Lignin Precursors Research Articles

CO2 capture in lignin-derived and nitrogen-doped hierarchical porous carbons

Nitrogen-doped and hierarchical porous carbons were synthesized from lignin precursor by KOH and NH3 activation with BET surface area of 1631–2922 m2/g. X-ray photoelectron spectroscopy (XPS) revealed 5.6–7.1 at.% nitrogen with predominantly pyridinic, amino and pyrrolic/pyridonic type of functionality. Maximum CO2 adsorption capacity at 298 K and 273 K and pressure upto 760 torr (1 bar) was found to be 5.48 and 8.6 mmol/g respectively, which is higher than that of majority of nitrogen-doped carbons, reported in literature. Ideal Adsorbed Solution Theory (IAST)-based selectivity for CO2/N2 revealed that selectivity increases with the increase in nitrogen content. IAST calculations were also employed to calculate the binary adsorption isotherms to simulate mixed gas adsorption for CO2 and N2 as a function of total pressure and mole fraction of CO2 in the gas mixture. Continuous CO2 adsorption-desorption studies for 10 cycles revealed that all the nitrogen-doped carbons retained its working capacity at the end of the cycles. Finally, a breakthrough study for (10/90) CO2/N2 mixture in a dynamic column experiment confirmed a physical separation of the two components. The overall results suggest that nitrogen-doped carbons from lignin can be used as potential adsorbents for CO2 capture purposes.

A 21 000-year record of fluorescent organic matter markers in the WAIS Divide ice core

Abstract. Englacial ice contains a significant reservoir of organic material (OM), preserving a chronological record of materials from Earth's past. Here, we investigate if OM composition surveys in ice core research can provide paleoecological information on the dynamic nature of our Earth through time. Temporal trends in OM composition from the early Holocene extending back to the Last Glacial Maximum (LGM) of the West Antarctic Ice Sheet Divide (WD) ice core were measured by fluorescence spectroscopy. Multivariate parallel factor (PARAFAC) analysis is widely used to isolate the chemical components that best describe the observed variation across three-dimensional fluorescence spectroscopy (excitation–emission matrices; EEMs) assays. Fluorescent OM markers identified by PARAFAC modeling of the EEMs from the LGM (27.0–18.0 kyr BP; before present 1950) through the last deglaciation (LD; 18.0–11.5 kyr BP), to the mid-Holocene (11.5–6.0 kyr BP) provided evidence of different types of fluorescent OM composition and origin in the WD ice core over 21.0 kyr. Low excitation–emission wavelength fluorescent PARAFAC component one (C1), associated with chemical species similar to simple lignin phenols was the greatest contributor throughout the ice core, suggesting a strong signature of terrestrial OM in all climate periods. The component two (C2) OM marker, encompassed distinct variability in the ice core describing chemical species similar to tannin- and phenylalanine-like material. Component three (C3), associated with humic-like terrestrial material further resistant to biodegradation, was only characteristic of the Holocene, suggesting that more complex organic polymers such as lignins or tannins may be an ecological marker of warmer climates. We suggest that fluorescent OM markers observed during the LGM were the result of greater continental dust loading of lignin precursor (monolignol) material in a drier climate, with lower marine influences when sea ice extent was higher and continents had more expansive tundra cover. As the climate warmed, the record of OM markers in the WD ice core changed, reflecting shifts in carbon productivity as a result of global ecosystem response.

Gene Duplication Leads to Altered Membrane Topology of a Cytochrome P450 Enzyme in Seed Plants.

Evolution of the phenolic metabolism was critical for the transition of plants from water to land. A cytochrome P450, CYP73, with cinnamate 4-hydroxylase (C4H) activity, catalyzes the first plant-specific and rate-limiting step in this pathway. The CYP73 gene is absent from green algae, and first detected in bryophytes. A CYP73 duplication occurred in the ancestor of seed plants and was retained in Taxaceae and most angiosperms. In spite of a clear divergence in primary sequence, both paralogs can fulfill comparable cinnamate hydroxylase roles both in vitro and in vivo. One of them seems dedicated to the biosynthesis of lignin precursors. Its N-terminus forms a single membrane spanning helix and its properties and length are highly constrained. The second is characterized by an elongated and variable N-terminus, reminiscent of ancestral CYP73s. Using as proxies the Brachypodium distachyon proteins, we show that the elongation of the N-terminus does not result in an altered subcellular localization, but in a distinct membrane topology. Insertion in the membrane of endoplasmic reticulum via a double-spanning open hairpin structure allows reorientation to the lumen of the catalytic domain of the protein. In agreement with participation to a different functional unit and supramolecular organization, the protein displays modified heme proximal surface. These data suggest the evolution of divergent C4H enzymes feeding different branches of the phenolic network in seed plants. It shows that specialization required for retention of gene duplicates may result from altered protein topology rather than change in enzyme activity.

Lignin accumulation and biosynthetic enzyme activities in relation to postharvest firmness of fresh waxy corn

Lignification and changes in the enzymes involved in lignin biosynthesis were investigated in relation to the reduced tenderness occurring during the postharvest storage of fresh waxy corn at 20 °C. The firmness of the corn and its lignin and polyphenol contents increased as storage time increased. The lignin content was significantly correlated (p .05) effect of polyphenol oxidase or cinnamyl alcohol dehydrogenase (CAD) on the lignin content was observed. Practical applications Tenderness is a main factor in sensory quality of fresh waxy corn. Secondary cell wall lignifications induced by phenylpropanoid pathway is the main reason of tenderness decline for postharvest fresh waxy corn. But little information of lignin accumulation of fresh waxy corn is obtained from the present studies. Analyzing the changes of lignin and related precursor materials, enzyme activities will help to comprehensively understand the regulatory mechanism of postharvest lignification of fresh waxy corn, which can provide suitable methods to delay postharvest lignin accumulation, and maintain the eating quality of fresh waxy corn.

Defining the Diverse Cell Populations Contributing to Lignification in Arabidopsis Stems

Many land plants evolved tall and sturdy growth habits due to specialized cells with thick lignified cell walls: tracheary elements that function in water transport and fibers that function in structural support. The objective of this study was to define how and when diverse cell populations contribute lignin precursors, monolignols, to secondary cell walls during lignification of the Arabidopsis (Arabidopsis thaliana) inflorescence stem. Previous work demonstrated that, when lignin biosynthesis is suppressed in fiber and tracheary element cells with thickened walls, fibers become lignin-depleted while vascular bundles still lignify, suggesting that nonlignifying neighboring xylem cells are contributing to lignification. In this work, we dissect the contributions of different cell types, specifically xylary parenchyma and fiber cells, to lignification of the stem using cell-type-specific promoters to either knock down an essential monolignol biosynthetic gene or to introduce novel monolignol conjugates. Analysis of either reductions in lignin in knockdown lines, or the addition of novel monolignol conjugates, directly identifies the xylary parenchyma and fiber cell populations that contribute to the stem lignification and the developmental timing at which each contribution is most important.

A “Double Click” for Illuminating Plant Cell Walls

In this issue of Cell Chemical Biology, Lion etal. (2017) report a multiplexed labeling method to visualize plant cell wall lignification invivo. This approach uses two different lignin precursor analogs tagged with azide and alkyne reporters that can be independently incorporated into cell walls, then differentially derivatized invivo via two bioorthogonal click reactions: strain-promoted and copper-assisted azide-alkyne cycloadditions.

Lignin-based polymeric surfactants for emulsion polymerization

A non-ionic surfactant system is synthesized by standard grafting of poly(ethylene oxide) from renewable lignin fragments and used for the emulsion polymerization of styrene. The lignin precursors are formed by hydrogenolysis and utilized as initiator for the oxyanionic polymerization of ethylene oxide leading to amphiphilic polymers, very similar to standard nonionic surfactant synthesis. Subsequently, the formed amphiphilic polymers are employed as stabilizers in the heterophase polymerization of styrene with various initiators. Poly (styrene) latexes with solids contents of up to 21% depending on stabilizer concentration have been obtained. Stabilizer efficiencies and performances were nicely comparable with those of nonylphenol-based, non-ionic industrial performance surfactants.

Effect of compressed liquid CO2 antisolvent treatment on the synthesis of hierarchically porous nanocarbon from kraft lignin

Carbon nanoparticles with hierarchical micro/mesoporous structure were developed from a commercial kraft lignin using a facile, one-pot green technology of a compressed liquid CO2 antisolvent treatment, followed by thermostablization and carbonization processes. The as-synthesized carbon nanoparticles were systematically characterized by investigating the surface properties of morphology, particle size, and chemical states with FESEM, HRTEM and XPS, respectively; surface areas and porous structures with BET analyzer; crystalline state with XRD and Raman spectroscopy; chemical features with FTIR; and elemental compositions with an elemental analyzer. The results indicated that the carbon nanoparticles fabricated from the smallest quasi-spherical lignin nanoparticles contained high nanocrystalline and amorphous carbons levels. The enhanced phenolic, carbonyl, and carboxylic functional groups on the particle surface promoted the dispersibility of the particles in deionized water. The size and morphology of the lignin precursor particles significantly affected the fabrication of the carbon nanoparticles by carbonization. The experiment suggested that such dispersible and porous carbon nanoparticles could be applicable in the fields of energy, biotechnology, and environmental pollution control.

The degradation of coniferyl alcohol and the complementary production of chlorogenic acids in the growth culture of Streptomyces albogriseolus KF977548 isolated from decaying wood residues

Abstract Coniferyl alcohol is one of the major precursors of lignin; the most abundant aromatic compound and a natural resource currently receiving attention because of the value-added metabolites resulting from its degradation. Growth study of Streptomyces albogriseolus KF977548 (strain AOB) isolated from decaying wood residues in a tropical estuarine ecosystem was carried out using coniferyl alcohol as a sole carbon source. Cell growth and metabolite production were monitored at 24 h interval by dry weight measurements and HPLC, LC–MS-DAD analyses. Biochemical and PCR assays were carried out to detect the major catabolic enzymes of interest. Strain AOB utilized coniferyl alcohol completely within 72 h (μ = 0.204 h −1 , T d = 3.4 h). Laccase and peroxidase were released into the growth medium up to 0.099 and 98 μmol/mL respectively. Protocatechuate 3, 4-dioxygenase and demethylase were detected in the genome whilst ortho -adipate pathway was clearly indicated. Growth on coniferyl alcohol or caffeic acid as mono substrates resulted in the production of secondary metabolites identified by HPLC–MS as 1-caffeoylquinic and 3,4,5-tricaffeoylquinic acids, known as chlorogenic acids, in the culture medium. The microbial production of chlorogenic acids from a lignin-related substrate base by strain AOB could arouse a plausible biotechnological process.

MdMyb93 is a regulator of suberin deposition in russeted apple fruit skins.

A comparison of the transcriptomes of russeted vs nonrusseted apple skins previously highlighted a tight relationship between a gene encoding an MYB-type transcription factor, MdMYB93, and some key suberin biosynthetic genes. The present work assesses the role of this transcription factor in the suberization process. A phylogenetic analysis of MdMYB93 and Arabidopsis thaliana MYBs was performed and the function of MdMYB93 was further investigated using Agrobacterium-mediated transient overexpression in Nicotiana benthamiana leaves. An RNA-Seq analysis was performed to highlight the MdMYB93-regulated genes. Ultraperformance liquid chromatography-triple time-of-flight (UPLC-TripleTOF) and GC-MS were used to investigate alterations in phenylpropanoid, soluble-free lipid and lipid polyester contents. A massive accumulation of suberin and its biosynthetic precursors in MdMYB93 agroinfiltrated leaves was accompanied by a remobilization of phenylpropanoids and an increased amount of lignin precursors. Gene expression profiling displayed a concomitant alteration of lipid and phenylpropanoid metabolism, cell wall development, and extracellular transport, with a large number of induced transcripts predicted to be involved in suberin deposition. The present work supports a major role of MdMYB93 in the regulation of suberin deposition in russeted apple skins, from the synthesis of monomeric precursors, their transport, polymerization, and final deposition as suberin in primary cell wall.

Investigating Biochemical and Developmental Dependencies of Lignification with a Click-Compatible Monolignol Analog in Arabidopsis thaliana Stems.

Lignin is a key structural component of plant cell walls that provides rigidity, strength, and resistance against microbial attacks. This hydrophobic polymer also serves a crucial role in water transport. Despite its abundance and essential functions, several aspects of lignin biosynthesis and deposition remain cryptic. Lignin precursors are known to be synthesized in the cytoplasm by complex biosynthetic pathways, after which they are transported to the apoplastic space, where they are polymerized via free radical coupling reactions into polymeric lignin. However, the lignin deposition process and the factors controlling it are unclear. In this study, the biochemical and developmental dependencies of lignification were investigated using a click-compatible monolignol analog, 3-O-propargylcaffeyl alcohol (3-OPC), which can incorporate into both in vitro polymerized lignin and Arabidopsis thaliana tissues. Fluorescence labeling of 3-OPC using click chemistry followed by confocal fluorescence microscopy enabled the detection and imaging of 3-OPC incorporation patterns. These patterns were consistent with endogenous lignification observed in different developmental stages of Arabidopsis stems. However, the concentration of supplied monolignols influenced where lignification occurred at the subcellular level, with low concentrations being deposited in cell corners and middle lamellae and high concentrations also being deposited in secondary walls. Experimental inhibition of multiple lignification factors confirmed that 3-OPC incorporation proceeds via a free radical coupling mechanism involving peroxidases/laccases and reactive oxygen species (ROS). Finally, the presence of peroxide-producing enzymes determined which cell walls lignified: adding exogenous peroxide and peroxidase caused cells that do not naturally lignify in Arabidopsis stems to lignify. In summary, 3-OPC accurately mimics natural lignification patterns in different developmental stages of Arabidopsis stems and allows for the dissection of key biochemical and enzymatic factors controlling lignification.

Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM.

To clarify the role of coniferin in planta, semi-quantitative cellular distribution of coniferin in quick-frozen Ginkgo biloba L. (ginkgo) was visualized by cryo time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) analysis. The amount and rough distribution of coniferin were confirmed through quantitative chromatography measurement using serial tangential sections of the freeze-fixed ginkgo stem. The lignification stage of the sample was estimated using microscopic observations. Coniferin distribution visualized at the transverse and radial surfaces of freeze-fixed ginkgo stem suggested that coniferin is stored in the vacuoles, and showed good agreement with the assimilation timing of coniferin to lignin in differentiating xylem. Consequently, it is suggested that coniferin is stored in the tracheid cells of differentiating xylem and is a lignin precursor.

Synthesis and Characterization of Biodegradable Lignin Nanoparticles with Tunable Surface Properties.

Lignin nanoparticles can serve as biodegradable carriers of biocidal actives with minimal environmental footprint. Here we describe the colloidal synthesis and interfacial design of nanoparticles with tunable surface properties using two different lignin precursors, Kraft (Indulin AT) lignin and Organosolv (high-purity lignin). The green synthesis process is based on flash precipitation of dissolved lignin polymer, which enabled the formation of nanoparticles in the size range of 45-250 nm. The size evolution of the two types of lignin particles is fitted on the basis of modified diffusive growth kinetics and mass balance dependencies. The surface properties of the nanoparticles are fine-tuned by coating them with a cationic polyelectrolyte, poly(diallyldimethylammonium chloride). We analyze how the colloidal stability and dispersion properties of these two types of nanoparticles vary as a function of pH and salinities. The data show that the properties of the nanoparticles are governed by the type of lignin used and the presence of polyelectrolyte surface coating. The coating allows the control of the nanoparticles' surface charge and the extension of their stability into strongly basic regimes, facilitating their potential application at extreme pH conditions.

Differential Responses of Vanilla Accessions to Root Rot and Colonization by Fusarium oxysporum f. sp. radicis-vanillae.

Root and stem rot (RSR) disease caused by Fusarium oxysporum f. sp. radicis-vanillae (Forv) is the most damaging disease of vanilla (Vanilla planifolia and V. × tahitensis, Orchidaceae). Breeding programs aimed at developing resistant vanilla varieties are hampered by the scarcity of sources of resistance to RSR and insufficient knowledge about the histopathology of Forv. In this work we have (i) identified new genetic resources resistant to RSR including V. planifolia inbreds and vanilla relatives, (ii) thoroughly described the colonization pattern of Forv into selected vanilla accessions, confirming its necrotic non-vascular behavior in roots, and (iii) evidenced the key role played by hypodermis, and particularly lignin deposition onto hypodermal cell walls, for resistance to Forv in two highly resistant vanilla accessions. Two hundred and fifty-four vanilla accessions were evaluated in the field under natural conditions of infection and in controlled conditions using in vitro plants root-dip inoculated by the highly pathogenic isolate Fo072. For the 26 accessions evaluated in both conditions, a high correlation was observed between field evaluation and in vitro assay. The root infection process and plant response of one susceptible and two resistant accessions challenged with Fo072 were studied using wide field and multiphoton microscopy. In susceptible V. planifolia, hyphae penetrated directly into the rhizodermis in the hairy root region then invaded the cortex through the passage cells where it induced plasmolysis, but never reached the vascular region. In the case of the resistant accessions, the penetration was stopped at the hypodermal layer. Anatomical and histochemical observations coupled with spectral analysis of the hypodermis suggested the role of lignin deposition in the resistance to Forv. The thickness of lignin constitutively deposited onto outer cell walls of hypodermis was highly correlated with the level of resistance for 21 accessions tested. The accumulation of p-coumaric and sinapic acids, two phenolic precursors of lignin, was observed in the resistant plants inoculated with Fo072, but not in the susceptible one. Altogether, our analyses enlightened the mechanisms at work in RSR resistant genotypes and should enhance the development of novel breeding strategies aimed at improving the genetic control of RSR of vanilla.

Engineering a Monolignol 4-O-Methyltransferase with High Selectivity for the Condensed Lignin Precursor Coniferyl Alcohol

Lignin, a rigid biopolymer in plant cell walls, is derived from the oxidative polymerization of three monolignols. The composition of monolignol monomers dictates the degree of lignin condensation, reactivity, and thus the degradability of plant cell walls. Guaiacyl lignin is regarded as the condensed structural unit. Polymerization of lignin is initiated through the deprotonation of the para-hydroxyl group of monolignols. Therefore, preferentially modifying the para-hydroxyl of a specific monolignol to deprive its dehydrogenation propensity would disturb the formation of particular lignin subunits. Here, we test the hypothesis that specific remodeling the active site of a monolignol 4-O-methyltransferase would create an enzyme that specifically methylates the condensed guaiacyl lignin precursor coniferyl alcohol. Combining crystal structural information with combinatorial active site saturation mutagenesis and starting with the engineered promiscuous enzyme, MOMT5 (T133L/E165I/F175I/F166W/H169F), we incrementally remodeled its substrate binding pocket by the addition of four substitutions, i.e. M26H, S30R, V33S, and T319M, yielding a mutant enzyme capable of discriminately etherifying the para-hydroxyl of coniferyl alcohol even in the presence of excess sinapyl alcohol. The engineered enzyme variant has a substantially reduced substrate binding pocket that imposes a clear steric hindrance thereby excluding bulkier lignin precursors. The resulting enzyme variant represents an excellent candidate for modulating lignin composition and/or structure in planta.

Coniferin β-glucosidase is ionically bound to cell wall in differentiating xylem of poplar

Lignin is crucial for the growth and persistence of vascular plants in diverse environments. Although biosynthesis and polymerization of lignin precursors have been subjected to in-depth studies, only limited information exists on the ways in which these precursors are transported. Recent research has demonstrated that V-ATPase-dependent transport of coniferyl alcohol glucoside (coniferin) is a major metabolic function in differentiating xylem of both angiosperms and gymnosperms. Localization of β-glucosidase involved in the cleavage of coniferin to coniferyl alcohol and glucose is important to clarify the role of coniferin in lignification; nevertheless, only little has been reported in angiosperms. Here, we demonstrated the presence of coniferin β-glucosidase activity in a cell wall ionically bound protein fraction extracted from differentiating xylem of poplar. Coniferin β-glucosidase localization is very likely in cell wall similar to previous reports in conifers. We identified a putative poplar coniferin β-glucosidase through phylogenetic analysis and named this protein “PtrBGL6”. Immunoprecipitation assays showed that the anti-PtrBGL6 antibody recognizes a coniferin β-glucosidase in a cell wall ionically bound protein fraction. Conserved coniferin β-glucosidases localized in cell wall in both angiosperm and gymnosperm implies their important roles in the formation of lignified cell wall.

Ancient origin of the biosynthesis of lignin precursors.

BackgroundLignin plays an important role in plant structural support and water transport, and is considered one of the hallmarks of land plants. The recent discovery of lignin or its precursors in various algae has raised questions on the evolution of its biosynthetic pathway, which could be much more ancient than previously thought. To determine the taxonomic distribution of the lignin biosynthesis genes, we screened all publicly available genomes of algae and their closest non-photosynthetic relatives, as well as representative land plants. We also performed phylogenetic analysis of these genes to decipher the evolution and origin(s) of lignin biosynthesis.ResultsEnzymes involved in making p-coumaryl alcohol, the simplest lignin monomer, are found in a variety of photosynthetic eukaryotes, including diatoms, dinoflagellates, haptophytes, cryptophytes as well as green and red algae. Phylogenetic analysis of these enzymes suggests that they are ancient and spread to some secondarily photosynthetic lineages when they acquired red and/or green algal endosymbionts. In some cases, one or more of these enzymes was likely acquired through lateral gene transfer (LGT) from bacteria.ConclusionsGenes associated with p-coumaryl alcohol biosynthesis are likely to have evolved long before the transition of photosynthetic eukaryotes to land. The original function of this lignin precursor is therefore unlikely to have been related to water transport. We suggest that it participates in the biological defense of some unicellular and multicellular algae.ReviewersThis article was reviewed by Mark Ragan, Uri Gophna, Philippe Deschamps.

Chlorogenic Acid Biosynthesis Appears Linked with Suberin Production in Potato Tuber (Solanum tuberosum).

Potato (Solanum tuberosum L.) is a good source of dietary antioxidants. Chlorogenic acid (CGA) and caffeic acid (CA) are the most abundant phenolic acid antioxidants in potato and are formed by the phenylpropanoid pathway. A number of CGA biosynthetic routes that involve hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT) and/or hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT) have been proposed, but little is known about their path in potato. CA production requires a caffeoyl shikimate esterase (CSE), and CA serves as a substrate of lignin precursor ferulic acid via the action of caffeic/5-hydroxyferulic acid O-methyltransferase (COMTI). CGA is precursor of caffeoyl-CoA and, via caffeoyl-CoA O-methyltransferase (CCoAOMT), of feruloyl-CoA. Feruloyl-CoA is required for lignin and suberin biosynthesis, crucial for tuber development. Here, metabolite and transcript levels of the mentioned and related enzymes, such as cinnamate 4-hydroxylase (C4H), were determined in the flesh and skin of fresh and stored tubers. Metabolite and transcript levels were higher in skin than in flesh, irrespective of storage. CGA and CA production appear to occur via p-coumaroyl-CoA, using HQT and CSE, respectively. HCT is likely involved in CGA remobilization toward suberin. The strong correlation between CGA and CA, the correspondence with C4H, HQT, CCoAOMT2, and CSE, and the negative correlation of HCT and COMTI in potato tubers suggest a major flux toward suberin.

Polyurethane Coatings Based on Chemically Unmodified Fractionated Lignin

New lignin-based thermoset polyurethane (PU) coatings with high lignin content are presented in this work. These materials were obtained by extracting a sol fraction from a raw kraft lignin sample with the bioderived solvent 2-methyltetrahydrofuran (MeTHF), and subsequently directly cross-linking the MeTHF-soluble lignin fraction with a toluene diisocyanate (TDI)-based polyisocyanate at different weight ratios. A thorough characterization of the PU materials highlighted their improved thermal stability, better film forming ability and higher hydrophobic character compared with the un-cross-linked lignin precursor. Additionally, force–distance curve measurements by atomic force microscopy were employed to determine the elastic modulus of the PU materials. Finally, the lignin-based PU materials were found to exhibit high adhesion on different substrates, including glass, wood and metals. The results of this study demonstrate that the direct reaction of chemically unmodified fractionated lignin with polyisoc...

A versatile click-compatible monolignol probe to study lignin deposition in plant cell walls.

Lignin plays important structural and functional roles in plants by forming a hydrophobic matrix in secondary cell walls that enhances mechanical strength and resists microbial decay. While the importance of the lignin matrix is well documented and the biosynthetic pathways for monolignols are known, the process by which lignin precursors or monolignols are transported and polymerized to form this matrix remains a subject of considerable debate. In this study, we have synthesized and tested an analog of coniferyl alcohol that has been modified to contain an ethynyl group at the C-3 position. This modification enables fluorescent tagging and imaging of this molecule after its incorporation into plant tissue by click chemistry-assisted covalent labeling with a fluorescent azide dye, and confers a distinct Raman signature that could be used for Raman imaging. We found that this monolignol analog is incorporated into in vitro-polymerized dehydrogenation polymer (DHP) lignin and into root epidermal cell walls of 4-day-old Arabidopsis seedlings. Incorporation of the analog in stem sections of 6-week-old Arabidopsis thaliana plants and labeling with an Alexa-594 azide dye revealed the precise locations of new lignin polymerization. Results from this study indicate that this molecule can provide high-resolution localization of lignification during plant cell wall maturation and lignin matrix assembly.