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1.
Leonard Blaschek Emiko Murozuka Henrik Serk Delphine Mnard Edouard Pesquet 《The Plant cell》2023,35(2):889
Vascular plants reinforce the cell walls of the different xylem cell types with lignin phenolic polymers. Distinct lignin chemistries differ between each cell wall layer and each cell type to support their specific functions. Yet the mechanisms controlling the tight spatial localization of specific lignin chemistries remain unclear. Current hypotheses focus on control by monomer biosynthesis and/or export, while cell wall polymerization is viewed as random and nonlimiting. Here, we show that combinations of multiple individual laccases (LACs) are nonredundantly and specifically required to set the lignin chemistry in different cell types and their distinct cell wall layers. We dissected the roles of Arabidopsis thaliana LAC4, 5, 10, 12, and 17 by generating quadruple and quintuple loss-of-function mutants. Loss of these LACs in different combinations led to specific changes in lignin chemistry affecting both residue ring structures and/or aliphatic tails in specific cell types and cell wall layers. Moreover, we showed that LAC-mediated lignification has distinct functions in specific cell types, waterproofing fibers, and strengthening vessels. Altogether, we propose that the spatial control of lignin chemistry depends on different combinations of LACs with nonredundant activities immobilized in specific cell types and cell wall layers.The spatial control of lignin chemistry, and thus of specific cellular functions, depends on combinations of laccases with nonredundant activities in specific cell types and cell wall layers.IN A NUTSHELL Background: Lignins are a diverse, complex group of aromatic polymers that accumulate in cell walls of vascular plants, reinforcing organs, and enabling long-distance water transport. The different cell wall layers of each cell type exhibit specific lignin chemistries with distinct proportions of specific aromatic substitutions and aliphatic functions. The spatial control of this lignin chemistry was supposed to depend exclusively on the chemical identity of the lignin monomers exported into the cell wall. However, monomer supply alone cannot fully explain the sharp spatial differences between each cell wall layer in the different cell types. We, therefore, investigated whether different paralogs of the lignin monomer-oxidizing LACCASE enzymes are responsible for spatially controlling lignin chemistry at the cell wall layer level for the different cell types in the vascular tissues of plants. Question: How are specific lignin chemistries spatially controlled by LACCASE paralogs in each cell wall layer and cell type? What are the roles of LACCASE-dependent lignin accumulation for the mechanical reinforcement and the waterproofing of different cell types in plant vascular tissues? Findings: We answered these questions by identifying the LACCASE paralogs specifically expressed in vascular cells undergoing lignin accumulation. We analyzed their functions using genetic engineering to switch off five of the six LACCASE paralog genes associated with lignin formation. Their importance in the cell wall layer and cell type lignin accumulation was determined by comparing plants sharing four of the five mutations in different LACCASE paralogs. We show that each LACCASE paralog exhibits specific substrate preference, pH optimum and localization differing between the cell wall layers of each cell type. Their lignin concentration and composition moreover depended on specific combinations of LACCASE paralogs, each enabling different aromatic substitutions and aliphatic functions to accumulate. Impairing these LACCASE-dependent lignin chemistries resulted in the loss of cell wall mechanical resistance of sap-conducting cells and the loss of cell wall waterproofing of organ-reinforcing fiber cells. Next steps: We are now pursuing research to understand the molecular mechanisms controlling the supply of lignin precursors as well as the temporal regulation activating lignification during the formation/maturation of each cell wall layer in the different cell types. 相似文献
2.
Samples of 1 kg of wheat straw, oat straw and paspalum hay were separated manually into botanical fractions, and the three largest fractions of each forage were analysed chemically for cell-wall constituents, silica and nitrogen. Proportions of digested dry matter, cellulose and hemicellulose, and potential digestibility for each of these major botanical fractions were determined when chaffed samples were placed in nylon bags and incubated for 12, 24, 48, 72 and 96 h in the rumen of sheep fed on lucerno. Cross-sections of botanical fractions were stained with safranin and fast green, and proportions of lignified tissue determined by light-microscopy and planimetry.Large differences in dry-matter digestibility between wheat straw and oat straw were attributed to the different proportions of botanical fractions. Within forages, stem was the largest fraction, the most lignified and had the lowest potential digestibility. Proportions of digested dry matter from botanical fractions at 12 h were poorly correlated with lignin content of dry matter (r = 0.25) but at 72 h were negatively correlated with lignin content of dry matter (r = ?0.84, P<0.01) and with proportions of lignified tissue (r = ?0.67, P<0.05) in the respective botanical fractions. Proportions of cellulose and hemicellulose digested at 72 h were strongly correlated with lignin content of cell walls (r = ?0.90, P<0.01; r = ?0.85, P<0.01, respectively). Proportions of lignified tissues were less closely correlated with all measurements of digestibility than were proportions of lignin in cell walls determined chemically. Development of a technique for measuring intensity of lignification might enhance the value of light-microscopy measurements. 相似文献
3.
Guang-Long Wang Ying Huang Xin-Yue Zhang Zhi-Sheng Xu Feng Wang Ai-Sheng Xiong 《Plant cell reports》2016,35(8):1743-1755
Key message
Carrot root development associates lignin deposition and regulation.Abstract
Carrot is consumed worldwide and is a good source of nutrients. However, excess lignin deposition may reduce the taste and quality of carrot root. Molecular mechanisms underlying lignin accumulation in carrot are still lacking. To address this problem, we collected taproots of wild and cultivated carrots at five developmental stages and analyzed the lignin content and characterized the lignin distribution using histochemical staining and autofluorescence microscopy. Genes involved in lignin biosynthesis were identified, and their expression profiles were determined. Results showed that lignin was mostly deposited in xylem vessels of carrot root. In addition, lignin content continuously decreased during root development, which was achieved possibly by reducing the expression of the genes involved in lignin biosynthesis. Carrot root may also prevent cell lignification to meet the demands of taproot growth. Our results will serve as reference for lignin biosynthesis in carrot and may also assist biologists to improve carrot quality.4.
5.
Background
Recent discoveries highlighting the metabolic malleability of plant lignification indicate that lignin can be engineered to dramatically alter its composition and properties. Current plant biotechnology efforts are primarily aimed at manipulating the biosynthesis of normal monolignols, but in the future apoplastic targeting of phenolics from other metabolic pathways may provide new approaches for designing lignins that are less inhibitory toward the enzymatic hydrolysis of structural polysaccharides, both with and without biomass pretreatment. To identify promising new avenues for lignin bioengineering, we artificially lignified cell walls from maize cell suspensions with various combinations of normal monolignols (coniferyl and sinapyl alcohols) plus a variety of phenolic monolignol substitutes. Cell walls were then incubated in vitro with anaerobic rumen microflora to assess the potential impact of lignin modifications on the enzymatic degradability of fibrous crops used for ruminant livestock or biofuel production. 相似文献6.
7.
Martin Schmidt Pradeep Perera Adam M. Schwartzberg Paul D. Adams P. James Schuck 《Journal of visualized experiments : JoVE》2010,(45)
Meeting growing energy demands safely and efficiently is a pressing global challenge. Therefore, research into biofuels production that seeks to find cost-effective and sustainable solutions has become a topical and critical task. Lignocellulosic biomass is poised to become the primary source of biomass for the conversion to liquid biofuels1-6. However, the recalcitrance of these plant cell wall materials to cost-effective and efficient degradation presents a major impediment for their use in the production of biofuels and chemicals4. In particular, lignin, a complex and irregular poly-phenylpropanoid heteropolymer, becomes problematic to the postharvest deconstruction of lignocellulosic biomass. For example in biomass conversion for biofuels, it inhibits saccharification in processes aimed at producing simple sugars for fermentation7. The effective use of plant biomass for industrial purposes is in fact largely dependent on the extent to which the plant cell wall is lignified. The removal of lignin is a costly and limiting factor8 and lignin has therefore become a key plant breeding and genetic engineering target in order to improve cell wall conversion.Analytical tools that permit the accurate rapid characterization of lignification of plant cell walls become increasingly important for evaluating a large number of breeding populations. Extractive procedures for the isolation of native components such as lignin are inevitably destructive, bringing about significant chemical and structural modifications9-11. Analytical chemical in situ methods are thus invaluable tools for the compositional and structural characterization of lignocellulosic materials. Raman microscopy is a technique that relies on inelastic or Raman scattering of monochromatic light, like that from a laser, where the shift in energy of the laser photons is related to molecular vibrations and presents an intrinsic label-free molecular "fingerprint" of the sample. Raman microscopy can afford non-destructive and comparatively inexpensive measurements with minimal sample preparation, giving insights into chemical composition and molecular structure in a close to native state. Chemical imaging by confocal Raman microscopy has been previously used for the visualization of the spatial distribution of cellulose and lignin in wood cell walls12-14. Based on these earlier results, we have recently adopted this method to compare lignification in wild type and lignin-deficient transgenic Populus trichocarpa (black cottonwood) stem wood15. Analyzing the lignin Raman bands16,17 in the spectral region between 1,600 and 1,700 cm-1, lignin signal intensity and localization were mapped in situ. Our approach visualized differences in lignin content, localization, and chemical composition. Most recently, we demonstrated Raman imaging of cell wall polymers in Arabidopsis thaliana with lateral resolution that is sub-μm18. Here, this method is presented affording visualization of lignin in plant cell walls and comparison of lignification in different tissues, samples or species without staining or labeling of the tissues.Download video file.(47M, mov) 相似文献
8.
9.
Aims
Rice is a well-known silica-accumulating plant. The dumbbell-shaped silica bodies in the silica cells in rice leaf epidermis are formed via biosilicification, but the underlying mechanisms are largely unknown.Methods
Leaves at different developmental stages were collected to investigate silica cell differentiation by analyzing structures and silicon localization in the silica cells.Results
Exogenous silicon application increased both shoot and root biomass. When silicon was supplied, silica cells in the leaf epidermis developed gradually into a dumbbell-shape and became increasingly silicified as leaves aged. Silicon deposition in the silica cells was not completed until the leaf was fully expanded. Multiple lines of evidence suggest that lignification of silica cell walls precedes silicon deposition in the lumen of silica cells. The organized needle-like silica microstructures were formed by moulding the inner cell walls and filling up the lumen of the silica cell following leaf maturation.Conclusions
Two processes were involved in silicon deposition: (1) the silica cell wall was lignified and silicified, and then (2) the silicon was deposited gradually in silica cells as leaves aged. Silica body formation was not completed until the leaf was fully mature. 相似文献10.
Attack on Lignified Grass Cell Walls by a Facultatively Anaerobic Bacterium 总被引:6,自引:6,他引:0
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Danny E. Akin 《Applied microbiology》1980,40(4):809-820
A filamentous, facultatively anaerobic microorganism that attacked lignified tissue in forage grasses was isolated from rumen fluid with a Bermuda grass-containing anaerobic medium in roll tubes. The microbe, designated 7-1, demonstrated various colony and cellular morphologies under different growth conditions. Scanning electron microscopy revealed that 7-1 attacked lignified cell walls in aerobic and anaerobic culture. 7-1 predominately degraded tissues reacting positively for lignin with the chlorine-sulfite stain (i.e., sclerenchyma in leaf blades and parenchyma in stems) rather than the more resistant acid phloroglucinol-positive tissues (i.e., lignified vascular tissue and sclerenchyma ring in stems), although the latter tissues were occasionally attacked. Turbidimetric tests showed that 7-1 in anaerobic culture grew optimally at 39°C at a pH of 7.4 to 8.0. Tests for growth on plant cell wall carbohydrates showed that 7-1 grew on xylan and pectin slowly in aerobic cultures but not with pectin and only slightly with xylan in anaerobic culture. 7-1 was noncellulolytic as shown by filter paper tests. The microbe used the phenolic acids sinapic, ferulic, and p-coumaric acids as substrates for growth; the more highly methoxylated acids were used more effectively. 相似文献
11.
Jaime Barros Him K Shrestha Juan C Serrani-Yarce Nancy L Engle Paul E Abraham Timothy J Tschaplinski Robert L Hettich Richard A Dixon 《The Plant cell》2022,34(9):3339
Lignin biosynthesis begins with the deamination of phenylalanine and tyrosine (Tyr) as a key branch point between primary and secondary metabolism in land plants. Here, we used a systems biology approach to investigate the global metabolic responses to lignin pathway perturbations in the model grass Brachypodium distachyon. We identified the lignin biosynthetic protein families and found that ammonia-lyases (ALs) are among the most abundant proteins in lignifying tissues in grasses. Integrated metabolomic and proteomic data support a link between lignin biosynthesis and primary metabolism mediated by the ammonia released from ALs that is recycled for the synthesis of amino acids via glutamine. RNA interference knockdown of lignin genes confirmed that the route of the canonical pathway using shikimate ester intermediates is not essential for lignin formation in Brachypodium, and there is an alternative pathway from Tyr via sinapic acid for the synthesis of syringyl lignin involving yet uncharacterized enzymatic steps. Our findings support a model in which plant ALs play a central role in coordinating the allocation of carbon for lignin synthesis and the nitrogen available for plant growth. Collectively, these data also emphasize the value of integrative multiomic analyses to advance our understanding of plant metabolism.Ammonia-lyases play a key role in coordinating the allocation of carbon for lignin synthesis and nitrogen availability for plant growth. 相似文献
12.
Rebecca A. Smith Mathias Schuetz Melissa Roach Shawn D. Mansfield Brian Ellis Lacey Samuels 《The Plant cell》2013,25(10):3988-3999
Lignin is a critical structural component of plants, providing vascular integrity and mechanical strength. Lignin precursors (monolignols) must be exported to the extracellular matrix where random oxidative coupling produces a complex lignin polymer. The objectives of this study were twofold: to determine the timing of lignification with respect to programmed cell death and to test if nonlignifying xylary parenchyma cells can contribute to the lignification of tracheary elements and fibers. This study demonstrates that lignin deposition is not exclusively a postmortem event, but also occurs prior to programmed cell death. Radiolabeled monolignols were not detected in the cytoplasm or vacuoles of tracheary elements or neighbors. To experimentally define which cells in lignifying tissues contribute to lignification in intact plants, a microRNA against CINNAMOYL CoA-REDUCTASE1 driven by the promoter from CELLULOSE SYNTHASE7 (ProCESA7:miRNA CCR1) was used to silence monolignol biosynthesis specifically in cells developing lignified secondary cell walls. When monolignol biosynthesis in ProCESA7:miRNA CCR1 lines was silenced in the lignifying cells themselves, but not in the neighboring cells, lignin was still deposited in the xylem secondary cell walls. Surprisingly, a dramatic reduction in cell wall lignification of extraxylary fiber cells demonstrates that extraxylary fibers undergo cell autonomous lignification. 相似文献
13.
Background
Currently the major barrier in biomass utilization is the lack of an effective pretreatment of plant cell wall so that the carbohydrates can subsequently be hydrolyzed into sugars for fermentation into fuel or chemical molecules. Termites are highly effective in degrading lignocellulosics and thus can be used as model biological systems for studying plant cell wall degradation.Results
We discovered a combination of specific structural and compositional modification of the lignin framework and partial degradation of carbohydrates that occurs in softwood with physical chewing by the termite, Coptotermes formosanus, which are critical for efficient cell wall digestion. Comparative studies on the termite-chewed and native (control) softwood tissues at the same size were conducted with the aid of advanced analytical techniques such as pyrolysis gas chromatography mass spectrometry, attenuated total reflectance Fourier transform infrared spectroscopy and thermogravimetry. The results strongly suggest a significant increase in the softwood cellulose enzymatic digestibility after termite chewing, accompanied with utilization of holocellulosic counterparts and an increase in the hydrolysable capacity of lignin collectively. In other words, the termite mechanical chewing process combines with specific biological pretreatment on the lignin counterpart in the plant cell wall, resulting in increased enzymatic cellulose digestibility in vitro. The specific lignin unlocking mechanism at this chewing stage comprises mainly of the cleavage of specific bonds from the lignin network and the modification and redistribution of functional groups in the resulting chewed plant tissue, which better expose the carbohydrate within the plant cell wall. Moreover, cleavage of the bond between the holocellulosic network and lignin molecule during the chewing process results in much better exposure of the biomass carbohydrate.Conclusion
Collectively, these data indicate the participation of lignin-related enzyme(s) or polypeptide(s) and/or esterase(s), along with involvement of cellulases and hemicellulases in the chewing process of C. formosanus, resulting in an efficient pretreatment of biomass through a combination of mechanical and enzymatic processes. This pretreatment could be mimicked for industrial biomass conversion. 相似文献14.
Delphine Mnard Leonard Blaschek Konstantin Kriechbaum Cheng Choo Lee Henrik Serk Chuantao Zhu Alexander Lyubartsev Nuoendagula Zoltn Bacsik Lennart Bergstrm Aji Mathew Shinya Kajita Edouard Pesquet 《The Plant cell》2022,34(12):4877
The biopolymer lignin is deposited in the cell walls of vascular cells and is essential for long-distance water conduction and structural support in plants. Different vascular cell types contain distinct and conserved lignin chemistries, each with specific aromatic and aliphatic substitutions. Yet, the biological role of this conserved and specific lignin chemistry in each cell type remains unclear. Here, we investigated the roles of this lignin biochemical specificity for cellular functions by producing single cell analyses for three cell morphotypes of tracheary elements, which all allow sap conduction but differ in their morphology. We determined that specific lignin chemistries accumulate in each cell type. Moreover, lignin accumulated dynamically, increasing in quantity and changing in composition, to alter the cell wall biomechanics during cell maturation. For similar aromatic substitutions, residues with alcohol aliphatic functions increased stiffness whereas aldehydes increased flexibility of the cell wall. Modifying this lignin biochemical specificity and the sequence of its formation impaired the cell wall biomechanics of each morphotype and consequently hindered sap conduction and drought recovery. Together, our results demonstrate that each sap-conducting vascular cell type distinctly controls their lignin biochemistry to adjust their biomechanics and hydraulic properties to face developmental and environmental constraints.During the development of each vascular cell, specific lignin chemistries control their biomechanics and water conduction properties to face environmental changes.IN A NUTSHELL Background: Lignin comprises multiple cell wall–localized aromatic polymers that are essential for vascular plants to conduct water and strengthen their organs. It has long been thought that lignin was randomly and nonspecifically assembled to provide mechanical strengthening and waterproofing to cells by filling-up the empty spaces in the cell walls. However, the different cell types and morphotypes forming the different sap-conducting pipes and their cell wall layers (inner vs. outer layer) exhibit specific lignin chemistries that are conserved among plant species. We, therefore, investigated the function of these specific lignin chemistries at the cell and cell wall layer levels for the different sap-conducting pipes in plants. Question: What is the function of a specific lignin chemistry for the different plant sap-conducting pipe cells? Can changes in the lignin chemistry of sap-conducting cells affect their hydraulic capacity when facing environmental conditions such as drought? Findings: We answered these questions by changing lignin levels and composition, using drugs to block lignin formation, and/or genetic engineering to switch off genes, in three complementary systems: (1) isolated cells grown in test tubes that we can trigger to become sap conduits, (2) annual plants, and (3) hardwood trees. We show that lignin chemistry is specific to each cell morphotype and changes during cell maturation, modifying the amount of lignin, the chemical composition of lignin units, and the position of these units in the longer polymer. These specific lignin chemistries are required for the proper function of each cell morphotype to properly conduct the sap and strengthen plant organs. Modifying the amount, the composition, and the time when specific units with distinct chemistry are incorporated in lignin of each cell morphotype has dramatic effects, causing defects in sap conduit hydraulic and biomechanical properties. The ratio between the different chemical units of lignin needs to be fine-tuned to adjust plant sap conduction and mechanical strengthening. Thus, changes in the proportion of lignin units with distinct chemistries confer different hydraulic and mechanical properties enabling plants to better resist and/or recover from drought. We also revealed that increases in the proportion of lignin units with aldehyde modulate plant pipe hydraulic and mechanical properties. Next steps: We are now working to identify and understand the molecular mechanisms that control the formation of specific lignin chemistries in distinct sites and times during the development of the different cell wall layers in each cell type and morphotype. 相似文献
15.
16.
Malate metabolism by NADP-malic enzyme in plant defense 总被引:8,自引:0,他引:8
Paula Casati María F. Drincovich Gerald E. Edwards Carlos S. Andreo 《Photosynthesis research》1999,61(2):99-105
Malate is involved in various metabolic pathways, and there are several enzymes that metabolize it. One important malate metabolizing enzyme is NADP-malic enzyme (NADP-ME). NADP-ME functions in many different pathways in plants, having an important role in C4 photosynthesis where it releases the CO2 to be used in carbon fixation by Rubisco. Apart from this specialized role, NADP-ME is thought to fulfill diverse housekeeping functions because of its universal presence in different plant tissues. NADP-ME is induced after wounding or exposure to UV-B radiation. In this way, the enzyme is implicated in defense-related deposition of lignin by providing NADPH for the two NADPH-dependent reductive steps in monolignol biosynthesis. On the other hand, it can supply NADPH for flavonoid biosynthesis as many steps in the flavonoid biosynthesis pathway require reductive power. Pyruvate, another product of NADP-ME reaction, can be used for obtaining ATP through respiration in the mitochondria; and may serve as a precursor for synthesis of phosphoenolpyruvate (PEP). PEP is utilized in the shikimate pathway, leading to the synthesis of aromatic amino acids including phenylalanine, the common substrate for lignin and flavonoid synthesis. Moreover, NADP-ME can be involved in mechanisms producing NADPH for synthesis of activated oxygen species that are produced in order to kill or damage pathogens. In conclusion, an increase in the levels of NADP-ME could provide building blocks and energy for biosynthesis of defense compounds, suggesting a role of malate metabolism in plant defense. 相似文献
17.
Background and Aims
The functions of plant latex have been discussed for a long time. Today, many studies support a defence mechanism as being its main function. A role as a self-healing mechanism was never attributed to the coagulation of latex. In this study we quantified the contribution of the coagulation of Ficus benjamina (weeping fig) latex to a restoration of the mechanical properties of the bark after external lesions.Methods
Tensile tests of F. benjamina bark were conducted either immediately after injury or at various latency times after injury.Key Results
A significant increase in the tensile strength of bark samples until 30 min after injury was found, and this effect could be attributed to the coagulation of plant latex alone. The tensile strength remains nearly constant until several hours or days after injury. Then, very probably due to other mechanisms such as cell growth and cell proliferation, the tensile strength begins to increase slightly again.Conclusions
The coagulation of latex seals lesions and serves as a quick and effective pre-step of subsequent, more effective, long-lasting self-healing mechanisms such as cell growth and proliferation. Thus, a fast self-healing effect can be included in the list of functions of plant latex. 相似文献18.
The use is reported of Mirande's reagent in epifluorescence microscopy which permits a clear distinction between cellulosic and lignified tissues. Homogeneous Prespermatophytae and gymnosperm xylem appeared entirely green with Mirande's reagent under ultraviolet excitation, whereas heteroxyled angiosperm wood showed a mixed pink and blue–green colour. This coloration was due to the fluorescence of cellulose, since certain elements in dicotyledonous wood (parenchyma, fibres, xylem rays) are not entirely lignified. Monocotyledonous (Poaceae) lignin showed an intense blue fluorescence due to hydroxycinnamic acids bound to the cell wall.The method showed that lignification occurs first in the middle lamella, and later in the secondary wall of xylem cells. In addition, this staining technique proved useful in the study of lignin and suberin deposition in response to various stress factors. 相似文献
19.