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At least 55 records · Page 3

Evaluating the Utility of Permethylated Polysaccharide Solution NMR Data for Characterization of Insoluble Plant Cell Wall Polysaccharides

Plant cell wall polysaccharide analysis encompasses the utilization of a variety of analytical tools, including gas and liquid chromatography, mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy. These methods provide complementary data, which enable confident structural proposals of the many complex polysaccharide structures that exist in the complex matrices of plant cell walls. However, cell walls contain fractions of varying solubilities, and a few techniques are available that can analyze all fractions simultaneously. We have discovered that permethylation affords the complete dissolution of both soluble and insoluble polysaccharide fractions of plant cell walls in organic solvents such as chloroform or acetonitrile, which can then be analyzed by a number of analytical techniques including MS and NMR. In this work, NMR structure analysis of 10 permethylated polysaccharide standards was undertaken to generate chemical shift data providing insights into spectral changes that result from permethylation of polysaccharide residues. This information is of especial relevance to the structure analysis of insoluble polysaccharide materials that otherwise are not easily investigated by solution-state NMR methodologies. The preassigned NMR chemical shift data is shown to be vital for NMR structure analysis of minor polysaccharide components of plant cell walls that are particularly difficult to assign by NMR correlation data alone. With the assigned chemical shift data, we analyzed the permethylated samples of destarched, alcohol-insoluble residues of switchgrass and poplar by two-dimensional NMR spectral profiling. Thus, we identified, in addition to the major polysaccharide components, two minor polysaccharides, namely, <5% 3-linked arabinoxylan (switchgrass) and <2% glucomannan (poplar). In particular, the position of the arabinose residue in the arabinoxylan of the switchgrass sample was confidently assigned based on chemical shift values, which are highly sensitive to local chemical environments. Furthermore, the high resolution afforded by the 1H NMR spectra of the permethylated switchgrass and poplar samples allowed facile relative quantitative analysis of their polysaccharide composition, utilizing only a few milligrams of the cell wall material. As a result, the concepts herein developed will thus facilitate NMR structure analysis of insoluble plant cell wall polysaccharides, more so of minor cell wall components that are especially challenging to analyze with current methods.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Methods for assessing wall interference in the 2- by 2-foot adaptive-wall wind tunnel

Discussed are two methods for assessing two-dimensional wall interference in the adaptive-wall test section of the NASA Ames 2 x 2-Foot Transonic Wind Tunnel: (1) a method for predicting free-air conditions near the walls of the test section (adaptive-wall methods); and (2) a method for estimating wall-induced velocities near the model (correction methods), both of which methods are based on measurements of either one or two components of flow velocity near the walls of the test section. Each method is demonstrated using simulated wind tunnel data and is compared with other methods of the same type. The two-component adaptive-wall and correction methods were found to be preferable to the corresponding one-component methods because: (1) they are more sensitive to, and give a more complete description of, wall interference; (2) they require measurements at fewer locations; (3) they can be used to establish free-stream conditions; and (4) they are independent of a description of the model and constants of integration.

Schairer, E. T.↗

Evaluation of a flexible wall testing technique to minimize wall interferences in the NASA Langley 0.3-m Transonic Cryogenic Tunnel

Free air simulations in conventional transonic wind tunnels require improvement which adaptive wall testing techniques can provide primarily by minimizing wall interferences. In addition, these techniques offer other substantial advantages such as increased Reynolds number capability and a reduction in tunnel drive power. The combination of an adaptive wall test section with a continuous flow cryogenic wind tunnel is unique. The test section has four solid walls with two flexible walls mounted between rigid sidewalls. This modification of an existing major facility stresses the practicalities of the testing technique. These practicalities were evaluated in terms of flexible wall test section design and operation. Increased hardware and operating complexity of the new test section is offset by a significant improvement in real-time data accuracy in two-dimensional testing. Validation testing has expanded the experience with flexible walled test sections into the realms of flight Reynolds numbers and high lift. Data accuracy has been assessed with regard to test section geometry and operating tolerances. The successful evaluation of the testing technique in two-dimensional testing has proved that a production type operation is possible, with suitable control system and test section design. The 0.3-m Transonic Cryogenic Tunnel with an adaptive wall test section currently represents the most advanced two-dimensional facility anywhere.

Wolf, Stephen W. D.↗

The role of wall calcium in the extension of cell walls of soybean hypocotyls

Calcium crosslinks are load-bearing bonds in soybean (Glycine max (L.) Merr.) hypocotyl cell walls, but they are not the same load-bearing bonds that are broken during acid-mediated cell elongation. This conclusion is reached by studying the relationship between wall calcium, pH and the facilitated creep of frozen-thawed soybean hypocotyl sections. Supporting data include the following observations: 1) 2-[(2-bis-[carboxymethyl]amino-5-methylphenoxy)methyl]-6-methoxy-8-bis[car boxymethyl]aminoquinoline (Quin 2) and ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) caused only limited facilitated creep as compared with acid, despite removal of comparable or larger amounts of wall calcium; 2) the pH-response curves for calcium removal and acid-facilitated creep were different; 3) reversible acid-extension occurred even after removal of almost all wall calcium with Quin 2; and 4) growth of abraded sections did not involve a proportional loss of wall calcium. Removal of wall calcium, however, increased the capacity of the walls to undergo acid-facilitated creep. These data indicate that breakage of calcium crosslinks is not a major mechanism of cell-wall loosening in soybean hypocotyl tissues.

Non-NASA Center↗

Comparison of Turbulent Heat-Transfer Results for Uniform Wall Heat Flux and Uniform Wall Temperature

The purpose of this note is to examine in a more precise way how the Nusselt numbers for turbulent heat transfer in both the fully developed and thermal entrance regions of a circular tube are affected by two different wall boundary conditions. The comparisons are made for: (a) Uniform wall temperature (UWT); and (b) uniform wall heat flux (UHF). Several papers which have been concerned with the turbulent thermal entrance region problem are given. 1 Although these analyses have all utilized an eigenvalue formulation for the thermal entrance region there were differences in the choices of eddy diffusivity expressions, velocity distributions, and methods for carrying out the numerical solutions. These differences were also found in the fully developed analyses. Hence when making a comparison of the analytical results for uniform wall temperature and uniform wall heat flux, it was not known if differences in the Nusselt numbers could be wholly attributed to the difference in wall boundary conditions, since all the analytical results were not obtained in a consistent way. To have results which could be directly compared, computations were carried out for the uniform wall temperature case, using the same eddy diffusivity, velocity distribution, and digital computer program employed for uniform wall heat flux. In addition, the previous work was extended to a lower Reynolds number range so that comparisons could be made over a wide range of both Reynolds and Prandtl numbers.

Siegel, R.↗

Bioenergy sorghum stem density increases threefold following internode elongation due to continued accumulation of lignified cell walls and complex regulation of genes involved in cell wall biosynthesis

Bioenergy sorghum is a highly productive drought tolerant C4 grass that accumulates ~ 80% of its harvested biomass in ~ 4 m long stems comprised of > 40 internodes that develop sequentially during an extended vegetative growth phase. Following elongation of each internode, internode density increases ~ threefold to fourfold primarily due to the accumulation of cell walls composed of cellulose, glucuronoarabinoxylan and lignin. Lignin accumulates initially on cell walls of sclerenchyma cells surrounding vascular bundles and later on cell walls of the stem rind and stem core pith parenchyma. Many genes involved in cell wall biosynthesis were expressed continuously during the stem internode densification process whereas others showed dynamic patterns of expression (high to low, low to high). Several CESA genes involved in primary cell wall cellulose synthesis were expressed in the stem rind and core throughout the stem densification phase. In contrast, CESA genes involved in secondary cell wall biogenesis were expressed continuously in the stem rind but downregulated in the stem core shortly after completion of internode elongation. Overall, accumulation of cell wall biomass in elongated internodes during stem densification increases stem mechanical strength and biomass bulk density while modifying biomass composition in ways that could impact the amount and release of cellulosic sugars and lignin-derived bioproducts.

09 BIOMASS FUELS↗

Distinguishing Mesoscale Polar Order (Unidirectional vs Bidirectional) of Cellulose Microfibrils in Plant Cell Walls Using Sum Frequency Generation Spectroscopy

Cellulose in plant cell walls are synthesized as crystalline microfibrils with diameters of 3–4 nm and lengths of around 1–10 μm. These microfibrils are known to be the backbone of cell walls, and their multiscale three-dimensional organization plays a critical role in cell wall functions including plant growth and recalcitrance to degradation. The mesoscale organization of microfibrils over a 1–100 nm range in cell walls is challenging to resolve because most characterization techniques investigating this length scale suffer from low spatial resolution, sample preparation artifacts, or inaccessibility of specific cell types. Here, we report a sum frequency generation (SFG) study determining the mesoscale polarity of cellulose microfibrils in intact plant cell walls. SFG is a nonlinear optical spectroscopy technique sensitive to the molecular-to-mesoscale order of noncentrosymmetric domains in amorphous matrices. However, the quantitative theoretical model to unravel the effect of polarity in packing of noncentrosymmetric domains on SFG spectral features has remained unresolved. In this work, we show how the phase synchronization principle of the SFG process is used to predict the relative intensities of vibrational modes with different polar angles from the noncentrosymmetric domain axis. Applying this model calculation for the first time and employing SFG microscopy, we found that cellulose microfibrils in certain xylem cell walls are deposited unidirectionally (or biased in one direction) instead of the bidirectional polarity which was believed to be dominant in plant cell walls from volume-averaged characterizations of macroscopic samples. Here, with this advancement in SFG analysis, one can now determine the relative polarity of noncentrosymmetric domains such as crystalline biopolymers interspersed in amorphous polymer matrices, which will open opportunities to study new questions that have not been conceived in the past.

36 MATERIALS SCIENCE↗

Molecular insights into the complex mechanics of plant epidermal cell walls

Computational analysis of cell walls Layers of intertwined fibers make up plant cell walls. The various types of fibers respond differently to deformation. Cellulose microfibrils, for example, can stretch or curve, changing their end-to-end length, and can also slide past each other, reorient relative directions, and bundle with neighboring microfibrils. Zhang et al. developed a computational model based on observations of onion skin epidermis that describes how these complex changes in space govern cell wall mechanics. The results inform ways to engineer multifunctional fibrous materials. Science , this issue p. 706

Science & Technology - Other Topics↗

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in β-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for β-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of β-(1,3)-glucan to the immune system occurs when the mannan layer is altered or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in this paper in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. Finally, AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system.

59 BASIC BIOLOGICAL SCIENCES↗

Transonic wall interference assessment and corrections for airfoil data from the 0.3-meter TCT adaptive wall test section

The wall interference assessment/correction code presented is nonlinear, involves four walls, and is applicable to transonic airfoil data from wind tunnels with shaped, solid top and bottom walls. Attention is given to its application to data from the NASA 0.3-m Transonic Cryogenic Tunnel Adaptive Test Section, for two sizes of a NACA 0012 airfoil and to simulated data for an inviscid two-dimensional full-potential code. This study indicates that while adaptive wall wind tunnels significantly reduce some aspects of wall-interference effects (by comparison to straight solid and slotted wall wind tunnels), residual wall and other interference effects are present.

Green, Lawrence L.↗

Wall shear stress measurement in blade end-wall corner region

The magnitude and the direction of wall shear stress and surface pressure in the blade end-wall corner region were investigated. The measurements were obtained on a specially designed Preston tube, the tip of which could be concentrically rotated about its axis of rotation at the measurement location. The magnitude of wall shear stress in the vicinity of the corner was observed to increase significantly (170 percent) compared to its far-upstream value; the increase was consistently higher on the blade surface compared to the value on the plate surface of the blade end-wall corner. On both surfaces in the blade end-wall corner, the variation of the wall shear stress direction was found to be more predominant in the vicinity of the blade leading-edge location. The trend of the measured wall shear stress direction showed good agreement with the limiting streamline directions obtained from the flow visualization studies.

Bhargava, R.↗

Simultaneous axial conduction in the fluid and the pipe wall for forced convective laminar flow with blowing and suction at the wall

Numerical solutions are reported for conjugate heat transfer in a porous pipe having an internal flow with blowing or suction at the inner surface of the pipe and constant heat flux at the outer surface. The effect of the simultaneous axial conduction through the wall and the fluid has been studied for the combined hydrodynamic and thermal entry lengths. The results show that the ratio of the thermal conductivities of the pipe wall to the fluid and the thickness of the pipe wall may become significant factors on the heat transfer when the Peclet number is small, especially for the case when fluid is injected into the pipe. It is also shown that the effect of axial wall conduction for the case of constant heat flux at the outer wall surface can be neglected when the wall thickness is small and the ratio of the conductivities of the wall to the fluid approaches unity.

Faghri, Amir↗

Effect of uneven wall temperature on local heat transfer in a rotating square channel with smooth walls and radial outward flow

The effect of uneven wall temperature on the local heat transfer coefficient in a rotating square channel with smooth walls and radial outward flow is investigated for Reynolds numbers from 2500 to 25,000 and rotation numbers from 0 to 0.352. Three cases of thermal boundary conditions are studied: (1) four walls uniform temperature, (2) four walls uniform heat flux, and (3) leading and trailing walls hot and two side walls cold. It is shown that the heat transfer coefficients on the leading surface are much lower than that of the trailing surface due to rotation. For case 1, the leading surface heat transfer coefficient decreases and then increases with increasing rotation numbers, and the trailing surface heat transfer coefficient increases monotonically with rotation numbers. The trailing surface heat transfer coefficients, as well as those for the side walls, for case 2 are higher than for case 1, and the leading surface heat transfer coefficients for cases 2 and 3 are significantly higher than for case 1.

Han, J.-C.↗

SECONDARY WALL ASSOCIATED MYB1 is a positive regulator of secondary cell wall thickening in Brachypodium distachyon and is not found in the Brassicaceae

Grass biomass is comprised chiefly of secondary walls that surround fiber and xylem cells. A regulatory network of interacting transcription factors in part regulates cell wall thickening. Here, we identified Brachypodium distachyon SECONDARY WALL ASSOCIATED MYB1 (SWAM1) as a potential regulator of secondary cell wall biosynthesis based on gene expression, phylogeny, and transgenic plant phenotypes. SWAM1 interacts with cellulose and lignin gene promoters with preferential binding to AC-rich sequence motifs commonly found in the promoters of cell wall-related genes. SWAM1 overexpression (SWAM-OE) lines had greater above-ground biomass with only a slight change in flowering time while SWAM1 dominant repressor (SWAM1-DR) plants were severely dwarfed with a striking reduction in lignin of sclerenchyma fibers and stem epidermal cell length. Cellulose, hemicellulose, and lignin genes were significantly down-regulated in SWAM1-DR plants and up-regulated in SWAM1-OE plants. There was no reduction in bioconversion yield in SWAM1-OE lines; however, it was significantly increased for SWAM1-DR samples. Phylogenetic and syntenic analyses strongly suggest that the SWAM1 clade was present in the last common ancestor between eudicots and grasses, but is not in the Brassicaceae. Collectively, these data suggest that SWAM1 is a transcriptional activator of secondary cell wall thickening and biomass accumulation in B. distachyon.

09 BIOMASS FUELS↗

Expression of S-adenosylmethionine Hydrolase in Tissues Synthesizing Secondary Cell Walls Alters Specific Methylated Cell Wall Fractions and Improves Biomass Digestibility

Plant biomass is a large source of fermentable sugars for the synthesis of bioproducts using engineered microbes. These sugars are stored as cell wall polymers, mainly cellulose and hemicellulose, and are embedded with lignin, which makes their enzymatic hydrolysis challenging. One of the strategies to reduce cell wall recalcitrance is the modification of lignin content and composition. Lignin is a phenolic polymer of methylated aromatic alcohols and its synthesis in tissues developing secondary cell walls is a significant sink for the consumption of the methyl donor S-adenosylmethionine (AdoMet). In this study, we demonstrate in Arabidopsis stems that targeted expression of AdoMet hydrolase (AdoMetase, E.C. 3.3.1.2) in secondary cell wall synthesizing tissues reduces the AdoMet pool and impacts lignin content and composition. In particular, both NMR analysis and pyrolysis gas chromatography mass spectrometry of lignin in engineered biomass showed relative enrichment of non-methylated p-hydroxycinnamyl (H) units and a reduction of dimethylated syringyl (S) units. This indicates a lower degree of methylation compared to that in wild-type lignin. Quantification of cell wall-bound hydroxycinnamates revealed a reduction of ferulate in AdoMetase transgenic lines. Biomass from transgenic lines, in contrast to that in control plants, exhibits an enrichment of glucose content and a reduction in the degree of hemicellulose glucuronoxylan methylation. We also show that these modifications resulted in a reduction of cell wall recalcitrance, because sugar yield generated by enzymatic biomass saccharification was greater than that of wild-type plants. Considering that transgenic plants show no important diminution of biomass yields, and that heterologous expression of AdoMetase protein can be spatiotemporally optimized, this novel approach provides a valuable option for the improvement of lignocellulosic biomass feedstock.

09 BIOMASS FUELS↗

The Impact of Cell Wall Feruloylation on Plant Growth, Responses to Environmental Stress, Plant Pathogens and Cell Wall Degradability

This article summarizes evolving concepts and scientific findings on cell wall feruloylation and ferulate oxidative coupling processes in grasses, and the effects these have on the wide range of cell wall properties and consequent plant responses to biotic and abiotic stress and tissue degradability. Updates of the different strategies that have been applied to genetically modifying cell wall feruloylation are presented. Special emphasis is given to the modification of cell wall feruloylation by heterologous expression of cell wall ferulic acid esterase, as this strategy has provided insights into the impact of feruloylation on the changes in the physicochemical properties of the cell wall with consequent effects on different plant processes. Emerging feruloyl transferase candidate genes codifying enzymes accounting for ferulate incorporation into grass arabinoxylans are also highlighted.

59 BASIC BIOLOGICAL SCIENCES↗