Artificial Bending Research Articles

Analysis of genes related to xylem cell wall development based on transcriptomics in Populus alba ‘Berolinensis’ tension wood

Populus alba ‘Berolinensis’ is a fast-growing, high-yielding species with strong biotic and abiotic stress resistance, and widely planted for timber, shelter belts and aesthetic purposes. In this study, molecular development is explored and the important genes regulating xylem formation in P. alba ‘Berolinensis’ under artificial bending treatments was identified. Anatomical investigation indicated that tension wood (TW) was characterized by eccentric growth of xylem and was enriched in cellulose; the degree of lignification was lower than for normal wood (NW) and opposite wood (OW). RNA-Seq-based transcriptome analysis was performed using developing xylem from three wood types (TW, OW and NW). A large number of differentially expressed genes (DEGs) were screened and 4889 counted. In GO and KEGG enrichment results, genes involved in plant hormone signal transduction, phenylpropanoid biosynthesis, and cell wall and secondary cell wall biogenesis play major roles in xylem development under artificial bending. Eight expansin (PalEXP) genes were identified from the RNA-seq data; four were differentially expressed during tension wood formation. Phylogenetic analysis indicated that PalEXLB1 belongs to the EXPB subfamily and that the other PalEXPs are members of the EXPA subfamily. A transcriptional regulatory network construction showed 10 transcription factors located in the first and second layers upstream of EXP, including WRKY, ERF and bHLH. RT‒qPCR analysis in leaves, stems and roots combined with transcriptome analysis suggests that PalEXPA2, PalEXPA4 and PalEXPA15 play significant regulatory roles in cell wall formation during tension wood development. The candidate genes involved in xylem cell wall development during tension wood formation marks an important step toward identifying the molecular regulatory mechanism of xylem development and wood property improvement in P. alba ‘Berolinensis’.

Protein profile analysis of tension wood development in response to artificial bending and gravitational stimuli in Betula platyphylla

Betula platyphylla Suk (birch) is an excellent short-term hardwood species with growth and wood characteristics well suited to wood industries. To investigate the molecular mechanism of wood development in birch, a tension wood (TW) induced system was used to explore the regulatory mechanism at the protein level and identify the key proteins involved in xylem development in birch. The results of dyeing with Safranin O-Fast Green indicated that the cellulose content of TW was significantly higher than that of opposite wood (OW) or normal wood (NW), and the lignin content in TW was significantly lower than that in OW and NW after artificial bending of birch stems. Protein profile analysis of TW, NW and OW by iTRAQ revealed that there were 639 and 460 differentially expressed proteins (DEPs) between TW/OW and TW/NW, respectively. The DEPs were mainly enriched in tyrosine metabolism, glycolysis/gluconeogenesis, phenylalanine and tyrosine metabolism, phenylpropanoid and pyruvate metabolism, the pentose phosphate pathway, the citrate cycle (TCA cycle), fructose and mannose metabolism, carbon fixation in photosynthetic organisms, fatty acid biosynthesis, photosynthesis proteins and other pathways. The proteins in the citrate cycle were upregulated. The expression levels of PGI, PGM and FRK proteins related to cellulose synthesis increased and the expression levels of PAL, 4CL and COMT related to lignin synthesis decreased, leading to an increase in cellulose content and decreased lignin levels in TW. PPI analysis revealed that key DEPs interact with each other, indicating that these proteins form complexes to implement this function, which may provide important insights for wood formation at the molecular level.

Tissue Metabolic Responses to Artificial Bending and Gravitation Stimuli in Betula platyphylla

Betula platyphylla Suk (Asian white birch) is an economically important tree species in the paper-pulping and biofuel industries. To investigate the mechanism of wood formation at the metabolic level, we evaluated metabolic responses associated with tension-wood formation. Four-year-old trees were subjected to artificial bending treatment for 6 weeks. The xylem growth rate of tension wood (TW) was significantly faster than that of opposite wood (OW), and it exhibited a higher cellulose content. Metabolomics analysis was performed on metabolites of TW, OW and normal wood (NW), and 183 metabolites were identified, of which levels of 142 were altered between groups. Metabolites related to fatty-acid and amino-acid metabolism, the glycolytic pathway, and the metabolism of fructose, mannose and starch sucrose were abundant in TW. Glucose 1-phosphoric acid, fructose and mannose associated with tension-wood development were elevated. Levels of xylitol and ribosol (related to the conversion of glucose), coniferol (the main monomer of lignin) and shikimic acid (an intermediate in lignin synthesis) were decreased in TW. These metabolites are likely involved in xylem development.

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.

Transcriptomic, Proteomic, and Metabolic Profiles of Catalpa bungei Tension Wood Reveal New Insight Into Lignin Biosynthesis Involving Transcription Factor Regulation.

Lignin is a complex polymer in plant cell walls whose proportion is second only to that of cellulose and plays an important role in the mechanical properties of wood and stress resistance of plants. Here, we induced tension wood (TW) formation in Catalpa bungei by artificial bending and analyzed the lignin metabolism of the TW. LC-MS analysis showed that a significantly higher content of coniferyl aldehyde was observed in the TW cell wall than in the opposite wood (OW) and normal wood (NW) cell walls. TW had significantly lower contents of coniferyl alcohol than OW and NW. Raman spectroscopy results indicated that TW had lower total lignin than OW and NW. The transcription and translation levels of most of the differentially expressed genes (DEGs) involved in lignin monomer biosynthesis indicated upregulation in TW/OW and TW/NW. We found no significant difference in the transcription levels of three collision gases (CADs) between TW and OW or between NW, but their translation levels were significantly downregulated in TW, suggesting post-transcriptional control for CAD. We predicted and analyzed transcription factors that could target DEGs involved in lignin monomer biosynthesis in TW. Based on the analysis of the relationships of targeting and coexpression, we found that NAC (evm.model.group1.695) could potentially target 4CLs and CCoAOMT, that HD-Zip (evm.model.group7.1157) had potential targeting relationships with CCoAOMT, F5H, and CCR, and that their expression levels were significantly positive. It is speculated that the upregulation of NAC and HD-ZIP transcription factors activates the expression of downstream target genes, which leads to a significant increase in coniferyl aldehyde in TW. However, the decrease in total lignin in TW may be caused by the significant downregulation of CAD translation and the significant decrease in precursors (coniferyl alcohol). Whether the expression of CAD genes is regulated by post-transcriptional control and affects TW lignin metabolism needs further study.

The BpMYB4 Transcription Factor From Betula platyphylla Contributes Toward Abiotic Stress Resistance and Secondary Cell Wall Biosynthesis.

The MYB (v-myb avian myeloblastosis viral oncogene homolog) family is one of the largest transcription factor families in plants, and is widely involved in the regulation of plant metabolism. In this study, we show that a MYB4 transcription factor, BpMYB4, identified from birch (Betula platyphylla Suk.) and homologous to EgMYB1 from Eucalyptus robusta Smith and ZmMYB31 from Zea mays L. is involved in secondary cell wall synthesis. The expression level of BpMYB4 was higher in flowers relative to other tissues, and was induced by artificial bending and gravitational stimuli in developing xylem tissues. The expression of this gene was not enriched in the developing xylem during the active season, and showed higher transcript levels in xylem tissues around sprouting and near the dormant period. BpMYB4 also was induced express by abiotic stress. Functional analysis indicated that expression of BpMYB4 in transgenic Arabidopsis (Arabidopsis thaliana) plants could promote the growth of stems, and result in increased number of inflorescence stems and shoots. Anatomical observation of stem sections showed lower lignin deposition, and a chemical contents test also demonstrated increased cellulose and decreased lignin content in the transgenic plants. In addition, treatment with 100 mM NaCl and 200 mM mannitol resulted in the germination rate of the over-expressed lines being higher than that of the wild-type seeds. The proline content in transgenic plants was higher than that in WT, but MDA content was lower than that in WT. Further investigation in birch using transient transformation techniques indicated that overexpression of BpMYB4 could scavenge hydrogen peroxide and O2.– and reduce cell damage, compared with the wild-type plants. Therefore, we believe that BpMYB4 promotes stem development and cellulose biosynthesis as an inhibitor of lignin biosynthesis, and has a function in abiotic stress resistance.

Genome-wide analysis of lncRNA and mRNA expression and endogenous hormone regulation during tension wood formation in Catalpa bungei

BackgroundPhytohormones are the key factors regulating vascular development in plants, and they are also involved in tension wood (TW) formation. Although the theory of hormone distribution in TW formation is widely supported, the effects of endogenous hormones on TW formation have not yet been assessed. In this study, TW formation was induced in Catalpa bungei by artificial bending. The phytohormone content of TW, opposite wood (OW) and normal wood (NW) was determined using liquid chromatography-mass spectrometry (LC-MS), and transcriptome sequencing was performed. The hormone content and related gene expression data were comprehensively analyzed.ResultsThe results of analyses of the plant hormone contents indicated significantly higher levels of cis-zeatin (cZ), indoleacetic acid (IAA) and abscisic acid (ABA) in TW than in OW. Genes involved in the IAA and ABA synthesis pathways, such as ALDH (evm.model.group5.1511) and UGT (evm.model.scaffold36.20), were significantly upregulated in TW. and the expression levels of ARF (evm.model.group5.1332), A-ARR (evm.model.group0.1600), and TCH4 (evm.model.group2.745), which participate in IAA, cZ and Brassinolide (BR) signal transduction, were significantly increased in TW. In particular, ARF expression may be regulated by long noncoding RNAs (lncRNAs) and the HD-ZIP transcription factor ATHB-15.ConclusionsWe constructed a multiple hormone-mediated network of C. bungei TW formation based on hormone levels and transcriptional expression profiles were identified during TW formation.

Sequence characterization and expression analysis of NAC genes from Betula platyphylla

The expression analysis of 21 BpNAC genes in stem, xylem from different growth season, and tension wood indicated that they might participate in xylem and tension wood development in birch. NAC transcription factors play important roles in the growth and development of plants. The NAC family has many members; however, the functions of the majority of them are unknown, especially in woody plants. In this study, to characterize the NAC members and explore NAC genes’ association with xylem development, 21 full-length NAC genes were identified in Betula platyphylla, a deciduous woody plant. Most of the 21 NAC proteins contain a complete NAC DNA-binding domain and a variable transcriptional regulation domain. Motif analysis showed that proteins within the same group mostly have similar motifs; however, there were no obvious similarities in gene structures among the identified BpNACs, based on exon/intron analysis. Cis-elements analysis indicated that promoters of the 21 BpNAC genes were enriched in ABRE and AC-box elements, suggesting that BpNAC genes might be related to ABA signal transduction pathways and xylem development. Quantitative real-time PCR analysis indicated that the 21 BpNACs were differentially expressed in the stem of B. platyphylla. In different stages of the growth season, BpNACs showed expression differences in the developing xylem. These genes also responded to artificial bending in tension wood and opposite wood, to varying degrees, and exhibited distinct expression patterns at different treatment times. The results indicated that these BpNAC genes might participate in tension wood development. Our study provided the basis for further functional investigations of BpNAC genes in birch.

Forest trees filter chronic wind‐signals to acclimate to high winds

Controlled experiments have shown that trees acclimate thigmomorphogenetically to wind-loads by sensing their deformation (strain). However, the strain regime in nature is exposed to a full spectrum of winds. We hypothesized that trees avoid overreacting by responding only to winds which bring information on local climate and/or wind exposure. Additionally, competition for light dependent on tree social status also likely affects thigmomorphogenesis. We monitored and manipulated quantitatively the strain regimes of 15 pairs of beech (Fagus sylvatica) trees of contrasting social status in an acclimated stand, and quantified the effects of these regimes on the radial growth over a vegetative season. Trees exposed to artificial bending, the intensity of which corresponds to the strongest wind-induced strains, enhanced their secondary growth by at least 80%. Surprisingly, this reaction was even greater - relatively - for suppressed trees than for dominant ones. Acclimated trees did not sense the different types of wind events in the same way. Daily wind speed peaks due to thermal winds were filtered out. Thigmomorphogenesis was therefore driven by intense storms. Thigmomorphogenesis is also likely to be involved in determining social status.

Stable constrained dynamics

We present a unification of the two main approaches to simulate deformable solids, namely elasticity and constraints. Elasticity accurately handles soft to moderately stiff objects, but becomes numerically hard as stiffness increases. Constraints efficiently handle high stiffness, but when integrated in time they can suffer from instabilities in the nullspace directions, generating spurious transverse vibrations when pulling hard on thin inextensible objects or articulated rigid bodies. We show that geometric stiffness, the tensor encoding the change of force directions (as opposed to intensities) in response to a change of positions, is the missing piece between the two approaches. This previously neglected stiffness term is easy to implement and dramatically improves the stability of inextensible objects and articulated chains, without adding artificial bending forces. This allows time step increases up to several orders of magnitude using standard linear solvers.

Comprehensive transcriptome analysis of developing xylem responding to artificial bending and gravitational stimuli in Betula platyphylla.

Betula platyphylla Suk (birch) is a fast-growing woody species that is important in pulp industries and the biofuels. However, as an important pulp species, few studies had been performed on its wood formation. In the present study, we investigated the molecular responses of birch xylem to artificial bending and gravitational stimuli. After trunks of birch trees were subjected to bending for 8 weeks, the cellulose content was significantly greater in tension wood (TW) than in opposite wood (OW) or normal wood (NW), whereas the lignin content in TW was significantly lower than that in OW and NW. In addition, TW grew more rapidly than OW and generated TW-specific fibers with an additional G-layer. Three transcriptome libraries were constructed from TW, OW and NW of B. platyphylla, respectively, after the plants were subjected to artificial bending. Overall, 80,909 nonredundant unigenes with a mean size of 768 nt were assembled. Expression profiles were generated, and 9,684 genes were found to be significantly differentially expressed among the TW, OW and NW libraries. These included genes involved in secondary cell wall structure, wood composition, and cellulose or lignin biosynthesis. Our study showed that during TW formation, genes involved in cellulose synthesis were induced, while the expression of lignin synthesis-related genes decreased, resulting in increased cellulose content and decreased lignin levels in TW. In addition, fasciclin-like arabinogalactan proteins play important role in TW formation. These findings may provide important insights into wood formation at the molecular level.

Transcriptome profiling of radiata pine branches reveals new insights into reaction wood formation with implications in plant gravitropism.

BackgroundFormation of compression (CW) and opposite wood (OW) in branches and bent trunks is an adaptive feature of conifer trees in response to various displacement forces, such as gravity, wind, snow and artificial bending. Several previous studies have characterized tracheids, wood and gene transcription in artificially or naturally bent conifer trunks. These studies have provided molecular basis of reaction wood formation in response to bending forces and gravity stimulus. However, little is known about reaction wood formation and gene transcription in conifer branches under gravity stress. In this study SilviScan® technology was used to characterize tracheid and wood traits in radiate pine (Pinus radiata D. Don) branches and genes differentially transcribed in CW and OW were investigated using cDNA microarrays.ResultsCW drastically differed from OW in tracheids and wood traits with increased growth, thicker tracheid walls, larger microfibril angle (MFA), higher density and lower stiffness. However, CW and OW tracheids had similar diameters in either radial or tangential direction. Thus, gravity stress largely influenced wood growth, secondary wall deposition, cellulose microfibril orientation and wood properties, but had little impact on primary wall expansion. Microarray gene transcription revealed about 29% of the xylem transcriptomes were significantly altered in CW and OW sampled in both spring and autumn, providing molecular evidence for the drastic variation in tracheid and wood traits. Genes involved in cell division, cellulose biosynthesis, lignin deposition, and microtubules were mostly up-regulated in CW, conferring its greater growth, thicker tracheid walls, higher density, larger MFA and lower stiffness. However, genes with roles in cell expansion and primary wall formation were differentially transcribed in CW and OW, respectively, implicating their similar diameters of tracheid walls and different tracheid lengths. Interestingly, many genes related to hormone and calcium signalling as well as various environmental stresses were exclusively up-regulated in CW, providing important clues for earlier molecular signatures of reaction wood formation under gravity stimulus.ConclusionsThe first comprehensive investigation of tracheid characteristics, wood properties and gene transcription in branches of a conifer species revealed more accurate and new insights into reaction wood formation in response to gravity stress. The identified differentially transcribed genes with diverse functions conferred or implicated drastic CW and OW variation observed in radiata pine branches. These genes are excellent candidates for further researches on the molecular mechanisms of reaction wood formation with a view to plant gravitropism.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-14-768) contains supplementary material, which is available to authorized users.

Electromechanical dynamics and optimization of pectoral fin–based ionic polymer–metal composite underwater propulsor

Ionic polymer–metal composites are soft artificial muscle-like bending actuators, which can work efficiently in wet environments such as water. Therefore, there is significant motivation for research on the development and design analysis of ionic polymer–metal composite based biomimetic underwater propulsion systems. Among aquatic animals, fishes are efficient swimmers with advantages such as high maneuverability, high cruising speed, noiseless propulsion, and efficient stabilization. Fish swimming mechanisms provide biomimetic inspiration for underwater propulsor design. Fish locomotion can be broadly classified into body and/or caudal fin propulsion and median and/or paired pectoral fin propulsion. In this article, the paired pectoral fin–based oscillatory propulsion using ionic polymer–metal composite for aquatic propulsor applications is studied. Beam theory and the concept of hydrodynamic function are used to describe the interaction between the beam and water. Furthermore, a quasi-steady blade element model that accounts for unsteady phenomena such as added mass effects, dynamic stall, and the cumulative Wagner effect is used to obtain hydrodynamic performance of the ionic polymer–metal composite propulsor. Dynamic characteristics of ionic polymer–metal composite fin are analyzed using numerical simulations. It is shown that the use of optimization methods can lead to significant improvement in performance of the ionic polymer–metal composite fin.

Active reinforcement of externally imposed folding in amphibians embryonic tissues

Although the folding of epithelial layers is one of the most common morphogenetic events, the underlying mechanisms of this process are still poorly understood. We aimed to determine whether an artificial bending of an embryonic cell sheet, which normally remains flat, is reinforced and stabilized by intrinsic cell transformations. We observed both reinforcement and stabilization in double explants of blastocoel roof tissue from Xenopus early gastrula embryos. The reinforcement of artificial bending occurred over the course of a few hours and was driven by the gradual apical constriction and radial elongation of previously compressed cells situated at the bending arch of the concave layer of explant. Apical constriction was associated with actomyosin contraction and endocytosis-mediated engulfing of the apical cell membranes. Cooperative apical constrictions of the concave layer of cells produced a tensile force that extended over the entire surface of the explant and correlated with apical contraction of the concave side cells. In the explants taken from the anterior regions of the embryo, this reinforcement was more stable and the bending better expressed than in those taken from suprablastoporal areas. The morphogenetic role of cell responses to the bending force is discussed.

EFFECTS OF PRUNING ON THE APPLE TREE: FROM TREE ARCHITECTURE TO MODELING

Arboricultural practices such as pruning, artificial bending or fruit thinning are crucial interventions in orchard management and are used for controlling tree size, penetration of light into the canopy and the equilibrium between vegetative and reproductive growth. The aim of this project is to explore the possibility of integrating such practices in a model of apple tree development. To this end we designed field experiments to study the effects of pruning (thinning or heading cuts) on two apple cultivars with contrasted architecture, 'Fuji' and 'Braeburn'. The results of these first experiments showed that the cultivars had significantly different reactions to pruning: 'Braeburn' trees showed a greater reduction in the total number of internodes compared to 'Fuji' trees, despite the fact that a greater number of internodes developed on 'Braeburn' trees. Thinning cuts of laterals tended to be compensated by an increase in lateral branching. Results also indicated that trees which had been pruned tended to develop trunks with similar number of internodes as control trees (which were not pruned). These experiments constitute a first step for assessing the rules underlying tree responses to pruning, which will be further integrated into a model of growth.

Changes in the shape of epithelial embryonic cells of the spur-toed frog upon deformation of the cell layer

A quantitative study was made of changes in the shape of cells in double explants of the blastocoel roof of the clawed frog gastrula within the first four hours after artificial bending of explants. It was found that, on the concave (contracted) side of explants, epithelial cells stretched out, and in many of them the apical surface contracted, whereas on the convex (stretched) side the cells remained isodiametric. The maximal difference in the apical index between epithelial cells located on the concave and convex sides was observed after 2 h of explant cultivation; by 2 h the artificially produced curvature of the explant further increased. Endocytosis on the concave side was more active than on the convex side. Experiments with inhibitors modulating the behavior of the actomyosin complex showed that unimpeded functioning of myosin II is more important for the apical contraction and elongation of cells than proper structural organization of the actin backbone.

Comparison of morphological and chemical properties between juvenile wood and compression wood of loblolly pine

Abstract In conifers, juvenile wood (JW) is always associated with compression wood (CW). Due to their similar properties, there is a common belief that JW is the same as CW. To resolve whether JW is identical to CW, 24 rooted cuttings of one loblolly pine clone were planted in growth chambers under normal, artificial bending, and windy environments. The results show that the morphology of JW is significantly different from CW. Furthermore, chemical analyses revealed that JW and CW are significantly different in chemical composition. Our results indicate that JW is different from CW, and the wood formed under a controlled windy environment is a mild type of compression wood.

Recent advances in the structural analysis of scientific balloons

The analysis of balloon envelopes by the finite element (FE) method is plagued by convergence problems. A practical FE analysis approach is based on the fact that in thin shells with non-zero Gaussian curvature the membrane solution component is essentially decoupled from the bending solution component. A proxy-problem is solved by using a small artificial bending stiffness that assures convergence without significantly affecting the membrane solution component. This approach has been previously validated on slightly overpressurized balloon envelopes. Extensions of this approach to more difficult problems in the structural analysis of balloon envelopes are presented. The convergence forcing modelling measures are discussed. Implications of the findings of the analysis results to future balloon designs are also discussed.

Finite element solution for the structural behavior of a scientific balloon

A finite element solution for the structural behavior of a scientific balloon has been obtained using a non-linear finite element code. The pneumatic skin is modelled by shell elements that are given a small artificial bending stiffness to overcome numerical problems yet the membrane solution remains unaffected. Validation of the analysis approach is provided through strain measurements on a small scale balloon that exhibits all essential features of a full scale balloon.

Simulation Study for Propulsion Mechanism Imitating Bending Motion of Organisms in Water. Swimming Snake's Bending Motion.

The application of dynamics in the living organism to the field of engineering is very instructive. To make an artificial bending wave propulsion mechanism, a simulation study of propulsion mechanisms imitating the bending motion of organisms in water was carried out. As the bending motion, a swimming snake's motion was employed. The bending wave of the organism was changed into a construction of the multilink mechanism. This simulation model was defined as the "multilink model". In addition, the simulation model based on the approximate theory of the organism's propulsion was defined as the "approximate organism model". This model is a prototype of the "multilink model". The thrust force and the swimming speed were calculated. The characteristics of the thrust force and the swimming speed with change in the frequency, the wave amplitude and the number of links were discussed.