Reduced Lignin Content Research Articles

Influence of drying temperature on the properties of Colombian banana fibers for its potential use as reinforcement in composite materials

This study investigated the impact of drying temperature on the physicochemical and mechanical properties of banana pseudostem fibers sourced from the Cordoba region in Colombia. Banana fibers (BFs) were extracted through mechanical decortication from the banana pseudostem (BP) of the plant and subsequently oven-dried at temperatures of 40 °C and 90 °C. Six mathematical models were employed to analyze the drying behavior of the fibers. The density of the BFs was determined using the apparent density method, and their chemical composition was evaluated via bromatological analysis. Fiber diameter was measured using optical microscopy (OM). The BF samples were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG), contact angle measurements, and tensile testing. The results indicated that noncellulosic materials were removed from the fibers when dried at 90 °C, as evidenced by alterations in thermal degradation and fiber surface morphology observed through TG and SEM, suggesting a reduction in lignin content. While drying temperature did not affect fiber stiffness or ductility, a correlation with fiber diameter was noted. Thinner fibers, ranging from 148 to 250 μm, exhibited increased tensile strength and Young’s modulus, attributed to a more compact microfibril arrangement.

Investigating the Variation between Lignin Content and the Fracture Characteristics in Capsicum annuum Mutant Stems

The cultivation of horticultural plants in controlled greenhouse environments is a pivotal practice in modern agriculture, offering the potential to enhance crop productivity and mitigate climate change effects. This study investigates the biomechanical properties and lignin content of various Capsicum annuum mutant lines—‘fragile-plant’ (frx), ‘tortuous internodi’ (tti), and ‘puffy-structured stem’ (pfi)—in comparison to a commercially established variety, ‘Garai Fehér’. We employed the acetyl bromide method to quantify lignin content and conducted three-point bending tests to assess rigidity in three distinct regions of the stem. Gene expression analysis of key lignin biosynthetic pathway genes (PAL, C4H, 4CL, CCoAOMT, CAD) was performed using qRT-PCR. The results revealed significant differences in lignin content and breaking force among the genotypes and stem regions. The tti mutants exhibited similar lignin content to the control but lower breaking strength, likely due to elongated internodes. The frx mutants showed uniformly reduced lignin content, correlating with their fragile stems. The pfi mutants displayed abnormally high lignin content in the top region yet demonstrated the lowest stem rigidity in every region. Overexpression of CAD and CCoAOMT was detected in the mutants in specific regions of the stem, suggesting alterations in lignin biosynthesis; however, we could not confirm the correlation between them. Our findings indicate that while lignin content generally correlates with stem rigidity, this trait is complex and influenced by more factors.

Impact of phenology, brown midrib (BMR), seed treatment, and herbivory on epicuticular wax content and fall armyworm (Spodoptera frugiperda) feeding in sorghum-sudangrass (Sorghum xdrummondii)

In crop production, management strategies have historically relied upon chemical control to mitigate insect pest pressures. As resistance to pesticides grows at an alarming rate, and as non-target effects are found more frequently, the search for sustainable, novel pest management strategies has become more important. Plants are equipped with diverse physical and chemical defenses against insect pests. Among these physical defenses, epicuticular waxes serve as an important first line of defense. Yet, how these chemical control methods such as seed treatment affects epicuticular waxes is underexplored. Brown midrib (BMR) is a host-plant trait from a recessive gene that affects the monolignol biosynthetic pathway, reducing lignin content, and presents as a gene of interest for evaluation as a form of pest management. Sorghum-sudangrass, Sorghum x drummondii, is an economically important forage crop with BMR; however, the effects of BMR have primarily been studied in animal agriculture and its effects on insects is largely unknown. In addition, how BMR affects epicuticular waxes is also unknown. This study aims to understand how the quantity of epicuticular wax in Sorghum x drummondii is affected by chemical control (seed treatments), BMR, phenological stages, and how wax affects a polyphagous, destructive pest, fall armyworm, Spodoptera frugiperda (FAW). Through wax quantification and FAW feeding induction and wax added artificial diet experiments, we show that while BMR and seed treatment does not affect wax content, plant phenology affects wax, and wax is induced by FAW feeding and has negative consequences for caterpillar growth.

Negative regulation of CcPAL2 gene expression by the repressor transcription factor CcMYB4-12 modulates lignin and capsaicin biosynthesis in Capsicum chinense fruits

Peppers globally renowned for their distinctive spicy flavor, have attracted significant research attention, particularly in understanding spiciness regulation. While the activator MYB's role in spiciness regulation is well-established, the involvement of repressor MYB factors remains unexplored. This study identified the MYB4 transcription factor through RNA-seq and genome-wide analysis as being associated with spiciness. Consequently, CcMYB4-2 and CcMYB4-12 were cloned from Hainan Huangdenglong peppers, both exhibiting nuclear subcellular localization. qRT-PCR analysis revealed that CcMYB4-2/4-12 had high expression levels during the accumulation period of capsaicin, but there were differences in their peak expression levels, which may be related to the formation of pepper spiciness. Heterologous expression in Arabidopsis thaliana resulted in significantly elevated CcMYB4-2/4-12 expression levels and reduced lignin content. In CcMYB4-2 silenced plants, PAL expression remained unchanged, while PAL expression significantly increased in CcMYB4-12 silenced plants, leading to elevated lignin content and reduced capsaicin content. Yeast one-hybrid (Y1H) and dual luciferase reporter assays (DLR) demonstrated that CcMYB4-2/4-12 inhibited the transcription of CcPAL2 by binding to its promoter. Notably, CcMYB4-12 exhibited more pronounced inhibition. Therefore, it is hypothesized that CcMYB4-12 plays a pivotal role in regulating lignin and capsaicin biosynthesis. This study elucidates the molecular mechanism of MYB4 binding to the PAL promoter, providing a foundational understanding for analyzing phenylpropanoid metabolism and its diverse branches. Key messageThrough functional verification analysis of the repressor CcMYB4, transcriptional regulation experiments revealed that CcMYB4 can bind to the CcPAL2 promoter, negatively regulating the capsaicin biosynthesis in Capsicum chinense fruits.

Overexpression of SlALC Increases Drought and Salt Tolerance and Affects Fruit Dehiscence in Tomatoes.

The bHLH transcription factors are important plant regulators against abiotic stress and involved in plant growth and development. In this study, SlALC, a gene coding for a prototypical DNA-binding protein in the bHLH family, was isolated, and SlALC-overexpression tomato (SlALC-OE) plants were generated by Agrobacterium-mediated genetic transformation. SlALC transgenic lines manifested higher osmotic stress tolerance than the wild-type plants, estimated by higher relative water content and lower water loss rate, higher chlorophyll, reducing sugar, starch, proline, soluble protein contents, antioxidant enzyme activities, and lower MDA and reactive oxygen species contents in the leaves. In SlALC-OE lines, there were more significant alterations in the expression of genes associated with stress. Furthermore, SlALC-OE fruits were more vulnerable to dehiscence, with higher water content, reduced lignin content, SOD/POD/PAL enzyme activity, and lower phenolic compound concentrations, all of which corresponded to decreased expression of lignin biosynthetic genes. Moreover, the dual luciferase reporter test revealed that SlTAGL1 inhibits SlALC expression. This study revealed that SlALC may play a role in controlling plant tolerance to drought and salt stress, as well as fruit lignification, which influences fruit dehiscence. The findings of this study have established a foundation for tomato tolerance breeding and fruit quality improvement.

Function Analysis of a Maize Endo-1,4-β-xylanase Gene ZmHSL in Response to High-Temperature Stress.

Rising temperature is a major threat to the normal growth and development of maize, resulting in low yield production and quality. The mechanism of maize in response to heat stress remains uncertain. In this study, a maize mutant Zmhsl-1 (heat sensitive leaves) with wilting and curling leaves under high temperatures was identified from maize Zheng 58 (Z58) mutant lines generated by ethyl methanesulfonate (EMS) mutagenesis. The Zmhsl-1 plants were more sensitive to increased temperature than Z58 in the field during growth season. The Zmhsl-1 plants had lower plant height, lower yield, and lower content of photosynthetic pigments. A bulked segregant analysis coupled with whole-genome sequencing (BSA-seq) enabled the identification of the corresponding gene, named ZmHSL, which encodes an endo-β-1,4-xylanase from the GH10 family. The loss-of-function of ZmHSL resulted in reduced lignin content in Zmhsl-1 plants, leading to defects in water transport and more severe leaf wilting with the increase in temperature. RNA-seq analysis revealed that the differentially expressed genes identified between Z58 and Zmhsl-1 plants are mainly related to heat stress-responsive genes and unfolded protein response genes. All these data indicated that ZmHSL plays a key role in lignin synthesis, and its defective mutation causes changes in the cell wall structure and gene expression patterns, which impedes water transport and confers higher sensitivity to high-temperature stress.

Identification and Analysis of the Mechanism of Stem Mechanical Strength Enhancement for Maize Inbred Lines QY1.

Enhancing stalk strength is a crucial strategy to reduce lodging. We identified a maize inbred line, QY1, with superior stalk mechanical strength. Comprehensive analyses of the microstructure, cell wall composition, and transcriptome of QY1 were performed to elucidate the underlying factors contributing to its increased strength. Notably, both the vascular bundle area and the thickness of the sclerenchyma cell walls in QY1 were significantly increased. Furthermore, analyses of cell wall components revealed a significant increase in cellulose content and a notable reduction in lignin content. RNA sequencing (RNA-seq) revealed changes in the expression of numerous genes involved in cell wall synthesis and modification, especially those encoding pectin methylesterase (PME). Variations in PME activity and the degree of methylesterification were noted. Additionally, glycolytic efficiency in QY1 was significantly enhanced. These findings indicate that QY1 could be a valuable resource for the development of maize varieties with enhanced stalk mechanical strength and for biofuel production.

Characterization of cellulose nanocrystals from Zhombwe (Neorautanenia brachypus (harms) CA Sm.) bagasse.

Increased awareness of environmental pollution has changed focus to the use of biodegradable materials because they lack persistence in the environment. This article focused on the production of cellulose nanocrystals from Zhombwe, Neorautanenia brachypus (Harms) CA Sm. bagasse using steam explosion, alkaline treatment, bleaching, purification, and acid hydrolysis. The chemical composition after the treatments was determined using TAPPI standards. Further characterization was done using x-ray Diffraction (XRD), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA). The nanoscale dimensions and morphology of the extracted nanocrystals was determined through field emission scanning electron microscopy (FE-SEM). FTIR spectroscopy and DSC confirmed the removal of noncellulosic compounds. XRD revealed that N. brachypus bagasse contained cellulose type I, which partly endured morphological change to polymorph II after purification and hydrolysis. FE-SEM revealed elliptical to rod-shaped structures after acid hydrolysis, which had a mean length and width of 1103 nm and 597 nm respectively. TAPPI tests revealed that successive chemical treatments increased crystallinity by 29.7%, enriched cellulose content by 74.2%, reduced lignin content by 21.7%, and reduced hemicellulose to less than 1%. The semicrystalline nature of the material produced in our work is a promising candidate for swelling hydrogel applications in areas such as wound dressing, heavy metal removal, controlled drug delivery, agriculture, and sanitary products. Future studies may focus on surface modification of nanocrystals to improve their thermal stability and therefore expand their range for potential industrial applications.

Expression and function analysis of phenylalanine ammonia-lyase genes involved in Bamboo lignin biosynthesis.

Bamboo, renowned as the fastest-growing plant globally, matures within an astonishingly short period of 40-50 days from shoots, reaching heights of 10-20 meters. Moreover, it can be harvested for various uses within 3-5 years. Bamboo exhibits exceptional mechanical properties, characterized by high hardness and flexibility, largely attributed to its lignin content. Phenylalanine ammonia-lyase (PAL) catalyzes the crucial initial step in lignin biosynthesis, but its precise role in bamboo lignification processes remains elusive. Thus, elucidating the functions of PAL genes in bamboo lignification processes is imperative for understanding its rapid growth and mechanical strength. Here, we systematically identified and classified PAL genes in Moso bamboo, ensuring nomenclature consistency across prior studies. Subsequently, we evaluated PAL gene expression profiles using publicly available transcriptome data. The downregulation of PePALs expression in Moso bamboo through in planta gene editing resulted in a decrease in PAL activity and a subsequent reduction in lignin content. In contrast, overexpression of PePAL led to enhanced PAL activity and an increase in lignin content. These findings highlight the critical role of PAL in the lignin biosynthesis process of Moso bamboo, which will help to unravel the mechanism underpinning bamboo's rapid growth and mechanical strength, with a specific emphasis on elucidating the functions of PAL genes.

Salt stress alters the cell wall components and structure in Miscanthus sinensis stems.

Miscanthus is a perennial grass suitable for the production of lignocellulosic biomass on marginal lands. The effects of salt stress on Miscanthus cell wall composition and its consequences on biomass quality have nonetheless received relatively little attention. In this study, we investigated how exposure to moderate (100 mM NaCl) or severe (200 mM NaCl) saline growing conditions altered the composition of both primary and secondary cell wall components in the stems of 15 Miscanthus sinensis genotypes. The exposure to stress drastically impacted biomass yield and cell wall composition in terms of content and structural features. In general, the observed compositional changes were more pronounced under severe stress conditions and were more apparent in genotypes with a higher sensitivity towards stress. Besides a severely reduced cellulose content, salt stress led to increased pectin content, presumably in the form of highly branched rhamnogalacturonan type I. Although salt stress had a limited effect on the total lignin content, the acid-soluble lignin content was strongly increased in the most sensitive genotypes. This effect was also reflected in substantially altered lignin structures and led to a markedly reduced incorporation of syringyl subunits and p-coumaric acid moieties. Interestingly, plants that were allowed a recovery period after stress ultimately had a reduced lignin content compared to those continuously grown under control conditions. In addition, the salt stress-induced cell wall alterations contributed to an improved enzymatic saccharification efficiency.

EuMYB308 regulates lignin accumulation by targeting EuLAC17 in Eucalyptus urophylla

It is known that laccase genes are involved in lignin synthesis. Eucalyptus is an important timber species; functional studies of specific laccase members and their upstream regulatory factors in Eucalyptus are incomplete, and the mechanism of lignin content variation in polyploid plants remains unclear. Therefore, in this study, we investigated the function of EuLAC17 by overexpressing it in tobaccos and the result showed the lignin content in transgenic tobacco increased significantly. Further, yeast one hybrid sequencing was used to identify a series of potential regulatory factors of EuLAC17, among which EuMYB308 showed the most significant correlation with EuLAC17. Yeast one hybrid, dual-luciferase reporter gene assay and electrophoretic mobility shift assay jointly verified that EuMYB308 could directly target the EuLAC17 promoter and repress its expression. In addition, overexpression of EuMYB308 resulted in significant reductions in plant growth, xylem cell lumen area, cell wall thickness and lignin content. Transcriptome sequencing showed that lignin biosynthesis genes including laccase genes were significantly differential expression in transgenic plants compared with wild-type plants. In triploid Eucalyptus urophylla, the expression of EuMYB308 was significantly upregulated, which may enhance the inhibition of EuLAC17 and thus reduce its expression level in triploid, which is one possible reason for the reduced lignin content in triploid E. urophylla. This study provides important insights into transcriptional regulation of lignin biosynthesis in Eucalyptus, and also contributes to revealing the genetic mechanism of lignin content variation in polyploid plants, which are of great significance for promoting the improvement of wood quality in forest trees and polyploid breeding.

Nutritional upgrade of olive mill stone waste, walnut shell and their mixtures by applying solid state fermentation initiated by Pleurotus ostreatus

Present study concerns the transformation of the agro-industrial by-products olive mill stone waste (OMSW) and walnut shell (WS) to a protein-enriched animal feedstuff utilizing the solid state fermentation (SSF) technique. For this purpose, various mixtures of these by-products were exploited as substrates of the SSF process which was initiated by the P. ostreatus fungus. The respective results indicated that the substrate consisted of 80% WS and 20% OMSW afforded the product with the highest increase in protein content, which accounted the 7.57% of its mass (69.35% increase). In addition, a 26.13% reduction of lignin content was observed, while the most profound effect was observed for their 1,3–1,6 β-glucans profile, which was increased by 3-folds reaching the 6.94% of substrate’s mass. These results are indicative of the OMSW and WS mixtures potential to act as efficient substrate for the development of novel proteinaceous animal feed supplements using the SSF procedure. Study herein contributes to the reintegration of the agro-industrial by-products aiming to confront the problem of proteinaceous animal feed scarcity and reduce in parallel the environmental footprint of the agro-industrial processes within the context of circular economy.

Plasmonics nanorod biosensor for in situ intracellular detection of gene expression biomarkers in intact plant systems

The intracellular developmental processes in plants, particularly concerning lignin polymer formation and biomass production are regulated by microRNAs (miRNAs). MiRNAs including miR397b are important for developing efficient and cost-effective biofuels. However, traditional methods of monitoring miRNA expression, like PCR, are time-consuming, require sample extraction, and lack spatial and temporal resolution, especially in real-world conditions. We present a novel approach using plasmonics nanosensing to monitor miRNA activity within living plant cells without sample extraction. Plasmonic biosensors using surface-enhanced Raman scattering (SERS) detection offer high sensitivity and precise molecular information. We used the Inverse Molecular Sentinel (iMS) biosensor on unique silver-coated gold nanorods (AuNR@Ag) with a high-aspect ratio to penetrate plant cell walls for detecting miR397b within intact living plant cells. MiR397b overexpression has shown promise in reducing lignin content. Thus, monitoring miR397b is essential for cost-effective biofuel generation. This study demonstrates the infiltration of nanorod iMS biosensors and detection of non-native miRNA 397b within plant cells for the first time. The investigation successfully demonstrates the localization of nanorod iMS biosensors through TEM and XRF-based elemental mapping for miRNA detection within plant cells of Nicotiana benthamiana. The study integrates shifted-excitation Raman difference spectroscopy (SERDS) to decrease background interference and enhance target signal extraction. In vivo SERDS testing confirms the dynamic detection of miR397b in Arabidopsis thaliana leaves after infiltration with iMS nanorods and miR397b target. This proof-of-concept study is an important stepping stone towards spatially resolved, intracellular miRNA mapping to monitor biomarkers and biological pathways for developing efficient renewable biofuel sources.

Expression of dehydroshikimate dehydratase in poplar induces transcriptional and metabolic changes in the phenylpropanoid pathway.

Modification of lignin in feedstocks via genetic engineering aims to reduce biomass recalcitrance to facilitate efficient conversion processes. These improvements can be achieved by expressing exogenous enzymes that interfere with native biosynthetic pathways responsible for the production of the lignin precursors. In planta expression of a bacterial 3-dehydroshikimate dehydratase in poplar trees reduced lignin content and altered the monomer composition, which enabled higher yields of sugars after cell wall polysaccharide hydrolysis. Understanding how plants respond to such genetic modifications at the transcriptional and metabolic levels is needed to facilitate further improvement and field deployment. In this work, we acquired fundamental knowledge on lignin-modified poplar expressing 3-dehydroshikimate dehydratase using RNA-seq and metabolomics. The data clearly demonstrate that changes in gene expression and metabolite abundance can occur in a strict spatiotemporal fashion, revealing tissue-specific responses in the xylem, phloem, or periderm. In the poplar line that exhibited the strongest reduction in lignin, we found that 3% of the transcripts had altered expression levels and ~19% of the detected metabolites had differential abundance in the xylem from older stems. The changes affected predominantly the shikimate and phenylpropanoid pathways as well as secondary cell wall metabolism, and resulted in significant accumulation of hydroxybenzoates derived from protocatechuate and salicylate.

Ectopic Production of 3,4-Dihydroxybenzoate in Planta Affects Cellulose Structure and Organization.

Lignocellulosic biomass is a highly sustainable and largely carbon dioxide neutral feedstock for the production of biofuels and advanced biomaterials. Although thermochemical pretreatment is typically used to increase the efficiency of cell wall deconstruction, genetic engineering of the major plant cell wall polymers, especially lignin, has shown promise as an alternative approach to reduce biomass recalcitrance. Poplar trees with reduced lignin content and altered composition were previously developed by overexpressing bacterial 3-dehydroshikimate dehydratase (QsuB) enzyme to divert carbon flux from the shikimate pathway. In this work, three transgenic poplar lines with increasing QsuB expression levels and different lignin contents were studied using small-angle neutron scattering (SANS) and wide-angle X-ray scattering (WAXS). SANS showed that although the cellulose microfibril cross-sectional dimension remained unchanged, the ordered organization of the microfibrils progressively decreased with increased QsuB expression. This was correlated with decreasing total lignin content in the QsuB lines. WAXS showed that the crystallite dimensions of cellulose microfibrils transverse to the growth direction were not affected by the QsuB expression, but the crystallite dimensions parallel to the growth direction were decreased by ∼20%. Cellulose crystallinity was also decreased with increased QsuB expression, which could be related to high levels of 3,4-dihydroxybenzoate, the product of QsuB expression, disrupting microfibril crystallization. In addition, the cellulose microfibril orientation angle showed a bimodal distribution at higher QsuB expression levels. Overall, this study provides new structural insights into the impact of ectopic synthesis of small-molecule metabolites on cellulose organization and structure that can be used for future efforts aimed at reducing biomass recalcitrance.

The DEP1 Mutation Improves Stem Lodging Resistance and Biomass Saccharification by Affecting Cell Wall Biosynthesis in Rice

BackgroundPlant cell walls have evolved precise plasticity in response to environmental stimuli. The plant heterotrimeric G protein complexes could sense and transmit extracellular signals to intracellular signaling systems, and activate a series of downstream responses. dep1 (Dense and Erect Panicles 1), the gain-of-function mutation of DEP1 encoding a G protein γ subunit, confers rice multiple improved agronomic traits. However, the effects of DEP1 on cell wall biosynthesis and wall-related agronomic traits remain largely unknown.ResultsIn this study, we showed that the DEP1 mutation affects cell wall biosynthesis, leading to improved lodging resistance and biomass saccharification. The DEP1 is ubiquitously expressed with a relatively higher expression level in tissues rich in cell walls. The CRISPR/Cas9 editing mutants of DEP1 (dep1-cs) displayed a significant enhancement in stem mechanical properties relative to the wild-type, leading to a substantial improvement in lodging resistance. Cell wall analyses showed that the DEP1 mutation increased the contents of cellulose, hemicelluloses, and pectin, and reduced lignin content and cellulose crystallinity (CrI). Additionally, the dep1-cs seedlings exhibited higher sensitivity to cellulose biosynthesis inhibitors, 2,6-Dichlorobenzonitrile (DCB) and isoxaben, compared with the wild-type, confirming the role of DEP1 in cellulose deposition. Moreover, the DEP1 mutation-mediated alterations of cell walls lead to increased enzymatic saccharification of biomass after the alkali pretreatment. Furthermore, the comparative transcriptome analysis revealed that the DEP1 mutation substantially altered expression of genes involved in carbohydrate metabolism, and cell wall biosynthesis.ConclusionsOur findings revealed the roles of DEP1 in cell wall biosynthesis, lodging resistance, and biomass saccharification in rice and suggested genetic modification of DEP1 as a potential strategy to develop energy rice varieties with high lodging resistance.

Hydrothermal enzyme pretreatment on structure and properties of apple branches

Abstract Pretreatment serves as an important step in maximizing the efficient utilization of woody biomass feedstock. This study focused on enhancing the component and structural properties of apple wood to optimize the utilization of apple wood resources effectively. This study presents the optimization of, the apple wood pretreatment system, combining bioenzyme and hydrothermal treatments. This study aimed to investigate the effect of different bioenzyme pretreatment conditions on the components and structural properties of apple wood extract and to analyze the feasibility of enzyme solution reuse. Fourier transform infrared spectroscopy analysis revealed an increase in cellulose content in the wood flour following enzyme treatment. Concurrently, changes in characteristic peaks indicated an increased hemicellulose content and a reduction in lignin content after the pretreatment, correlating with fraction analysis results. Scanning electron microscopy illustrated that the dense structure of the fiber surface was disrupted after treatment. Notably, the results indicated that the recovered biocomposite enzymes exhibited excellent activity, suggesting their potential for reuse.

One-pot extraction of nanocellulose from raw durian husk fiber using carboxylic acid-based deep eutectic solvent with in situ ultrasound assistance

Nanocellulose (CNF) has emerged as a promising alternative to synthetic petroleum-based polymers, but the conventional preparation process involves multiple tedious steps, heavily dependent on chemical input, and proves cost-inefficient. This study presented an, in situ ultrasound-assisted extraction using deep eutectic solvent (DES) based on choline chloride and oxalic acid for more facile production of CNF from raw durian husk fibers. FESEM analysis confirmed the successful extraction of web-like nanofibril structure with width size ranging from 18 to 26 nm. Chemical composition analysis and FTIR revealed the selective removal of lignin and hemicellulose from the raw fiber. As compared to post-ultrasound treatment, in situ ultrasound-assisted extraction consistently outperforms, yielding a higher CNF yield with finer fiber width and significantly reduced lignin content. Integrating this eco-friendly in situ ultrasonication-assisted one-pot extraction method with a 7.5 min interval yielded the highest CNF yield of 58.22 % with minimal lignin content. The superior delignification ability achieved through the proposed in situ ultrasound-assisted protocol surpasses the individual efficacy of DES and ultrasonication processes, neither of which yielded CNF in our experimental setup. This single-step fabrication process significantly reduces chemical usage and streamlines the production steps yielding web-structured CNF that is ideal for sustainable application in membrane and separator.

Effect of partial delignification and densification on chemical, morphological, and mechanical properties‏ of wood: Structural property evolution

Efforts have been dedicated to developing sustainable alternatives for conventional construction materials, resulting in the development of densified wood as an innovative, environmentally friendly, and high-performance material. This study focuses on investigating the effects of treatment parameters (e.g., partial delignification boiling time, chemical concentrations, and hot-pressing pressures) on the chemical, morphological, and mechanical properties of the densified wood. Results revealed that the partial delignification process of low-density Balsa wood reduced lignin content by 50–80%, which enhanced the crystallinity of the wood by 21–54%. However, prolonged periods of partial delignification caused a degradation in the crystalline regions in wood. The modulus of rupture (MOR) and modulus of elasticity (MOE) of the densified wood samples increased by 90–680% and 10–810% compared with those of the raw wood samples, respectively. The MOR of the densified wood also showed a dependency on lignin content and porosity. An optimal lignin content of approximately 10% was associated with the highest MOR in densified wood. Moreover, the relationship between the MOR and porosity suggested that the MOR improvement occurred in two stages. The first stage of MOR improvement is controlled by the reduction in the porosity and the second stage is controlled by the improved hydrogen bonding among cellulose nanofibrils.

Genome-wide analysis of the PME gene family reveals its role in suppressing fruit lignification in pear

In pears, the presence of stone cells adversely affects fruit quality. Pectin methylesterase (PME) plays various roles in plant biology, including lignin biosynthesis. However, only a limited fraction has been functionally characterized, and the distribution and function of PME in many Rosaceae trees remain unexplored. In this study, we identified 396 putative PME family candidate genes, with 81 in Pyrus bretschneideri, 92 in Malus domestica, 62 in Fragaria vesca, 65 in Prunus mume, 15 in Pyrus communis, and 81 in Pyrus pyrifolia. Leveraging insights from model plants, we categorized PME family genes into four groups. Additionally, the evolution of the PME gene family was shaped by various gene duplication events, primarily dispersed duplication, influenced by purifying selection. A specific gene, Pbr031522.1, designated PbPME35, emerged as a candidate associated with lignin biosynthesis in pear fruits, supported by RNA-seq data. The role of PbPME35 in repressing lignification was validated through its overexpression in pear callus and Arabidopsis. Overall, our findings highlight the ability of PbPME35 to reduce lignin content in pear fruit by downregulating the expression levels of lignin biosynthesis genes. These findings provide new insights into the characteristics of PME genes and their role in regulating lignification in pear fruits.