Differences In Lignin Content Research Articles

Investigating the response of soil nitrogen cycling to grass invasion

In heathlands, high mineral N input causes replacement of Calluna vulgaris, the dominant plant, by fast-growing grasses such as Molinia caerulea. The vegetation shift signifies altered litter quality from low- to high-quality litter due to differences in lignin content. Litter quality usually affects decomposition processes, which can, in turn, alter nutrient cycling. Therefore, the change in plant dominance in this ecosystem possibly alters soil carbon and nutrient cycles, and consequently, ecosystem services (e.g. biodiversity conservation, groundwater recharge, …). We hypothesise that, because of its higher litter quality, nutrient turnover becomes faster with grass encroachment. We tested this hypothesis in a field set-up consisting of 14 plots presenting a gradient of increasing grass dominance (from 0% to 100%). We measured nine soil parameters and assessed possible associations between grass dominance and the soil parameters using multivariate analysis and linear mixed models. We found that grass dominance significantly impacted net N mineralisation and the root biomass. Our results showed very low net N mineralisation rates (0.09 ± 0.04 mg N (kg soil)−1 day−1) and relative nitrification rates (1.99 ± 0.62%). At high grass levels, acid phosphatase activity was significantly lower than at lower grass percentages. These results show that grass encroachment has a minimal impact on heathland soil biochemistry at this point. Still, we consider that it may take many years to translate a change in litter quality and dynamics into a change in soil functioning.

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.

Lignin Distribution on Cell Wall Micro-Morphological Regions of Fibre in Developmental Phyllostachys pubescens Culms.

Bamboo is a natural fibre reinforced composite with excellent performance which is, to a certain extent, an alternative to the shortage of wood resources. The heterogeneous distribution and molecular structure of lignin is one of the factors that determines its performance, and it is the key and most difficult component in the basic research into the chemistry of bamboo and in bamboo processing and utilization. In this study, the distribution of lignin components and lignin content in micro-morphological regions were measured in semi-quantitative level by age and radial location by means of visible-light microspectrophotometry (VLMS) coupled with the Wiesner and Maule reaction. There as guaiacyl lignin and syringyl lignin in the cell wall of the fibre. Lignin content of the secondary cell wall and cell corner increased at about 10 days, reached a maximum at 1 year, and then decreased gradually. From 17 days to 4 years, the lignin content of the secondary cell wall in the outer part of bamboo is higher than that in the middle part (which is, in turn, higher than that in the inner part of the bamboo). VLSM results of the micro-morphological regions showed that bamboo lignification developed by aging. Guaiacyl and syringl lignin units can be found in the cell wall of the fibre, parenchyma, and vessel. There was a difference in lignin content among different ages, different radial location, and different micro-morphological regions of the cell wall. The fibre walls were rich in guaiacyl lignin in the early stage of lignification and rich in syringyl units in the later stage of lignification. The guaiacyl and syringyl lignin deposition of bamboo green was earlier than that of the middle part of bamboo culm, and that of the middle part of bamboo culm was earlier than that of bamboo yellow. The single molecule lignin content of the thin layer is higher than that of thick layers, while the primary wall is higher than the secondary cell wall, showing that lignin deposition is consistent with the rules of cell wall formation. The obtained cytological information is helpful to understand the origin of the anisotropic, physical, mechanical, chemical, and machining properties of bamboo.

Influence of juvenile and mature wood on anatomical and chemical properties of early and late wood from Chinese fir plantation

The proportion of juvenile wood affects the utilization of wood seriously, and the transition year of juvenile wood (JW) and mature wood (MW) plays a decisive role in the rotation and the modification of wood. To find out the demarcation of JW and MW, the tracheid length (TL) and microfibril angle (MFA) of early wood (EW) and late wood (LW) from four Chinese fir clones were measured by optical microscopy and X-ray diffraction. Then the data were analyzed by the k-means clustering method. The correlation and the differences among wood properties between JW and MW were compared. Results indicated that the LW showed better properties than that of EW, but the anatomical differences between EW and LW did not influence the demarcation of JW and MW. The cluster analysis of TL and MFA showed that the transition year was in the 16th year and the transition zone of EW and LW was different among clones. The MW has longer and wider tracheid, thicker cell walls, and smaller MFA. In terms of chemistry, MW had a higher content of holocellulose, α-cellulose, less content of extract, but no significant difference in lignin content compared with JW. The stabilization of chemical components was earlier than that of the anatomic properties. Correlation analysis showed that there were strong correlations between the chemical composition and anatomical characteristics in JW and MW. In general, compared with chemical components, anatomical indicators were more suitable for JW and MW demarcation. The differences and correlations between JW and MW properties provide a theoretical basis for wood rotation and planting.

Simultaneous manipulation of lignin structure and secondary cell wall formation in transgenic poplar

Increasing the wood density to a preferred range contributes to upgrading the value of wood as raw material. Lignin manipulation can also improve wood quality. This study attempted to accelerate secondary cell wall formation in transgenic poplar with an altered lignin structure. To achieve this, OsSWN1, a rice master switch for secondary cell wall formation in fiber cells, was overexpressed in poplar plants in which monolignol biosynthesis was suppressed by the expression of an RNA-interference construct targeted to the gene for cinnamyl alcohol dehydrogenase 1. The generated transgenic poplars successfully overexpressed the chimeric OsSWN1 construct and monolignol biosynthesis remained altered in these plants. Secondary cell wall thickness was increased in the transgenic plants and their wood density was higher compared to the background line. No difference in lignin content was observed, except in one transgenic line. High saccharification characteristics observed in the background line were preserved in the transgenic line with lower OsSWN1 overexpression, but higher OsSWN1 overexpression had a slight negative impact on enzyme saccharification. Our data suggested that fine-tuning of accelerated deposition of the secondary cell wall combined with alteration of monolignol biosynthesis should improve the lignocellulose quality for conventional and future biorefinery uses.

Identification of enzymatic genes with the potential to reduce biomass recalcitrance through lignin manipulation in Arabidopsis

BackgroundDuring the chemical and biochemical decomposition of lignocellulosic biomasses, lignin is highly recalcitrant. Genetic transformation of plants to qualitatively and/or quantitatively modify lignin may reduce these recalcitrant properties. Efficient discovery of genes to achieve lignin manipulation is thus required.ResultsTo screen for new genes to reduce lignin recalcitrance, we heterologously expressed 50 enzymatic genes under the control of a cinnamate 4-hydroxylase (C4H) gene promoter, derived from a hybrid aspen, which is preferentially active in tissues with lignified cell walls in Arabidopsis plants. These genes encode enzymes that act on metabolites in shikimate, general phenylpropanoid, flavonoid, or monolignol biosynthetic pathways. Among these genes, 30, 18, and 2 originated from plants, bacteria, and fungi, respectively. In our first screening step, 296 independent transgenic plants (T1 generation) harboring single or multiple transgenes were generated from pools of seven Agrobacterium strains used for conventional floral-dip transformation. Wiesner and Mäule staining patterns in the stems of the resultant plants revealed seven and nine plants with apparent abnormalities in the two respective staining analyses. According to genomic PCR and subsequent direct sequencing, each of these 16 plants possessed a gene encoding either coniferaldehyde dehydrogenase (calB), feruloyl-CoA 6′-hydroxylase (F6H1), hydroxycinnamoyl-CoA hydratase/lyase (couA), or ferulate 5-hydroxylase (F5H), with one transgenic plant carrying both calB and F6H1. The effects of these genes on lignin manipulation were confirmed in individually re-created T1 transgenic Arabidopsis plants. While no difference in lignin content was detected in the transgenic lines compared with the wild type, lignin monomeric composition was changed in the transgenic lines. The observed compositional change in the transgenic plants carrying calB, couA, and F5H led to improved sugar release from cell walls after alkaline pretreatment.ConclusionsSimple colorimetric characterization of stem lignin is useful for simultaneous screening of many genes with the potential to reduce lignin recalcitrance. In addition to F5H, the positive control, we identified three enzyme-coding genes that can function as genetic tools for lignin manipulation. Two of these genes (calB and couA) accelerate sugar release from transgenic lignocelluloses.

Light absorption and emissions inventory of humic-like substances from simulated rainforest biomass burning in Southeast Asia

Humic-like substances (HULIS) are complex mixtures that are highly associated with brown carbon (BrC) and are important components of biomass burning (BB) emissions. In this study, we investigated the light absorption, emission factors (EFs), and amounts of HULIS emitted from the simulated burning of 27 types of regionally important rainforest biomass in Southeast Asia. We observed that HULIS had a high mass absorption efficiency at 365 nm (MAE365), with an average value of 2.6 ± 0.83 m2 g−1 C. HULIS emitted from BB accounted for 65% ± 13% of the amount of water-soluble organic carbon (WSOC) and 85% ± 10% of the light absorption of WSOC at 365 nm. The EFs of HULIS from BB averaged 2.3 ± 2.1 g kg−1 fuel, and the burning of the four vegetation subtypes (herbaceous plants, shrubs, evergreen trees, and deciduous trees) exhibited different characteristics. The differences in EFs among the subtypes were likely due to differences in lignin content in the vegetation, the burning conditions, or other factors. The light absorption characteristics of HULIS were strongly associated with the EFs. The annual emissions (minimum–maximum) of HULIS from BB in this region in 2016 were 200–371 Gg. Furthermore, the emissions from January to April accounted for 99% of the total annual emissions of HULIS, which is likely the result of the burning activities during this season. The most significant emission regions were Cambodia, Burma, Thailand, and Laos. This study, which evaluated emissions of HULIS by simulating open BB, contributes to a better understanding of the light-absorbing properties and regional budgets of BrC in this region.

Effect of Lignin Content on Cellulolytic Saccharification of Liquid Hot Water Pretreated Sugarcane Bagasse.

Lignin contributes to the rigid structure of the plant cell wall and is partially responsible for the recalcitrance of lignocellulosic materials to enzymatic digestion. Overcoming this recalcitrance is one the most critical issues in a sugar-flat form process. This study addresses the effect of low lignin sugarcane bagasse on enzymatic hydrolysis after liquid hot water pretreatment at 190 °C and 20 min (severity factor: 3.95). The hydrolysis of bagasse from a sugarcane line selected for a relatively low lignin content, gave an 89.7% yield of cellulose conversion to glucose at 40 FPU/g glucan versus a 68.3% yield from a comparably treated bagasse from the high lignin bred line. A lower enzyme loading of 5 FPU/g glucan (equivalent to 3.2 FPU/g total solids) resulted in 31.4% and 21.9% conversion yields, respectively, for low and high lignin samples, suggesting the significance of lignin content in the saccharification process. Further increases in the enzymatic conversion of cellulose to glucose were achieved when the bagasse sample was pre-incubated with a lignin blocking agent, e.g., bovine serum albumin (50 mg BSA/g glucan) at 50 °C for 1 h prior to an actual saccharification. In this work, we have demonstrated that even relatively small differences in lignin content can result in considerably increased sugar production, which supports the dissimilarity of bagasse lignin content and its effects on cellulose digestibility. The increased glucose yields with the addition of BSA helped to decrease the inhibition of non-productive absorption of cellulose enzymes onto lignin and solid residual lignin fractions.

Transcriptome profiling of Brassica napus stem sections in relation to differences in lignin content

BackgroundBrassica crops are cultivated widely for human consumption and animal feed purposes, and oilseed rape/canola (Brassica napus and rapa) is the second most important oilseed worldwide. Because of its natural diversity and genetic complexity, genomics studies on oilseed rape will be a useful resource base to modify the quantity and quality of biomass in various crops, and therefore, should have a positive impact on lignocellulosic biofuel production. The objective of this study was to perform microarray analysis on two variable lignin containing oilseed rape cultivars to target novel genes and transcription factors of importance in Brassica lignin regulation for applied research.ResultsTo gain insight into the molecular networks controlling cell wall biosynthetic and regulatory events, we conducted lignin and microarray analysis of top and basal stem sections of brown seeded Brassica napus DH12075 and yellow seeded YN01–429 cultivars. A total of 9500 genes were differentially expressed 2-fold or higher in the stem between the cultivars, with a higher number of expressed genes in the basal section. Of the upregulated genes, many were transcription factors and a considerable number of these were associated with secondary wall synthesis and lignification in B. napus and other plant species. The three largest groups of transcription factors with differential expression were C2H2 and C3HC4 zinc fingers and bHLH. A significant number of genes related to lignin and carbohydrate metabolism also showed differential expression patterns between the stem sections of the two cultivars. Within the same cultivar, the number of upregulated genes was higher in the top section relative to the basal one.ConclusionIn this study, we identified and established expression patterns of many new genes likely involved in cell wall biosynthesis and regulation. Some genes with known roles in other biochemical pathways were also identified to have a potential role in cell wall biosynthesis. This stem transcriptome profiling will allow for selecting novel regulatory and structural genes for functional characterization, a strategy which may provide tools for modifying cell wall composition to facilitate fermentation for biofuel production.

Contribution of both lignin content and sinapyl monomer to disease resistance in tobacco

As a major component of the cell wall, lignin has been suggested to play an important role in the plant defence response to various pathogens. However, how lignin is involved in plant pathogen interaction is still unclear. Here, a series of transgenic tobacco lines were cultivated with a range of differences in lignin content and composition. Evaluation of pathogen resistance in these plants indicated that lower total lignin content aggravated the severity of tobacco black shank and bacterial wilt diseases, while increased sinapyl lignin (S) alleviated the disease symptoms. The regression analysis indicated both lignin content and S lignin were positively correlated with disease resistance. These two factors had additive effects, exhibiting stronger correlation with disease resistance when they were combined. Neither guaiacyl lignin (G) nor S/G ratio showed close correlation with disease resistance. The expression of pathogenesis‐related protein genes PR2 and PR3 was induced after pathogen inoculation. However, the up‐regulation of PR2 and PR3 was not associated with a disease resistance‐induced increase in lignin content. These data collectively suggest that both total lignin content and S lignin are main factors that contribute to the basic defence response in tobacco.

Parenchyma cell wall structure in twining stem of Dioscorea balcanica

Anatomical adaptation of liana plants includes structural changes in cell walls of different tissues: fibers, vessel elements and tracheids. However, the contribution of parenchyma cells to stem twining in liana plants is mostly unknown. The aim of this investigation is to determine changes in stem parenchyma cell walls that are correlated with the twinning process in liana plants. Parenchyma cell wall structure was studied on the stem cross sections of straight and twisted internodes of monocotyledonous liana Dioscorea balcanica, by different microscopy techniques: light microscopy, scanning electron microscopy, fluorescence detected linear dichroism microscopy and Fourier transform infrared microspectrometry. In addition, chemical analysis of the entire stem internodes was performed using photometric and chromatographic methods. Parenchyma cell walls of twisted D. balcanica internodes are characterized by: lower amounts of cellulose (obtained by FTIR microspectrometry) with different cellulose microfibril orientation (shown by Scanning electron microscopy), but no changes in “cellulose fibril order” (obtained by Differential polarization laser scanning microscopy); lower amounts of xyloglucan, higher amounts of xylan, higher amounts of lignin with modified organization—less condensed lignin (obtained by FTIR microspectrometry). At the same time, chemical analysis of the entire internodes did not show significant differences in lignin content and cell wall bound phenols related to stem twining, except for the presence of diferulate cross-links exclusively in twisted internodes. Our results indicate that adaptations to mechanical strain in D. balcanica stems involve modifications in parenchyma cell wall structure and chemistry, which provide decreased stiffness, higher strength and increased elasticity of twisted internodes.

Lignin Content and Resistance to Sclerotinia minor in Peanut

ABSTRACT Lignin has been shown to be an important component for plant defense in several pathosystems, but the relationship between peanut stem lignin content and resistance in the field to Sclerotinia blight has not been investigated. Stem lignin was quantified from twenty runner, six virginia, and ten spanish genotypes grown in the greenhouse using the acetyl bromide method. Significant differences in lignin content were found within the runner and spanish entries, but not among the virginia genotypes. Disease data collected in the field over two to three years were used to test correlations between lignin content and Sclerotinia blight resistance for a subset of the runner and virginia entries. No significant correlations were found. Within the runner entries, the highest and lowest stem lignin content was found in entries with the most disease resistance. These results indicate that preformed stem lignin content is not a reliable predictor for resistance to Sclerotinia blight in peanut. In addition, commercial peanut cultivars appear to vary considerably in lignin content, and the genotypes with lower levels of stem lignin may be useful to producers who can use peanut haulm for animal feed. Southwest Runner, a cultivar with high resistance to Sclerotinia blight, had the lowest stem lignin content of the 36 peanut lines tested.

Bark coverage and insects influence wood decomposition: Direct and indirect effects

Rates of terrestrial wood decomposition are known to vary widely depending on regional and local climatic conditions, substrate characteristics and the organisms involved but the influence of many factors remain poorly quantified. We sought to determine how bark and insects contribute to decomposition in a southeastern U.S. forest. Open-topped stainless steel pans with screened bottoms were used to prevent subterranean termite (Rhinotermitidae: Reticulitermes spp.) colonization from “protected” logs. After a 20-month study period, we compared mass loss and lignin content between these and logs assigned to “unprotected” treatments that permitted termite colonization. The experiment was repeated for 1) logs from which bark had or had not been initially removed and 2) logs with sealed or unsealed ends. Logs with bark lost significantly (∼2.4-fold) more mass than those without bark, likely due to the moisture-conserving properties of bark. Logs with unsealed ends lost significantly more mass than those with sealed ends. There was no significant difference in mass loss between protected and unprotected logs but logs with visible termite activity lost significantly more mass than those without termite damage. Few differences in lignin content were detected in this study but when logs with bark were analyzed separately, those with visible damage from long-horned beetles (Coleoptera: Cerambycidae) had lower lignin content than those without cerambycid activity. This suggests that cerambycids may promote decomposition indirectly through interactions with fungi or other organisms capable of degrading lignin. Our results suggest that insects can have significant direct and indirect effects on wood decomposition and clearly demonstrate the importance of bark in determining wood decay rates and insect activity.

Impact of Different Lignin Fractions on Saccharification Efficiency in Diverse Species of the Bioenergy Crop Miscanthus

Lignin is a key factor limiting saccharification of lignocellulosic feedstocks. In this comparative study, various lignin methods—including acetyl bromide lignin (ABL), acid detergent lignin (ADL), Klason lignin (KL), and modified ADL and KL determination methods—were evaluated for their potential to assess saccharification efficiency. Six diverse accessions of the bioenergy crop miscanthus were used for this analysis, which included accessions of Miscanthus sinensis, Miscanthus sacchariflorus, and hybrid species. Accessions showed large variation in lignin content. Lignin estimates were different between methods, but (highly) correlated to each other (0.54 ≤ r ≤ 0.94). The strength of negative correlations to saccharification efficiency following either alkaline or dilute acid pretreatment differed between lignin estimates. The strongest and most consistent correlations (−0.48 ≤ r ≤ −0.85) were obtained with a modified Klason lignin method. This method is suitable for high throughput analysis and was the most effective in detecting differences in lignin content (p < 0.001) between accessions.

Chemical and ultrastructural changes of ash wood thermally modified using the thermo-vacuum process: I. Histo/cytochemical studies on changes in the structure and lignin chemistry

AbstractChanges in structure and lignin chemistry were investigated in ash wood thermally modified (TMW) by the thermo-vacuum (Termovuoto) process for 3 h at 190–220°C by means of light, fluorescence, and transmission electron (TEM) microscopy combined with histo/cytochemistry. Variation in changes in native cell color in TMWs was positively correlated with differences in lignin content between cell types and cell wall regions in the reference wood. Histochemical staining showed increasing amounts of acidic groups in TMWs with different response to ethanol extraction between secondary cell walls and CMLcc(compound middle lamella/middle lamella cell corner) regions. Fluorescence microscopy of TMWs and references showed a difference in intensity and color emission of lignin autofluorescence, reflecting modification of lignin in TMWs. Changes in histochemistry and fluorescence were prominent at and above 200°C. With TEM, increased intensity of lignin staining and distortion of fiber S1layers were detected in TMW treated for 3 h at 220°C (TMW3 h, 220°C). TMW3 h, 220°Cdiffered significantly in molecular ultrastructure of fiber cell walls compared to references, such as loss of the lamellar structure and size and distribution of lignin aggregates. The modification in CMLccstructure in ash TMW3 h, 220°Cis different from that of softwoods.

Investigation into the deterioration process of archaeological bamboo strips of China from four different periods by chemical and anatomical analysis

The chemical and the anatomical features of the archeological bamboo strips of China from four different periods were compared with the non-degraded fresh bamboo to understand the deterioration process. Unprocessed bamboo strips, isolated hemicelluloses and lignin fractions were analyzed with wet chemical analysis, GC, FTIR, UV and XRD to investigate the chemical degradation. It was found that the crystalline cellulose and the polyxylose degraded with increasing age, leaving the amorphous cellulose and the lignin with syringyl moieties modified as the main residual cell wall materials. Serious degradation of p-coumaric acid and ferulic acid were detected and indicated the cleavage of the linkage between the lignin and the carbohydrate, which might facilitate the degradation of carbohydrate. The anatomical analysis with SEM and TEM revealed the vanishing of the parenchymatous ground tissue and vessels and the cellulose microfibrils in the multilayered secondary wall. The differences in lignin content and in tissue density (cell thickness/cell size) between the parenchyma cells and the sclerenchymatous fiber cells fundamentally led to the formation of the unique anatomical structure of the archeological bamboo strips which was composed of parallel solid bundles. The effects of the chemical and the anatomical degradation on the mechanical properties were also discussed.

Reactivity of syringyl and guaiacyl lignin units and delignification kinetics in the kraft pulping of Eucalyptus globulus wood using Py-GC–MS/FID

Eucalyptus globulus sapwood and heartwood showed no differences in lignin content (23.0% vs. 23.7%) and composition: syringyl-lignin (17.9% vs. 18.0%) and guaiacyl-lignin (4.8% vs. 5.2%). Delignification kinetics of S- and G-units in heartwood and sapwood was investigated by Py-GC–MS/FID at 130, 150 and 170°C and modeled as double first-order reactions. Reactivity differences between S and G-units were small during the main pulping phase and the higher reactivity of S over G units was better expressed in the later pulping stage. The residual lignin composition in pulps was different from wood or from samples in the initial delignification stages, with more G and H-units. S/G ratio ranged from 3 to 4.5 when pulp residual lignin was higher than 10%, decreasing rapidly to less than 1. The S/H was initially around 20 (until 15% residual lignin), decreasing to 4 when residual lignin was about 3%.

Transcript abundance of enzymes involved in lignin biosynthesis of Eucalyptus globulus genotypes with contrasting levels of pulp yield and wood density

One hundred genotypes of Eucalyptus globulus were ranked according to specific consumption of wood (cubic meters of wood needed to produce 1 ton of pulp). Ten of the most contrasting genotypes were separated in two groups of five clones each; group 1 (G1) with high wood density, high pulp yield, and low specific consumption, and group 2 (G2) with low density, low pulp yield, and high specific consumption. The contrasting genotypes also had significant differences in lignin content, percent syringyl unit composition, and frequency of β-O-4 linkages. Gene expression for phenylalanine ammonia-lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), 4-coumarate:CoA ligase (4CL) and ferulate 5-hydrolase (F5H) was analyzed in the contrasting genotypes. In both groups, transcript abundance for CAD, PAL, and 4CL were similar and only F5H presented significant differences between groups, with high values in the best ranked genotypes G1 in comparison to G2. Correlations between traits were estimated for lignin content vs. pulp yield (R2 = 0.97), pulp yield vs. syringyl units (R2 = 0.82), β-O-4 linkages vs. pulp yield (R2 = 0.84), and β-O-4 linkages vs. syringyl units (R2 = 0.97). Correlations between chemical composition and transcript abundance for F5H were calculated, finding correlation values with lignin content (R2 = 0.81), syringyl units (R2 = 0.83), and pulp yield (R2 = 0.81). The measurement of transcript abundance of F5H represents a potential genomic tool for tree improvement programs to select trees with high pulp yield.

Label-free &lt;em&gt;in situ&lt;/em&gt; Imaging of Lignification in Plant Cell Walls

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 biofuels. 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 chemicals. 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 fermentation. 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 factor 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 modifications. 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 walls. 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 wood. Analyzing the lignin Raman bands 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-μm. 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.

The relationship between the form of dead bark and lignin content in Scots pine (Pinus sylvestris L.)

In this study the proportion of lignin in wood tissue was analysed in 36 Scots pines differing in their form of dead bark. Material for analyses came from pure pine stands aged 89-91 years located in northern Poland. Chemical analyses were performed in the mature wood zone comprising the last 10 diameter increments located at a height of 1.3 m (DBH). Results indicate differences in lignin content and growth rate in pines whose stems were characterized by different forms of dead bark. The study showed significant differences in lignin contents between trees with shell and scaly dead bark and trees with ropy bark form. Pines with ropy bark were characterized by the lowest lignin content in relation to trees having the other 2 forms of dead bark. Moreover, a relationship was observed between diameter growth rate and lignin content in wood tissue.