scholarly journals Overexpression of the rice BAHD acyltransferase AT10 increases xylan-bound p-coumarate and reduces lignin in Sorghum bicolor

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yang Tian ◽  
Chien-Yuan Lin ◽  
Joon-Hyun Park ◽  
Chuan-Yin Wu ◽  
Ramu Kakumanu ◽  
...  
Keyword(s):  
Cell Wall ◽  
Sorghum Bicolor ◽  
Cell Walls ◽  
Lignin Content ◽  
Complex Structure ◽  
Enzymatic Saccharification ◽  
Cellulose Microfibrils ◽  
Biomass Composition ◽  
Bioenergy Crops ◽  
Transgenic Lines

Abstract Background The development of bioenergy crops with reduced recalcitrance to enzymatic degradation represents an important challenge to enable the sustainable production of advanced biofuels and bioproducts. Biomass recalcitrance is partly attributed to the complex structure of plant cell walls inside which cellulose microfibrils are protected by a network of hemicellulosic xylan chains that crosslink with each other or with lignin via ferulate (FA) bridges. Overexpression of the rice acyltransferase OsAT10 is an effective bioengineering strategy to lower the amount of FA involved in the formation of cell wall crosslinks and thereby reduce cell wall recalcitrance. The annual crop sorghum represents an attractive feedstock for bioenergy purposes considering its high biomass yields and low input requirements. Although we previously validated the OsAT10 engineering approach in the perennial bioenergy crop switchgrass, the effect of OsAT10 expression on biomass composition and digestibility in sorghum remains to be explored. Results We obtained eight independent sorghum (Sorghum bicolor (L.) Moench) transgenic lines with a single copy of a construct designed for OsAT10 expression. Consistent with the proposed role of OsAT10 in acylating arabinosyl residues on xylan with p-coumarate (pCA), a higher amount of p-coumaroyl-arabinose was released from the cell walls of these lines upon hydrolysis with trifluoroacetic acid. However, no major changes were observed regarding the total amount of pCA or FA esters released from cell walls upon mild alkaline hydrolysis. Certain diferulate (diFA) isomers identified in alkaline hydrolysates were increased in some transgenic lines. The amount of the main cell wall monosaccharides glucose, xylose, and arabinose was unaffected. The transgenic lines showed reduced lignin content and their biomass released higher yields of sugars after ionic liquid pretreatment followed by enzymatic saccharification. Conclusions Expression of OsAT10 in sorghum leads to an increase of xylan-bound pCA without reducing the overall content of cell wall FA esters. Nevertheless, the amount of total cell wall pCA remains unchanged indicating that most pCA is ester-linked to lignin. Unlike other engineered plants overexpressing OsAT10 or a phylogenetically related acyltransferase with similar putative function, the improvements of biomass saccharification efficiency in sorghum OsAT10 lines are likely the result of lignin reductions rather than reductions of cell wall-bound FA. These results also suggest a relationship between xylan-bound pCA and lignification in cell walls.

scholarly journals Identifying transcription factors that reduce wood recalcitrance and improve enzymatic degradation of xylem cell wall in Populus

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Chiaki Hori ◽  
Naoki Takata ◽  
Pui Ying Lam ◽  
Yuki Tobimatsu ◽  
Soichiro Nagano ◽  
...  
Keyword(s):  
Cell Wall ◽  
Populus Tremuloides ◽  
Cell Walls ◽  
Enzymatic Degradation ◽  
Plant Biomass ◽  
Lignin Content ◽  
Enzymatic Saccharification ◽  
Wall Structure ◽  
Xylem Cell

AbstractDeveloping an efficient deconstruction step of woody biomass for biorefinery has been drawing considerable attention since its xylem cell walls display highly recalcitrance nature. Here, we explored transcriptional factors (TFs) that reduce wood recalcitrance and improve saccharification efficiency in Populus species. First, 33 TF genes up-regulated during poplar wood formation were selected as potential regulators of xylem cell wall structure. The transgenic hybrid aspens (Populus tremula × Populus tremuloides) overexpressing each selected TF gene were screened for in vitro enzymatic saccharification. Of these, four transgenic seedlings overexpressing previously uncharacterized TF genes increased total glucan hydrolysis on average compared to control. The best performing lines overexpressing Pt × tERF123 and Pt × tZHD14 were further grown to form mature xylem in the greenhouse. Notably, the xylem cell walls exhibited significantly increased total xylan hydrolysis as well as initial hydrolysis rates of glucan. The increased saccharification of Pt × tERF123-overexpressing lines could reflect the improved balance of cell wall components, i.e., high cellulose and low xylan and lignin content, which could be caused by upregulation of cellulose synthase genes upon the expression of Pt × tERF123. Overall, we successfully identified Pt × tERF123 and Pt × tZHD14 as effective targets for reducing cell wall recalcitrance and improving the enzymatic degradation of woody plant biomass.

Silicification of wood-cell walls

1994 ◽  
Vol 52 ◽  
pp. 428-429
Author(s):  
S. E. Keckler ◽  
D. M. Dabbs ◽  
N. Yao ◽  
I. A. Aksay
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Cell Walls ◽  
Lignin Content ◽  
Resin Composite ◽  
Middle Lamella ◽  
Cellulose Fibers ◽  
Complex Structures ◽  
Rich Region ◽  
Inorganic Precursors

Cellular organic structures such as wood can be used as scaffolds for the synthesis of complex structures of organic/ceramic nanocomposites. The wood cell is a fiber-reinforced resin composite of cellulose fibers in a lignin matrix. A single cell wall, containing several layers of different fiber orientations and lignin content, is separated from its neighboring wall by the middle lamella, a lignin-rich region. In order to achieve total mineralization, deposition on and in the cell wall must be achieved. Geological fossilization of wood occurs as permineralization (filling the void spaces with mineral) and petrifaction (mineralizing the cell wall as the organic component decays) through infiltration of wood with inorganics after growth. Conversely, living plants can incorporate inorganics into their cells and in some cases into the cell walls during growth. In a recent study, we mimicked geological fossilization by infiltrating inorganic precursors into wood cells in order to enhance the properties of wood. In the current work, we use electron microscopy to examine the structure of silica formed in the cell walls after infiltration of tetraethoxysilane (TEOS).

The effects of thermal treatment on the nanomechanical behavior of bamboo (Phyllostachys pubescens Mazel ex H. de Lehaie) cell walls observed by nanoindentation, XRD, and wet chemistry

Holzforschung ◽  
2017 ◽  
Vol 71 (2) ◽  
pp. 129-135 ◽  
Author(s):  
Yanjun Li ◽  
Chengjian Huang ◽  
Li Wang ◽  
Siqun Wang ◽  
Xinzhou Wang
Keyword(s):  
Cell Wall ◽  
Thermal Treatment ◽  
Cell Walls ◽  
Treatment Time ◽  
Lignin Content ◽  
Phyllostachys Pubescens ◽  
X Ray Diffraction ◽  
Wet Chemistry ◽  
X Ray ◽  
Wet Chemical

Abstract The effects of thermal treatment of bamboo at 130, 150, 170, and 190°C for 2, 4, and 6 h were investigated in terms of changes in chemical composition, cellulose crystallinity, and mechanical behavior of the cell-wall level by means of wet chemical analysis, X-ray diffraction (XRD), and nanoindentation (NI). Particularly, the reduced elastic modulus (Er), hardness (H), and creep behavior were in focus. Both the temperature and treatment time showed significant effects. Expectedly, the hemicelluloses were degraded and the relative lignin content was elevated, while the crystallinity of the cellulose moiety was increased upon thermal treatment. The Er and H data of the cell wall were increased after 6 h treatment at 190°C, from 18.4 to 22.0 GPa and from 0.45 to 0.65 GPa, respectively. The thermal treatment led to a decrease of the creep ratio (CIT) under the same conditions by ca. 28%. The indentation strain state (εi) also decreased significantly after thermal treatment during the load-holding stage.

Lignification of spruce tracheid secondary cell walls related to longitudinal hardness and modulus of elasticity using nano-indentation

2002 ◽  
Vol 80 (10) ◽  
pp. 1029-1033 ◽  
Author(s):  
W Gindl ◽  
H S Gupta ◽  
C Grünwald
Keyword(s):  
Cell Wall ◽  
Modulus Of Elasticity ◽  
Cell Walls ◽  
Secondary Cell Wall ◽  
Lignin Content ◽  
Wood Formation ◽  
Nano Indentation ◽  
Secondary Cell Walls ◽  
Cell Wall Development ◽  
The One

The lignin content and the mechanical properties of lignifying and fully lignified spruce tracheid secondary cell walls were determined using UV microscopy and nano-indentation, respectively. The average lignin content of developing tracheids was 0.10 g·g–1, as compared with 0.21 g·g–1 in mature tracheids. The modulus of elasticity of developing cells was on average 22% lower than the one measured in mature, fully lignified cells. For the longitudinal hardness, a larger difference of 26% was observed. As lignifying cells in the cambial zone are undergoing cell wall development, spaces in the cellulose–hemicellulose structure are filled with lignin and the density of the cell wall is believed to increase. It is therefore suggested that the observed difference in modulus of elasticity between developing and fully lignified cell walls is due to the filling of spaces with lignin and an increase of the packing density of the cell wall during lignification. Although remarkably less stiff than the composite polysaccharide structure in the secondary cell wall, lignin may be considered equally hard. Therefore, the observed increase in lignin content may contribute directly to the measured increase of hardness.Key words: secondary cell wall, hardness, lignin, modulus of elasticity, wood formation.

High-Resolution Field Emission Scanning Electron Microscopy (FESEM) Imaging of Cellulose Microfibril Organization in Plant Primary Cell Walls

2017 ◽  
Vol 23 (5) ◽  
pp. 1048-1054 ◽  
Author(s):  
Yunzhen Zheng ◽  
Daniel J. Cosgrove ◽  
Gang Ning
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Wall ◽  
High Resolution ◽  
Field Emission ◽  
Cell Walls ◽  
Cellulose Microfibrils ◽  
Critical Point Drying ◽  
Sputter Coating ◽  
Scanning Electron

AbstractWe have used field emission scanning electron microscopy (FESEM) to study the high-resolution organization of cellulose microfibrils in onion epidermal cell walls. We frequently found that conventional “rule of thumb” conditions for imaging of biological samples did not yield high-resolution images of cellulose organization and often resulted in artifacts or distortions of cell wall structure. Here we detail our method of one-step fixation and dehydration with 100% ethanol, followed by critical point drying, ultrathin iridium (Ir) sputter coating (3 s), and FESEM imaging at a moderate accelerating voltage (10 kV) with an In-lens detector. We compare results obtained with our improved protocol with images obtained with samples processed by conventional aldehyde fixation, graded dehydration, sputter coating with Au, Au/Pd, or carbon, and low-voltage FESEM imaging. The results demonstrated that our protocol is simpler, causes little artifact, and is more suitable for high-resolution imaging of cell wall cellulose microfibrils whereas such imaging is very challenging by conventional methods.

scholarly journals Cellulose synthase complexes act in a concerted fashion to synthesize highly aggregated cellulose in secondary cell walls of plants

2016 ◽  
Vol 113 (40) ◽  
pp. 11348-11353 ◽  
Author(s):  
Shundai Li ◽  
Logan Bashline ◽  
Yunzhen Zheng ◽  
Xiaoran Xin ◽  
Shixin Huang ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Cellulose Synthase ◽  
Cellulose Microfibrils ◽  
Critical Component ◽  
Distinct Structure ◽  
Secondary Cell Walls ◽  
Composition Structure ◽  
Membrane Spanning

Cellulose, often touted as the most abundant biopolymer on Earth, is a critical component of the plant cell wall and is synthesized by plasma membrane-spanning cellulose synthase (CESA) enzymes, which in plants are organized into rosette-like CESA complexes (CSCs). Plants construct two types of cell walls, primary cell walls (PCWs) and secondary cell walls (SCWs), which differ in composition, structure, and purpose. Cellulose in PCWs and SCWs is chemically identical but has different physical characteristics. During PCW synthesis, multiple dispersed CSCs move along a shared linear track in opposing directions while synthesizing cellulose microfibrils with low aggregation. In contrast, during SCW synthesis, we observed swaths of densely arranged CSCs that moved in the same direction along tracks while synthesizing cellulose microfibrils that became highly aggregated. Our data support a model in which distinct spatiotemporal features of active CSCs during PCW and SCW synthesis contribute to the formation of cellulose with distinct structure and organization in PCWs and SCWs of Arabidopsis thaliana. This study provides a foundation for understanding differences in the formation, structure, and organization of cellulose in PCWs and SCWs.

The fate of acetyl groups and sugar components during the digestion of grass cell walls in sheep

1977 ◽  
Vol 89 (2) ◽  
pp. 327-340 ◽  
Author(s):  
E. Jane Morris ◽  
J. S. D. Bacon
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Lignin Content ◽  
Sodium Ethoxide ◽  
Mesophyll Cell ◽  
Cell Types ◽  
Gas Liquid Chromatography ◽  
Cell Wall Preparation ◽  
The Relationship ◽  
Isolated Cell

SummaryThe digestibilities of grass cell wall constituents determined in a digestion trial were compared with those obtained by suspending various isolated cell wall preparations in nylon bags in the rumen of a sheep. Particular attention was paid to acetyl groups and to individual sugars, which were determined in both cases by gas liquid chromatography.For dried grass and hay in the digestion trial the cell wall constituents showed digestibilities decreasing in the following order: arabinose, galactose, glucose, xylose, acetyl, lignin.For a leaf cell wall preparation derived from all cell types except mesophyll, the nylon bag technique allowed the same order of digestibilities; rhamnose and uronic acids were also measured and found to be rapidly digested. Mesophyll cell walls placed in nylon bags were more readily digested than non-mesophyll. All the sugars, and also acetyl groups, were digested to the same extent.In a grass cell wall preparation isolated from sheep faeces, tested similarly, xylose and glucose were digested to the same extent, but acetyl groups were less digested.Removal of acetyl groups, using sodium ethoxide, which left the sugar composition and lignin content unchanged, increased the digestibility particularly of the cell walls from faeces.The results are discussed with reference to the relationship between cell wall composition and digestibility.

scholarly journals Cell wall composition determines handedness reversal in helicoidal cellulose architectures of Pollia condensata fruits

2021 ◽  
Vol 118 (51) ◽  
pp. e2111723118
Author(s):  
Yin Chang ◽  
Rox Middleton ◽  
Yu Ogawa ◽  
Tom Gregory ◽  
Lisa M. Steiner ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Hierarchical Structures ◽  
Cell Wall Composition ◽  
Cellulose Microfibrils ◽  
Plant Cell Walls ◽  
Mechanical Responses ◽  
Structural Colors ◽  
Left Handed ◽  
Helicoidal Structure

Chiral asymmetry is important in a wide variety of disciplines and occurs across length scales. While several natural chiral biomolecules exist only with single handedness, they can produce complex hierarchical structures with opposite chiralities. Understanding how the handedness is transferred from molecular to the macroscopic scales is far from trivial. An intriguing example is the transfer of the handedness of helicoidal organizations of cellulose microfibrils in plant cell walls. These cellulose helicoids produce structural colors if their dimension is comparable to the wavelength of visible light. All previously reported examples of a helicoidal structure in plants are left-handed except, remarkably, in the Pollia condensata fruit; both left- and right-handed helicoidal cell walls are found in neighboring cells of the same tissue. By simultaneously studying optical and mechanical responses of cells with different handednesses, we propose that the chirality of helicoids results from differences in cell wall composition. In detail, here we showed statistical substantiation of three different observations: 1) light reflected from right-handed cells is red shifted compared to light reflected from left-handed cells, 2) right-handed cells occur more rarely than left-handed ones, and 3) right-handed cells are located mainly in regions corresponding to interlocular divisions. Finally, 4) right-handed cells have an average lower elastic modulus compared to left-handed cells of the same color. Our findings, combined with mechanical simulation, suggest that the different chiralities of helicoids in the cell wall may result from different chemical composition, which strengthens previous hypotheses that hemicellulose might mediate the rotations of cellulose microfibrils.

scholarly journals Genetic Modification of KNAT7 Transcription Factor Expression Enhances Saccharification and Reduces Recalcitrance of Woody Biomass in Poplars

2021 ◽  
Vol 12 ◽  
Author(s):  
Yogesh Kumar Ahlawat ◽  
Akula Nookaraju ◽  
Anne E. Harman-Ware ◽  
Crissa Doeppke ◽  
Ajaya K. Biswal ◽  
...  
Keyword(s):  
Lignin Content ◽  
Woody Biomass ◽  
Bioenergy Crops ◽  
Transgenic Lines ◽  
Saccharification Efficiency ◽  
Transgenic Poplars ◽  
Specific Alteration ◽  
Guaiacyl Lignin ◽  
Transcription Factor Expression ◽  
The Impact

The precise role of KNAT7 transcription factors (TFs) in regulating secondary cell wall (SCW) biosynthesis in poplars has remained unknown, while our understanding of KNAT7 functions in other plants is continuously evolving. To study the impact of genetic modifications of homologous and heterologous KNAT7 gene expression on SCW formation in transgenic poplars, we prepared poplar KNAT7 (PtKNAT7) overexpression (PtKNAT7-OE) and antisense suppression (PtKNAT7-AS) vector constructs for the generation of transgenic poplar lines via Agrobacterium-mediated transformation. Since the overexpression of homologous genes can sometimes result in co-suppression, we also overexpressed Arabidopsis KNAT7 (AtKNAT7-OE) in transgenic poplars. In all these constructs, the expression of KNAT7 transgenes was driven by developing xylem (DX)-specific promoter, DX15. Compared to wild-type (WT) controls, many SCW biosynthesis genes downstream of KNAT7 were highly expressed in poplar PtKNAT7-OE and AtKNAT7-OE lines. Yet, no significant increase in lignin content of woody biomass of these transgenic lines was observed. PtKNAT7-AS lines, however, showed reduced expression of many SCW biosynthesis genes downstream of KNAT7 accompanied by a reduction in lignin content of wood compared to WT controls. Syringyl to Guaiacyl lignin (S/G) ratios were significantly increased in all three KNAT7 knockdown and overexpression transgenic lines than WT controls. These transgenic lines were essentially indistinguishable from WT controls in terms of their growth phenotype. Saccharification efficiency of woody biomass was significantly increased in all transgenic lines than WT controls. Overall, our results demonstrated that developing xylem-specific alteration of KNAT7 expression affects the expression of SCW biosynthesis genes, impacting at least the lignification process and improving saccharification efficiency, hence providing one of the powerful tools for improving bioethanol production from woody biomass of bioenergy crops and trees.

scholarly journals Bundling of cellulose microfibrils in native and polyethylene glycol-containing wood cell walls revealed by small-angle neutron scattering

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Paavo A. Penttilä ◽  
Michael Altgen ◽  
Muhammad Awais ◽  
Monika Österberg ◽  
Lauri Rautkari ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neutron Scattering ◽  
Plant Cell ◽  
Small Angle ◽  
Cell Walls ◽  
Small Angle Neutron Scattering ◽  
Plant Cell Wall ◽  
Raw Materials ◽  
Cellulose Microfibrils ◽  
Wall Structure

AbstractWood and other plant-based resources provide abundant, renewable raw materials for a variety of applications. Nevertheless, their utilization would greatly benefit from more efficient and accurate methods to characterize the detailed nanoscale architecture of plant cell walls. Non-invasive techniques such as neutron and X-ray scattering hold a promise for elucidating the hierarchical cell wall structure and any changes in its morphology, but their use is hindered by challenges in interpreting the experimental data. We used small-angle neutron scattering in combination with contrast variation by poly(ethylene glycol) (PEG) to identify the scattering contribution from cellulose microfibril bundles in native wood cell walls. Using this method, mean diameters for the microfibril bundles from 12 to 19 nm were determined, without the necessity of cutting, drying or freezing the cell wall. The packing distance of the individual microfibrils inside the bundles can be obtained from the same data. This finding opens up possibilities for further utilization of small-angle scattering in characterizing the plant cell wall nanostructure and its response to chemical, physical and biological modifications or even in situ treatments. Moreover, our results give new insights into the interaction between PEG and the wood nanostructure, which may be helpful for preservation of archaeological woods.

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