Syringyl Monomers Research Articles

Thinning shelter forest reduced the soil priming effect and soil microbial respiration by altering the contribution of soil organic carbon from different sources

Thinning affects the understorey microclimate and the material input into the shelter forest soil, and it alters the contribution of the plant-derived and microbial-derived carbon (C). This difference effects the utilization of C by soil microbes and ultimately soil C emission. There is still no adequate characterization for the contribution of plant-derived and microbial-derived C to soil organic C (SOC) under thinning. Moreover, how this difference affects microbially driven C emissions (soil microbial respiration, SMR) and the contribution to the soil priming effect in this process are still unknown. The contribution of plant-derived and microbial-derived C to SOC were quantified by comparing lignin phenols and amino sugars under thinning and control treatments. Meanwhile, a 30-day indoor cultivation experiment was conducted. Soil priming effect and SMR were tested by adding 13C-labelled glucose to the soil of the thinning and control treatments. The purpose was to investigate the contribution of C from different sources to SOC, as well as its influence mechanism on soil priming effect and SMR.The results indicate that thinning significantly increased the fungal-derived (10.19 and 7.34 g kg−1) and microbial-derived C (11.05 and 8.19 g kg−1) of Quercus acutissima Carruth (QAC) and Pinus thunbergii Parl (PTP). However, the bacterial-derived C (0.86 and 0.85 g kg−1) decreased. Compared to the control treatment, the plant-derived, fungal-derived and microbial-derived C increased by 6.6 % and 1.1 %, 17.9 % and 12.2 %, and 16.6 % and 10.1 %, respectively, while bacterial-derived C decreased by 1.3 % and 2.1 %. Thinning reduced the soil priming effect of QAC by lowering the pH and increasing F/B and fungal-derived C. At the same time, this phenomenon was not evident in PTP, mainly attributed to the content of SOC and vanillyl and syringyl monomers. Thinning reduced the SMR, which was mainly related to the suppression of soil priming effect. In summary, this study has demonstrated that thinning reduced the soil priming effect and SMR of shelter forests by altering the contribution of plant-derived and microbial-derived C to SOC.

Highly catalytic supramolecular host–guest complex for high value directional conversion of lignin to syringyl monomer

In order to meet the challenge of enzyme catalysis of waste lignin, laccase (LAC)- guaiacyl(G)-type monomers noncovalent supramolecular system (LGS) were constructed for conversion of lignin. In this contribution, the catalytic effect of LGS formed by LAC and G-type monomers was studied. LAC changes the secondary structure conformation of its binding site to accommodate the G-type monomer, which is bound by hydrogen bonding and hydrophobic interactions. A mechanistic study highlights that the non-covalent complexation accelerates the internal electron transfer rate of LGS and syringol substrate for subsequent coupling reactions. In the presence of guaiacol/4-ethylguaiacol/vanillin-LAC, the conversion of dealkali lignin were 16.44, 29.12 and 22.72, respectively, higher than that in the presence of LAC alone. And the product of syringyl monomer was significantly increased in the actual lignin catalysis. Our work explains the mechanisms underlying existing enzyme-substrate interactions and enhanced catalytic system can be used for efficient utilization of waste.

A tailored fast thioacidolysis method incorporating multi-reaction monitoring mode of GC-MS for higher sensitivity on lignin monomer quantification

Abstract Thioacidolysis is widely used for lignin structural characterization by cleaving β-aryl ethers to release syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H) monomers followed by GC analysis. However, the traditional thioacidolysis method requires tedious extraction steps with chlorinated solvent underlying harmful to health, limiting its efficiency and application. Herein, an improved thioacidolysis method with high sensitivity for the quantitation of lignin-derived monomers was developed. The improved protocol used a quick, streamlined procedure to recover the monomeric products using ethyl acetate as extracting solvent and MS detector in multi-reaction monitoring mode to enhance its ability to detect extremely low concentration (0.1 ppb with signal-to-noise higher than 2) of monomeric products. Additionally, a fast GC program was established to speed up the GC quantitation. Several representative lignocellulose samples, including gymnosperm, angiosperm, and poaceae, were used to test this tailored method. The results demonstrated that the ratios of lignin monomer compositions determined by this method were consistent with that of traditional procedure despite the slightly higher monomer yields measured. More importantly, this method uses non-chlorinated solvent for microscale extraction and requires no evaporation step for workup, which is a green and efficient way for the quantification of lignin monomer compositions.

Lignin properties and cell wall response to deconstruction by alkaline pretreatment and enzymatic hydrolysis in brown midrib sorghums

Lignin has an adverse impact on the deconstruction of plant cell wall biopolymers in biorefining processes and its reduction and/or alteration during biosynthesis is one target for decreasing plant cell wall recalcitrance. In this work, the impact of two brown midrib mutations (bmr6 and bmr12) in two sorghum background lines (the commercial hybrid Atlas and near-isogenic BTx623) on lignin properties and the plants’ response to cell wall deconstruction to monomeric sugars via alkaline pretreatment and enzymatic hydrolysis is investigated with the goal of assessing how differences in lignin content and properties impact the plant’s response to pretreatment. We identify that both bmr sorghum lines show significantly lower abundance of water-extractable sugars (glucose, sucrose, and fructose) and alkali-saponifiable p-coumarate. Furthermore, both these properties exhibited identical trends across both background lines. Next, both untreated and mild alkali-pretreated bmr sorghums were shown to exhibit higher glucose hydrolysis yields following enzymatic hydrolysis than the control lines. Following pretreatment, the Atlas bmr sorghums exhibited more lignin solubilization and the solubilized lignin was of lower molar mass than the background control line suggesting that differences in the lignin response to pretreatment resulted these differences. Finally, significant differences were observed in the lignin content, lignin monomer distribution, and inter-unit linkages in the Atlas bmr line relative to the control line with key differences including lower syringyl monomer content in both bmr lines, higher relative abundance of β-O-4 linkages in the bmr6 line, and the presence of 5-hydroxy guaiacyl monomers and benzodioxane (α-O-5/β-O-4) linkages in the bmr12 line.

Survey of wheat straw stem characteristics for enhanced resistance to lodging

Lodging is one of the major constraints that threaten crop productivity. Although the relationship between cell walls and straw strength has been well recognized, little relevant research has been done in wheat, particularly on the monomer composition and structural characteristics of cell wall polymers and the arrangement of vascular bundles. In this study, we systematically investigated cell wall- and straw-related traits in a range of wheat germplasm resources and culm mutants using a high-throughput platform for cell wall analysis. We found that varieties with higher breaking force exhibited higher levels of crystalline cellulose but fewer hemicellulose components than other varieties. The lignin content was not consistent with the breaking force; instead, the lignin monomer constitution might be important because a significantly higher proportion of p-hydroxyphenyl (H) and guaiacyl (G) but a lower proportion of syringyl (S) monomers of lignin was found in the higher breaking force group. The crystallinity detected by X-ray diffraction was positively correlated with breaking force, indicating that the physical/chemical properties of polysaccharides also deserve attention. In terms of anatomical characteristics, the varieties with higher breaking force had a lower number and area of smaller vascular bundles in the peripheral sclerenchyma than other varieties. These results, together with the finding of a highly significant correlation between stem breaking force and straw fresh weight, 2nd internode width, flag leaf width and SiO2 content, should provide systematic information for breeding for lodging resistance.

Regulation of lignin composition by nitrogen rate and density and its relationship with stem mechanical strength of wheat

Increasing wheat yield is one of the effective ways to guarantee food security. In the current field production, dense planting with reasonable nitrogen is the most direct and efficient technique to achieve high yield. However, lodging, caused by overly large population with improper nitrogen management, has been a major limiting factor of grain production. In the present study, we comprehensively evaluated the composition and distribution of lignin monomers in stems and its relationship with mechanical strength. The results showed that syringyl (S) monomers were the predominant lignin monomeric units with enhancing mechanical strength. The reduction in the content of S subunits and total lignin, and the increased proportion of p-hydroxyphenyl (H) subunits were the crucial factors of low breaking strength of stems at high planting density and nitrogen fertilizer consumption. Furthermore, we speculated that bioengineering transformation and cultivation measures could be used to modify lignin composition and achieve high yield and high efficiency of wheat.

Molecular insights into the inhibitory effect of nitrogen fertilization on manure decomposition

Nitrogen (N) input rapidly increases available N in natural and managed ecosystems, potentially altering key ecosystem processes like decomposition. The effect of N enrichment on decomposition rates may be affected by the chemical quality of organic matter (OM), and the extent of N increment. Manure decomposition is an important process in agricultural systems, releasing nutrients and contributing to carbon (C) cycling. However, its response to N fertilization is poorly understood. To help address this knowledge gap, we decomposed pig manure (PM) and chicken manure (CM) under two rates of N fertilization (N1, 75 kg N ha−1; N2, 112.5 kg N ha−1) in a cropland in northeast China. We used litterbags to determine the dynamics of manure decomposition, while monitoring changes in the molecular composition with solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. After one-year, the decomposition rate of PM was significantly greater than CM (0.516 vs. 0.483 year−1). Spectra of 13C NMR indicated that PM initially contained more O-alkyl C and di-O-alkyl C (representing cellulose). In contrast, the contents of alkyl C (representing lipids) and aromatic C (representing lignin) were less in PM than CM, such that PM was overall more easily degradable. There was no N rate effect on CM decomposition. However, the decomposition rate of PM was significantly lower under high N than low N (0.410 vs. 0.622 year−1), apparently related to suppressed degradation of O-alkyl C and di-O-alkyl C. This result was surprising, since N enrichment is generally expected to promote degradation of more labile compounds like cellulose. At the same time, the loss of syringyl monomer of lignin in PM was reduced by high N fertilization. Together, these results suggest that decreased losses of O-alkyl C and di-O-alkyl C may have resulted from physical association of cellulose with more resistant lignin compounds. Net N mineralization was observed from manure decomposition and was greater for CM than PM, and high N fertilization suppressed N release from PM. Overall, our findings suggest that high rate of N fertilization may slow the decomposition of otherwise labile manure, potentially promoting greater C retention in soils.

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.).

Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense, is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated) and infected (inoculated) seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi)] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD), growth (weight and height), and disease severity (DS). Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5–34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%.

Enzymatic activities for lignin monomer intermediates highlight the biosynthetic pathway of syringyl monomers in Robinia pseudoacacia.

Most of the known 4-coumarate:coenzyme A ligase (4CL) isoforms lack CoA-ligation activity for sinapic acid. Therefore, there is some doubt as to whether sinapic acid contributes to sinapyl alcohol biosynthesis. In this study, we characterized the enzyme activity of a protein mixture extracted from the developing xylem of Robinia pseudoacacia. The crude protein mixture contained at least two 4CLs with sinapic acid 4-CoA ligation activity. The crude enzyme preparation displayed negligible sinapaldehyde dehydrogenase activity, but showed ferulic acid 5-hydroxylation activity and 5-hydroxyferulic acid O-methyltransferase activity; these activities were retained in the presence of competitive substrates (coniferaldehyde and 5-hydroxyconiferaldehyde, respectively). 5-Hydroxyferulic acid and sinapic acid accumulated in the developing xylem of R. pseudoacacia, suggesting, in part at least, sinapic acid is a sinapyl alcohol precursor in this species.

Formaldehyde stabilization facilitates lignin monomer production during biomass depolymerization

Practical, high-yield lignin depolymerization methods could greatly increase biorefinery productivity and profitability. However, development of these methods is limited by the presence of interunit carbon-carbon bonds within native lignin, and further by formation of such linkages during lignin extraction. We report that adding formaldehyde during biomass pretreatment produces a soluble lignin fraction that can be converted to guaiacyl and syringyl monomers at near theoretical yields during subsequent hydrogenolysis (47 mole % of Klason lignin for beech and 78 mole % for a high-syringyl transgenic poplar). These yields were three to seven times those obtained without formaldehyde, which prevented lignin condensation by forming 1,3-dioxane structures with lignin side-chain hydroxyl groups. By depolymerizing cellulose, hemicelluloses, and lignin separately, monomer yields were between 76 and 90 mole % for these three major biomass fractions.

Manipulation of Guaiacyl and Syringyl Monomer Biosynthesis in an Arabidopsis Cinnamyl Alcohol Dehydrogenase Mutant Results in Atypical Lignin Biosynthesis and Modified Cell Wall Structure.

Modifying lignin composition and structure is a key strategy to increase plant cell wall digestibility for biofuel production. Disruption of the genes encoding both cinnamyl alcohol dehydrogenases (CADs), including CADC and CADD, in Arabidopsis thaliana results in the atypical incorporation of hydroxycinnamaldehydes into lignin. Another strategy to change lignin composition is downregulation or overexpression of ferulate 5-hydroxylase (F5H), which results in lignins enriched in guaiacyl or syringyl units, respectively. Here, we combined these approaches to generate plants enriched in coniferaldehyde-derived lignin units or lignins derived primarily from sinapaldehyde. The cadc cadd and ferulic acid hydroxylase1 (fah1) cadc cadd plants are similar in growth to wild-type plants even though their lignin compositions are drastically altered. In contrast, disruption of CAD in the F5H-overexpressing background results in dwarfism. The dwarfed phenotype observed in these plants does not appear to be related to collapsed xylem, a hallmark of many other lignin-deficient dwarf mutants. cadc cadd, fah1 cadc cadd, and cadd F5H-overexpressing plants have increased enzyme-catalyzed cell wall digestibility. Given that these CAD-deficient plants have similar total lignin contents and only differ in the amounts of hydroxycinnamaldehyde monomer incorporation, these results suggest that hydroxycinnamaldehyde content is a more important determinant of digestibility than lignin content.

Peroxidase 4 is involved in syringyl lignin formation in Arabidopsis thaliana

Syringyl lignins result from the oxidative polymerization of sinapyl alcohol in a reaction mediated by syringyl (basic) peroxidases. Several peroxidases have been identified in the genome of Arabidopsis thaliana as close homologues to ZePrx, the best characterized basic peroxidase so far, but none of these has been directly involved in lignification. We have used a knock-out mutant of AtPrx4, the closest homologue to ZePrx, to study the involvement of this basic peroxidase in the physiology of the plant under both long- and short-day light conditions. Our results suggest that AtPrx4 is involved in cell wall lignification, especially in syringyl monomer formation. The disruption of AtPrx4 causes a decrease in syringyl units proportion, but only when light conditions are optimal. Moreover, the effect of AtPrx4 disruption is age-dependent, and it is only significant when the elongation process of the stem has ceased and lignification becomes active. In conclusion, AtPrx4 emerges as a basic peroxidase regulated by day length with an important role in lignification.

Enhanced Lignin Monomer Production Caused by Cinnamic Acid and Its Hydroxylated Derivatives Inhibits Soybean Root Growth

Cinnamic acid and its hydroxylated derivatives (p-coumaric, caffeic, ferulic and sinapic acids) are known allelochemicals that affect the seed germination and root growth of many plant species. Recent studies have indicated that the reduction of root growth by these allelochemicals is associated with premature cell wall lignification. We hypothesized that an influx of these compounds into the phenylpropanoid pathway increases the lignin monomer content and reduces the root growth. To confirm this hypothesis, we evaluated the effects of cinnamic, p-coumaric, caffeic, ferulic and sinapic acids on soybean root growth, lignin and the composition of p-hydroxyphenyl (H), guaiacyl (G) and syringyl (S) monomers. To this end, three-day-old seedlings were cultivated in nutrient solution with or without allelochemical (or selective enzymatic inhibitors of the phenylpropanoid pathway) in a growth chamber for 24 h. In general, the results showed that 1) cinnamic, p-coumaric, caffeic and ferulic acids reduced root growth and increased lignin content; 2) cinnamic and p-coumaric acids increased p-hydroxyphenyl (H) monomer content, whereas p-coumaric, caffeic and ferulic acids increased guaiacyl (G) content, and sinapic acid increased sinapyl (S) content; 3) when applied in conjunction with piperonylic acid (PIP, an inhibitor of the cinnamate 4-hydroxylase, C4H), cinnamic acid reduced H, G and S contents; and 4) when applied in conjunction with 3,4-(methylenedioxy)cinnamic acid (MDCA, an inhibitor of the 4-coumarate:CoA ligase, 4CL), p-coumaric acid reduced H, G and S contents, whereas caffeic, ferulic and sinapic acids reduced G and S contents. These results confirm our hypothesis that exogenously applied allelochemicals are channeled into the phenylpropanoid pathway causing excessive production of lignin and its main monomers. By consequence, an enhanced stiffening of the cell wall restricts soybean root growth.

Chemical responses to modified lignin composition in tension wood of hybrid poplar (Populus tremula x Populus alba)

The effect of altering the expression level of the F5H gene was investigated in three wood tissues (normal, opposite and tension wood) in 1-year-old hybrid poplar clone 717 (Populus tremula × Populus alba L.), containing the F5H gene under the control of the C4H promoter. Elevated expression of the F5H gene in poplar has been previously reported to increase the percent syringyl content of lignin. The wild-type and three transgenic lines were inclined 45° for 3 months to induce tension wood formation. Tension and opposite wood from inclined trees, along with normal wood from control trees, were analyzed separately for carbohydrates, lignin, cellulose crystallinity and microfibril angle (MFA). In the wild-type poplar, the lignin in tension wood contained a significantly higher percentage of syringyl than normal wood or opposite wood. However, there was no significant difference in the percent syringyl content of the three wood types within each of the transgenic lines. Increasing the F5H gene expression caused an increase in the percent syringyl content and a slight decrease in the total lignin in normal wood. In tension wood, the addition of a gelatinous layer in the fiber walls resulted in a consistently lower percentage of total lignin in the tissue. Acid-soluble lignin was observed to increase by up to 2.3-fold in the transgenic lines. Compared with normal wood and opposite wood, cell wall crystallinity in tension wood was higher and the MFA was smaller, as expected, with no evidence of an effect from modifying the syringyl monomer ratio. Tension wood in all the lines contained consistently higher total sugar and glucose percentages when compared with normal wood within the respective lines. However, both sugar and glucose percentages were lower in the tension wood of transgenic lines when compared with the tension wood of wild-type trees. Evaluating the response of trees with altered syringyl content to gravity will improve our understanding of the changes in cell wall chemistry and ultrastructural properties of normal, opposite and tension wood in plants.

Characterization of Woody and Herbaceous Biomasses Lignin Composition with 1064 nm Dispersive Multichannel Raman Spectroscopy

Biomass representing different classes of bioenergy feedstocks, including woody and herbaceous species, was measured with 1064 nm Raman spectroscopy. Pine, oak, poplar, kenaf, miscanthus, pampas grass, switchgrass, alfalfa, orchard grass, and red clover were included in this study. Spectral differences have been identified with an emphasis on lignin guaiacyl and syringyl monomer content and carotenoid compounds. The interpretation of the Raman spectra was correlated with (13)C-nuclear magnetic resonance cross-polarization/magic-angle spinning spectra of select biomass samples. Thioacidolysis quantification of guaiacyl and syringyl monomer composition and the library of Raman spectra were used as a training set to develop a principal component analysis model for classifying plant samples and a principal component regression model for quantifying lignin guaiacyl and syringyl composition. Raman spectroscopy with 1064 nm excitation offers advantages over alternative techniques for biomass characterization, including low spectral backgrounds, higher spectral resolution, short analysis times, and nondestructive analyses.

Exogenous caffeic acid inhibits the growth and enhances the lignification of the roots of soybean ( Glycine max)

The allelopathic effect of caffeic acid was tested on root growth, phenylalanine ammonia-lyase (PAL) and peroxidase (POD) activities, hydrogen peroxide (H 2O 2) accumulation, lignin content and monomeric composition of soybean ( Glycine max) roots. We found that exogenously applied caffeic acid inhibited root growth, decreased the PAL activity and H 2O 2 content and increased the soluble and cell wall-bound POD activities. The p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) monomers and total lignin (H + G + S) increased in the caffeic acid-exposed roots. When applied in conjunction with piperonylic acid (PIP, an inhibitor of the cinnamate 4-hydroxylase, C4H), caffeic acid equalized the inhibitory effect of PIP, whereas the application of methylene dioxocinnamic acid (MDCA, an inhibitor of the 4-coumarate:CoA ligase, 4CL) plus caffeic acid decreased lignin production. These results indicate that exogenously applied caffeic acid can be channeled into the phenylpropanoid pathway via the 4CL reaction, resulting in an increase of lignin monomers that solidify the cell wall and inhibit root growth.

Root Growth Inhibition and Lignification Induced by Salt Stress in Soybean

AbstractSalt stress was evaluated on root growth, enzyme activities (phenylalanine ammonia‐lyase or PAL and soluble plus cell wall‐bound peroxidase or POD), hydrogen peroxide (H2O2) production, total phenolic content and lignin content and composition in soybean (Glycine max L. Merrill) roots. Three‐day‐old seedlings were cultivated in half‐strength Hoagland’s solution (pH 6.0), with or without addition of 50–200 mm of NaCl, into a growth chamber (25 °C, 12/12 h light/dark photoperiod, irradiance of 280 μmol m−2 s−1) for 24 h. In general, root length and fresh and dry weights decreased after NaCl treatment. PAL activity decreased, soluble and cell wall‐bound POD activities increased, and H2O2 content significantly decreased after NaCl exposure. Consequently, total phenolic and lignin contents and p‐hydroxyphenyl (H) and syringyl (S) monomers of lignin increased in NaCl‐treated roots. Altogether, these results suggest that the effects caused by NaCl may be owing to the enhanced lignin production that solidifies the cell wall and restricts root growth.

The Effects on Lignin Structure of Overexpression of Ferulate 5-Hydroxylase in Hybrid Poplar1

Poplar (Populus tremula x alba) lignins with exceedingly high syringyl monomer levels are produced by overexpression of the ferulate 5-hydroxylase (F5H) gene driven by a cinnamate 4-hydroxylase (C4H) promoter. Compositional data derived from both standard degradative methods and NMR analyses of the entire lignin component (as well as isolated lignin fraction) indicated that the C4HF5H transgenic's lignin was comprised of as much as 97.5% syringyl units (derived from sinapyl alcohol), the remainder being guaiacyl units (derived from coniferyl alcohol); the syringyl level in the wild-type control was 68%. The resultant transgenic lignins are more linear and display a lower degree of polymerization. Although the crucial beta-ether content is similar, the distribution of other interunit linkages in the lignin polymer is markedly different, with higher resinol (beta-beta) and spirodienone (beta-1) contents, but with virtually no phenylcoumarans (beta-5, which can only be formed from guaiacyl units). p-Hydroxybenzoates, acylating the gamma-positions of lignin side chains, were reduced by >50%, suggesting consequent impacts on related pathways. A model depicting the putative structure of the transgenic lignin resulting from the overexpression of F5H is presented. The altered structural features in the transgenic lignin polymer, as revealed here, support the contention that there are significant opportunities to improve biomass utilization by exploiting the malleability of plant lignification processes.

Ozone affects shikimate pathway transcripts and monomeric lignin composition in European beech (Fagus sylvatica L.)

The shikimate pathway plays a pivotal role in the formation of aromatic secondary compounds in plants. It finally leads to the formation of, for example, stilbenes, flavonoids, and lignins. Ozone effects on transcript levels of the shikimate pathway were studied in leaves of European beech saplings in the greenhouse (300 nl l−1, 8 h/day, 30 days), and in leaves of adult beech trees at the Kranzberg Forest free-air ozone fumigation site (ambient and twice ambient ozone levels) between June and October 2004. Quantitative real-time RT-PCR (qRT-PCR) with RNA isolated from beech saplings showed a strong induction of 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase3 (DAHPS3), 3-dehydroquinate dehydratase/shikimate dehydrogenase (DHQD/SD), 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS), and chorismate mutase (CM) starting at day seven from the onset of ozone treatment. In contrast, sun leaves of adult European beech showed only a weak elevation of shikimate pathway transcripts throughout the vegetation period studied. In addition, we examined lignin content and monomeric structure in leaves of European beech saplings from the greenhouse experiment. Leaves exhibiting highest amount of ozone-dependent lesions had the highest lignin content as determined according to Klason. Moreover, with increasing leaf damage, the syringyl (S) monomer content decreased, whereas the content of guaiacyl (G) and p-hydroxyphenyl (H) units increased. These results overall suggest that the composition of lignin is affected by ozone and is already regulated at early steps of the shikimate pathway.

‘Defence lignin’ and hydroxycinnamyl alcohol dehydrogenase activities in wounded Eucalyptus gunnii

‘Defence lignin’ and hydroxycinnamyl alcohol dehydrogenase activities in wounded <i>Eucalyptus gunnii</i>