has no influence on Cd sorption on sediments. The addition of BCM distinctly decreased both SMX (by 51.4-87.2%) and Cd concentrations (by 56.2-91.3%) in overlying water, as well as in TCLP extracts (by 55.6-86.1% and 58.2-91.9% for SMX and Cd, respectively), as compared with sediments without biochar. Both germination rate and root length of pakchoi increased with increasing doses of BCM in contaminated sediments, 5.0% BCM showed greater promotion on pakchoi growth than 5.0% BC. Overall, BCM in the sediments does not only decrease the bioavailability of SMX and Cd, but it also diminishes the phytotoxicity, and thereby shows great application potential for in situ remediation of sediments polluted with antibiotics and heavy metals.

Role of Vertical Transmission of Shoot Endophytes in Root-Associated Microbiome Assembly and Heavy Metal Hyperaccumulation in Sedum alfredii.

The transmission mode of shoot-associated endophytes in hyperaccumulators and their roles in root microbiome assembly and heavy metal accumulation remain unclear. Using 16S rRNA gene profiling, we investigated the vertical transmission of shoot-associated endophytes in relation to growth and Cd/Zn accumulation of Sedum alfredii ( Crassulaceae). Endophytes were transmitted from shoot cuttings to the rhizocompartment of new plants in both sterilized (γ-irradiated) and native soils. Vertical transmission was far more efficient in the sterile soil, and the transmitted endophytes have become a dominant component of the newly established root-associated microbiome. Based on 16S rRNA genes, the vertically transmitted taxa were identified as the families of Streptomycetaceae, Nocardioidaceae, Pseudonocardiaceae, and Rhizobiaceae. Abundances of Streptomycetaceae, Nocardioidaceae, and Pseudonocardiaceae were strongly correlated with increased shoot biomass and total Cd/Zn accumulation. Inoculation of S. alfredii with the synthetic bacterial community sharing the same phylogenetic relatedness with the vertically transmitted endophytes resulted in significant improvements in plant biomass, root morphology, and Cd/Zn accumulation. Our results demonstrate that successful vertical transmission of endophytes from shoots of S. alfredii to its rhizocompartments is possible, particularly in soils with attenuated microbiomes. Furthermore, the endophyte-derived microbiome plays an important role in metal hyperaccumulation.

Ion-mediated increases in xylem hydraulic conductivity: seasonal differences between coexisting ring- and diffuse-porous temperate tree species.

Ion-mediated changes in hydraulic conductivity (ΔKh) represent a mechanism allowing plants to regulate the rate of xylem transport. However, the significance of ΔKh for ring-porous (RPS) and diffuse-porous tree species (DPS) remains unknown. Here, we examined ΔKh in young branches of three coexisting, temperate RPS (Fraxinus excelsior, Quercus robur, Robinia pseudoacacia) and three DPS (Acer pseudoplatanus, Carpinus betulus, Fagus sylvatica) across the whole year, and assessed the relationships of ΔKh to branch anatomy. Ring-porous species exhibited twice as high ΔKh (10.3% vs 5.3%) within the growing season (i.e., during wood production) compared with DPS, and the production of the annual ring was identified as a crucial process affecting maximum ΔKh within the season. In addition, xylem in branches of RPS generally contained more axial parenchyma (AP; 18% vs 7%) and was characterized by a greater relative contact fraction between vessels and parenchyma (FVP; 59% vs 18%) than xylem in DPS. Simultaneously, ΔKh measured within the growing season was positively correlated with AP, FVP and bark proportions, suggesting that parenchyma in branches may be important for high ΔKh. Significant increase in ΔKh observed during the growing season may help RPS to restore conductive capacity after winter, better compensate transport loss by drought-induced embolism and thereby improve water delivery to leaves.

Abscisic acid-mediated modifications of radial apoplastic transport pathway play a key role in cadmium uptake in hyperaccumulator Sedum alfredii.

Abscisic acid (ABA) is a key phytohormone underlying plant resistance to toxic metals. However, regulatory effects of ABA on apoplastic transport in roots and consequences for uptake of metal ions are poorly understood. Here, we demonstrate how ABA regulates development of apoplastic barriers in roots of two ecotypes of Sedum alfredii and assess effects on cadmium (Cd) uptake. Under Cd treatment, increased endogenous ABA level was detected in roots of nonhyperaccumulating ecotype (NHE) due to up-regulated expressions of ABA biosynthesis genes (SaABA2, SaNCED), but no change was observed in hyperaccumulating ecotype (HE). Simultaneously, endodermal Casparian strips (CSs) and suberin lamellae (SL) were deposited closer to root tips of NHE compared with HE. Interestingly, the vessel-to-CSs overlap was identified as an ABA-driven anatomical trait. Results of correlation analyses and exogenous applications of ABA/Abamine indicate that ABA regulates development of both types of apoplastic barriers through promoting activities of phenylalanine ammonialyase, peroxidase, and expressions of suberin-related genes (SaCYP86A1, SaGPAT5, and SaKCS20). Using scanning ion-selected electrode technique and PTS tracer confirmed that ABA-promoted deposition of CSs and SL significantly reduced Cd entrance into root stele. Therefore, maintenance of low ABA levels in HE minimized deposition of apoplastic barriers and allowed maximization of Cd uptake via apoplastic pathway.

Mechanisms underlying the long-term survival of the monocot Dracaena marginata under drought conditions.

Efficient water management is essential for the survival of vascular plants under drought stress. While interrelations among drought stress, plant anatomy and physiological functions have been described in woody dicots, similar research is very limited for non-palm arborescent and shrubby monocots despite their generally high drought tolerance. In this study, potted transplants of Dracaena marginata Lam. in primary growth stage were exposed to several short- and long-term drought periods. Continuous measurements of sap flow and stem diameter, the evaluation of capacitance and leaf conductance, the quantification of non-structural carbohydrates (NSC), and organ-specific anatomical analyses were performed to reveal the mechanisms promoting plant resistance to limited soil moisture. The plants showed sensitive stomata regulation in the face of drying soil, but only intermediate resistance to water loss through cuticular transpiration. The water losses were compensated by water release from stem characterized by densely interconnected, parenchyma-rich ground tissue and considerable hydraulic capacitance. Our results suggest that the high concentration of osmotically active NSC in aboveground organs combined with the production of root pressures supported water uptake and the restoration of depleted reserves after watering. The described anatomical features and physiological mechanisms impart D. marginata with high resistance to irregular watering and long-term water scarcity. These findings should help to improve predictions with respect to the impacts of droughts on this plant group.

The apoplasmic pathway via the root apex and lateral roots contributes to Cd hyperaccumulation in the hyperaccumulator Sedum alfredii.

Although the significance of apoplasmic barriers in roots with regards to the uptake of toxic elements is generally known, the contribution of apoplasmic bypasses (ABs) to cadmium (Cd) hyperaccumulation is little understood. Here, we employed a combination of stable isotopic tracer techniques, an ABs tracer, hydraulic measurements, suberin lamellae staining, metabolic inhibitors, and antitranspirants to investigate and quantify the impact of the ABs on translocation of Cd to the xylem in roots of a hyperaccumulating (H) ecotype and a non-hyperaccumulating (NH) ecotype of Sedum alfredii. In the H ecotype, the Cd content in the xylem sap was proportional to hydrostatic pressure, which was attributed to pressure-driven flow via the ABs. The contribution of the ABs to Cd transportation to the xylem was dependent on the Cd concentration applied to the H ecotype (up to 37% at the highest concentration used). Cd-treated H ecotype roots showed significantly higher hydraulic conductance compared with the NH ecotype (76 vs 52 × 10­8 m s­1MPa­1), which is in accordance with less extensive suberization due to reduced expression of suberin-related genes. The main entry sites of apoplasmically transported Cd were localized in the root apexes and lateral roots of the H ecotype, where suberin lamellae were not well developed. These findings highlight the significance of the apoplasmic bypass in Cd hyperaccumulation in hyperaccumulating ecotypes of S. alfredii.

Partitioning of vessel resistivity in three liana species.

Vessels with simple perforation plates, found in the majority of angiosperms, are considered the evolutionarily most advanced conduits, least impeding the xylem sap flow. Nevertheless, when measured, their hydraulic resistivity (R, i.e., inverse value of hydraulic conductivity) is significantly higher than resistivity predicted using Hagen-Poiseuille equation (RHP). In our study we aimed (i) to quantify two basic components of the total vessel resistivity - vessel lumen resistivity and end wall resistivity, and (ii) to analyze how the variable inner diameter of the vessel along its longitudinal axis affects resistivity. We measured flow rates through progressively shortened stems of hop (Humulus lupulus L.), grapevine (Vitis vinifera L.), and clematis (Clematis vitalba L.) and used elastomer injection for identification of open vessels and for measurement of changing vessel inner diameters along its axis. The relative contribution of end wall resistivity to total vessel resistivity was 0.46 for hop, 0.55 for grapevine, and 0.30 for clematis. Vessel lumen resistivity calculated from our measurements was substantially higher than theoretical resistivity - about 43% for hop, 58% for grapevine, and 52% for clematis. We identified variation in the vessel inner diameter as an important source of vessel resistivity. The coefficient of variation of vessel inner diameter was a good predictor for the increase of the ratio of integral RHP to RHP calculated from the mean value of inner vessel diameter. We discuss the fact that we dealt with the longest vessels in a given stem sample, which may lead to the overestimation of vessel lumen resistivity, which consequently precludes decision whether the variable vessel inner diameter explains fully the difference between vessel lumen resistivity and RHP we observed.

Linking xylem water storage with anatomical parameters in five temperate tree species.

The release of water from storage compartments to the transpiration stream is an important functional mechanism that provides the buffering of sudden fluctuations in water potential. The ability of tissues to release water per change in water potential, referred to as hydraulic capacitance, is assumed to be associated with the anatomy of storage tissues. However, information about how specific anatomical parameters determine capacitance is limited. In this study, we measured sapwood capacitance (C) in terminal branches and roots of five temperate tree species (Fagus sylvatica L., Picea abies L., Quercus robur L., Robinia pseudoacacia L., Tilia cordata Mill.). Capacitance was calculated separately for water released mainly from capillary (CI; open vessels, tracheids, fibres, intercellular spaces and cracks) and elastic storage compartments (CII; living parenchyma cells), corresponding to two distinct phases of the moisture release curve. We found that C was generally higher in roots than branches, with CI being 3-11 times higher than CII Sapwood density and the ratio of dead to living xylem cells were most closely correlated with C In addition, the magnitude of CI was strongly correlated with fibre/tracheid lumen area, whereas CII was highly dependent on the thickness of axial parenchyma cell walls. Our results indicate that water released from capillary compartments predominates over water released from elastic storage in both branches and roots, suggesting the limited importance of parenchyma cells for water storage in juvenile xylem of temperate tree species. Contrary to intact organs, water released from open conduits in our small wood samples significantly increased CI at relatively high water potentials. Linking anatomical parameters with the hydraulic capacitance of a tissue contributes to a better understanding of water release mechanisms and their implications for plant hydraulics.

An improved method for the visualization of conductive vessels in Arabidopsis thaliana inflorescence stems.

Dye perfusion is commonly used for the identification of conductive elements important for the study of xylem development as well as precise hydraulic estimations. The tiny size of inflorescence stems, the small amount of vessels in close arrangement, and high hydraulic resistivity delimit the use of the method for quantification of the water conductivity of Arabidopsis thaliana, one of the recently most extensively used plant models. Here, we present an extensive adjustment to the method in order to reliably identify individual functional (conductive) vessels. Segments of inflorescence stems were sealed in silicone tubes to prevent damage and perfused with a dye solution. Our results showed that dyes often used for staining functional xylem elements (safranin, fuchsine, toluidine blue) failed with Arabidopsis. In contrast, Fluorescent Brightener 28 dye solution perfused through segments stained secondary cell walls of functional vessels, which were clearly distinguishable in native cross sections. When compared to identification based on the degree of development of secondary cell walls, identification with the help of dye perfusion revealed a significantly lower number of functional vessels and values of theoretical hydraulic conductivity. We found that lignified but not yet functional vessels form a substantial portion of the xylem in apical and basal segments of Arabidopsis and, thus, significantly affect the analyzed functional parameters of xylem. The presented methodology enables reliable identification of individual functional vessels, allowing thus estimations of hydraulic conductivities to be improved, size distributions and vessel diameters to be refined, and data variability generally to be reduced.

A comparative structural and functional study of leaf traits and sap flow in Dracaena cinnabari and Dracaena draco seedlings.

Water relations for two remote populations of Dracaena tree species from the dragon tree group, Dracaena cinnabari Balfour f. and Dracaena draco (L.) L., were studied to test our hypothesis that morphological and anatomical differences in leaf structure may lead to varied functional responses to changing environmental conditions. Sap flow measurements were performed using the heat field deformation method for four Dracaena seedlings grown in one glasshouse and two greenhouses, and leaf traits related to plant-water relationships were characterised. All traits studied confirmed that D. cinnabari leaves are more xeric in their morpho-anatomical structure compared with D. draco leaves. No radial sap flow variability was detected in D. draco plant stems, whereas sap flow was found to be higher in the inner part of D. cinnabari stems. The regular occurrence of reverse sap flow at night in both Dracaena species was consistent with a staining experiment. Vapour pressure deficit (VPD) was found to be the main driver for transpiration for both Dracaena species. However, the relationship between VPD and sap flow appeared to be different for each species, with a clockwise or no hysteresis loop for D. draco and a counter-clockwise hysteresis loop for D. cinnabari. This resulted in a shorter transpiration cycle in D. cinnabari. The observed superior water-saving strategy of D. cinnabari corresponds to its more xeric morpho-anatomical leaf structure compared with D. draco.