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At least 73 records · Page 4

A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield

Lignification of secondary cell walls is a major factor conferring recalcitrance of lignocellulosic biomass to deconstruction for fuels and chemicals. Genetic modification can reduce lignin content and enhance saccharification efficiency, but usually at the cost of moderate-to-severe growth penalties. We have developed a method, using a single DNA construct that uses CRISPR–Cas9 gene editing to knock-out expression of an endogenous gene of lignin monomer biosynthesis while at the same time expressing a modified version of the gene’s open reading frame that escapes cutting by the Cas9 system and complements the introduced mutation in a tissue-specific manner. Expressing the complementing open reading frame in vessels allows for the regeneration of Arabidopsis plants with reduced lignin, wild-type biomass yield, and up to fourfold enhancement of cell wall sugar yield per plant. The above phenotypes are seen in both homozygous and bi-allelic heterozygous T1 lines, and are stable over at least four generations. The method provides a rapid approach for generating reduced lignin trees or crops with one single transformation event, and, paired with a range of tissue-specific promoters, provides a general strategy for optimizing loss-of-function traits that are associated with growth penalties. This method should be applicable to any plant species in which transformation and gene editing are feasible and validated vessel-specific promoters are available.

59 BASIC BIOLOGICAL SCIENCES↗

Overexpression of the rice BAHD acyltransferase AT10 increases xylan-bound p-coumarate and reduces lignin in Sorghum bicolor

Abstract Background The development of bioenergy crops with reduced recalcitrance to enzymatic degradation represents an important challenge to enable the sustainable production of advanced biofuels and bioproducts. Biomass recalcitrance is partly attributed to the complex structure of plant cell walls inside which cellulose microfibrils are protected by a network of hemicellulosic xylan chains that crosslink with each other or with lignin via ferulate (FA) bridges. Overexpression of the rice acyltransferase OsAT10 is an effective bioengineering strategy to lower the amount of FA involved in the formation of cell wall crosslinks and thereby reduce cell wall recalcitrance. The annual crop sorghum represents an attractive feedstock for bioenergy purposes considering its high biomass yields and low input requirements. Although we previously validated the OsAT10 engineering approach in the perennial bioenergy crop switchgrass, the effect of OsAT10 expression on biomass composition and digestibility in sorghum remains to be explored. Results We obtained eight independent sorghum ( Sorghum bicolor (L.) Moench) transgenic lines with a single copy of a construct designed for OsAT10 expression. Consistent with the proposed role of OsAT10 in acylating arabinosyl residues on xylan with p -coumarate ( p CA), a higher amount of p -coumaroyl-arabinose was released from the cell walls of these lines upon hydrolysis with trifluoroacetic acid. However, no major changes were observed regarding the total amount of p CA or FA esters released from cell walls upon mild alkaline hydrolysis. Certain diferulate (diFA) isomers identified in alkaline hydrolysates were increased in some transgenic lines. The amount of the main cell wall monosaccharides glucose, xylose, and arabinose was unaffected. The transgenic lines showed reduced lignin content and their biomass released higher yields of sugars after ionic liquid pretreatment followed by enzymatic saccharification. Conclusions Expression of OsAT10 in sorghum leads to an increase of xylan-bound p CA without reducing the overall content of cell wall FA esters. Nevertheless, the amount of total cell wall p CA remains unchanged indicating that most p CA is ester-linked to lignin. Unlike other engineered plants overexpressing OsAT10 or a phylogenetically related acyltransferase with similar putative function, the improvements of biomass saccharification efficiency in sorghum OsAT10 lines are likely the result of lignin reductions rather than reductions of cell wall-bound FA. These results also suggest a relationship between xylan-bound p CA and lignification in cell walls.

09 BIOMASS FUELS↗

ToF-SIMS imaging reveals that p -hydroxybenzoate groups specifically decorate the lignin of fibres in the xylem of poplar and willow

Esterified phenolics occur as γ-linked lignin acylations in many plant taxa but little is known about the distribution or function of such groups. In the family Salicaceae, p-hydroxybenzoate groups are present in the lignin of poplars (Populus spp.) and willows (Salix spp.). In this study, the distribution of cell wall-bound p-hydroxybenzoate was examined in different tissue and cell types of poplar and willow trees. This analysis showed that p-hydroxybenzoate groups were most prevalent in juvenile stems, and were elevated in tension wood and following nitrogen fertilisation. Closer examination of stem cross-sections using time-of-flight secondary-ion mass spectrometry (ToF-SIMS) directly showed that p-hydroxybenzoate groups occur predominantly in the cell walls of fibres and are largely absent from vessel elements. Furthermore, these results point to a role for ester-linked groups in modifying the syringyl-rich lignin of fibres, perhaps to strengthen the cell walls of these structural elements or to increase lignification rates.

59 BASIC BIOLOGICAL SCIENCES↗

Synthesis of hydroxycinnamoyl shikimates and their role in monolignol biosynthesis

Hydroxycinnamoyl shikimates were reported in 2005 to be intermediates in monolignol biosynthesis. 3-Hydroxylation of p-coumarate, originally thought to occur via coumarate 3-hydroxylase (C 3 H) from p-coumaric acid or its CoA thioester, was revealed to be via the action of coumaroyl shikimate 3'-hydroxylase (C 3 'H) utilizing p-coumaroyl shikimate as the substrate, itself derived from p-coumaroyl-CoA via hydroxycinnamoyl-CoA: shikimate hydroxycinnamoyltransferase (HCT). The same HCT was conjectured to convert the product, caffeoyl shikimate, to caffeoyl-CoA to continue on the pathway starting with its 3-O-methylation. At least in some plants, however, a more recently discovered caffeoyl shikimate esterase (CSE) enzyme hydrolyzes caffeoyl shikimate to caffeic acid from which it must again produce its CoA thioester to continue on the monolignol biosynthetic pathway. HCT and CSE are therefore monolignol biosynthetic pathway enzymes that have provided new opportunities to misregulate lignification. To facilitate studies into the action and substrate specificity of C 3 H/C 3 'H, HCT, and CSE enzymes, as well as for metabolite authentication and for enzyme characterization, including kinetics, a source of authentic substrates and products was required. A synthetic scheme starting from commercially available shikimic acid and the four key hydroxycinnamic acids (p-coumaric, caffeic, ferulic, and sinapic acid) has been developed to provide this set of hydroxycinnamoyl shikimates for researchers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Genetic Modification of KNAT7 Transcription Factor Expression Enhances Saccharification and Reduces Recalcitrance of Woody Biomass in Poplars

The precise role of KNAT7 transcription factors (TFs) in regulating secondary cell wall (SCW) biosynthesis in poplars has remained unknown, while our understanding of KNAT7 functions in other plants is continuously evolving. To study the impact of genetic modifications of homologous and heterologous KNAT7 gene expression on SCW formation in transgenic poplars, we prepared poplar KNAT7 ( PtKNAT7 ) overexpression ( PtKNAT7 -OE) and antisense suppression ( PtKNAT7 -AS) vector constructs for the generation of transgenic poplar lines via Agrobacterium -mediated transformation. Since the overexpression of homologous genes can sometimes result in co-suppression, we also overexpressed Arabidopsis KNAT7 ( AtKNAT7-OE ) in transgenic poplars. In all these constructs, the expression of KNAT7 transgenes was driven by developing xylem (DX)-specific promoter, DX15. Compared to wild-type (WT) controls, many SCW biosynthesis genes downstream of KNAT7 were highly expressed in poplar PtKNAT7-OE and AtKNAT7-OE lines. Yet, no significant increase in lignin content of woody biomass of these transgenic lines was observed. PtKNAT7-AS lines, however, showed reduced expression of many SCW biosynthesis genes downstream of KNAT7 accompanied by a reduction in lignin content of wood compared to WT controls. Syringyl to Guaiacyl lignin (S/G) ratios were significantly increased in all three KNAT7 knockdown and overexpression transgenic lines than WT controls. These transgenic lines were essentially indistinguishable from WT controls in terms of their growth phenotype. Saccharification efficiency of woody biomass was significantly increased in all transgenic lines than WT controls. Overall, our results demonstrated that developing xylem-specific alteration of KNAT7 expression affects the expression of SCW biosynthesis genes, impacting at least the lignification process and improving saccharification efficiency, hence providing one of the powerful tools for improving bioethanol production from woody biomass of bioenergy crops and trees.

59 BASIC BIOLOGICAL SCIENCES↗

Downregulation of barley ferulate 5-hydroxylase dramatically alters straw lignin structure without impact on mechanical properties

Barley is a major cereal crop for temperate climates, and a diploid genetic model for polyploid wheat. Cereal straw biomass is an attractive source of feedstock for green technologies but lignin, a key determinant of feedstock recalcitrance, complicates bio-conversion processes. However, manipulating lignin content to improve the conversion process could negatively affect agronomic traits. An alternative approach is to manipulate lignin composition which influences the physical and chemical properties of straw. This study validates the function of a barley ferulate 5-hydroxylase gene and demonstrates that its downregulation using the RNA-interference approach substantially impacts lignin composition. We identified five barley genes having putative ferulate 5-hydroxylase activity. Downregulation of HvF5H1 substantially reduced the lignin syringyl/guaiacyl (S/G) ratio in straw while the lignin content, straw mechanical properties, plant growth habit, and grain characteristics all remained unaffected. Metabolic profiling revealed significant changes in the abundance of 173 features in the HvF5H1-RNAi lines. The drastic changes in the lignin polymer of transgenic lines highlight the plasticity of barley lignification processes and the associated potential for manipulating and improving lignocellulosic biomass as a feedstock for green technologies. On the other hand, our results highlight some differences between the lignin biosynthetic pathway in barley, a temperate climate grass, and the warm climate grass, rice, and underscore potential diversity in the lignin biosynthetic pathways in grasses.

59 BASIC BIOLOGICAL SCIENCES↗

Validation of a metabolite–GWAS network for Populus trichocarpa family 1 UDP-glycosyltransferases

Metabolite genome-wide association studies (mGWASs) are increasingly used to discover the genetic basis of target phenotypes in plants such as Populus trichocarpa , a biofuel feedstock and model woody plant species. Despite their growing importance in plant genetics and metabolomics, few mGWASs are experimentally validated. Here, we present a functional genomics workflow for validating mGWAS-predicted enzyme–substrate relationships. We focus on uridine diphosphate–glycosyltransferases (UGTs), a large family of enzymes that catalyze sugar transfer to a variety of plant secondary metabolites involved in defense, signaling, and lignification. Glycosylation influences physiological roles, localization within cells and tissues, and metabolic fates of these metabolites. UGTs have substantially expanded in P. trichocarpa , presenting a challenge for large-scale characterization. Using a high-throughput assay, we produced substrate acceptance profiles for 40 previously uncharacterized candidate enzymes. Assays confirmed 10 of 13 leaf mGWAS associations, and a focused metabolite screen demonstrated varying levels of substrate specificity among UGTs. A substrate binding model case study of UGT-23 rationalized observed enzyme activities and mGWAS associations, including glycosylation of trichocarpinene to produce trichocarpin, a major higher-order salicylate in P. trichocarpa. We identified UGTs putatively involved in lignan, flavonoid, salicylate, and phytohormone metabolism, with potential implications for cell wall biosynthesis, nitrogen uptake, and biotic and abiotic stress response that determine sustainable biomass crop production. Our results provide new support for in silico analyses and evidence-based guidance for in vivo functional characterization.

59 BASIC BIOLOGICAL SCIENCES↗

Spacelab mission 2: Experimental descriptions

The second Spacelab Mission and the 12 multidisciplinary experiments selected to fly on board are described. These experiments include the following: vitamin D metabolities and bone demineralization; interaction of oxygen and gravity influenced lignification; ejectable plasma diagnostics package; plasma depletion experiments for ionospheric and radio astronomical studies; small helium cooled IR telescope; elemental composition and energy spectra of cosmic ray nuclei; hard X-ray imaging of clusters of galaxies and other extended X-ray sources; solar magnetic and velocity field measurement system; solar coronal helium abundance Spacelab experiment; solar UV high resolution telescope and spectroraph; solar UV spectral irradiance monitor; and properties of superfluid helium in zero-G.

Clifton, K. S.↗

Science on the Space Shuttle

On the Space Shuttle's third flight, scientific instruments will study the electromagnetic environment with charging and electron beams. Beam plasma discharge will be studied. The plasma diagnostics package contains electromagnetic and particle sensors to study the ionosphere. An attempt will be made to establish a more accurate base of solar UV irradiance measurements with an absolute error of 10 percent or less over the wavelength region 120-400 nm. The solar flare X-ray polarimeter will observe flare X-rays emitted between 5 and 30 keV and measure their polarization as a function of time and photon energy. A photopolarimeter will help study zodiacal light, and interplanetary dust will be sampled by a section of thick aluminum foil. Plant seedlings will be grown to research the effect of near-zero gravity on lignification. A thermal canister experiment will help determine whether instruments can be maintained at a fixed temperature under varying thermal loads.

Neupert, W. M.↗

OSS-1 - A pathfinder mission for space science on Shuttle

On the third Shuttle flight (STS-3), the Orbiter carried a payload of nine scientific instruments. The payload was designated OSS-1 because the program was originaly managed by the Office of Space Science. The OSS-1 objectives are discussed, taking into account the Plasma Diagnostics Package, a study concerned with vehicle charging and potential, the Thermal Canister Experiment, the Solar Flare X-ray Polarimeter, the Solar Ultraviolet Spectral Irradiance Monitor, the study of the influence of weightlessness on lignification in developing plant seedlings, the Microabrasion Foil Experiment, the Contamination Monitor Package, and a study of the characteristics of the Shuttle/Spacelab induced atmosphere. The OSS-1 payload was launched on STS-3 on March 22, 1982.

Neupert, W. M.↗

Aromatic biosynthesis in pine tissues

Pinus elliotti is a woody plant species responsive to gravity and capable of synthesizing large quantities of lignin. Lignification begins very quickly after germination; lignin is detected in the vascular region within 4 days after germination and rapidly progresses up the hypocotyl. Young pine seedlings bend in response to geostimulation for about 10 days after germination, with the most rapid response time occurring in 4- to 5-day-old seedlings. Various chemicals were used to establish their effects on the geotropic response in this gymnosperm species. IAA completely arrests the geotropic response for 18 to 24 hr. Afterward the seedlings respond geostimulation as if they were not treated. The same pattern of response will occur with a second IAA treatment. If the synthetic auxin, 2-4,D, is used, the georesponse is permanently blocked. The method of application does not appear to be critical; addition of auxin to only one side of the seedling gave results similar to those obtained by treating the entire seedling.

Cowles, J. R.↗

4-Coumarate:coenzyme A ligase and isoperoxidase expression in Zinnia mesophyll cells induced to differentiate into tracheary elements

When cultured in inductive medium containing adequate auxin and cytokinin, isolated mesophyll cells of Zinnia elegans L. cv Envy differentiate into tracheary elements with lignified secondary wall thickenings. Differentiation does not occur when cells are cultured in control medium, which has reduced levels of auxin and/or cytokinin. The activities of two enzymes involved in lignin synthesis, 4-coumarate:coenzyme A ligase and peroxidase, were examined. An induction-specific cationic isoperoxidase, visualized by low pH polyacrylamide gel electrophoresis, is detectable in soluble and wall fractions of cultured Zinnia cells long before tracheary elements visibly differentiate and is thus an early marker of differentiation. Compounds (such as antiauxins, anticytokinins, and tunicamycin) that inhibit or delay differentiation alter the expression of this isoperoxidase. 4-Coumarate:coenzyme A ligase activity increases dramatically only as cells differentiate. Together, these results suggest that the onset of lignification in differentiating Zinnia cells might be controlled by the availability of precursors synthesized by way of 4-coumarate:coenzyme A ligase. These precursors would then be polymerized into lignin in the cell wall by the induction-specific isoperoxidase.

NASA Discipline Plant Biology↗

Towards the specification of consecutive steps in macromolecular lignin assembly

When Pinus taeda cell suspension cultures are exposed to 8% sucrose solution, the cells undergo significant intracellular disruption, irregular wall thickening/lignification with concomitant formation of an 'extracellular lignin precipitate. However, addition of potassium iodide (KI), an H202 scavenger, inhibits this lignification response, while the ability to synthesize the monolignols, p-coumaryl and coniferyl alcohols, is retained. Lignin synthesis (i.e. polymerization) is thus temporarily correlated with H202 generation, strongly implying a regulatory role for the latter. Time course analyses of extracellular metabolites leading up to polymer formation reveal that coniferyl alcohol, but not p-coumaryl alcohol, undergoes substantial coupling reactions to give various lignans. Of these, the metabolites, dihydrodehydrodiconiferyl alcohol, shonanin (divanillyl tetrahydrofuran) and its apparent aryl tetralin derivative, cannot be explained simply on the basis of phenolic coupling. It is proposed that these moieties are the precursors of so-called reduced substructures in the lignin macromolecule. This adds a new perspective to the lignin assembly mechanism.

Non-NASA Center↗

Dirigent proteins and dirigent sites in lignifying tissues

Tissue-specific dirigent protein gene expression and associated dirigent (site) localization were examined in various organs of Forsythia intermedia using tissue printing, in situ mRNA hybridization and immunolabeling techniques, respectively. Dirigent protein gene expression was primarily noted in the undifferentiated cambial regions of stem sections, whereas dirigent protein sites were detected mainly in the vascular cambium and ray parenchyma cell initials. Immunolocalization also revealed cross-reactivity with particular regions of the lignified cell walls, these being coincident with the known sites of initiation of lignin deposition. These latter regions are considered to harbor contiguous arrays of dirigent (monomer binding) sites for initiation of lignin biopolymer assembly. Dirigent protein mRNA expression was also localized in the vascular regions of roots and petioles, whereas in leaves the dirigent sites were primarily associated with the palisade layers and the vascular bundle. That is, dirigent protein mediated lignan biosynthesis was initiated primarily in the cambium and ray cell initial regions of stems as well as in the leaf palisade layers, this being in accordance with the occurrence of the lignans for defense purposes. Within lignified secondary xylem cell walls, however, dirigent sites were primarily localized in the S(1) sublayer and compound middle lamella, these being coincident with previously established sites for initiation of macromolecular lignin biosynthesis. Once initiation occurs, lignification is proposed to continue through template polymerization.

NASA Discipline Plant Biology↗

Curvature in Arabidopsis inflorescence stems is limited to the region of amyloplast displacement

Gravitropic sensing in stems and stem-like organs is hypothesized to occur in the endodermis. However, since the endodermis runs the entire length of the stem, the precise site of gravisensing has been difficult to define. In this investigation of gravisensitivity in inflorescence stems of Arabidopsis, we positioned stems in a high gradient magnetic field (HGMF) on a rotating clinostat. Approximately 40% of the young, wild-type (WT) inflorescences, for all positions tested, curved toward the HGMF in the vicinity of the stem exposed to the field. In contrast, when the wedge was placed in the basal region of older inflorescence stems, no curvature was observed. As a control, the HGMF was applied to a starchless mutant, and 5% of the stems curved toward the field. Microscopy of the endodermis in the WT showed amyloplast displacement in the vicinity of the HGMF. Additional structural studies demonstrated that the basal region of WT stems experienced amyloplast displacement and, therefore, suggest this region is capable of gravity perception. However, increased lignification likely prevented curvature in the basal region. The lack of apical curvature after basal amyloplast displacement indicates that gravity perception in the base is not transmitted to the apex. Thus, these results provide evidence that the signal (and thus, response) resulting from perception in Arabidopsis inflorescence stems is spatially restricted.

NASA Discipline Plant Biology↗

Regiochemical control of monolignol radical coupling: a new paradigm for lignin and lignan biosynthesis

BACKGROUND: Although the lignins and lignans, both monolignol-derived coupling products, account for nearly 30% of the organic carbon circulating in the biosphere, the biosynthetic mechanism of their formation has been poorly understood. The prevailing view has been that lignins and lignans are produced by random free-radical polymerization and coupling, respectively. This view is challenged, mechanistically, by the recent discovery of dirigent proteins that precisely determine both the regiochemical and stereoselective outcome of monolignol radical coupling. RESULTS: To understand further the regulation and control of monolignol coupling, leading to both lignan and lignin formation, we sought to clone the first genes encoding dirigent proteins from several species. The encoding genes, described here, have no sequence homology with any other protein of known function. When expressed in a heterologous system, the recombinant protein was able to confer strict regiochemical and stereochemical control on monolignol free-radical coupling. The expression in plants of dirigent proteins and proposed dirigent protein arrays in developing xylem and in other lignified tissues indicates roles for these proteins in both lignan formation and lignification. CONCLUSIONS: The first understanding of regiochemical and stereochemical control of monolignol coupling in lignan biosynthesis has been established via the participation of a new class of dirigent proteins. Immunological studies have also implicated the involvement of potential corresponding arrays of dirigent protein sites in controlling lignin biopolymer assembly.

NASA Discipline Plant Biology↗

Paleo-Environment and C-14 Dating: The Key to the Depositional Age of the Tha Chang and Related Sand Pits, Northeastern Thailand

Tha Chang sand pits, Nakhon Ratchasima Province and many other sand pits in the area adjacent to the Mun River are characterized by their fluviatile environment in association with mass wasting deposits, along the paleo-river channel and the flood plain of the Mun River. Sediments of these deposits are characterized by clasts of various rock types especially the resistant ones with frequent big tree trunks, logs and wood fragments in different sizes and various stages of transformation from moldering stage to lignification and petrification. Widespread pyritization of the lower horizon suggests strongly reducing environment during burial. The Tha Chang deposits have been received much attention from geoscientists especially paleontologist communities, as they contain fragments of some distinct vertebrate species such as Stegadon sp., hominoid primate, rhinoceros Aceratherium and others. Based on the associated mammal fauna and hominoid fossils, the late Miocene ( 9 - 6 Ma) was given for the time of deposition of this sand and gravel unit. Some other reports believed that sediments and materials of these sand and gravel quarries (pits) were deposited by high-energy flood pulses contemporaneous with the tektites forming event during mid-Pleistocene at c. 0.8 Ma. Interpretation from Palynostratigraphical study suggested that the lower horizon of Tha Chang sand pit was deposited during Pliocene/Pleistocene period and the upper horizons are Pleistoncene/Holocene. It is crystal clear that all the fluviatile sediments including tektites and almost all fossil fragments being deposited in these sand pits were, likely a multiple times reworked materials. Only some old bamboo trees, some old crowling trees and fossils grasses observed on the old river bank are considered in situ. C-14 dating of 5 old wood specimens from Tha Chang Sand Pits, 15 old wood specimens from Chumpuang Sand Pits and one sample of old pottery from a Chumpuang Sand Pit were carried out in the NSF- Arizona AMS Laboratory. Although, there is no sharp boundary between the unconsolidated sedimentary horizons in the pits, C-14 ages obtained from the Tha Chang vary from 34,340 BP at the middle horizon (approx 10 m below ground zero) to >49,900 BP at the lower horizon with unknown basal formation (highly pyritized zone approx 20 - 25 m below ground zero). The ages for the Chumpuang vary from 41,700 BP, >45,900 BP and >49,900 BP from the upper most to the lower most of a broad horizon (approx 8 m to approx 12 m below ground zero). The C-14 age of the pottery collected from layer approximately 5 m below ground zero is 2,514 BP. The nature of fluviatile together with occasional mass wasting characteristics of all sand pits studies suggest the relatively faster depositional rate of the lower horizon which involved more flooding and mass wasting deposits than those of the upper horizons. The apparent of some mixing of the wood ages may indicate reworking and lag deposits nature of the area. The depositional rate of the upper most sand and soil horizon (5 m thick) is approximately 1 m per 500 years which mean both erosion and deposition had played a significant role during that time period. In term of the true age of the formation, we argue that since most of the materials deposited are reworked materials, all ages obtained from fossil fragments could not be the age of sand and gravel formation. Furthermore, the maximum age of all the tektite bearing horizons cannot be older than 0.8 Ma. The oldest C-14 age of 49,900 BP is interpreted as the minimum age of the Tha Chang and related sand pits formation when geomorphology of the area was a lot more hilly and much higher gradient than that of the present day.

Putthapiban, P.↗

Enhancing monolignol ferulate conjugate levels in poplar lignin via OsFMT1

The phenolic polymer lignin is one of the primary chemical constituents of the plant secondary cell wall. Due to the inherent plasticity of lignin biosynthesis, several phenolic monomers have been shown to be incorporated into the polymer, as long as the monomer can undergo radicalization so it can participate in coupling reactions. In this study, we significantly enhance the level of incorporation of monolignol ferulate conjugates into the lignin polymer to improve the digestibility of lignocellulosic biomass. Overexpression of a rice Feruloyl-CoA Monolignol Transferase (FMT), OsFMT1, in hybrid poplar (Populus alba x grandidentata) produced transgenic trees clearly displaying increased cell wall-bound ester-linked ferulate, p-hydroxybenzoate, and p-coumarate, all of which are in the lignin cell wall fraction, as shown by NMR and DFRC. We also demonstrate the use of a novel UV–Vis spectroscopic technique to rapidly screen plants for the presence of both ferulate and p-hydroxybenzoate esters. Lastly we show, via saccharification assays, that the OsFMT1 transgenic poplars have significantly improved processing efficiency compared to wild-type and Angelica sinensis-FMT-expressing poplars. The findings demonstrate that OsFMT1 has a broad substrate specificity and a higher catalytic efficiency compared to the previously published FMT from Angelica sinensis (AsFMT). Importantly, enhanced wood processability makes OsFMT1 a promising gene to optimize the composition of lignocellulosic biomass.

09 BIOMASS FUELS↗