Reaction Wood Formation Research Articles

Cortical microtubule dynamics during reaction wood formation ensures context-appropriate cellulose microfibril angle in woody trees

Key messageCortical microtubule arrays are the primary mechanism for guiding the re-orientation of cellulose microfibrils and determining MFA in secondary cell wall of wood fibre and tracheid cells in reaction wood.Microtubules are directly and indirectly involved in guiding cellulose synthase complexes (CSCs) through the plasma membrane. The angle of cellulose deposition is a critical response to environmental signals and/or stress conditions, and particularly crucial during reaction wood formation, a process in which woody plants deposit additional cell wall material to counteract gravitational forces. Tubulin genes are upregulated in response to gravitational stimulus during reaction wood formation, which can result in changes to microtubule assembly. In this study, microtubules were visualised in three woody tree species (two angiosperms: Eucalyptus globulus Labill., Populus alba L., and one gymnosperm: Pinus radiata D. Don.) using immunofluorescence to quantitatively evaluate microtubule organisation during reaction wood formation. Our results suggest that reorientation of the cortical microtubule array affects secondary cell wall deposition, even across different types of reaction wood, by ensuring context-appropriate orientation of cellulose microfibrils and determining MFA in wood cells. Pharmacological studies conducted on in vitro cultured stem segments or in vivo during reaction wood formation corroborated these important roles for microtubules during wood development. This study starts to unveil the role of tubulins during wood formation by exploring cortical microtubule array organisation in trees subjected to gravitational stimulus and it sheds light on cellular and molecular mechanisms behind cellulose deposition in tree species.

Usporedba reakcijskoga i normalnog drva nekih komercijalnih vrsta drva

This study aims to analyze the reaction wood samples for some industrial tree species naturally grown in Kastamonu province in Turkey and compare them with the relevant standards. Some anatomical, chemical, fiber morphological, optical properties, and color changes before and after drying were analyzed for the reaction wood (RW) samples. While the holocellulose content of fir and pine compression wood (CW) was found to be lesser (~3-4 %), the lignin content was higher than those of the opposite wood (OW) (~34 % for pine and 12 % for fir). On the contrary, the amount of holocellulose was found to be higher (~1-4 %), and the lignin was lower (at about 6-15 %) in the tension wood (TW) samples. It was observed that average lengths are more extended in TW(~50-54 %) and shorter in CW (~13-17 %) than those of OW. Significant differences were observed between the anatomical structures of the coniferous and deciduous species studied. Although, the greatest color differences in wet and oven-dried samples of coniferous trees were measured in CW (~15-17 %), it has been found as about 0.7-3 % in TW for deciduous species. Some differences were observed in the anatomical, optical, fiber morphological, and chemical properties of the RW for the studied wood species. Due to its higher lignin content and better physical properties, CW can be used for producing small households and hand tools, ornaments, toys, etc. It will also be appropriate for use in milling and turning work. It is recommended that, because of the lower lignin content and higher polysaccharide ratio, TW should be primarily used for the cellulose, pulp, and paper industries, where high mechanical resistance values are required. Consequently, RW formation causes some physical, chemical, mechanical, anatomical, and optical differences compared to OW in deciduous and coniferous species.

Study of antioxidant enzymes’ activity in reaction wood of poplar tree (Populus alba L.)

Summary Studying biochemical variation in reaction wood is needed to understand the nature of reaction wood formation and the physiology of trees. The activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (GPX)) was studied in normal wood (NW) and reaction wood (RW) (opposite wood (OW) and tension wood (TW)) in the bole of poplar trees (Populus alba L.). Four-year-old trees were induced to produce reaction wood by forced bending. The activity of antioxidant enzymes was studied with repeated sampling during a growing season. The results showed that total protein content and antioxidant enzymes are sensitive to mechanically induced stress. TW and OW showed higher activity of enzymes than NW as well as higher amounts of total protein. OW showed a higher concentration of CAT and SOD than TW. APX was more active than GPX in both TW and OW. In addition to mechanical stress, the effect of other climate factors was observed in the reactive oxygen species accumulation.

Multiomics studies with co-transformation reveal microRNAs via miRNA-TF-mRNA network participating in wood formation in Hevea brasiliensis.

MicroRNAs (miRNAs) are small endogenous non-coding RNAs that play an important role in wood formation in plants. However, the significance of the link between miRNAs and their target transcripts in wood formation remains unclear in rubber tree (Hevea brasiliensis). In this study, we induced the formation of reaction wood by artificially bending rubber trees for 300 days and performed small RNA sequencing and transcriptome deep sequencing (RNA-seq) to describe the complement of miRNAs and their targets contributing to this process. We identified 5, 11, and 2 differentially abundant miRNAs in normal wood (NW) compared to tension wood (TW), in NW relative to opposite wood (OW), and between TW and OW, respectively. We also identified 12 novel miRNAs and 39 potential miRNA-mRNA pairs with different accumulation patterns in NW, TW, and OW. We noticed that many miRNAs targeted transcription factor genes, which were enriched in KEGG pathways associated with phenylpropanoid biosynthesis, phenylalanine metabolism, and pyruvate metabolism. Thus, miRNA-TF-mRNA network involved in wood formation via tension wood model were constructed. We validated the differential accumulation of miRNAs and their targets by RT-qPCR analysis and overexpressed miRNA in Nicotiana benthamiana with its potential target gene. These results will provide a reference for a deep exploration of growth and development in rubber tree.

Structure of cellulose in birch phloem fibres in tension wood: an X-ray nanodiffraction study

BackgroundTo gain a better understanding of bark layer structure and function, especially of the phloem fibres and their contribution to the posture control of trees, it is important to map the structural properties of these cells. The role of bark can also be linked to the reaction wood formation and properties which are essential when it comes to studying the questions related to tree growth. To offer new insights into the role of bark in the postural control of trees, we studied the micro- and nanoscale structures of the phloem and its nearest layers. This study is the first time, in which phloem fibres in trees have been extensively examined using X-ray diffraction (XRD).We determined the orientation of cellulose microfibrils in phloem fibres of Silver birch saplings by using scanning synchrotron nanodiffraction. The samples consisted of phloem fibres extracted from tension, opposite and normal wood (TW, OW, NW).ResultsUsing scanning XRD, we were able to obtain new information about the mean microfibril angle (MFA) in cellulose microfibrils in phloem fibres connected to reaction wood. A slight but consistent difference was detected in the average MFA values of phloem fibres between the TW and OW sides of the stem. Using scanning XRD, different contrast agents (intensity of the main cellulose reflection or calcium oxalate reflection, mean MFA value) were used to produce 2D images with 200 nm spatial resolution.ConclusionsBased on our results, the tension wood formation in the stem might be related to the structure and properties of phloem fibres. Thus, our results suggest that the nanostructure of phloem fibres is involved in the postural control of trees containing tension and opposite wood.

Wood quality of residual branches of Hymenaea courbaril L. from logging in the Amazon rainforest

Abstract Branches remain in the forest environment as logging activity residue. Considering the large size of many Amazonian trees, their branches have considerable dimensions and can contribute to a sustainable wood alternative for various applications. Due to the formation of reaction wood in the branches, relevant macro and ultrastructural changes can occur in its characteristics in relation to the trunk wood. However, the woods of the branches are not technologically well known. Therefore, this work aimed to evaluate the wood quality of branches of Hymenaea courbaril L., comparing it with wood of trunks. The extractives and lignin contents, basic density, shrinkage, fiber biometry, microfibril angle and mechanical resistance to compression parallel to the fibers were analyzed. The branch wood had smaller fiber dimensions and a higher microfibril angle than the trunk wood. The basic density was similar between these materials. The linear and volumetric shrinkages were smaller in the branch wood than in the trunk, while the axial shrinkage was higher in the branch. The parallel compressive strength was also lower in the branch wood than in the trunk. The branch wood has properties suitable for products with higher added value such as furniture, decorative objects, floors and utensils in general.

Gene expression analysis revealed Hbr-miR396b as a key piece participating in reaction wood formation of Hevea brasiliensis (rubber tree)

Rubber tree (Hevea brasiliensis) is a perennial tree, which is important for wood-related industrial applications. However, the molecular basis of wood formation in H. brasiliensis is largely unknown. In this study, we induced the formation of reaction wood by artificial bending three 3−year−old rubber trees for 7 or 100 days, and collected xylem samples from tension wood, opposite wood, and normal wood, and subjected them to small RNA sequencing to identify the microRNAs (miRNAs) participated in rubber tree wood formation. We identified 25 known microRNAs (miRNAs) belonging to 21 miRNA families in 7-day wood and 24 known miRNAs belonging to 20 miRNA families in 100-day wood. Three known miRNAs, hbr-miR396b, hbr-miR6168, and hbr-miR6485, that accumulated in 100-day wood targeted gene members of the three transcription factor families C3H, SQUAMOSA BINDING PROTEIN (SBP), and AUXIN RESPONSE FACTOR (ARF). Gene Ontology analysis showed genes co-expressed with transcription factor genes targeted by miRNAs were associated with cellulose metabolism, cellulose biosynthesis, and cellulose synthase in 100-day but not 7-day wood, likely reflecting the longer duration of artificial bending treatment. We also constructed a co-expression network that includes miRNAs, their target transcription factor genes, and the genes co-expressed with these genes, revealing that cellulose biosynthesis was regulated by 3 known and 13 novel miRNAs, which may all participate in reaction wood formation. In summary, this study identified potential key miRNA−transcription factor−mRNA networks involved in reaction wood formation of rubber tree and laid a theoretical foundation for further elucidating the mechanism of wood formation in this species.

RNA Sequencing Reveals Phenylpropanoid Biosynthesis Genes and Transcription Factors for Hevea brasiliensis Reaction Wood Formation.

Given the importance of wood in many industrial applications, much research has focused on wood formation, especially lignin biosynthesis. However, the mechanisms governing the regulation of lignin biosynthesis in the rubber tree (Hevea brasiliensis) remain to be elucidated. Here, we gained insight into the mechanisms of rubber tree lignin biosynthesis using reaction wood (wood with abnormal tissue structure induced by gravity or artificial mechanical treatment) as an experimental model. We performed transcriptome analysis of rubber tree mature xylem from tension wood (TW), opposite wood (OW), and normal wood (NW) using RNA sequencing (RNA-seq). A total of 214, 1,280, and 32 differentially expressed genes (DEGs) were identified in TW vs. NW, OW vs. NW, and TW vs. OW, respectively. GO and KEGG enrichment analysis of DEGs from different comparison groups showed that zeatin biosynthesis, plant hormone signal transduction, phenylpropanoid biosynthesis, and plant–pathogen interaction pathways may play important roles in reaction wood formation. Sixteen transcripts involved in phenylpropanoid biosynthesis and 129 transcripts encoding transcription factors (TFs) were used to construct a TF–gene regulatory network for rubber tree lignin biosynthesis. Among them, MYB, C2H2, and NAC TFs could regulate all the DEGs involved in phenylpropanoid biosynthesis. Overall, this study identified candidate genes and TFs likely involved in phenylpropanoid biosynthesis and provides novel insights into the mechanisms regulating rubber tree lignin biosynthesis.

Modern reactivations of a thick colluvial landslide reconstructed from dendrochronology: the example of the Xiakou landslide, China

AbstractIn this study, we investigated two types of growth disturbance (GD) in tree‐ring sequences, namely, the abrupt growth reduction in annual tree‐ring width, and the formation of reaction wood, in tree stems growing on the thick colluvial Xiakou landslide in China. In total, 96 GDs were identified in 183 core samples that were acquired from the 81 trees on the Xiakou landslide, revealing nine reactivation phases of the landslide body over the past 30 years. These reactivations were found to have occurred in the years of 1987, 1988, 1989, 1992, 1993, 1995, 1997, 2006, and 2007. Further hazard analysis indicated that the probability of landslide reactivation in 20 years was highest in the upper body of the landslide and on the boundaries where the new and old landslides meet. In contrast, the most stable region was the lower landslide body. Under the circumstances of an extreme lack of historical data, this work demonstrated the usefulness of dendrochronology in reconstructing modern reactivations over the past 30 years for a giant, deep‐seated, colluvial landslide with a volume exceeding approximately 10 million m3. This study has broader impacts because landslides in colluvium are a globally common types of land degradation.

Early transcriptional response to gravistimulation in poplar without phototropic confounding factors.

In response to gravistimulation under anisotropic light, tree stems showing an active cambium produce reaction wood that redirects the axis of the trees. Several studies have described transcriptomic or proteomic models of reaction wood relative to the opposite wood. However, the mechanisms leading to the formation of reaction wood are difficult to decipher because so many environmental factors can induce various signalling pathways leading to this developmental reprogramming. Using an innovative isotropic device where the phototropic response does not interfere with gravistimulation we characterized the early molecular responses occurring in the stem of poplar after gravistimulation in an isotropic environment, and without deformation of the stem. After 30 min tilting at 35° under anisotropic light, we collected the upper and lower xylems from the inclined stems. Controls were collected from vertical stems. We used a microarray approach to identify differentially expressed transcripts. High-throughput real-time PCR allowed a kinetic experiment at 0, 30, 120 and 180 min after tilting at 35°, with candidate genes. We identified 668 differentially expressed transcripts, from which we selected 153 candidates for additional Fluidigm qPCR assessment. Five candidate co-expression gene clusters have been identified after the kinetic monitoring of the expression of candidate genes. Gene ontology analyses indicate that molecular reprogramming of processes such as ‘wood cell expansion’, ‘cell wall reorganization’ and ‘programmed cell death’ occur as early as 30 min after gravistimulation. Of note is that the change in the expression of different genes involves a fine regulation of gibberellin and brassinosteroid pathways as well as flavonoid and phosphoinositide pathways. Our experimental set-up allowed the identification of genes regulated in early gravitropic response without the bias introduced by phototropic and stem bending responses.

Reaction Wood Anatomical Traits and Hormonal Profiles in Poplar Bent Stem and Root.

Reaction wood (RW) formation is an innate physiological response of woody plants to counteract mechanical constraints in nature, reinforce structure and redirect growth toward the vertical direction. Differences and/or similarities between stem and root response to mechanical constraints remain almost unknown especially in relation to phytohormones distribution and RW characteristics. Thus, Populus nigra stem and root subjected to static non-destructive mid-term bending treatment were analyzed. The distribution of tension and compression forces was firstly modeled along the main bent stem and root axis; then, anatomical features, chemical composition, and a complete auxin and cytokinin metabolite profiles of the stretched convex and compressed concave side of three different bent stem and root sectors were analyzed. The results showed that in bent stems RW was produced on the upper stretched convex side whereas in bent roots it was produced on the lower compressed concave side. Anatomical features and chemical analysis showed that bent stem RW was characterized by a low number of vessel, poor lignification, and high carbohydrate, and thus gelatinous layer in fiber cell wall. Conversely, in bent root, RW was characterized by high vessel number and area, without any significant variation in carbohydrate and lignin content. An antagonistic interaction of auxins and different cytokinin forms/conjugates seems to regulate critical aspects of RW formation/development in stem and root to facilitate upward/downward organ bending. The observed differences between the response stem and root to bending highlight how hormonal signaling is highly organ-dependent.

Autofluorescence in Plants.

Plants contain abundant autofluorescent molecules that can be used for biochemical, physiological, or imaging studies. The two most studied molecules are chlorophyll (orange/red fluorescence) and lignin (blue/green fluorescence). Chlorophyll fluorescence is used to measure the physiological state of plants using handheld devices that can measure photosynthesis, linear electron flux, and CO2 assimilation by directly scanning leaves, or by using reconnaissance imaging from a drone, an aircraft or a satellite. Lignin fluorescence can be used in imaging studies of wood for phenotyping of genetic variants in order to evaluate reaction wood formation, assess chemical modification of wood, and study fundamental cell wall properties using Förster Resonant Energy Transfer (FRET) and other methods. Many other fluorescent molecules have been characterized both within the protoplast and as components of cell walls. Such molecules have fluorescence emissions across the visible spectrum and can potentially be differentiated by spectral imaging or by evaluating their response to change in pH (ferulates) or chemicals such as Naturstoff reagent (flavonoids). Induced autofluorescence using glutaraldehyde fixation has been used to enable imaging of proteins/organelles in the cell protoplast and to allow fluorescence imaging of fungal mycelium.

ELIMÄKI Locus Is Required for Vertical Proprioceptive Response in Birch Trees.

Tree architecture has evolved to support a top-heavy above-ground biomass, but this integral feature poses a weight-induced challenge to trunk stability. Maintaining an upright stem is expected to require vertical proprioception through feedback between sensing stem weight and responding with radial growth. Despite its apparent importance, the principle by which plant stems respond to vertical loading forces remains largely unknown. Here, by manipulating the stem weight of downy birch (Betula pubescens) trees, we show that cambial development is modulated systemically along the stem. We carried out a genetic study on the underlying regulation bycombining an accelerated birch flowering program with a recessive mutation at the ELIMÄKI locus(EKI), which causes a mechanically defective response to weight stimulus resulting in stem collapse after just 3months. We observed delayed wood morphogenesis in eki compared with WT, along with a more mechanically elastic cambial zone and radial compression of xylem cell size, indicating that rapid tissue differentiation is critical for cambial growth under mechanical stress. Furthermore, the touch-induced mechanosensory pathway was transcriptionally misregulated in eki, indicating that the ELIMÄKI locus is required to integrate the weight-growth feedback regulation. By studying this birch mutant, we were able to dissect vertical proprioception from the gravitropic response associated with reaction wood formation. Our study provides evidence for both local and systemic responses to mechanical stimuli during secondary plant development.

Soil conditions required for reaction wood formation of drunken trees in a continuous permafrost region

ABSTRACT Black spruce trees lean to form “drunken” forest on degrading permafrost; however, the causes of tree leaning on continuous permafrost remain unclear. Leaning events are recorded by reaction wood formation in tree rings, and it remains unclear what soil conditions are required for reaction wood formation of drunken trees. Tree disk morphology and soil hummock properties were examined for fifty tree–mound combinations in Northwest Territories, Canada. Spruce trees growing on mound edges form reaction wood on the downslope sides of their trunks. Reaction wood formation in mature trees was greatest in stem tissues between ground level and 30 cm aboveground. Reaction wood formation occurred only in trees growing on mound edges. The extent of reaction wood formation was higher in trees growing in clayey soils than in trees on sandy soils. For trees growing on clayey mound edges, the extent of reaction wood formation decreased with increasing permafrost table depth. Black spruce tree rings formed between ground level and 30 cm aboveground could record movement of clayey soil hummocks over shallow, underlying permafrost tables. A combination of clayey soil texture and shallow permafrost table is likely required for development of hummocks and drunken forests on the continuous permafrost region studied.

The sensitivity of dendrogeomorphic approaches to assessing landslide movements

An understanding of past landslide behaviour is crucial for the estimation of future landslide activity. In this respect, methods of absolute dating are frequently used for landslide history assessment. Dendrogeomorphic (tree ring-based) approaches are standard methods for the spatio-temporal reconstruction of landslide activity with annual to even sub-annual resolution and timespans of several centuries (depending on the tree age). Nevertheless, the verification of the effectiveness, sensitivity and precision of these dendrogeomorphic approaches has not been addressed in more detail despite their wide spectrum of limitations. This study makes an assessment of tree ring-based chronologies of landslides based on a comparison with the independent data of past landslide activity from instrumental monitoring. The chronology of past landslide events was created from 528 tree ring series from 132 individuals of Norway spruce (Picea abies (L.) Karst.) growing on a monitored landslide in the Outer Western Carpathians (the Ondřejník Mt.) and was expressed as the yearly variation in the standard event-response index. The landslide was monitored at two sites via technical levelling (since 2007) and an automated wire extensometer (since 2014). The obtained results show a high sensitivity of reaction wood formation to movements on the order of several mm (e.g., the abrupt acceleration of movements in February 2017, as recorded by the extensometer). Moreover, a one-year delay in the growth response to movement occurring in the middle part of the growing season of the previous year was detected. This fact fully supports the introduction of the dendrological year to dendrogeomorphic research. The partial lack of correspondence between tree ring records and data from more distant monitoring sites suggests the need for separate dendrogeomorphic analyses of the individual homogenous parts of landslide areas. Finally, the use of four instead of two increment cores per tree did not provide any extra event years but increased the values of the event-response index of the dated events.

Traumatic Resin Ducts in Alaska Mountain Hemlock Trees Provide a New Proxy for Winter Storminess

AbstractWinter is a critical season for land‐surface feedbacks and ecosystem processes; however, most high‐latitude paleo‐environmental reconstructions are blind to cold season conditions. Here we introduce a winter‐sensitive, paleo‐proxy record that is based on the relative frequency of tangential rows of traumatic resin ducts (TRDs) in the annual growth rings of mountain hemlocks (Tsuga mertensiana) growing near treeline in Southeast Alaska. Hemlocks produce a row of TRDs in the earlywood portion of their annual rings in response to cambial damage incurred during winter. Multidecadal bouts of TRD production were followed by growth‐leader replacement, reaction wood formation, and divergence in radial growth between storm‐damaged trees and less exposed mountain hemlock forests. These patterns are consistent with TRDs being a response to tree damage caused by ice and snowstorms, a conclusion supported by the krummholz tree architecture at these sites. This relationship is further corroborated by significant correlations between our TRD record and the strength of the wintertime Aleutian Low (AL) pressure system that is linked to tree‐damaging agents like wind, precipitation, and ice storm strength in Southeast Alaska. The combined TRD/krummholz architecture record indicates that abrupt shifts between strong and weak AL phases occurred every several decades since CE 1700 and that the 1800s had relatively long AL phases with heavy snowpacks. In addition to describing the magnitude and tempo of wintertime climate change in Northwestern North America, these results suggest that North Pacific Decadal Variability underlies the long‐term dynamics of treeline ecosystems along the northeast Pacific coast.

The <i>ELIMÄKI</i> Locus is Required for Vertical Proprioception in Birch Trees

Tree architecture has evolved to support a top-heavy above-ground biomass, but this integral feature poses a weight-induced challenge to trunk stability. Maintaining an upright stem is expected to require vertical proprioception through feedback between sensing stem weight and modulating radial growth. Despite its apparent importance, the principle by which plant stems respond to vertical loading forces remains largely unknown. Here, by manipulating the stem weight of downy birch (Betula pubescens) trees, we show that cambial development is modulated systemically along the stem. We carried out a genetic study on the underlying regulation by combining an accelerated birch flowering program with a recessive mutation at the ELIMAKI locus (EKI), which causes a mechanically defective response to weight stimulus resulting in stem collapse after just three months. We observed delayed wood morphogenesis in eki compared with WT, along with a more mechanically elastic cambial zone and radial compression of xylem cell size, indicating that timely tissue differentiation is critical for cambial growth under mechanical stress. Furthermore, the touch-induced mechanosensory pathway was transcriptionally misregulated in eki, indicating that the ELIMAKI locus is required to integrate the weight-growth feedback regulation. By studying this birch mutant, we were able to dissect vertical proprioception from the gravitropic response associated with reaction wood formation. Taken together, our study provides evidence for both local and systemic responses to mechanical stimuli during secondary plant development.

Reaction wood anatomy and lignin distribution in Gnetum gnemon branches

This study investigated the anatomical and chemical characteristics of the reaction wood of a gymnpsperm species, Gnetum gnemon, and discussed on contributing factor for the type of reaction wood in this species. Cell morphology, microfibril angle (MFA) of the S2 layer and lignin distribution in secondary walls of tracheary elements, and lignin content were examined on three branches. Observations included no G-layer formation, significant decreases in vessel frequency, and altered MFA, and visible-light absorbance after lignin colour reactions in tracheid and fiber tracheid walls on the upper side in almost all samples. These results suggest that reaction wood in G. gnemon was similar to that in ‘tension-wood-like-reaction wood’ in angiosperms. On the other hand, reaction wood showed decrease in the lignin concentration in the fiber tracheid walls compared to the tracheid walls. In addition, the lignin in the tracheid and fiber tracheid walls was originally rich in syringyl units, suggesting that changes in the anatomical and chemical characteristics of secondary xylem due to reaction wood formation might relate to the ratio of the syringyl to guaiacyl units in lignin in the cell walls which function for mechanical support.

Relationships between tree size and reaction wood formation in 23 Japanese angiosperms

The objective of this study is to clarify the relationships between tree sizes and the anatomical, chemical, and physical characteristics of reaction wood. Naturally inclined stems or branches in 23 Japanese angiosperms, including 3 tree sizes at adult trees, 7 arbor, 7 subarbor, and 9 shrub species, were used. The presence of a gelatinous (G-) layer, basic density, vessel frequency, and lignin content were examined on the upper and lower sides of each sample. Almost all the species showed a decrease in vessel frequency and lignin content on the upper side, which are typical characteristics of reaction wood in angiosperms. A G-layer was formed in four arbor, one subarbor, and two shrub species. The ratio of the upper and lower sides in vessel frequency and lignin content gradually decreased with an increase in tree size. Based on the results of the principal component and cluster analyses, shrub species were classified into the different groups: from arbor and subarbor groups. These results indicate that the magnitude of alteration in the characteristics due to reaction wood formation of arbor and subarbor might differ from that of shrub, although the characteristics changed by reaction wood formation might be the same in the three tree sizes at adult trees.

Poplar woody taproot under bending stress: the asymmetric response of the convex and concave sides.

Background and Aims Progress has been made in understanding the physiological and molecular basis of root response to mechanical stress, especially in the model plant Arabidopsis thaliana, in which bending causes the initiation of lateral root primordia toward the convex side of the bent root. In the case of woody roots, it has been reported that mechanical stress induces an asymmetric distribution of lateral roots and reaction wood formation, but the mechanisms underlying these responses are largely unknown. In the present work, the hypothesis was tested that bending could determine an asymmetric response in the two sides of the main root axis as cells are stretched on the convex side and compressed on the concave side. Methods Woody taproots of 20 seedlings were bent to an angle of 90° using a steel net. Changes in the anatomy, lignin and phytohormone content and proteome expression in the two sides of the bent root were analysed; anatomical changes, including dissimilarities and similarities to those found in poplar bent woody stem, were also considered. Key Results Compression forces at the concave side of poplar root induced the formation of reaction wood which presented a high lignin content and was associated with the induction of cambium cell activity. Auxin seemed to be the main hormone triggering lignin deposition and cell wall strengthening in the concave sides. Abscisic acid appeared to function in the water stress response induced by xylem structures and/or osmotic alterations in the compression sides, whereas gibberellins may control cell elongation and gravitropisms. Conclusions Poplar root reaction wood showed characteristics different from those produced in bent stem. Besides providing biomechanical functions, a bent root ensures water uptake and transport in the deforming condition induced by tension and compression forces by two different strategies: an increase in xylem thickness in the compressed side, and lateral root formation in the tension side.