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At least 37 records · Page 2

Poplar and shrub willow energy crops in the United States: field trial results from the multiyear regional feedstock partnership and yield potential maps based on the PRISM‐ELM model

Abstract To increase the understanding of poplar and willow perennial woody crops and facilitate their deployment for the production of biofuels, bioproducts, and bioenergy, there is a need for broadscale yield maps. For national analysis of woody and herbaceous crops production potential, biomass feedstock yield maps should be developed using a common framework. This study developed willow and poplar potential yield maps by combining data from a network of willow and poplar field trials and the modeling power of PRISM‐ELM. Yields of the top three willow cultivars across 17 sites ranged from 3.60 to 14.6 Mg ha −1 yr −1 dry weight, while the yields from 17 poplar trials ranged from 7.5 to 15.2 Mg ha −1 yr −1 . Relationships between the environmental suitability estimates from the PRISM‐ELM model and results from field trials had an R 2 of 0.60 for poplar and 0.81 for willow. The resulting potential yield maps reflected the range of poplar and willow yields that have been reported in the literature. Poplar covered a larger geographic range than willow, which likely reflects the poplar breeding efforts that have occurred for many more decades using genotypes from a broader range of environments than willow. While the field trial data sets used to develop these models represent the most complete information at the time, there is a need to expand and improve the model by monitoring trials over multiple cutting cycles and across a broader range of environmental gradients. Despite some limitations, the results of these models represent a dramatic improvement in projections of potential yield of poplar and willow crops across the United States.

09 BIOMASS FUELS↗

The Populus Superoxide Dismutase Gene Family and Its Responses to Drought Stress in Transgenic Poplar Overexpressing a Pine Cytosolic Glutamine Synthetase (GS1a)

Glutamine synthetase (GS) plays a central role in plant nitrogen assimilation, a process intimately linked to soil water availability. We previously showed that hybrid poplar (Populus tremula X alba, INRA 717-1B4) expressing ectopically a pine cytosolic glutamine synthetase gene (GS1a) display enhanced tolerance to drought. Preliminary transcriptome profiling revealed that during drought, members of the superoxide dismutase (SOD) family were reciprocally regulated in GS poplar when compared with the wild-type control, in all tissues examined. SOD was the only gene family found to exhibit such patterns. In silico analysis of the Populus genome identified 12 SOD genes and two genes encoding copper chaperones for SOD (CCSs). The poplar SODs form three phylogenetic clusters in accordance with their distinct metal co-factor requirements and gene structure. Nearly all poplar SODs and CCSs are present in duplicate derived from whole genome duplication, in sharp contrast to their predominantly single-copy Arabidopsis orthologs. Drought stress triggered plant-wide down-regulation of the plastidic copper SODs (CSDs), with concomitant up-regulation of plastidic iron SODs (FSDs) in GS poplar relative to the wild type; this was confirmed at the activity level. We also found evidence for coordinated downregulation of other copper proteins, including plastidic CCSs and polyphenol oxidases, in GS poplar under drought conditions. Both gene duplication and expression divergence have contributed to the expansion and transcriptional diversity of the Populus SOD/CCS families. Coordinated down-regulation of major copper proteins in drought-tolerant GS poplars supports the copper cofactor economy model where copper supply is preferentially allocated for plastocyanins to sustain photosynthesis during drought. Our results also extend previous findings on the compensatory regulation between chloroplastic CSDs and FSDs, and suggest that this copper-mediated mechanism represents a common response to oxidative stress and other genetic manipulations, as in GS poplars, that affect photosynthesis.

59 BASIC BIOLOGICAL SCIENCES↗

Poplar as Biofiber Reinforcement in Composites for Large-Scale 3D Printing

The economic viability of the biofuel industry could be improved by adding a high-value revenue stream for biomass supply chains: bioderived composites for the rapidly expanding large-scale additive manufacturing industry (i.e., 3D printing). Using fibrillated fibers derived from biomass (e.g., Populus ) to reinforce polymers for 3D printing applications would be less expensive compared to using conventional carbon fibers. Poplar fibers of different mesh sizes (<180, 180–425, 425–850, and 850–2360 μm) were used to prepare poplar–polylactic acid (PLA) composites. The poplar/PLA composites were successfully printed using a large-scale 3D printer to create a podium support. The tensile strength of the composites increased from 34 to 54 MPa as the poplar fiber size decreased. Here, the fracture surfaces of composites derived from smaller poplar fibers (<180 μm) were more compact with fewer voids compared with the composites made with larger poplar fibers. Because of the porous and hollow microstructures, smaller poplar fibers contained more pores on their outer surfaces, which were available for the access and penetration of PLA. Poplar has potential for use as a thermoplastic reinforcement for large-scale 3D printing.

36 MATERIALS SCIENCE↗

Bark morphological and chemical features are differentially correlated with disease resistance and yield in hybrid poplar taxa

In the southeastern United States, the establishment of short-rotation intensively cultured plantations of hybrid poplar has been hindered by its susceptibility to stem cankers. We evaluated the tradeoffs between biomass yield and disease tolerance in hybrid poplar genotypes belonging to P. deltoides × P. maximowiczii (DM), P. deltoides × P. nigra (DN), P. trichocarpa × P. maximowiczii (TM), and P. deltoides × P. deltoides (DD) taxa. We hypothesized that canker resistant genotypes will have thicker bark but bark thickness and biomass yield will be negatively correlated. After two growing seasons, the DD genotypes developed thicker bark compared to the genotypes of other taxa and bark thickness was not correlated with biomass yield in the DD genotypes (R 2 = 0.002). However, in the TM, DM, and DN genotypes, bark thickness was negatively correlated with biomass yield (R 2 = 0.33–0.77). Disease incidence studies revealed that the DM genotypes were most susceptible to canker whereas no disease was detected in DD genotypes. Furthermore, bark analysis conducted by Fourier transform infrared spectroscopy coupled with multivariate analysis showed that that DD genotypes to be chemically separate from the three hybrid genotypes and that bark chemistry was correlated with canker disease incidence. Taken together, these results reveal that it is possible to generate hybrid poplar genotypes with thicker bark, disease resistance, and higher biomass yields. This insight should guide further efforts to develop genetically improved hybrid poplar genotypes, both in terms of biomass yield and disease tolerance, for cultivation in the southeastern United States. Hybrid poplar cultivation in southeastern United States is hindered by its susceptibility to stem cankers. We evaluated tradeoffs between yield and canker disease resistance in various hybrid poplar genotypes. After two growing seasons, the DD genotypes showed disease resistance and developed thicker bark that was chemically distinct from the other genotypes. Bark thickness was not correlated with yield in the DD genotypes but was negatively correlated with yield in the other genotypes. These results will guide the development of hybrid poplar genotypes that are both disease resistant and high yielding for cultivation in the southeastern United States.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Inorganic acid-catalyzed hydrogen peroxide-acetic acid pretreatment for selective delignification of poplar

Effects of different inorganic acids as catalysts on the delignification ability of hydrogen peroxide-acetic acid (HPAA) pretreatment were unclear. Herein, the oxidizability of HCl, HBr, H 2 SO 4 , and H 3 PO 4 -catalyzed HPAA on the delignification of poplar were investigated. The ionized H + from inorganic acids catalyzed the synthesis of peracetic acid and improved the oxidizability of HPAA. The anion of inorganic acids caused ineffective decomposition of peroxides and inorganic acids. After the addition of poplar, the H 2 SO 4 -catalyzed HPAA pretreatment exhibited the lowest pH than other inorganic acids catalyzed systems. After the catalysis by inorganic acid, HPAA removed 78.1–91.4 % poplar lignin. Compared with other inorganic acids, the H + of H 2 SO 4 exhibited superior retention in HPAA pretreatment of poplar. After poplar pretreated with the same initial pH value of inorganic acid-catalyzed HPAA, the H 2 SO 4 -catalyzed HPAA showed the strongest delignification ability of poplar. The presence of free radicals in inorganic acid-catalyzed HPAA was unnecessary for the delignification of poplar. Furthermore, this paper elucidated that the ionized H + of inorganic acid mainly enhanced the oxidizability of HPAA for the selectively delignification of ability of HPAA pretreatment.

In-situ synthesis↗

The effects of expression of a hyperthermophilic endoglucanase and nutrient conditions on poplar growth and physiology

Abstract Efficient production and processing of poplar biomass feedstock requires costly pretreatments and enzyme additives. Transgenic alterations of poplar can reduce the need for these inputs by increasing biomass, improving lignocellulose quality and enhancing nutrient uptake. Previously, a transgenic line of poplar expressing a bacterial hyperthermophilic endoglucanase (TnCelB) in Populus alba × grandidentata (P39) was developed and characterized. This study reports the effects on the TnCelB transgenic poplar line under a reduced nutrient treatment. Overall, the nutrient treatment was the source of more observed significant differences than the genotype. Wild type and TnCelB poplar had similar responses in biomass allocation and net photosynthesis. TnCelB trees had a wrinkled leaf phenotype and relative to wild type, had reduced total biomass, reduced water-use efficiency, and a decreased proportion of cellulose to hemicellulose and lignin. In low nutrient conditions, TnCelB trees had increased structural carbohydrates with stable lignin values. The TnCelB line presents a viable option for poplar biomass feedstock, offering biomass comparable to wild type poplar and more efficient processing, with only mild negative phenotypes.

Natalie, Bethanie M. (ORCID:0009000843856585)↗

Salicylic acid activates poplar defense against the biotrophic rust fungus Melampsora larici-populina via increased biosynthesis of catechin and proanthocyanidins

Poplar trees synthesize flavan-3-ols (catechin and proanthocyanidins) as a defense against foliar rust fungi, but the regulation of this defense response is poorly understood. Here, we investigated the role of hormones in regulating flavan-3-ol accumulation in poplar during rust infection. We profiled levels of defense hormones, signaling genes, and flavan-3-ol metabolites in black poplar leaves at different stages of rust infection. Hormone levels were manipulated by external sprays, genetic engineering, and drought to reveal their role in rust fungal defenses. Levels of salicylic acid (SA), jasmonic acid, and abscisic acid increased in rust-infected leaves and activated downstream signaling, with SA levels correlating closely with those of flavan-3-ols. Pretreatment with the SA analog benzothiadiazole increased flavan-3-ol accumulation by activating the MYB–bHLH–WD40 complex and reduced rust proliferation. Furthermore, transgenic poplar lines overproducing SA exhibited higher amounts of flavan-3-ols constitutively via the same transcriptional activation mechanism. These findings suggest a strong association among SA, flavan-3-ol biosynthesis, and rust resistance in poplars. Abscisic acid also promoted poplar defense against rust infection, but likely through stomatal immunity independent of flavan-3-ols. Jasmonic acid did not confer any apparent defense responses to the fungal pathogen. We conclude that SA activates flavan-3-ol biosynthesis in poplar against rust infection.

Plant Sciences↗

Understanding Multiscale Structural Changes During Dilute Acid Pretreatment of Switchgrass and Poplar

Biofuels produced from lignocellulosic biomass hold great promise as a renewable alternative energy and fuel source. To realize a cost and energy efficient approach, a fundamental understanding of the deconstruction process is critically necessary to reduce biomass recalcitrance. Here, the structural and morphological changes over multiple scales (5–6000 Å) in herbaceous (switchgrass) and woody (hybrid poplar) biomass during dilute sulfuric acid pretreatment were explored using neutron scattering and X-ray diffraction. Switchgrass undergoes a larger increase (20–84 Å) in the average diameter of the crystalline core of the elementary cellulose fibril than hybrid poplar (19–50 Å). Switchgrass initially forms lignin aggregates with an average size of 90 Å that coalesce to 200 Å, which is double that observed for hybrid poplar, 55–130 Å. Switchgrass shows a smooth-to-rough transition in the cell wall surface morphology unlike the diffuse-to-smooth transition of hybrid poplar. Yet, switchgrass and hybrid poplar pretreated under the same experimental conditions result in pretreated switchgrass producing higher glucose yields (~76 wt %) than pretreated hybrid poplar (~60 wt %). This observation shows that other aspects like cellulose allomorph transitions, cellulose accessibility, cellular biopolymer spatial distribution, and enzyme–substrate interactions may be more critical in governing the enzymatic hydrolysis efficiency.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar

Abstract Background Biological conversion of lignocellulosic biomass is significantly hindered by feedstock recalcitrance, which is typically assessed through an enzymatic digestion assay, often preceded by a thermal and/or chemical pretreatment. Here, we assay 17 lines of unpretreated transgenic black cottonwood ( Populus trichocarpa ) utilizing a lignocellulose-degrading, metabolically engineered bacterium, Caldicellulosiruptor bescii . The poplar lines were assessed by incubation with an engineered C. bescii strain that solubilized and converted the hexose and pentose carbohydrates to ethanol and acetate. The resulting fermentation titer and biomass solubilization were then utilized as a measure of biomass recalcitrance and compared to data previously reported on the transgenic poplar samples. Results Of the 17 transgenic poplar lines examined with C. bescii , a wide variation in solubilization and fermentation titer was observed. While the wild type poplar control demonstrated relatively high recalcitrance with a total solubilization of only 20% and a fermentation titer of 7.3 mM, the transgenic lines resulted in solubilization ranging from 15 to 79% and fermentation titers from 6.8 to 29.6 mM. Additionally, a strong inverse correlation ( R 2 = 0.8) between conversion efficiency and lignin content was observed with lower lignin samples more easily converted and solubilized by C. bescii . Conclusions Feedstock recalcitrance can be significantly reduced with transgenic plants, but finding the correct modification may require a large sample set to identify the most advantageous genetic modifications for the feedstock. Utilizing C. bescii as a screening assay for recalcitrance, poplar lines with down-regulation of coumarate 3-hydroxylase 3 (C3H3) resulted in the highest degrees of solubilization and conversion by C. bescii . One such line, with a growth phenotype similar to the wild-type, generated more than three times the fermentation products of the wild-type poplar control, suggesting that excellent digestibility can be achieved without compromising fitness of the tree.

09 BIOMASS FUELS↗

Field evaluation of transgenic hybrid poplars with desirable wood properties and enhanced growth for biofuel production by bicistronic expression of PdGA20ox1 and PtrMYB3 in wood-forming tissue

Background: To create an ideotype woody bioenergy crop with desirable growth and biomass properties, we utilized the viral 2A-meidated bicistronic expression strategy to express both PtrMYB3 ( MYB46 ortholog of Populus trichocarpa , a master regulator of secondary wall biosynthesis) and PdGA20ox1 (a GA20-oxidase from Pinus densiflora that produces gibberellins) in wood-forming tissue (i.e., developing xylem). Results: Transgenic Arabidopsis plants expressing the gene construct DX15::PdGA20ox1-2A-PtrMYB3 showed a significant increase in both stem fresh weight (threefold) and secondary wall thickening (1.27-fold) relative to wild-type (WT) plants. Transgenic poplars harboring the same gene construct grown in a greenhouse for 60 days had a stem fresh weight up to 2.6-fold greater than that of WT plants. In a living modified organism (LMO) field test conducted for 3 months of active growing season, the stem height and diameter growth of the transgenic poplars were 1.7- and 1.6-fold higher than those of WT plants, respectively, with minimal adverse growth defects. Although no significant changes in secondary wall thickening of the stem tissue of the transgenic poplars were observed, cellulose content was increased up to 14.4 wt% compared to WT, resulting in improved saccharification efficiency of the transgenic poplars. Moreover, enhanced woody biomass production by the transgenic poplars was further validated by re-planting in the same LMO field for additional two growing seasons. Conclusions: Taken together, these results show considerably enhanced wood formation of our transgenic poplars, with improved wood quality for biofuel production.

09 BIOMASS FUELS↗

Genetic identification of 91 poplar cultivars based on SSR markers

Accurate genetic identification and relationship analysis of poplar cultivars is necessary to establish commercial poplar plantations and select suitable breeding strategies. In this study, 91 poplar cultivars belonging to four sections (Aigeiros, Tacamahaca, Populus and Turanga) and inter/intra-sectional hybrids were genotyped using 18 polymorphic simple sequence repeat (SSR) markers. In total, 222 alleles were amplified with an average of 12.3 alleles per marker. The mean polymorphic information content and power of discrimination were 0.706 and 0.813, respectively. Five SSR markers (ORPM_103, ORPM_247, GCPM_1048, GCPM_1255 and LG_X_19) constituted a core fingerprint and were sufficient to identify all the tested cultivars. With some notable exceptions, cultivars of the same species generally clustered together in cluster (UPGMA) and ordination (PCO) analyses. Flow cytometry indicated that 11 poplar cultivars were triploid. Among these, seven had three alleles at some loci, suggesting that SSR markers could indicate the ploidy level to some extent. This study provides useful genetic information for the identification and protection of poplar cultivars in China and offers a guideline for the selection of poplar crossing parents based on ploidy level and genetic relationships.

59 BASIC BIOLOGICAL SCIENCES↗

Elucidating Mechanisms of Rust Pathogenesis for Engineering Resistance in Poplar

The overall objective of this research was to elucidate the molecular basis for the virulence of Melampsora larici-populina towards Populus spp. in order to address the formidable challenge of engineering durable resistance in poplar against leaf rust. Our original research plan was built on the technically innovative approach of using dual host systems and known host defense proteins and pathogen avirulent effectors along with genome-wide approaches to identify key pathogenesis effectors of Melampsora larici-populina that target poplar defense and nutrient acquisition. Our efforts focused on developing high-throughput experimental approaches and systems to enable us to address our overarching hypothesis that rust effectors with homology to other fungal effectors will either bypass the poplar immune system and/or suppress effector-triggered immunity through conserved mechanisms, and that an analysis of these interaction networks would provide for new approaches to develop rust-resistant poplar. Three main objectives were originally proposed: (1) mine key rust effectors that suppress host immunity; (2) elucidate interaction networks in poplar targeted by key rust effectors; (3) generate transgenic poplar resources for identification of genes involved in pathogen infection or host defense triggered by rust. Generally, we worked towards all three objectives by developing and applying complementary molecular, genomic, biochemical and plant transformation approaches.

54 ENVIRONMENTAL SCIENCES↗

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↗

Genetically engineered poplar wood effectively enhances the efficiency of deep eutectic solvent-mediated one-pot processing

Although lignocellulosic biomass is a renewable resource with the potential to replace fossil-derived fuels and chemicals, its recalcitrance, largely due to lignin, limits its utilization. Recent advancements in genetic engineering have produced transgenic trees with reduced lignin content and/or modified lignin structure without compromising growth traits. Here, three engineered poplar varieties are evaluated as feedstocks using a biocompatible one-pot deep eutectic solvent-mediated process that integrates biomass fractionation and enzymatic saccharification within a single reactor, eliminating water washing and reconditioning. All transgenic poplars exhibit higher fermentable sugar yields than wild-type (WT) trees. Notably, QsuB poplar, incorporating 3,4-dihydroxybenzoate in lignin, achieves the highest glucose conversion yield of 91.3% (vs. 73.0% from WT). AT5 and MdCHS3 poplars, incorporating ferulate esters and naringenin, also demonstrate improved glucose yields (86.7 and 84.7%, respectively), confirming reduced biomass recalcitrance. Additionally, residual lignins are valorized via hydrogenolysis into phenolic compounds, with comparable alkylphenol production across all lines. These findings demonstrate that the transgenic poplar lines not only serve as superior feedstocks for sugar conversion but also provide a rich resource for phenolic compound production, enhancing the operational and economic viability of integrated biorefinery processes.

biomasses↗

Effect of Dilute Acetic Acid Hydrolysis on Xylooligosaccharide Production and the Inhibitory Effect of Cellulolytic Enzyme Lignin from Poplar

Acetic acid (AC) hydrolysis has been reported to prepare xylooligosaccharides (XOS) from poplar. However, the influence of AC hydrolysis on the lignin structure changes is not clear, which is important for the following enzymatic hydrolysis of poplar. Herein, AC was used to produce XOS, and cellulase adsorption on cellulolytic enzyme lignin (CEL) from AC-hydrolyzed poplar and its inhibitory effect on two commercial cellulase preparations were investigated. AC hydrolysis gave a XOS yield of 39.8% from poplar. After AC hydrolysis at 170 °C, the hydrophobicity and ζ-potential of CEL decreased to 2.3 L/g and 14.8 mV, respectively. The adsorption strength of CTec2 on CEL samples did not increase by AC hydrolysis, and the inhibitory effect of CEL on Celluclast 1.5L and β-glucosidase was observed, but not on CTec2. CEL samples improved the lytic polysaccharide monooxygenase (LPMO) activity of the enzymatic hydrolysis by CTec2. After CEL samples were added in enzymatic hydrolysis, the free filter paper activity of Celluclast 1.5L and β-G retained in the enzymatic hydrolysate decreased from 60.5 to 29.3–42.9%. The addition of CEL samples in enzymatic hydrolysis could not decrease the free filter paper activity of CTec2 retained in the enzymatic hydrolysate. In the enzymatic hydrolysis with CEL samples, higher glucose yields were obtained by CTec2 than those by Celluclast 1.5L and β-glucosidase. This work will help to understand the structure and inhibitory effects of AC-CELs and guide the development of AC hydrolysis for the production of XOS and monosaccharides from poplar.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Investigating the Role of Extensin Proteins in Poplar Biomass Recalcitrance

The biological conversion of cellulosic biomass to biofuel is hindered by cell wall recalcitrance, which can limit the ability of cellulases to access and break down cellulose. The purpose of this study was to investigate whether hydroxyproline-rich cell wall proteins (extensins) are present in poplar stem biomass, and whether these proteins may contribute to recalcitrance. Three classical extensin genes were identified in Populus trichocarpa through bioinformatic analysis of poplar genome sequences, with the following proposed names: PtEXTENSIN1 (Potri.001G019700); PtEXTENSIN2 (Potri.001G020100); PtEXTENSIN3 (Potri.018G050100). Tissue print immunoblots localized the extensin proteins in poplar stems to regions near the vascular cambium. Different thermochemical pretreatments reduced but did not eliminate hydroxyproline (Hyp, a proxy for extensins) from the biomass. Protease treatment of liquid hot water-pretreated poplar biomass reduced Hyp content by a further 16% and increased subsequent glucose yield by 20%. These data suggest that extensins may contribute to recalcitrance in pretreated poplar biomass, and that incorporating protease treatment into pretreatment protocols could result in a small but significant increase in the yield of fermentable glucose.

09 BIOMASS FUELS↗

Two-dimensional heteronuclear single quantum coherence (HSQC) NMR spectra of lignin isolated from field grown transgenic poplar

Here we present a curated dataset of a series of two-dimensional heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectra of lignin isolated from a field grown transgenic poplar engineered with a monolignol 4-O-methyltransferase (MOMT4). The poplar was collected from a 2-year-old rotation trees within a three-year field trial experiment. The poplar was Soxhlet-extracted with toluene/ethanol and the extractives-free poplar was then ball-milled in a Retsch PM100 planetary ball mill using a porcelain jar with ceramic balls at 600 rpm for 2 h (in 5 min on and 5 min off cycles to avoid excessive sample heating). The ball-milled materials were then subjected to enzymatic hydrolysis for 48 h followed by centrifugation and washing with deionized water. The solid residue was extracted twice with 96:4 (v/v) 1,4-dioxane/water mixture at room temperature overnight. The extracts were combined, rotary evaporated, and freeze-dried to recover the lignin. The dry lignin samples were dissolved in deuterated dimethyl sulfoxide for NMR experiments. 13C–1H HSQC experiments were performed in a Bruker Avance III HD 500 MHz NMR spectrometer operating at a frequency of 125.12 MHz for the 13C nucleus using a standard Bruker pulse sequence (hsqcetgpsisp2.2) on a Prodigy platform cryoprobe. The NMR spectra were acquired under the following acquisition conditions: 220 ppm spectral width in F1 (13C) dimension with 256 data points and 12 ppm spectral width in F2 (1H) dimension with 1024 data points, a 90° pulse, a one bond C–H coupling constant of 145 Hz, a 1.0 s pulse delay, and 64 scans. All the data was processed using the Bruker’s TopSpin 3.6 software. The NMR spectra provides structural characteristics information about lignin in field grown transgenic MOMT4 poplar. Additional meta data is embedded in the raw spectra figures.

Lignin structure, HSQC, poplar, field trial, MOMT4↗

Proton NMR spectra of lignin isolated from field grown transgenic poplar

Here we present a curated dataset of a series of 1H nuclear magnetic resonance (NMR) spectra of lignin isolated from transgenic monolignol 4-O-methyltransferase (MOMT4) engineered poplar. The transgenic poplar was collected from a 2-year-old rotation trees within a three-year field trial experiment. Two replicates were collected for each transgenic poplar for the 1H NMR analysis. The poplar samples were Soxhlet-extracted with toluene/ethanol to remove the extractives and the extractives-free poplar was then ball-milled in a Retsch PM100 planetary ball mill using a porcelain jar with ceramic balls at 600 rpm for 2 h. The ball-milled materials were subjected to enzymatic hydrolysis for 48 h followed by centrifugation and washing with deionized water. The solid residue was extracted twice with 96:4 (v/v) 1,4-dioxane/water mixture at room temperature overnight. The extracts were combined, rotary evaporated, and freeze-dried to recover lignin. The dry lignin samples were dissolved in deuterated dimethyl sulfoxide and transferred into a 5 mm NMR tube. 1H NMR experiments were performed in a Bruker Avance III HD 500 MHz NMR spectrometer operating at a frequency of 125.12 MHz for the 13C nucleus using a standard Bruker pulse sequence (zg) on a Prodigy platform cryoprobe. The NMR spectra were acquired with 16 ppm spectra width, 32k data points, 3s pulse delay, and 16 scans. All the data was processed using the Bruker’s TopSpin 3.6 software. Additional meta data is embedded in the raw spectra files.

1H NMR, lignin, poplar, field trial, MOMT4, CBI↗