Putative involvement of Thioredoxin h in early response to gravitropic stimulation of poplar stems

Gravity perception and gravitropic response are essential for plant development. In herbaceous species, it is widely accepted that one of the primary events in gravity perception involves the displacement of amyloplasts within specialized cells. However, the early signaling events leading to stem reorientation are not fully known, especially in woody species in which primary and secondary growth occur. Thirty-six percent of the identified proteins that were differentially expressed after gravistimulation were established as potential Thioredoxin targets. In addition, Thioredoxin h expression was induced following gravistimulation. In situ immunolocalization indicated that Thioredoxin h protein co-localized with the amyloplasts located in the endodermal cells. These investigations suggest the involvement of Thioredoxin h in the first events of signal transduction in inclined poplar stems, leading to reaction wood formation.     

<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> as a model for gelatinous fiber formation

Trees and herbaceous plants continuously monitor their position to maintain vertical stem growth and regulate branch orientation. When orientation is altered from the vertical, they form a special type of wood called reaction wood that differs chemically and structurally from normal wood and forces reorientation of the organ or whole plant. The reaction wood of dicotyledons is called tension wood and is characterized by nonlignified gelatinous fibers. The altered chemical and mechanical properties of tension wood reduce wood quality and represent a major problem for the timber and pulping industries. Repeated clipping of the emerging inflorescence stems of Arabidopsis thaliana augments wood formation in organs, including those inflorescence stems that are allowed to develop later. Gravistimulation of such inflorescence stems induces tension wood formation, allowing the use of A. thaliana for a molecular and genetic analysis of the mechanisms of tension wood formation.     

SOME OBSERVATIONS ON THE MECHANISM OF ORIENTATION MOVEMENT OF WOODY STEMS

The force that induces orientation movement of inclined or bent woody stems is generated in reaction wood even in young terminal stems with a large proportion of soft tissues to secondary xylem. Compression wood formed in pine as a response to inclination expands longitudinally after the stresses are released by sawing it from the stem. The increment of length of compression wood when sawed is equal to the decrement of its length which occurs during drying. This suggests that stresses developed by compression wood in the stem are related to imbibition of water by its cell walls. Not all compression wood develops tensile forces in the stem. Neutral compression wood was observed in the lower portion of inclined stems of pine. Tension wood in poplar develops contractile forces in the stem during its aestival maturation. However, when harvested before developing contractile forces in situ, it develops such forces during drying. This suggests that in poplar the mechanism which produces forces responsible for orientation bending also involves changes in cell wall hydration.     

Proteomic analysis of root meristems and the effects of acetohydroxyacid synthase-inhibiting herbicides in the root of Medicago truncatula

Quantitative proteome analyses of meristematic and nonmeristematic tissues from Medicago truncatula primary and lateral roots and meristem tissues from plants treated with acetohydroxyacid synthase-inhibiting herbicides were made. The accumulation of 81 protein spots changed in meristematic and nonmeristematic tissues and 51 protein spots showed significant changes in accumulation in herbicide-treated meristems. Identified proteins indicate two trends, (i) increased accumulation of cell division and redox-mediating proteins in meristems compared to nonmeristematic tissues and (ii) increased accumulation of pathogenesis-related and decreased accumulation of metabolic proteins in herbicide-treated roots.     

Study of tension wood in the artificially inclined seedlings of <Emphasis Type="Italic">Koelreuteria henryi</Emphasis> Dummer and its biomechanical function of negative gravitropism

Key message

Stem reorientation is critical to tree survival. With anatomical observation and strain measurement, the tension wood formation and biomechanical behavior were studied to gain insights into tree uprighting process.

Abstract

Tension wood plays a role in maintaining the mechanical stability of angiosperm trees. Both biological and physical aspects of tension wood are essential in understanding the mechanism of trunk or branch reorientation. In this study, we worked on both tension wood formation and its biomechanical function in artificially inclined 2-year-old Koelreuteria henryi seedlings. The tension wood formation and reorientation process of the trunk last for about 3 months. With pinning method, we confirmed that at the beginning of inclination the cambial zone including the vascular cambium and the developing normal wood fibers on the upper side of the inclined trunk perceives the onset of mechanical change and starts to produce G-fibers that generate a strong contractile released growth strain (RGS) for gravitropic correction. Stronger contractile RGS and more tension wood were found at the trunk base than at the half-height, suggesting that the trunk base plays a key role in trunk uprighting of K. henryi seedlings. The eccentric cambial growth in the tension wood side increases the efficiency of gravitropic correction and the compressive strains measured in the opposite wood of some inclined seedlings also help the upright movement.

     

Growth and posture control strategies in Fagus sylvatica and Acer pseudoplatanus saplings in response to canopy disturbance

Background and Aims

Forest tree saplings that grow in the understorey undergo frequent changes in their light environment to which they must adapt to ensure their survival and growth. Crown architecture, which plays a critical role in light capture and mechanical stability, is a major component of sapling adaptation to canopy disturbance. Shade-adapted saplings typically have plagiotropic stems and branches. After canopy opening, they need to develop more erect shoots in order to exploit the new light conditions. The objective of this study was to test whether changes in sapling stem inclination occur after canopy opening, and to analyse the morphological changes associated with stem reorientation.

Methods

A 4-year canopy-opening field experiment with naturally regenerated Fagus sylvatica and Acer pseudoplatanus saplings was conducted. The appearance of new stem axes, stem basal diameter and inclination along the stem were recorded every year after canopy opening.

Key Results

Both species showed considerable stem reorientation resulting primarily from uprighting (more erect) shoot movements in Fagus, and from uprighting movements, shoot elongation and formation of relay shoots in Acer. In both species, the magnitude of shoot uprighting movements was primarily related to initial stem inclination. Both the basal part and the apical part of the stem contributed to uprighting movements. Stem movements did not appear to be limited by stem size or by stem growth.

Conclusions

Stem uprighting movements in shade-adapted Fagus and Acer saplings following canopy disturbance were considerable and rapid, suggesting that stem reorientation processes play a significant role in the growth strategy of the species.     

Proteomic analysis of rice leaf, stem and root tissues during growth course

Rice proteins were isolated from leaf, stem and root tissues, harvesting at 1, 2, 4, 8 and 10 weeks after budding. Each tissue of each age was separately pulverized in liquid nitrogen, and the resulted tissue powders were suspended in 10% TCA-acetone and followed by acetone suspension to precipitate at low temperature, which resulted in the tissue-specific and age-specific protein mixture. The protein mixtures were separated by 2-DE using polyacrylamide gels (26 x 20 cm). The protein spots were identified by N-terminal sequence analysis and by MALDI and LC-MS/MS analyses after in-gel tryptic digestion. From a total of 4532 spots, 676 unique proteins were identified, of which 80 proteins (12%) were observed in all three tissues: leaf, stem and root. In addition, 45 (7%) were common in leaf and stem, 57 (8%) in stem and root, and 10 (2%) proteins in root and leaf. Also 141 unique proteins (21%) were observed only for leaf, 96 (14%) for stem, and 247 (36%) for root tissue. Proteins playing a role for photosynthesis and energy production were most abundant in leaf and stem, and those for cell defense were rich in roots.     

Stem-righting mechanism in gymnosperm trees deduced from limitations in compression wood development

BACKGROUND AND AIMS: In response to inclination stimuli, gymnosperm trees undergo corrective growth during which compression wood develops on the lower side of the inclined stem. High compressive growth stress is generated in the compression wood region and is an important factor in righting the stem. The aims of the study were to elucidate how the generation of compressive growth stress in the compression wood region is involved in the righting response and thus to determine a righting mechanism for tree saplings. METHODS: Cryptomeria japonica saplings were grown at inclinations of 0 degrees (vertical) to 50 degrees. At each inclination angle, the growth stress on the lower side of the inclined stem was investigated, together with the degree of compression-wood development such as the width of the current growth layer and lignin content, and the upward bending moment. KEY RESULTS: Growth stress, the degree of compression wood development, and the upward moment grew as the stem inclination angle increased from 0 to 30 degrees, but did not rise further at inclinations > 30 degrees. CONCLUSIONS: The results suggest the following righting mechanism for gymnosperm saplings. As the stem inclination is elevated from 0 to 30 degrees, the degree of compression wood development increases to force the sapling back to its original orientation; at inclinations > 30 degrees, the maximum degree of compression wood is formed and additional time is needed for the stem to reorient itself.     

Proteome analysis of Arabidopsis thaliana by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry

In the present study we show results of a large-scale proteome analysis of the recently sequenced plant Arabidopsis thaliana. On the basis of a previously published sequential protein extraction protocol, we prepared protein extracts from eight different A. thaliana tissues (primary leaf, leaf, stem, silique, seedling, seed, root, and inflorescence) and analysed these by two-dimensional gel electrophoresis. A total of 6000 protein spots, from three of these tissues, namely primary leaf, silique and seedling, were excised and the contained proteins were analysed by matrix assisted laser desorption/ionisation time of flight mass spectrometry peptide mass fingerprinting. This resulted in the identification of the proteins contained in 2943 spots, which were found to be products of 663 different genes. In this report we present and discuss the methodological and biological results of our plant proteome analysis.     

2-DE proteome analysis of a proliferating and differentiating human neuronal stem cell line (ReNcell VM)

The proteome of a proliferating human stem cell line was analyzed and then utilized to detect stem cell differentiation-associated changes in the protein profile. The analysis was conducted with a stable human fetal midbrain stem cell line (ReNcell VM) that displays the properties of a neural stem cell. Therefore, acquisition of proteomic data should be representative of cultured human neural stem cells (hNSCs) in general. Here we present a 2-DE protein-map of this cell line with annotations of 402 spots representing 318 unique proteins identified by MS. The subsequent proteome profiling of differentiating cells of this stem cell line at days 0, 4 and 7 of differentiation revealed changes in the expression of 49 identified spots that could be annotated to 45 distinct proteins. This differentiation-associated expression pattern was validated by Western blot analysis for transgelin-2, proliferating cell nuclear antigen, as well as peroxiredoxin 1 and 4. The group of regulated proteins also included NudC, ubiquilin-1, STRAP, stress-70 protein, creatine kinase B, glial fibrillary acidic protein and vimentin. Our results reflect the large rearrangement of the proteome during the differentiation process of the stem cells to terminally differentiated neurons and offer the possibility for further characterization of specific targets driving the stem cell differentiation.     

Patterns of auxin distribution during gravitational induction of reaction wood in poplar and pine

Gravistimulation of tree stems affects wood development by unilaterally inducing wood with modified properties, called reaction wood. Commonly, it also stimulates cambial growth on the reaction wood side. Numerous experiments involving applications of indole-3-acetic acid (IAA) or IAA-transport inhibitors have suggested that reaction wood is induced by a redistribution of IAA around the stem. However, in planta proof for this model is lacking. Therefore, we have mapped endogenous IAA distribution across the cambial region tissues in both aspen (Populus tremula, denoted poplar) and Scots pine (Pinus sylvestris) trees forming reaction wood, using tangential cryosectioning combined with sensitive gas chromatography-mass spectrometry analysis. Moreover, we have documented the kinetics of IAA during reaction wood induction in these species. Our analysis of endogenous IAA demonstrates that reaction wood is formed without any obvious alterations in IAA balance. This is in contrast to gravitropic responses in roots and shoots where a redistribution of IAA has been documented. It is also of interest that cambial growth on the tension wood side was stimulated without an increase in IAA. Taken together, our results suggest a role for signals other than IAA in the reaction wood response, or that the gravitational stimulus interacts with the IAA signal transduction pathway.     

Differential protein expression in the corpus callosum (splenium) of human alcoholics: a proteomics study

It is widely accepted that the chronic use of alcohol induces metabolic abnormalities and neuronal damage in the brain, which can lead to cognitive dysfunction. Neuroimaging studies reveal that alcohol-induced brain damage is region specific and prominent damage has been observed in both gray and white matter of the prefrontal cortex, and a wide range of white matter structures including the corpus callosum. Molecular mechanisms underlying these structural changes are largely unknown. Using proteomics we have analysed the changes in protein expression in the splenium of the corpus callosum in two different alcoholic groups. Protein extracts from splenium of 22 human brains (nine controls, seven uncomplicated alcoholics and six complicated alcoholics with hepatic cirrhosis-designated complicated) were separated using two-dimensional gel electrophorosis. Image analysis revealed that there were significant alterations in protein expression for 25 protein spots in the uncomplicated alcoholic group and 45 in the complicated group compared to control (P<0.05; ANOVA). In a total of 72 spots (identified as 36 proteins), 15 (identified as 14 proteins) spots overlapped between two alcoholic groups. Another 32 protein spots (26 different proteins) were identified only in the complicated alcoholics. It is therefore possible that these 26 proteins in the complicated group are likely to be the results of hepatic compromise. When compared with our previous data of white matter from the prefrontal cortex in alcoholics, large numbers of identified proteins in the splenium are different. This suggests that there may be different mechanisms causing alcohol-induced brain damage in different regions of the white matter. Our data also indicate the importance of other pathways including oxidative stress, lipid peroxidation and apoptosis as potential causes of alcohol-induced brain damage.     

水稻稻瘟菌抗性相关蛋白的双向电泳分析

建立抗感水稻品种受稻瘟菌侵染和未侵染蛋白质的双向凝胶电泳图谱,分析其差异表达的蛋白,寻找稻瘟病的抗性相关蛋白,以阐明稻瘟病的发病机制。采用TCA/丙酮沉淀法提取四种愈伤组织材料的总蛋白并采用固相pH梯度( immobilized pH gradient, IPG)双向凝胶电泳( two-dimensional gel electrophshiya, oresis, 2-DE)分离四种材料总蛋白质, 凝胶经银染显色后,用PDQuest图像分析软件进行比较分析、识别差异表达的蛋白质。成功获得抗感水稻品种受稻瘟菌侵染和未侵染蛋白质的双向凝胶电泳图谱。获得未侵染内香优2号平均蛋白点数为447个,汕优63平均蛋白点数为440个;侵染后抗性品种内香优2号平均蛋白数为523个,感性品种汕优63平均蛋白质点数为326个。内香优2号未经侵染和侵染后图谱匹配率达 89%和 87%,汕优63未经侵染和侵染后图谱匹配率达86％和85％。内香优2号的差异表达蛋白点数为76个,汕优63的差异表达蛋白点数为114个。两者间存在一些差异表达的蛋白质,为阐明稻瘟病的发病机制打下了坚实的基础。