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Reaction mechanisms in delignification of pine Kraft-AQ pulp with hydrogen peroxide using Mn(IV)-Me4DTNE as catalyst.

Pine Kraft-AQ pulp was bleached with hydrogen peroxide catalyzed by [LMn(IV)(2) (mu-O)(3)](ClO(4))(2) at 80 degrees C for 120 min under optimum reaction conditions. The resulting bleached pulp was hydrolyzed with cellulase to obtain insoluble and soluble residual lignins. The alkaline effluent from the bleaching was acidified to precipitate alkaline soluble lignin. These lignin preparations were purified, and then analyzed by 2D HMQC NMR spectroscopic techniques. The results showed that biphenyl (5-5) and stilbene structures are preferentially degraded in the bleaching process, while beta-O-4, beta-5, and beta-beta structures undergo degradation only to a lesser extent. This implies that hydrogen peroxide bleaching using the catalyst is more effective in delignification of softwood pulps than hardwood pulps. The possible reaction mechanisms for the delignification of residual lignin in the pine Kraft-AQ pulp in the bleaching process are discussed on the basis of the 2D HMQC NMR spectroscopic data and the model compound experiments.

Hydrogen Peroxide↗

Comparative study of crude and purified cellulose from wheat straw.

A sequential totally chlorine-free procedure for isolation of cellulose from wheat straw was proposed in this study. The dewaxed straw was pretreated with 0.5 M NaOH in 60% methanol at 60 degrees C for 2.5 h under ultrasonic irradiation for 0-35 min and sequentially posttreated with 2% H(2)O(2)-0.2% TAED at pH 11.8 for 12 h at 48 degrees C, which together solubilized 85.3-86.1% of the original hemicelluloses and 91.7-93.2% of the original lignin, respectively. The yield of crude cellulose ranged between 46.2 and 49.2% on a dry weight basis related to wheat straw, which contained 11.2-12.2% residual hemicelluloses and 2.5-2.9% remaining lignin. Further treatment of the corresponding crude cellulosic preparations with 80% acetic acid-70% nitric acid under the condition given yielded 36.8-37.7% of the purified cellulose, which contained minor amounts of bound hemicelluloses (2.5-2.8%) and was relatively free of associated lignin (0.1-0.2%). The isolated crude and purified cellulose samples were comparatively studied by FT-IR and CP/MAS (13)C NMR spectroscopy, and the relative crystallinity was also estimated. The final stage treatment with 80% acetic acid-70% nitric acid decreased the hemicelluloses and lignin associated in the crude cellulose but led to 3.1-5.4% degradation of the original cellulose; in addition, the purity of the obtained cellulose was high. However, it was found that the final stage treatment is not severe enough to cause decrystallization of cellulose. The thermal stability of the purified cellulose is higher than that of the corresponding crude cellulose.

Cellulose↗

Behavior of monolignol glucosides in angiosperms.

To examine the behavior of cinnamaldehyde and cinnamyl alcohol glucosides in lignifying tissues, angiosperms were subjected to tracer experiments using radioisotope-labeled and stable isotope-labeled glucosides. The aglycone from coniferaldehyde glucoside was efficiently incorporated into guaiacyl and syringyl lignin as a cinnamyl alcohol unit. The aglycone from coniferin was also incorporated into guaiacyl and syringyl lignin. However, some of the coniferin-derived aglycone that was incorporated into lignin passed through a cinnamaldehyde form prior to dehydrogenative polymerization. When coniferin was administered together with coniferaldehyde glucoside, syringyl units were rarely synthesized from coniferin via the cinnamyl alcohol stage, whereas numerous syringyl units were synthesized from coniferaldehyde glucoside. These observations suggest that the coniferaldehyde form is crucial for the biosynthesis of syringyl lignin in angiosperms.

Acrolein↗

Estimation of wood density and chemical composition by means of diffuse reflectance mid-infrared Fourier transform (DRIFT-MIR) spectroscopy.

Sitka spruce (Picea sitchensis) samples (491) from 50 different clones as well as 24 different tropical hardwoods and 20 Scots pine (Pinus sylvestris) samples were used to construct diffuse reflectance mid-infrared Fourier transform (DRIFT-MIR) based partial least squares (PLS) calibrations on lignin, cellulose, and wood resin contents and densities. Calibrations for density, lignin, and cellulose were established for all wood species combined into one data set as well as for the separate Sitka spruce data set. Relationships between wood resin and MIR data were constructed for the Sitka spruce data set as well as the combined Scots pine and Sitka spruce data sets. Calibrations containing only five wavenumbers instead of spectral ranges 4000-2800 and 1800-700 cm(-1) were also established. In addition, chemical factors contributing to wood density were studied. Chemical composition and density assessed from DRIFT-MIR calibrations had R2 and Q2 values in the ranges of 0.6-0.9 and 0.6-0.8, respectively. The PLS models gave residual mean squares error of prediction (RMSEP) values of 1.6-1.9, 2.8-3.7, and 0.4 for lignin, cellulose, and wood resin contents, respectively. Density test sets had RMSEP values ranging from 50 to 56. Reduced amount of wavenumbers can be utilized to predict the chemical composition and density of a wood, which should allow measurements of these properties using a hand-held device. MIR spectral data indicated that low-density samples had somewhat higher lignin contents than high-density samples. Correspondingly, high-density samples contained slightly more polysaccharides than low-density samples. This observation was consistent with the wet chemical data.

Cellulose↗

Chitosan treatment of wheat seeds induces resistance to Fusarium graminearum and improves seed quality.

Chitosan treatment (2-8 mg/mL) of wheat seeds significantly improved seed germination to recommended seed certification standards (>85%) and vigor at concentrations >4 mg/mL, in two cultivars of spring wheat (Norseman and Max), by controlling seed-borne Fusarium graminearum infection. The germination was <80% in the control and >85% in benomyl- and chitosan-treated seeds. Seed-borne F. graminearum was reduced to >50% at higher chitosan treatments compared to the control. Synthesis of phenolic acids was stimulated in primary leaves following chitosan treatment, and levels of these phenolic acids, especially ferulic acid, increased significantly with increasing chitosan concentration. Lignin content of primary leaves also showed a similar pattern. The synthesis of precursors of lignin such as p-coumaric, ferulic, and sinapic acids and phenolic acids having antimicrobial activity such as benzoic, p-coumaric, caffeic, protocatechuic, chlorogenic, ferulic, and gallic acids was also stimulated by chitosan treatment. The induction of phenolic acids and lignin was significantly lower in cv. Max compared to Norseman. Chitosan also inhibited fungal transmission to the primary roots of germinating seedlings. Results suggest that chitosan controlled seed-borne F. graminearum infection and increased the resistance in seedlings by stimulating the accumulation of phenolics and lignin. Thus, chitosan has a potential for improvement of seed quality and enhancement of crop yields as well as increased value of stored grains for food and feed.

Benomyl↗

Substrate profiles and expression of caffeoyl coenzyme A and caffeic acid O-methyltransferases in secondary xylem of aspen during seasonal development.

Seasonal expression of caffeoyl-CoA O-methyltransferase (EC 2.1.1.104) was analyzed in aspen developing secondary xylem in parallel with caffeate O-methyltransferase (EC 2.1.1.68). Enzyme activity and mRNA levels for both enzymes peaked in the middle of the growing season. These results strongly suggest that both forms of O-methyltransferase were actively participating in lignin precursor biosynthesis during the growing season. To determine the role of each enzyme form, xylem extracts from two days in the growing season were assayed with four substrates: caffeoyl-CoA, 5-hydroxyferuloyl-CoA, caffeate acid and 5-hydroxyferulic acid. Recombinant forms of caffeoyl-CoA and caffeate O-methyltransferase were also assayed with these substrates. The recombinant enzymes have different substrate specificity with the caffeoyl-CoA O-methyltransferase being essentially specific for CoA ester substrates with a preference for caffeoyl-CoA, while caffeate O-methyltransferase utilized all four substrates with a preference for the free acid forms. We suggest that caffeoyl-CoA O-methyltransferase is likely to be responsible for biosynthesis of lignin precursors in the guaiacyl pathway and may represent a more primitive enzyme form leftover from very early land plant evolution. Caffeate O-methyltransferase is more likely to be responsible for lignin precursor biosynthesis in the syringyl pathway, especially since it can catalyze methylation of 5-hydroxyferuloyl-CoA quite effectively. This latter enzyme form then may be considered a more recently evolved component of the lignin biosynthetic pathways of the evolutionarily advanced plants such as angiosperms.

Base Sequence↗

Experimental model for in vivo determination of dietary fibre and its effect on the absorption of nutrients in the small intestine.

1. An experimental model for the determination of dietary fibre according to the definition of Trowell et al. (1976) is described. Food was subjected to in vivo digestion in ileostomy patients, and the ileostomy contents were collected quantitatively, the polysaccharide components of which were analysed by gas--liquid chromatography and the Klason lignin by gravimetric determination. The model was used for the determination of dietary fibre in AACC (American Association of Cereal Chemists), wheat bran and for studies on the extent of hydrolysis of wheat-bran fibre in the stomach and small intestine. The effect of wheat bran on ileostomy losses of nitrogen, starch and electrolytes was also investigated. 2. Nine patients with established ileostomies were studied during two periods while on a constant low-fibre diet. In the second period 16 g AACC wheat bran/d was added to the diet. The ileostomy contents and duplicate portions of the diet were subjected to determinations of wet weight, dry weight, water content, fibre components, starch, N, sodium and potassium. 3. The wet weight of ileostomy contents increased by 94 g/24 h and dry weight by 10 g/24 h after consumption of bran. The dietary fibre of AACC bran, determined as the increase in polysaccharides and lignin of ileostomy contents after consumption of bran, was 280 g/kg fresh weight (310 g/kg dry matter). Direct analysis of polysaccharides and lignin in bran gave a value of 306 g/kg fresh weight. Of the added bran hemicellulose and cellulose 80--100% and 75--100% respectively were recovered in ileostomy contents. There was no significant difference between the two periods in amount of N, starch and K found in the ileostomy contents. The Na excretion increased during the 'bran' period and correlated well with the wet weight of ileostomy contents. 4. In conclusion, it seems probable that determination of dietary fibre by in vivo digestion in ileostomy patients comes very close to the theoretical definition of dietary fibre, as the influence of bacteria in the ileum seems small. Bacterial growth should be avoided by using a technique involving the change of ileostomy bags every 2 h and immediate deep-freezing of the ileostomy contents. True dietary fibre can be determined by direct analysis of polysaccharides and lignin in the food, at least in bran. Very little digestion of hemicellulose and cellulose from bran occurs in the stomach and small bowel. The 10--20% loss in some patients may be due to digestion by the gastric juice or to bacterial fermentation in the ileum, or both. The extra amount of faecal N after consumption of bran, reported by others, is probably produced in the large bowel.

Adult↗

Effects of ruminant digestion and metabolism on phenolic monomers of forages.

Immature and mature lucerne (Medicago sativa) and tall fescue (Festuca arundinacea) hays were analysed for their lignin and phenolic monomer (hydroxybenzoic and hydroxycinnamic acid derivatives) contents. These hays were given to four sheep with rumen, duodenal and ileal cannulas, and to a steer with rumen and abomasal cannulas, to investigate the extent and sites of digestion of lignin and phenolic monomers. The hays and digesta samples were analysed for alkali and nitrobenzene-extractable phenolic monomers. Lanthanum oxide was used as an external marker in the digestion studies. Urine was also collected for estimates of total phenolic balance. Grass hays contained greater concentrations of alkali-labile phenolic monomers than did legume hays, whereas legume hays had higher levels of nitrobenzene oxidation products. Mature tall fescue hay had greater concentrations of phenolic monomers than did immature tall fescue hay, but corresponding differences with maturity were not observed with lucerne hays. There were quantitative differences between diets in the digestibilities and in the sites of digestion of phenolic monomers. Digestibilities ranged from 0.191 to 1.004 for alkali-labile compounds and from -1.137 to 0.868 for nitrobenzene oxidation products. Composition of lignin was altered during its passage through the digestive tract. The proportion of phenolic monomers not recovered in urine and faeces varied from -1.014 to 0.871 for individual compounds and differed between diets. Soluble phenolic monomers decreased in concentration as digesta passed from the rumen to the abomasum and duodenum, but increased again in the ileum; levels varied with diet. Phenolics were rapidly solubilized in the rumen after feeding. Results indicate that forages differ in their phenolic monomer content and in the digestibility of these compounds. Therefore, lignin is not an inert compound in the digestive tract of ruminants as has been previously reported.

Animals↗

Biodegradability enhancement by wet oxidation in alkaline media: delignification as a case study.

Nowadays many industries are considering the recycling of process waters as a way of improving environmental safety, preventing pollution, and avoiding the loss of valuable production materials. One industry in the forefront of this trend is the pulp and paper industry. Lignin is a pollutant present in the mill process waters and such macromolecules can cause problems during biological treatment of process waters. Wet oxidation (WO) is a process that can be used as a pre-treatment method for lignin fragmentation and improvement of biodegradability. Wet oxidation (WO) under alkaline conditions permits faster lignin fragmentation than the conventional WO process and, therefore, should favour biodegradability improvement. In this study, the experiments were carried out in a high-pressure batch reactor with an alkali lignin solution at temperatures up to 438 K, an alkali concentration of 1.5-3.5 g l(-1) and an oxygen partial pressure of 0.4 to 1.5 MPa. At an alkali concentration of 3.5 g l(-1)1 and 0.4 MPa of oxygen partial pressure, an increase in BOD/COD ratio was achieved from an initial 11% to 71%. The experiments also showed that the amount of small molecules in the solution measured by Immediately Available BOD (IA BOD) depends on the amount of alkali added and the operating temperature.

Biodegradation, Environmental↗

Lignofibrils on the external cell wall surface of cultured plant cells.

Small strands and bundles of strands extend from the outside surface of suspension-cultured cells of Daucus, Ipomoea, and Phaseolus into the medium. This fibrous cell coat is present in all samples from various growth stages but appears to increase in quantity in the order Ipomoea < Phaseolus < Daucus. The bundles are often many microns in length and display great variation in frequency, size, and form. Identification of the composition of the strands and bundles as lignin is consistent with the following observations: alkaline nitrobenzene oxidation of the strands to compounds which resemble monomers of wood lignin; resistance of the strands to pronase, trypsin, pectinase, and lipase; strong irreversible adsorption of heavy metals; deposition of silver granules by treatment with silver nitrate-hexamine reagent; extraction of the bundles with aqueous dioxane (Björkman procedure); presence in quantity of a structured form of Klason lignin; and existence of material giving a positive test with the Wiesner reagent. Large individual strands (lignofibrils) from Phaseolus show the form of a flat ribbon with very thin branches at irregular intervals. This form does not vary with preparatory techniques, although its electron opacity does. Intercellular spaces display considerable structure and sometimes contain sheets of fibrillar material merging with both the middle lamella between the cells and the surface bundles facing the medium. These sheets are probably another form of association of the lignofibrils. It is suggested that natural fibrous lignin may be a much commoner component of plant tissue than suspected hitherto.

Cell Fractionation↗

Effects of tomato pomace and mixed-vegetable pomace on serum and liver cholesterol in rats.

The effects of tomato pomace and mixed-vegetable pomace, vegetable processing by-products, on serum and liver cholesterol were compared against pectin, wheat bran, cellulose, and lignin. Male Sprague-Dawley rats (100 g) were fed high sucrose, semi-purified diets containing basal (0), 5% or 10% of the fiber source and either 0, 0.5% cholesterol or 0.5% cholesterol + 0.125% sodium cholate. After 28 days, there were highly significant differences (P less than 0.001) in serum and liver cholesterols due to fiber source, dietary cholesterol and the interaction. The effect of fiber source level was significant (P less than 0.05) for liver cholesterol only. Pectin groups had serum and liver cholesterol levels lower (P less than 0.05) than the others. At the 10% level, tomato pomace, wheat bran and lignin (Indulin AT) groups had serum cholesterols higher (P less than 0.05) than basal; the mixed-vegetable pomace and cellulose groups were not different from basal. Liver cholesterols of tomato pomace, mixed-vegetable pomace, wheat bran and lignin groups were not different at the 10% level, but were higher than the pectin, cellulose and basal groups (P less than 0.05). In a second experiment, serum cholesterols of rats fed pectin were lower (P less than 0.05) than for those fed cellulose or two types of lignin (Indulin AT, Reax 27).

Animals↗

Influence of dietary fiber on lipid metabolism in meal-fed rats.

The effects of the four major components of dietary fiber--cellulose, hemicellulose, pectin and lignin--on lipid metabolism were studied in meal-fed Wistar rats maintained on a cholesterol-free semipurified diet for 21 days. Transit time was decreased and fecal weight increased compared to rats fed a fiber-free diet. Rats fed cellulose or lignin gained significantly less weight than rats fed hemicellulose or pectin. The fibers had no effect on serum cholesterol levels, but serum triacylglycerol levels were significantly higher in rats fed cellulose. Liver cholesterol levels were higher in cellulose-fed rats but liver triacylglycerol levels were highest in rats fed hemicellulose or pectin. Hepatic cholesterol 7 alpha-hydroxylase and HMG-CoA reductase activities were highest in rats fed cellulose and pectin, respectively. Epididymal fat cell size was similar in all groups but fat cell number was highest in pectin-fed rats. Perirenal fat cell size was greatest in rats fed cellulose or lignin and fat cell number in rats fed cellulose or hemicellulose. Lipoprotein lipase activity (per 10(6) cells) was elevated in epididymal fat of rats fed pectin and in perirenal fat of rats fed hemicellulose. Carcass lipid accumulation was highest in rats fed cellulose or lignin. Rats fed cellulose accumulated significantly higher levels of carcass cholesterol and triacylglycerol. Of the fibers fed, cellulose led to an accumulation of serum, liver and carcass lipid. The four fibers fed represent purified and altered forms of cellular components and the observed effects cannot be extrapolated to diets containing foods rich in one or another of these components.

Adipose Tissue↗

Down-regulation of cinnamoyl-CoA reductase in tomato (Solanum lycopersicum L.) induces dramatic changes in soluble phenolic pools.

Health-beneficial properties of many secondary plant metabolites have created much interest into the control of their biosynthesis in crop species. Phenolic compounds, including flavonoids, hydroxycinnamates, and tannins, make up an important group of such phytonutrients. They are formed via the phenylpropanoid pathway and share common precursors with lignin, an insoluble cell wall-associated polymer. In this study, the aim was to reduce lignin biosynthesis so as to enhance the availability of these precursors and, thereby, stimulate the production of soluble, potentially health-promoting, phenolic compounds in tomato (Solanum lycopersicum L.). First two tomato genes encoding cinnamoyl-CoA reductase (CCR), a key enzyme in the formation of lignin monomers, were identified and characterized. Transgenic plants exhibiting a reduced lignin content were subsequently obtained through an RNAi strategy targeting one of these genes. As anticipated, the total level of soluble phenolics was higher in stems and leaves of the transformants as compared with control plants. This was correlated with an increased antioxidant capacity of the corresponding plant extracts. Analysis of the soluble phenolic fraction by HPLC-MS revealed that vegetative organs of CCR down-regulated plants contained higher amounts of chlorogenic acid and rutin, and accumulated new metabolites undetectable in the wild type, such as N-caffeoyl putrescine and kaempferol rutinoside. In fruits, CCR down-regulation triggered the moderate accumulation of two new compounds in the flesh, but the total phenolic content was not affected. Although the prospects of exploiting such a strategy for crop improvement are limited, the results provide further insight into the control of the phenylpropanoid pathway in the Solanaceae.

Aldehyde Oxidoreductases↗

Nematode infection and reproduction in transgenic and mutant Arabidopsis and tobacco with an altered phenylpropanoid metabolism.

Transgenic and mutant Arabidopsis and tobacco plants with altered phenylpropanoid metabolism were infected with the plant parasitic root knot nematode Meloidogyne incognita to assess the effect of the transgene or mutation on nematode infection and reproduction. Modifications in the lignin biosynthetic pathway which alter lignin composition in roots affected reproduction. In Arabidopsis with increased levels of syringyl lignin, reproduction was lower than in wild-type plants, while in tobacco with reduced levels of syringyl lignin, life cycle progression was stimulated. Overexpression of a MYB transcription factor of phenylpropanoid metabolism in tobacco significantly stimulated reproduction of M. incognita, while overexpression of L-phenylalanine ammonia-lyase had no effect. Arabidopsis transparent testa mutants with deficiencies in flavonoid pathway enzymes did not affect reproduction of M. incognita in the present infection tests.

Animals↗

Profiling of oligolignols reveals monolignol coupling conditions in lignifying poplar xylem.

Lignin is an aromatic heteropolymer, abundantly present in the walls of secondary thickened cells. Although much research has been devoted to the structure and composition of the polymer to obtain insight into lignin polymerization, the low-molecular weight oligolignol fraction has escaped a detailed characterization. This fraction, in contrast to the rather inaccessible polymer, is a simple and accessible model that reveals details about the coupling of monolignols, an issue that has raised considerable controversy over the past years. We have profiled the methanol-soluble oligolignol fraction of poplar (Populus spp.) xylem, a tissue with extensive lignification. Using liquid chromatography-mass spectrometry, chemical synthesis, and nuclear magnetic resonance, we have elucidated the structures of 38 compounds, most of which were dimers, trimers, and tetramers derived from coniferyl alcohol, sinapyl alcohol, their aldehyde analogs, or vanillin. All structures support the recently challenged random chemical coupling hypothesis for lignin polymerization. Importantly, the structures of two oligomers, each containing a gamma-p-hydroxybenzoylated syringyl unit, strongly suggest that sinapyl p-hydroxybenzoate is an authentic precursor for lignin polymerization in poplar.

Lignin↗

Water deficits affect caffeate O-methyltransferase, lignification, and related enzymes in maize leaves. A proteomic investigation.

Drought is a major abiotic stress affecting all levels of plant organization and, in particular, leaf elongation. Several experiments were designed to study the effect of water deficits on maize (Zea mays) leaves at the protein level by taking into account the reduction of leaf elongation. Proteomic analyses of growing maize leaves allowed us to show that two isoforms of caffeic acid/5-hydroxyferulic 3-O-methyltransferase (COMT) accumulated mostly at 10 to 20 cm from the leaf point of insertion and that drought resulted in a shift of this region of maximal accumulation toward basal regions. We showed that this shift was due to the combined effect of reductions in growth and in total amounts of COMT. Several other enzymes involved in lignin and/or flavonoid synthesis (caffeoyl-CoA 3-O-methyltransferase, phenylalanine ammonia lyase, methylenetetrahydrofolate reductase, and several isoforms of S-adenosyl-l-methionine synthase and methionine synthase) were highly correlated with COMT, reinforcing the hypothesis that the zone of maximal accumulation corresponds to a zone of lignification. According to the accumulation profiles of the enzymes, lignification increases in leaves of control plants when their growth decreases before reaching their final size. Lignin levels analyzed by thioacidolysis confirmed that lignin is synthesized in the region where we observed the maximal accumulation of these enzymes. Consistent with the levels of these enzymes, we found that the lignin level was lower in leaves of plants subjected to water deficit than in those of well-watered plants.

Gene Expression Regulation, Enzymologic↗

A coumaroyl-ester-3-hydroxylase insertion mutant reveals the existence of nonredundant meta-hydroxylation pathways and essential roles for phenolic precursors in cell expansion and plant growth.

Cytochromes P450 monooxygenases from the CYP98 family catalyze the meta-hydroxylation step in the phenylpropanoid biosynthetic pathway. The ref8 Arabidopsis (Arabidopsis thaliana) mutant, with a point mutation in the CYP98A3 gene, was previously described to show developmental defects, changes in lignin composition, and lack of soluble sinapoyl esters. We isolated a T-DNA insertion mutant in CYP98A3 and show that this mutation leads to a more drastic inhibition of plant development and inhibition of cell growth. Similar to the ref8 mutant, the insertion mutant has reduced lignin content, with stem lignin essentially made of p-hydroxyphenyl units and trace amounts of guaiacyl and syringyl units. However, its roots display an ectopic lignification and a substantial proportion of guaiacyl and syringyl units, suggesting the occurrence of an alternative CYP98A3-independent meta-hydroxylation mechanism active mainly in the roots. Relative to the control, mutant plantlets produce very low amounts of sinapoyl esters, but accumulate flavonol glycosides. Reduced cell growth seems correlated with alterations in the abundance of cell wall polysaccharides, in particular decrease in crystalline cellulose, and profound modifications in gene expression and homeostasis reminiscent of a stress response. CYP98A3 thus constitutes a critical bottleneck in the phenylpropanoid pathway and in the synthesis of compounds controlling plant development. CYP98A3 cosuppressed lines show a gradation of developmental defects and changes in lignin content (40% reduction) and structure (prominent frequency of p-hydroxyphenyl units), but content in foliar sinapoyl esters is similar to the control. The purple coloration of their leaves is correlated to the accumulation of sinapoylated anthocyanins.

Arabidopsis↗

Lignification in transgenic poplars with extremely reduced caffeic acid O-methyltransferase activity.

Transgenic poplars (Populus tremula x Populus alba) were obtained by introduction of a sense homologous transgene encoding caffeic acid O-methyltransferase (COMT) under the control either of the cauliflower mosaic virus double 35S promoter or of the eucalyptus cinnamyl alcohol dehydrogenase promoter. Although these constructs conferred a moderate overexpression of COMT in some lines, a transgenic line with the double 35S promoter was found where COMT activity in woody tissues was close to zero due to a gene-silencing phenomenon. For the first time in COMT down-regulated trees, this alteration substantially reduced lignin level in 6-month-old trees (17% decrease). Lignin structure was found to be strongly altered, with a two times higher content in condensed bonds, an almost complete lack of syringyl units, and the incorporation of 5-hydroxyguaiacyl units to the most remarkable extent reported so far. Consistent with the higher cellulose content and with the higher condensation degree of the lignin, the impact of the transformation on the kraft-pulping performances of the poplar trees positively affected the pulp yield (10% relative increase), but made lignins less amenable to industrial degradations.

Cellulose↗