Seasonal dynamics of phloem and xylem formation in silver fir and Norway spruce as affected by drought

The dynamics of phloem growth ring formation in silver fir (Abies alba Mill.) and Norway spruce (Picea abies Karst.) at different sites in Slovenia during the droughty growing season of 2003 was studied. We also determined the timing of cambial activity, xylem and phloem formation, and counted the number of cells in the completed phloem and xylem growth rings. Light microscopy of cross-sections revealed that cambial activity started on the phloem and xylem side simultaneously at all three plots. However, prior to this, 1–2 layers of phloem derivatives near the cambium were differentiated without previous divisions. The structure of the early phloem was similar in silver fir and Norway spruce. Differences in the number of late phloem cells were found among sites. Phloem growth rings were the widest in Norway spruce growing at the lowland site. In all investigated trees, the cambium produced 5–12 times more xylem cells than phloem ones. In addition, the variability in the number of cells in the 2003 growth ring around the stem circumference of the same tree and among different trees was higher on the xylem side than on the phloem side. Phloem formation is presumably less dependent on environmental factors but is more internally driven than xylem formation.     

Comparative axial widening of phloem and xylem conduits in small woody plants

Key message

Along the stem axis phloem’s sieve elements increase in diameter basally at rates comparable to those of xylem conduits and in agreement with principles of hydraulic optimization.

Abstract

Plant physiology relies on the efficiency of the two long-distance transport systems of xylem and phloem. Xylem architecture comprises conduits of small dimensions towards the stem apex, where transpiration-induced tensions are the highest along the root-to-leaves hydraulic pathway, and widen basally to minimize the path length resistance to water flow. Instead, information on phloem anatomy and allometry is extremely scarce, although potentially relevant for the efficiency of sugar transportation. We measured the hydraulic diameter (Dh) of both xylem conduits and phloem sieve elements in parallel at different heights along the stem of a small tree of Picea abies, Fraxinus excelsior and Salix eleagnos. Dh increased from the stem apex to base in both xylem and phloem, with a higher scaling exponent (b) of sieve elements than that of tracheids in the conifer (0.19 vs. 0.14) and lower than that of vessels in the angiosperms (0.14–0.22 vs. 0.19–0.40). In addition, sieve elements were larger than tracheids in P. abies and narrower than angiosperms vessels at any height along the stem. In conclusion, axial conduit widening would seem to be a key feature of both xylem and phloem long-distance transport architectures.     

Seasonal variation in formation,structure, and chemical properties of phloem in <Emphasis Type="Italic">Picea abies</Emphasis> as studied by novel microtechniques

Main conclusion

Phloem production and structural development were interlinked with seasonal variation in the primary and secondary metabolites of phloem. Novel microtechniques provided new perspectives on understanding phloem structure and chemistry. To gain new insights into phloem formation in Norway spruce (Picea abies), we monitored phloem cell production and seasonal variation in the primary and secondary metabolites of inner bark (non-structural carbohydrates and phenolic stilbene glucosides) during the 2012 growing season in southern and northern Finland. The structure of developing phloem was visualised in 3D by synchrotron X-ray microtomography. The chemical features of developing phloem tissues isolated by laser microdissection were analysed by chemical microanalysis. Within-year phloem formation was associated with seasonal changes in non-structural carbohydrates and phenolic extractive contents of inner bark. The onset of phloem cell production occurred in early and mid-May in southern and northern Finland, respectively. The maximal rate of phloem production and formation of a tangential band of axial phloem parenchyma occurred in mid-June, when total non-structural carbohydrates peaked (due to the high amount of starch). In contrast, soluble sugar content dropped during the most active growth period and increased in late summer and winter. The 3D visualisation showed that the new axial parenchyma clearly enlarged from June to August. Sub-cellular changes appeared to be associated with accumulation of stilbene glucosides and soluble sugars in the newest phloem. Stilbene glucosides also increased in inner bark during late summer and winter. Our findings may indicate that stilbene biosynthesis in older phloem predominantly occurs after the formation of the new band(s) of axial parenchyma. The complementary use of novel microtechniques provides new perspectives on the formation, structure, and chemistry of phloem.

     

The growth and nutrients status of conifers on ash-treated cutaway peatland

Key message

Use of wood ash or a mixture of wood and oil shale ashes increases the concentrations of P and K in the assimilation organs of conifers and stimulates tree growth.

Abstract

The effect of fertilization with wood ash (10 and 15 t ha^?1) and a mixture of wood ash (10 t ha^?1) and oil shale ash (8 t ha^?1) on the growth (height, root collar diameter, biomass, biomass production) and nutrient concentrations in subsoil and needles of young Pinus sylvestris and Picea abies plants on the Puhatu (Northeast Estonia) cutaway peatland in the first 2 years were studied. After the second growing year differences in the average height growth of P. abies and P. sylvestris were statistically significantly higher on ash-treated plots than on the control plots (p < 0.05), being respectively 1.4–1.6 and 1.5–1.7 times greater than height growth of the control trees. The best results on root collar diameter were observed on mixture ash treatments: the root collars were 1.9 (P. abies) and 2.2 (P. sylvestris) times larger than of the control trees. The biomass of the two conifer species and the biomass production of P. sylvestris in 2012 was the greatest on the mixture ash treatments. Five months after fertilization with ashes the concentrations of P, K, Ca and Mg were higher on the treated plots than on the control plot. Although the concentrations of P and K in P. sylvestris needles rose after the treatment with ash, seedlings suffered from P and K deficiency. The concentrations of P and K in P. abies needles were on optimum. The P/N and the K/N ratios in needles were also improved compared to control trees needles.     

Hydrological properties of bark of selected forest tree species. Part I: the coefficient of development of the interception surface of bark

Key message

The coefficient of development of the interception surface of bark allows for objective assessment of the degree of bark surface differentiation between different species.

Abstract

Inter-species differentiation of bark morphology and its variability progressing with tree age suggest that the hydrological properties of the bark of particular species depend on the degree of development of the outer bark surface of trees. The aim of the present research was to develop a method of calculating the actual bark surface with the use of the coefficient of development of the interception surface of bark, describing the degree of development of the outer bark surface of trees. The primary aim was to show inter-species differentiation of the coefficient of development of the interception surface of bark at breast height, as well as its variability within a single species, progressing with tree age. The present study shows the results obtained for 77 bark samples collected at the breast height of the following tree species: Pinus sylvestris L., Larix decidua Mill., Abies alba Mill., Picea abies L., Quercus robur L., Fagus sylvatica L., Acer pseudoplatanus L. and Betula pendula Ehrh. In all of the examined species, the coefficient of development of the interception surface of bark shows a distinct relation to the breast-height diameter. The highest values of coefficient of development of the interception surface of bark among the thickest trees are reached by: L. decidua—2.56, Pinus sylvestris—2.28 and B. pendula—2.44, whereas the lowest values are reached by the bark of European beech F. sylvatica—1.07. The coefficient of development of the interception surface of bark describes the morphological differentiation of the outer bark surface of trees in an objective way. Owing to its mathematical form, the coefficient of development of the interception surface of bark may be useful in the modelling of hydrological processes occurring in forest ecosystems.     

A novel insight into the structure of amphivasal secondary bundles on the example of Dracaena draco L. stem

Key message

Pattern of tracheids found along the bundles extends understanding of their cross - sectional anatomy and sheds a new light on the issue of radial transport in monocotyledons with secondary growth.

Abstract

Secondary growth of Dracaena draco L. stem is connected with the formation of amphivasal vascular bundles in which a centrally located phloem is surrounded by a ring of xylem cells (tracheids). However, as visible in a single transverse section, there is a tendency towards variation among the secondary bundles from such with a xylem ring to ones in which the tracheids do not completely surround the phloem, i.e., are separated by vascular parenchyma cells. We aimed to elucidate the cross-sectional anatomy of amphivasal secondary bundles using the method of serial sectioning (with sections 3 μm thick), which allowed us to follow very precisely the bundle structure along its length. The analysis revealed that the xylem arrangement in these bundles depends on the position of a section in the bundle path. Each amphivasal bundle is composed of sectors where tracheids form a ring, as well as of such where tracheids are separated by vascular parenchyma cells. We hypothesize that this structure of amphivasal vascular bundles facilitates radial transport of assimilates to the sink tissues. The result of the anatomical analysis is discussed in a physiological context.     

Comparing growth phenology of co-occurring deciduous and evergreen conifers exposed to drought

Plant phenological events are influenced by climate factors such as temperature and rainfall. To evaluate phenological responses to water availability in a Spring Heath-Pine wood (Erico-Pinetum typicum), the focus of this study was to determine intra-annual dynamics of apical and lateral growth of co-occurring early successional Larix decidua and Pinus sylvestris and late successional Picea abies exposed to drought. The effect of reduced plant water availability on growth phenology was investigated by conducting a rainfall exclusion experiment. Timing of key phenological dates (onset, maximum rate, end, duration) of growth processes were compared among species at the rain-sheltered and control plot during 2011 and 2012. Shoot and needle elongation were monitored on lateral branches in the canopy at c. 16 m height and radial growth was recorded by automatic dendrometers at c. 1.3 m height of >120 yr old trees. Different sequences in aboveground growth phenology were detected among the three species under the same growing conditions. While onset of radial growth in April through early May was considerably preceded by onset of needle growth in Larix decidua (5–6 weeks) and shoot growth in Pinus sylvestris (c. 3 weeks), it occurred quite simultaneously with onset of shoot growth in Picea abies. Low water availability had a minor impact on onset of aboveground growth, which is related to utilization of stored water, but caused premature cessation of aboveground growth. At the control plot mean growing season length was 130 days in Pinus sylvestris, 95 days in Larix decidua and 73 days in Picea abies, supporting the hypothesis that early successional species are resource expenders, while late successional species are more efficient in utilizing resources and develop safer life strategies. High synchronicity found in culmination of radial growth in late spring (mid-May through early June) prior to occurrence of more favourable environmental conditions in summer might indicate sink competition for carbohydrates to belowground organs. This is supported by completion of apical growth in mid June in all species, except for needle growth of Pinus sylvestris which lasted until early August. Phenological observations of conifers exposed to drought revealed that tree water status early during the growing season determines total annual aboveground growth and, besides temperature, species-specific endogenous and/or environmental factors (most likely photoperiod and/or different threshold temperatures) are involved in controlling apical and lateral growth resumption after winter dormancy.     

SEASONAL DEVELOPMENT OF THE SECONDARY PHLOEM IN PINUS

Functional sieve cells are present at all times in the secondary phloem of Pinus banksiana Lamb., P. resinosa Ait., and P. strobus L. With regard to a given year's growth increment, all but the last-formed sieve cells (2-4 layers) cease functioning the same season they are derived from the cambium. The former overwinter and remain functional until new sieve cells differentiate in spring. Toward the end of March undifferentiated cells in the outer margin of the cambial zone begin to differentiate into sieve cells. About a week later, cambial activity (cell division) commences. All early phloem is produced by early May before new xylem differentiation begins. Most sieve cells are differentiated by late August, but a few not until late September. Cessation of function begins in late May or June with formation of definitive callose on sieve areas of the sieve cells which overwintered and continues slowly to sieve cells of the current season's early phloem. By mid-December all but the last-formed sieve cells (i.e., those which will overwinter in a functional state) are devoid of contents. Phloem differentiation precedes xylem differentiation by approximately 1 1/2 months. Xylem and phloem production cease more or less simultaneously in August, xylem and phloem differentiation in September.     

Identification of three wheat globulin genes by screening a Triticum aestivum BAC genomic library with cDNA from a diabetes-associated globulin

Background

Plant systemic signaling characterized by the long distance transport of molecules across plant organs involves the xylem and phloem conduits. Root-microbe interactions generate systemic signals that are transported to aerial organs via the xylem sap. We analyzed the xylem sap proteome of soybean seedlings in response to pathogenic and symbiotic interactions to identify systemic signaling proteins and other differentially expressed proteins.

Results

We observed the increase of a serine protease and peroxidase in the xylem sap in response to Phytophthora sojae elicitor treatment. The high molecular weight fraction of soybean xylem sap was found to promote the growth of Neurospora crassa in vitro at lower concentrations and inhibit growth at higher concentrations. Sap from soybean plants treated with a P. sojae elicitor had a significantly higher inhibitory effect than sap from control soybean plants. When soybean seedlings were inoculated with the symbiont Bradyrhizobium japonicum, the abundance of a xyloglucan transendoglycosyl transferase protein increased in the xylem sap. However, RNAi-mediated silencing of the corresponding gene did not significantly affect nodulation in soybean hairy root composite plants.

Conclusion

Our study identified a number of sap proteins from soybean that are differentially induced in response to B. japonicum and P. sojae elicitor treatments and a majority of them were secreted proteins.     

Phloem unloading via the apoplastic pathway is essential for shoot distribution of root-synthesized cytokinins

Root-synthesized cytokinins are transported to the shoot and regulate the growth, development, and stress responses of aerial tissues. Previous studies have demonstrated that Arabidopsis (Arabidopsis thaliana) ATP binding cassette (ABC) transporter G family member 14 (AtABCG14) participates in xylem loading of root-synthesized cytokinins. However, the mechanism by which these root-derived cytokinins are distributed in the shoot remains unclear. Here, we revealed that AtABCG14-mediated phloem unloading through the apoplastic pathway is required for the appropriate shoot distribution of root-synthesized cytokinins in Arabidopsis. Wild-type rootstocks grafted to atabcg14 scions successfully restored trans-zeatin xylem loading. However, only low levels of root-synthesized cytokinins and induced shoot signaling were rescued. Reciprocal grafting and tissue-specific genetic complementation demonstrated that AtABCG14 disruption in the shoot considerably increased the retention of root-synthesized cytokinins in the phloem and substantially impaired their distribution in the leaf apoplast. The translocation of root-synthesized cytokinins from the xylem to the phloem and the subsequent unloading from the phloem is required for the shoot distribution and long-distance shootward transport of root-synthesized cytokinins. This study revealed a mechanism by which the phloem regulates systemic signaling of xylem-mediated transport of root-synthesized cytokinins from the root to the shoot.

Phloem unloading via the apoplastic pathway is essential for shoot distribution and long-distance translocation of root-synthesized cytokinins from the root to the shoot through the xylem.     

Shoot development of Norway spruce (Picea abies) involves changes in piperidine alkaloids and condensed tannins

Key Message

Secondary chemistry of P. abies peaks early in shoot development. Condensed tannins accumulate already in late buds while piperidine alkaloid biosynthesis take place in early stage shoots.

Abstract

Plants protect new vegetative parts with defensive secondary metabolite compounds. We investigated how concentrations of piperidine alkaloids and condensed tannins change during bud burst and shoot growth in adult Picea abies. We detected 12 individual piperidine compounds, of which epipinidinone and 1,2-dehydropinidinone and two tentatively identified 1,6-imines are reported for the first time in P. abies. In addition three piperidine alkaloid compounds remain partly identified. We found that concentrations of both total piperidine alkaloids and condensed tannins were highest immediately after bud burst. While concentrations of condensed tannins started to increase during bud opening, the dilution effect decreased concentrations in the developing needles of mature branches. By contrast, the decrease of total alkaloid concentrations in mature shoots was not due to the dilution effect, but was connected to the disappearance of precursor components of biosynthesis. The concentrations of major alkaloid components remain stable from dormant buds to mature needles and twigs, underlining their importance for P. abies, although their real ecological significance is yet to be solved. Based on the structural features and timing of appearance of individual compounds, we also propose a hypothetical biosynthesis route for trans-substituted coniferous piperidine alkaloids.     

Development and morphology of stone cells in phloem of Toxicodendron vernicifluum

Key message

The morphology and development of sumac phloem sclereid were observed, sclereid was developed from phloem parenchyma and lignin was deposited in the cell wall of parenchyma and formation sclereid.

Abstract

Sumac [Toxicodendron vernicifluum (Stokes) F.A.Barkley] is a unique economic tree species in China. Raw lacquer is the sap flowing from the phloem of sumac. Stone cell clusters exist in the secondary phloem of sumac stem. In the present study, the morphology and development of stone cell clusters in sumac phloem were observed with optical microscope and transmission electron microscope. The distribution of lignin in the composition molecules of secondary phloem was observed with histochemistry method and fluorescence microscope. The results showed that phloem stone cells of sumac were developed from phloem parenchyma cells, and that lignin was deposited in layers in the cell wall of phloem parenchyma cells which cause the formation of stone cell clusters and which have the secondary wall. Studies on the ultrastructure of stone cells indicated that there was an obvious stratification and pits during the process of lignin deposition.     

Model-assisted evaluation of crop load effects on stem diameter variations and fruit growth in peach

Key message

The paper identifies and quantifies how crop load influences plant physiological variables that determine stem diameter variations to better understand the effect of crop load on drought stress indicators.

Abstract

Stem diameter (D ^stem) variations have extensively been applied in optimisation strategies for plant-based irrigation scheduling in fruit trees. Two D ^stem derived water status indicators, maximum daily shrinkage (MDS) and daily growth rate (DGR), are however influenced by other factors such as crop load, making it difficult to unambiguously use these indicators in practical irrigation applications. Furthermore, crop load influences the growth of individual fruits, because of competition for assimilates. This paper aims to explain the effect of crop load on DGR, MDS and individual fruit growth in peach using a water and carbon transport model that includes simulation of stem diameter variations. This modelling approach enabled to relate differences in crop load to differences in xylem and phloem water potential components. As such, crop load effects on DGR were attributed to effects on the stem phloem turgor pressure. The effect of crop load on MDS could be explained by the plant water status, the phloem carbon concentration and the elasticity of the tissue. The influence on fruit growth could predominantly be explained by the effect on the early fruit growth stages.     

Mistletoes and mutant albino shoots on woody plants as mineral nutrient traps

Background and Aims

Potassium, sulphur and zinc contents of mistletoe leaves are generally higher than in their hosts. This is attributed to the fact that chemical elements which are cycled between xylem and phloem in the process of phloem loading of sugars are trapped in the mistletoe, because these parasites do not feed their hosts. Here it is hypothesized that mutant albino shoots on otherwise green plants should behave similarly, because they lack photosynthesis and thus cannot recycle elements involved in sugar loading.

Methods

The mineral nutrition of the mistletoe Scurrula elata was compared with that of albino shoots on Citrus sinensis and Nerium oleander. The potential for selective nutrient uptake by the mistletoe was studied by comparing element contents of host leaves on infected and uninfected branches and by manipulation of the haustorium–shoot ratio in mistletoes. Phloem anatomy of albino leaves was compared with that of green leaves.

Key Results

Both mistletoes and albino leaves had higher contents of potassium, sulphur and zinc than hosts or green leaves, respectively. Hypothetical discrimination of nutrient elements during the uptake by the haustorium is not supported by our data. Anatomical studies of albino leaves showed characteristics of release phloem.

Conclusions

Both albino shoots and mistletoes are traps for elements normally recycled between xylem and phloem, because retranslocation of phloem mobile elements into the mother plant or the host is low or absent. It can be assumed that the lack of photosynthetic activity in albino shoots and thus of sugars needed in phloem loading is responsible for the accumulation of elements. The absence of phloem loading is reflected in phloem anatomy of these abnormal shoots. In mistletoes the evolution of a parasitic lifestyle has obviously eliminated substantial feeding of the host with photosynthates produced by the mistletoe.     

Ascorbic acid and development of xylem and phloem cells in the pine trunk

Changes in the levels of ascorbic acid (AA), its oxidized form, dehydroascorbic acid (DHA), and uronic acids as initial precursors for the AA synthesis were studied as related to the degree of xylem and phloem cell development in the course of early and late wood formation in the trunks of Scots pine (Pinus sylvestris L.). The cells of mature and conducting phloem, cambial zone, differently developed cells in the zones of cell enlargement and maturation were obtained by successive scraping tissue layers from trunk segments of 20–25-year-old trees; tissue identification was checked anatomically and histochemically. The contents of compounds tested were calculated per dry weight and per cell basis. We found great differences in the contents of AA and DHA and also in their ratio in dependence of the wood type developing in the pine trunks during growth period and on the stage of differentiation of xylem and phloem cells. Changes in the AA content during xylem cell differentiation were accompanied by changes in the content of uronic acids. The amounts of AA, DHA, and uronic acids were the highest at the stage of early lignification and reduced with tracheid maturation. The AA to DHA ratio changed differently in the course of early and late xylem lignification. It reduced from the start of lignification to the formation of early mature xylem and, in contrast, increased in mature late wood; this indicates a difference in the level of redox processes in these tissues.     

Seasonal development of phloem in scots pine stems

The formation of phloem was studied for two years in stems of 50 to 60 year old trees of Scots pine (Pinus sylvestris L.) growing in nature. The development of phloem of the current year begins 10 to 20 days before the xylem formation and is completed with the termination of shoot growth in the end of June. Observations over the seasonal activity of cambium producing sieve cells of phloem and duration of their differentiation as compared to the xylem derivatives of cambium have shown that the maxima of formation of phloem and xylem cells could coincide or not coincide by season, while the activities of their differentiation were always in antiphase. The sieve cells of early phloem were separated from those of late phloem by a layer of tannin-containing cells, which are formed simultaneously with the formation of late xylem cells by the cambium. Seasonal dynamics of accumulation of starch grain in structural elements of the phloem is related to the xylem development. The content of metabolites in differentiating and mature phloem elements, in the cambium zone, and in the xylem cells growing in the radial direction depended on cell specificity, stage of their development, and type of forming wood, early or late, which differ in the cell wall parameters and, hence, requirement of assimilates. Significant differences were described between the content of low molecular weigh carbohydrates, amino acids, organic acids, and phenol compounds using two methods of calculation: per dry weight and per cell.     

Phloem as Capacitor: Radial Transfer of Water into Xylem of Tree Stems Occurs via Symplastic Transport in Ray Parenchyma

The transfer of water from phloem into xylem is thought to mitigate increasing hydraulic tension in the vascular system of trees during the diel cycle of transpiration. Although a putative plant function, to date there is no direct evidence of such water transfer or the contributing pathways. Here, we trace the radial flow of water from the phloem into the xylem and investigate its diel variation. Introducing a fluorescent dye (0.1% [w/w] fluorescein) into the phloem water of the tree species Eucalyptus saligna allowed localization of the dye in phloem and xylem tissues using confocal laser scanning microscopy. Our results show that the majority of water transferred between the two tissues is facilitated via the symplast of horizontal ray parenchyma cells. The method also permitted assessment of the radial transfer of water during the diel cycle, where changes in water potential gradients between phloem and xylem determine the extent and direction of radial transfer. When injected during the morning, when xylem water potential rapidly declined, fluorescein was translocated, on average, farther into mature xylem (447 ± 188 µm) compared with nighttime, when xylem water potential was close to zero (155 ± 42 µm). These findings provide empirical evidence to support theoretical predictions of the role of phloem-xylem water transfer in the hydraulic functioning of plants. This method enables investigation of the role of phloem tissue as a dynamic capacitor for water storage and transfer and its contribution toward the maintenance of the functional integrity of xylem in trees.Physiological and hydraulic functioning of the two long-distance transport systems in trees, xylem and phloem, have intrigued plant researchers for more than a century. Since the pioneering work of Dixon and Joly (1895; cohesion-tension theory for xylem) and Münch (1930; pressure flow hypothesis for phloem), the majority of studies have investigated these systems independently of each other. Although the work of Stout and Hoagland (1939) as well as Biddulph and Markle (1944) laid the foundation for the physiological nexus between xylem and phloem, it is only recently that we have begun to understand the importance of the hydraulic nexus (Hölttä et al., 2006, 2009; Sevanto et al., 2011, 2014). Processes related to both nexus occur in parallel, and here the term physiological nexus covers all metabolite exchange, including the bidirectional flow of amino acids, minerals, and carbohydrates (Wardlaw, 1974; Ferrier et al., 1975; Pfautsch et al., 2009, 2015; De Schepper et al., 2013; for review, see van Bel, 1990, 2003). The term hydraulic nexus refers to the function of phloem as a capacitor, where water stored in phloem moves into xylem vessels to maintain the integrity of the transpiration stream (Zweifel et al., 2000, and refs. therein). Throughout this article, we use the term phloem collectively for cells that make up the transport phloem of woody plants (including sieve element/companion cell complexes, parenchyma cells, etc.), as opposed to collection and release phloem tissue, which differ in structure and function. Transport phloem is characterized by the retention of “high hydrostatic pressure by retrieval of leaked osmotica accompanied by water flux” (Patrick, 2013).According to the cohesion-tension theory, water in xylem vessels is constantly under tension and moves in a metastable state from roots to leaves along a hydrostatic pressure gradient. Depending on both the availability of soil moisture and the vapor pressure deficit of the atmosphere, this tension can exceed the cohesive forces that bind water molecules, resulting in the formation of a gas void that, after expanding, can lead to rupture of the water column inside individual vessels (termed cavitation; Zimmermann, 1983). Once cavitated, vessels become dysfunctional, and the transport of water and nutrients to leaves declines. However, water stored in woody tissues of trees can be mobilized to alleviate the risk of cavitation, and recent theory suggests that both water and carbohydrates from phloem may aid in the reversal of vessel embolism (i.e. air intrusion), although the evidence is indirect (Salleo et al., 2009; Brodersen et al., 2010; Nardini et al., 2011).All parts of plants have a water storage capacity (symplastic and apoplastic), and this capacitance increases with tree size and age (Phillips et al., 2003). The ability to mobilize stored water varies according to the force required to drag it out of storage (Holbrook, 1995). One-half century ago, Reynolds (1965) highlighted the importance of the volume of internally stored water to support the transpiration of trees. Since then, studies have quantified the fraction of stored water in total daily transpiration for a range of tree species. This fraction varies between 2% and 20% (Tyree and Yang, 1990; Čermák et al., 2007, and refs. therein; Barnard et al., 2011; Pfautsch and Adams, 2013) and is generally smaller in angiosperms compared with gymnosperms, where a maximum fraction of 50% was reported for Pinus sylvestris (Waring et al., 1979). Given that the daily water use of large adult trees can easily reach 260 to 380 L (Čermák et al., 2007; Pfautsch et al., 2011), considerable volumes of stored water must be mobilized from and restored back into capacitors on a daily basis. Remobilization of stored water also can prolong stomatal opening and thus increase carbon gain (Goldstein et al., 1998).The volume of stored water released depends on the elasticity of the storage tissue, its connectivity to xylem vessels, and the gradient of water potential (ψ) between the storage tissue and vessels. Cells with elastic walls represent ideal capacitors because they can change their volume as a consequence of small changes in ψ. Thus, phloem, cambium, and juvenile xylem cells are well suited for water storage and release (Yang and Tyree, 1992; Zweifel et al., 2014). The magnitude of release and refill of stored water in trees can be approximated by separately measuring the change in thickness of phloem and xylem during a diel cycle using high-precision dendrometers (Zweifel et al., 2014). Whitehead and Jarvis (1981) have calculated that around 90% of the diurnal change in stem radius can be attributed to changes in the water content of cambial and phloem tissues. To date, it is commonly accepted that tree bark, independent of the wood below, swells during the night and shrinks during the day (Zweifel et al., 2000), reflecting the water flow dynamics that characterize the dynamic exchange of water between phloem and xylem.Phloem and xylem tissues are separated by rows of intermediary cambial cells. However, depending on the species, phloem and xylem are connected through uniseriate or multiseriate strands of radially aligned ray parenchyma cells, commonly termed wood rays. These rays have been shown to be capable of symplastic water transport through plasmodesmata (Höll, 1975). Based on measurements of radial conductance, Sevanto et al. (2011) suggested that aquaporins also might be involved in the radial transfer of water. Both theoretical and experimental approaches have been developed to better understand the dynamic exchange of water between xylem and phloem. Hölttä et al. (2006, 2009) developed a model based on Münch’s hypothesis and included a term that represents the hydraulic connection between the two tissue types. Through incorporating this term, model outputs suggest the occurrence of a constant exchange of water between xylem and phloem along gradients of ψ. However, some authors suggested that changes in ψ of xylem alone might be insufficient to account for the observed diurnal shrinkage and swelling of bark (Sevanto et al., 2003). Along this line of argument, loading and unloading of carbohydrates in phloem tissue has been suggested to further impact the radial transfer of water and associated changes in bark thickness (Mencuccini et al., 2013).Nevertheless, to date, all approaches remain indirect, and the routes of water transfer between phloem and xylem have yet to be determined. Here, we present a technique that enables the visualization of water transfer from phloem to xylem tissues and resolves the apoplastic and symplastic pathways and cell types. The method involves the injection of an aqueous solution that contains fluorescent dye into phloem followed by analyses of woody tissues using confocal laser scanning microscopy. We assess the effectiveness of three different dyes to stain possible transfer pathways. We also introduce dye during different time intervals of the diel transpiration cycle to test the effect of predicted dynamic changes in ψ between phloem and xylem on the transfer processes. We hypothesized that radial transfer of water would be most pronounced during periods where conditions of the hydraulic nexus between phloem and xylem differ the most. These differences are expected during high rates of transpiration that cause a steep decline in xylem ψ, commonly observed during morning hours. We use leaf water potential (ψ_L) and high-precision dendrometer measurements to identify relevant time intervals. The simultaneous assessment of ψ_L and the independent diameter fluctuation of phloem and xylem may provide empirical evidence for the role of phloem as a water storage capacitor that helps mitigate increasing tension in the transpiration stream.     

Temperature thresholds for the onset of xylogenesis in alpine shrubs on the Tibetan Plateau

Key message

The threshold minimum air temperature driving xylem growth of alpine  Rhododendron aganniphum is lower than that commonly observed at the treeline of conifers.

Abstract

Understanding how alpine shrubs grow and which environmental factors drive their biomass gain could help to functionally differentiate trees and shrubs. The cambium is the main meristem responsible for wood formation in trees and shrubs. Thus, a better knowledge of cambium growth dynamics in alpine shrubs would allow explaining why shrubs displace trees above the treeline. Here, we aim to investigate the timings and dynamics of xylogenesis and to identify the thermal thresholds controlling the onset of xylem growth of Rhododendron aganniphum, a tall shrub growing above the alpine treeline on the Tibetan Plateau. Timings of xylogenesis and radial growth rates were assessed from anatomical observations of the developing xylem during three growing seasons (2011, 2012, and 2013). The threshold temperature at which xylogenesis had a 0.5 probability of being active was calculated with logistic regressions. The onset of xylogenesis was observed between mid and late June, whereas the end of xylogenesis lasted from mid to late September. Overall, the duration of xylem growth lasted 88–101 days, and 94 % of the ring was formed from June to August. The threshold for the onset of xylem growth was observed at 2.0 ± 0.6 °C for the minimum air temperature, lower than that commonly observed for treeline conifers (ca. 6 °C). This low thermal threshold allows alpine shrubs to have a growing season long enough to complete xylem production and maturation during the warmest summer months. Our results suggest that the time required to complete xylogenesis is critical to understand why shrubs displace trees above the treeline.

     

Declining hydraulic performances and low carbon investments in tree rings predate Scots pine drought-induced mortality

Key message

The retrospective analysis of wood anatomical features evidences how a long-term deterioration of hydraulic performance and carbon use portend drought-induced mortality in Scots pine.

Abstract

Widespread episodes of drought-induced tree mortality are predicted to become more frequent as climate becomes warmer and drier. Nevertheless, growth trends and their links to changes in wood anatomy before tree dies are still poorly understood. Wood anatomical features provide valuable information that can be extracted to infer the mechanisms leading to tree death. In this study, we characterize drought-induced mortality affecting two Scots pine (Pinus sylvestris) sites (Prades and Arcalís) located in the North Eastern Iberian Peninsula. Co-occurring now-dead and living Scots pine trees were sampled and their wood anatomical features were measured and compared. We aimed to detect differences in anatomical features between living and dead trees, and to infer past physiological performances that might have determined their subsequent death or survival. Now-dead trees showed lower tracheid and resin duct production, and smaller radial lumen diameters than co-occurring living trees. At the more xeric Prades site, these anatomical differences were larger and chronic, i.e. were observed over the three studied decades, whilst they were less pronounced at the other, more mesic Arcalís site, where tree mortality episodes were more recent. This indicates that dead trees’ hydraulic conductivity was severely affected and that carbon investment in xylem formation and resin duct production was constrained prior to tree death. Our findings show that both hydraulic deterioration and low carbon allocation to xylem formation were associated to drought-induced mortality in Scots pine. Nevertheless, the temporal dynamics of these processes differed between populations as a function of site climatic conditions.