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Grass lignins contain substantial amounts of p-coumarate (pCA) that acylate the side-chains of the phenylpropanoid polymer backbone. An acyltransferase, named p-coumaroyl-CoA:monolignol transferase (OsPMT), that could acylate monolignols with pCA in vitro was recently identified from rice. In planta, such monolignol-pCA conjugates become incorporated into lignin via oxidative radical coupling, thereby generating the observed pCA appendages; however p-coumarates also acylate arabinoxylans in grasses. To test the authenticity of PMT as a lignin biosynthetic pathway enzyme, we examined Brachypodium distachyon plants with altered BdPMT gene function. Using newly developed cell wall analytical methods, we determined that the transferase was involved specifically in monolignol acylation. A sodium azide-generated Bdpmt-1 missense mutant had no (

Numerous evolutionary innovations were required to enable freshwater green algae to colonize terrestrial habitats and thereby initiate the evolution of land plants (embryophytes). These adaptations probably included changes in cell-wall composition and architecture that were to become essential for embryophyte development and radiation. However, it is not known to what extent the polymers that are characteristic of embryophyte cell walls, including pectins, hemicelluloses, glycoproteins and lignin, evolved in response to the demands of the terrestrial environment or whether they pre-existed in their algal ancestors. Here we show that members of the advanced charophycean green algae (CGA), including the Charales, Coleochaetales and Zygnematales, but not basal CGA (Klebsormidiales and Chlorokybales), have cell walls that are comparable in several respects to the primary walls of embryophytes. Moreover, we provide both chemical and immunocytochemical evidence that selected Coleochaete species have cell walls that contain small amounts of lignin or lignin-like polymers derived from radical coupling of hydroxycinnamyl alcohols. Thus, the ability to synthesize many of the components that characterize extant embryophyte walls evolved during divergence within CGA. Our study provides new insight into the evolutionary window during which the structurally complex walls of embryophytes originated, and the significance of the advanced CGA during these events.

Lignin engineering is a promising strategy to optimize lignocellulosic plant biomass for use as a renewable feedstock for agro-industrial applications. Current efforts focus on engineering lignin with monomers that are not normally incorporated into wild-type lignins. Here we describe an Arabidopsis line in which the lignin is derived to a major extent from a non-traditional monomer. The combination of mutation in the gene encoding caffeic acid O-methyltransferase (comt) with over-expression of ferulate 5-hydroxylase under the control of the cinnamate 4-hydroxylase promoter (C4H:F5H1) resulted in plants with a unique lignin comprising almost 92% benzodioxane units. In addition to biosynthesis of this particular lignin, the comt C4H:F5H1 plants revealed massive shifts in phenolic metabolism compared to the wild type. The structures of 38 metabolites that accumulated in comt C4H:F51 plants were resolved by mass spectral analyses, and were shown to derive from 5-hydroxy-substituted phenylpropanoids. These metabolites probably originate from passive metabolism via existing biochemical routes normally used for 5-methoxylated and 5-unsubstituted phenylpropanoids and from active detoxification by hexosylation. Transcripts of the phenylpropanoid biosynthesis pathway were highly up-regulated in comt C4H:F5H1 plants, indicating feedback regulation within the pathway. To investigate the role of flavonoids in the abnormal growth of comt C4H:F5H1 plants, a mutation in a gene encoding chalcone synthase (chs) was crossed in. The resulting comt C4H:F5H1 chs plants showed partial restoration of growth. However, a causal connection between flavonoid deficiency and this restoration of growth was not demonstrated; instead, genetic interactions between phenylpropanoid and flavonoid biosynthesis could explain the partial restoration. These genetic interactions must be taken into account in future cell-wall engineering strategies.

BACKGROUND: We used a biomimetic model system to ascertain how reductions in ferulate – lignin cross-linking and shifts in lignin composition influence ruminal cell wall fermentation. Primary walls from maize cell suspensions with normal or reduced feruloylation were artificially lignified with various monolignols previously identified in normal, mutant, and transgenic plants. Cell wall fermentability was determined from gas production during invitro incubation with rumen microflora and by analysis of non-fermented polysaccharides. RESULTS: Hemicellulose fermentation lag time increased by 37%, rate decreased by 37%, and the extent declined by 18% as cell wall lignin content increased from 0.5 to 124 mg g −1 . Lignification increased lag time for cellulose fermentation by 12-fold. Ferulate – lignin cross-linking accounted for at least one-half of the inhibitory effect of lignin on cell wall fermentation. Incorporating sinapyl p-coumarate, a precursor of p-coumaroylated grass lignin, increased the extent of hemicellulose fermentation by 5%. Polymerizing varying ratios of coniferyl and sinapyl alcohols or incorporating 5-hydroxyconiferyl alcohol, coniferaldehyde, sinapyl acetate, or dihydroconiferyl alcohol into lignin did not alter the kinetics of cell wall fermentation. CONCLUSION: The results indicate that selection or engineering of plants for reduced lignification or ferulate – lignin cross-linking will improve fiber fermentability more than current approaches for shifting lignin composition. c � 2008 Society of Chemical Industry

Biodegradation by brown-rot fungi is quantitatively one of the most important fates of lignocellulose in nature. It has long been thought that these basidiomycetes do not degrade lignin significantly, and that their activities on this abundant aromatic biopolymer are limited to minor oxidative modifications. Here we have applied a new technique for the complete solubilization of lignocellulose to show, by one-bond (1)H-(13)C correlation nuclear magnetic resonance spectroscopy, that brown rot of spruce wood by Gloeophyllum trabeum resulted in a marked, non-selective depletion of all intermonomer side-chain linkages in the lignin. The resulting polymer retained most of its original aromatic residues and was probably interconnected by new linkages that lack hydrogens and are consequently invisible in one-bond (1)H-(13)C correlation spectra. Additional work is needed to characterize these linkages, but it is already clear that the aromatic polymer remaining after extensive brown rot is no longer recognizable as lignin.

*Summary A molecular marker compound, derived from lignin by the thioacidolysis degradative method, for structures produced when ferulic acid is incorporated into lignin in angiosperms (poplar, Arabidopsis, tobacco), has been structurally identified as 1,2,2-trithioethyl ethylguaiacol [1-(4-hydroxy-3-methoxyphenyl)-1,2,2-tris(ethylthio)ethane]. Its truncated side chain and distinctive oxidation state suggest that it derives from ferulic acid that has undergone bis-8-O-4 (cross) coupling during lignification, as validated by model studies. A diagnostic contour for such structures is found in two-dimensional 13 C‐ 1 H correlated (HSQC) NMR spectra of lignins isolated from cinnamoyl CoA reductase (CCR)-deficient poplar. As low levels of the marker are also released from normal (i.e. non-transgenic) plants in which ferulic acid may be present during lignification, notably in grasses, the marker is only an indicator for CCR deficiency in general, but is a reliable marker in woody angiosperms such as poplar. Its derivation, together with evidence for 4-O-etherified ferulic acid, strongly implies that ferulic acid is incorporated into angiosperm lignins. Its endwise radical coupling reactions suggest that ferulic acid should be considered an authentic lignin precursor. Moreover, ferulic acid incorporation provides a new mechanism for producing branch points in the polymer. The findings sharply contradict those reported in a recent study on CCR-deficient Arabidopsis.

Summary Two solvent systems for fully dissolving, and optionally derivatizing, finely ground plant cell wall material at room temperature are described: dimethylsulfoxide (DMSO) and tetrabutylammonium fluoride (TBAF) or N-methylimidazole (NMI). In situ acetylation produces acetylated cell walls (Ac-CWs) that are fully soluble in chloroform. Lignin structures tested remain fully intact. The dispersion of 13C–1H correlations afforded by two-dimensional (2D) nuclear magnetic resonance (NMR) experiments reveals the major lignin units, allowing the whole lignin fraction to be analyzed by high-resolution solution-state NMR methods for the first time. Non-degradative cell wall dissolution offers the potential to analyze polysaccharide components, and improve current cell wall analytical methods by using standard homogeneous solution-state chemistry.

Underwater vibration detection is of great importance in personal safety, environmental protection, and military defense. Sealing layers are required in many underwater sensor architectures, leading to limited working-life and reduced sensitivity. Here, a flexible, superhydrophobic, and conductive tungsten disulfide (WS2) nanosheets-wrapped sponge (SCWS) is reported for the high-sensitivity detection of tiny vibration from the water surfaces and from the grounds. When the SCWS is immersed in water, a continuous layer of bubbles forms on its surfaces, providing the sensor with two special abilities. One is sealing-free feature due to the intrinsic water-repellent property of SCWS. The other is functioning as a vibration-sensitive medium to convert mechanical energy into electric signals through susceptible physical deformation of bubbles. Therefore, the SCWS can be used to precisely detect tiny vibration of water waves, and even sense those caused by human footsteps, demonstrating wide applications of this amphibious (water/ground) vibration sensor. Results of this study can initiate the exploration of superhydrophobic materials with elastic and conductive properties for underwater flexible electronic applications.