RESUMO
The woody stems of coniferous gymnosperms produce specialised compression wood to adjust the stem growth orientation in response to gravitropic stimulation. During this process, tracheids develop a compression-wood-specific S2 L cell wall layer with lignins highly enriched with p-hydroxyphenyl (H)-type units derived from H-type monolignol, whereas lignins produced in the cell walls of normal wood tracheids are exclusively composed of guaiacyl (G)-type units from G-type monolignol with a trace amount of H-type units. We show that laccases, a class of lignin polymerisation enzymes, play a crucial role in the spatially organised polymerisation of H-type and G-type monolignols during compression wood formation in Japanese cypress (Chamaecyparis obtusa). We performed a series of chemical-probe-aided imaging analysis on C. obtusa compression wood cell walls, together with gene expression, protein localisation and enzymatic assays of C. obtusa laccases. Our data indicated that CoLac1 and CoLac3 with differential oxidation activities towards H-type and G-type monolignols were precisely localised to distinct cell wall layers in which H-type and G-type lignin units were preferentially produced during the development of compression wood tracheids. We propose that, not only the spatial localisation of laccases, but also their biochemical characteristics dictate the spatial patterning of lignin polymerisation in gymnosperm compression wood.
Assuntos
Lignina , Madeira , Cycadopsida , Lacase , PolímerosRESUMO
The distribution of lignin, 8-5' and 8-8' linked lignin substructure, and noncellulosic polysaccharides in hemp (Cannabis sativa L.) phloem fibers were explored based on histochemical and immunological methods. Ultraviolet absorption and potassium permanganate staining were observed mainly in the compound middle lamella (CML) and S1 layers, and rarely in the G-layer of phloem fibers, suggesting that lignin concentration is high at the CML and S1 layers, and very low at the G-layer of hemp fibers. Acriflavine staining, uniform KM1 labeling (8-5' linked lignin substructure), and no KM2 labeling (8-8' linked structure) were observed in the G-layer, suggesting that there is a small amount of lignin-like compound with 8-5' linked structure in the G-layer. In addition, some fiber cells showed a multilayered structure. Uniform arabinogalactan protein (AGP) labeling was observed on the S1 layers and G-layers using JIM14, but little appeared in the CML of hemp fibers, indicating that these layers of the phloem fibers contain AGP. Immunogold labeling of xylan (LM11) and glucomannan (LM21) showed that xylan and glucomannan were mainly present in the S1 layers and the G-layers, respectively. In some phloem fibers, LM21 immunofluorescence labeling showed multilayered structure, suggesting the heterogeneous distribution of glucomannan.
Assuntos
Cannabis/química , Parede Celular/química , Galactanos/metabolismo , Lignina/metabolismo , Floema/química , Polissacarídeos/metabolismo , Anticorpos Monoclonais/imunologia , Imunofluorescência , Lignina/imunologia , Mananas/análise , Coloração e Rotulagem , Xilanos/análiseRESUMO
Histochemical screening of a flax ethyl methanesulfonate population led to the identification of 93 independent M2 mutant families showing ectopic lignification in the secondary cell wall of stem bast fibers. We named this core collection the Linum usitatissimum (flax) lbf mutants for lignified bast fibers and believe that this population represents a novel biological resource for investigating how bast fiber plants regulate lignin biosynthesis. As a proof of concept, we characterized the lbf1 mutant and showed that the lignin content increased by 350% in outer stem tissues containing bast fibers but was unchanged in inner stem tissues containing xylem. Chemical and NMR analyses indicated that bast fiber ectopic lignin was highly condensed and rich in G-units. Liquid chromatography-mass spectrometry profiling showed large modifications in the oligolignol pool of lbf1 inner- and outer-stem tissues that could be related to ectopic lignification. Immunological and chemical analyses revealed that lbf1 mutants also showed changes to other cell wall polymers. Whole-genome transcriptomics suggested that ectopic lignification of flax bast fibers could be caused by increased transcript accumulation of (1) the cinnamoyl-CoA reductase, cinnamyl alcohol dehydrogenase, and caffeic acid O-methyltransferase monolignol biosynthesis genes, (2) several lignin-associated peroxidase genes, and (3) genes coding for respiratory burst oxidase homolog NADPH-oxidases necessary to increase H2O2 supply.
Assuntos
Parede Celular/química , Linho/genética , Regulação da Expressão Gênica de Plantas , Lignina/metabolismo , Proteínas de Plantas/genética , Oxirredutases do Álcool/genética , Oxirredutases do Álcool/metabolismo , Aldeído Oxirredutases/genética , Aldeído Oxirredutases/metabolismo , Parede Celular/ultraestrutura , Biologia Computacional , Linho/química , Linho/enzimologia , Linho/ultraestrutura , Perfilação da Expressão Gênica , Peróxido de Hidrogênio/metabolismo , Lignina/química , Metiltransferases/genética , Metiltransferases/metabolismo , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Especificidade de Órgãos , Filogenia , Proteínas de Plantas/metabolismo , Caules de Planta/química , Caules de Planta/enzimologia , Caules de Planta/genética , Caules de Planta/ultraestrutura , Plantas Geneticamente Modificadas , Transcriptoma , Xilema/química , Xilema/enzimologia , Xilema/genética , Xilema/ultraestruturaRESUMO
Cereal crop by-products are a promising source of renewable raw material for the production of biofuel from lignocellulose. However, their enzymatic conversion to fermentable sugars is detrimentally affected by lignins. Here the characterization of the Brachypodium Bd5139 mutant provided with a single nucleotide mutation in the caffeic acid O-methyltransferase BdCOMT6 gene is reported. This BdCOMT6-deficient mutant displayed a moderately altered lignification in mature stems. The lignin-related BdCOMT6 gene was also found to be expressed in grains, and the alterations of Bd5139 grain lignins were found to mirror nicely those evidenced in stem lignins. The Bd5139 grains displayed similar size and composition to the control. Complementation experiments carried out by introducing the mutated gene into the AtCOMT1-deficient Arabidopsis mutant demonstrated that the mutated BdCOMT6 protein was still functional. Such a moderate down-regulation of lignin-related COMT enzyme reduced the straw recalcitrance to saccharification, without compromising the vegetative or reproductive development of the plant.
Assuntos
Brachypodium/fisiologia , Lignina/genética , Metiltransferases/genética , Proteínas de Plantas/genética , Biocombustíveis/análise , Brachypodium/genética , Parede Celular/química , Grão Comestível/fisiologia , Lignina/metabolismo , Metiltransferases/metabolismo , Mutação , Fenóis/metabolismo , Proteínas de Plantas/metabolismo , Caules de Planta/fisiologiaRESUMO
MAIN CONCLUSION: Immunolabeling by using monoclonal antibodies showed that xylan deposition precedes the formation of 8-5'-linked structure of lignin in normal and compression woods of Chamaecyparis obtusa. Xylan deposition and formation of 8-5'-linked lignin structure in differentiating xylems from normal and compression woods in Chamaecyparis obtusa were examined by immunoelectron microscopy using monoclonal antibodies (LM10 or LM11) to detect xylan localization. The 8-5'-linked lignin structure was immunolocalized using KM1 antibody. Xylan and 8-5'-linked lignin double immunolabeling was performed using secondary antibodies labeled with colloidal gold particles of different diameters. In normal wood, KM1 labeling occurred in the compound middle lamella (CML) and S1 layer during S1 layer formation and increased as S2 and S3 layers formed, with labeling occurring at the outer part of the previous layer. In compression wood, mild KM1 labeling occurred in the CML and outer part of the S1 layer at the later S1 layer formation stage, with increased labeling as the S2 layer formed. Minor labeling occurred in the outer part of the S2 layer during helical cavity formation. Comparison between KM1 labeling and KMnO4 staining suggested that lignin other than 8-5'-linked structure was formed during early lignification, and the proportion of 8-5'-linked lignin structure increased at later stages of lignification in both normal and compression woods. LM10 and LM11 labeling occurred slightly earlier than KM1 labeling, suggesting that xylan deposition preceded the formation of 8-5'-linked lignin in normal and compression woods. Less labeling by KM1, LM10, and LM11 occurred in the outer part of the S2 layer in compression wood, which has abundant lignin. Thus, lignin in these parts is composed of lignin substructures other than the 8-5' linkage.
Assuntos
Chamaecyparis/metabolismo , Lignina/análise , Madeira/metabolismo , Xilanos/análise , Anticorpos Monoclonais/imunologia , Chamaecyparis/ultraestrutura , Imuno-Histoquímica/métodos , Lignina/química , Lignina/imunologia , Microscopia Eletrônica de Transmissão , Microscopia Imunoeletrônica , Estrutura Molecular , Madeira/ultraestrutura , Xilanos/imunologiaRESUMO
INTRODUCTION: Hesperidin, a flavonoid known to have important pharmacological effects, accumulates particularly in the peels of satsuma mandarin (Citrus unshiu). Although histochemical studies have suggested that hesperidin forms crystals in some tissues of the Rutaceae and Umbelliferae, there has been no rigorous in situ detection or identification of hesperidin crystals in C. unshiu. OBJECTIVE: To characterise the chemical component of the crystals found in C. unshiu peels using Raman microscopy. METHODS: Sections of C. unshiu peels were made. The distribution and morphology of crystals in the sections were analysed microscopically. Raman microscopy was used to detect hesperidin in the sections directly. RESULTS: The crystals were more abundant in immature peel and were observed particularly in areas surrounding vascular bundles, around the border between the flavedo and albedo layers and just below the epidermal cells. In the morphological analysis by scanning electron microscopy, needle-shaped crystals aggregated and formed clusters of spherical crystals. Spectra obtained by Raman microscopy of the crystals in the peel sections were consistent with those of the hesperidin standard. CONCLUSION: This study showed the detailed distribution of crystals in C. unshiu peels and their main component was identified using Raman microscopy to be hesperidin for the first time.
Assuntos
Citrus/química , Hesperidina/isolamento & purificação , Extratos Vegetais/isolamento & purificação , Citrus/ultraestrutura , Frutas/química , Frutas/ultraestrutura , Hesperidina/química , Microscopia Eletrônica de Varredura , Extratos Vegetais/químicaRESUMO
Lignin biosynthesis is an essential physiological activity of vascular plants if they are to survive under various environmental stresses on land. The biosynthesis of lignin proceeds in the cell wall by polymerization of precursors; the initial step of lignin polymerization is the transportation of lignin monomers from the cytosol to the cell wall, which is critical for lignin formation. There has been much debate on the transported form of the lignin precursor, either as free monolignols or their glucosides. In this study, we performed biochemical analyses to characterize the membrane transport mechanism of lignin precursors using angiosperms, hybrid poplar (Populus sieboldii × Populus grandidentata) and poplar (Populus sieboldii), as well gymnosperms, Japanese cypress (Chamaecyparis obtusa) and pine (Pinus densiflora). Membrane vesicles prepared from differentiating xylem tissues showed clear ATP-dependent transport activity of coniferin, whereas less than 4% of the coniferin transport activity was seen for coniferyl alcohol. Bafilomycin A1 and proton gradient erasers markedly inhibited coniferin transport in hybrid poplar membrane vesicles; in contrast, vanadate had no effect. Cis-inhibition experiments suggested that this transport activity was specific for coniferin. Membrane fractionation of hybrid poplar microsomes demonstrated that transport activity was localized to the tonoplast- and endomembrane-rich fraction. Differentiating xylem of Japanese cypress exhibited almost identical transport properties, suggesting the involvement of a common endomembrane-associated proton/coniferin antiport mechanism in the lignifying tissues of woody plants, both angiosperms and gymnosperms.
Assuntos
Cinamatos/metabolismo , Plantas/metabolismo , Xilema/metabolismo , Trifosfato de Adenosina/metabolismo , Transporte Biológico , Membrana Celular/metabolismo , Quimera , Cupressus/metabolismo , Cycadopsida/metabolismo , Lignina/metabolismo , Microssomos/metabolismo , Pinus/metabolismo , Proteínas de Plantas/metabolismo , Populus/genética , Populus/metabolismo , PrótonsRESUMO
Mouse monoclonal antibodies were generated against dehydrodiconiferyl alcohol- or pinoresinol-p-aminohippuric acid (pAHA)-bovine serum albumin (BSA) conjugate as probes that specifically react with 8-5' or 8-8' linked structure of lignin in plant cell walls. Hybridoma clones were selected that produced antibodies that positively reacted with dehydrodiconiferyl alcohol- or pinoresinol-pAHA-BSA and negatively reacted with pAHA-BSA and guaiacylglycerol-beta-guaiacyl ether-pAHA-BSA conjugates containing 8-O-4' linkage. Eight clones were established for each antigen and one of each clone that positively reacted with wood sections was selected. The specificity of these antibodies was examined by competitive ELISA tests using various lignin dimers with different linkages. The anti-dehydrodiconiferyl alcohol antibody reacted specifically with dehydrodiconiferyl alcohol and did not react with other model compounds containing 8-O-4', 8-8', or 5-5' linkages. The anti-pinoresinol antibody reacted specifically with pinoresinol and syringaresinol and did not react with the other model compounds containing 8-O-4', 8-5', or 5-5' linkages. The antibodies also did not react with dehydrodiconiferyl alcohol acetate or pinoresinol acetate, indicating that the presence of free phenolic or aliphatic hydroxyl group was an important factor in their reactivity. In sections of Japanese cypress (Chamaecyparis obtusa), labeling by the anti-dehydrodiconiferyl alcohol antibody was found in the secondary walls of phloem fibers and in the compound middle lamellae, and secondary walls of tracheids. Weak labeling by the anti-pinoresinol antibody was found in secondary walls of phloem fibers and secondary walls and compound middle lamellae of developed tracheids. These labelings show the localization of 8-5' and 8-8' linked structure of lignin in the cell walls.
Assuntos
Anticorpos Monoclonais/imunologia , Parede Celular/imunologia , Chamaecyparis/citologia , Chamaecyparis/metabolismo , Lignina/química , Lignina/imunologia , Animais , Especificidade de Anticorpos/imunologia , Antígenos/imunologia , Chamaecyparis/ultraestrutura , Ensaio de Imunoadsorção Enzimática , Feminino , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB C , Floema/metabolismo , Floema/ultraestrutura , Coloração pela Prata , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The ultrastructure of the innermost surface of Cryptomeria japonica differentiating normal wood (NW) and compression wood (CW) was comparatively investigated by field emission electron microscopy (FE-SEM) combined with enzymatic degradation of hemicelluloses. Cellulose microfibril (CMF) bundles were readily observed in NW tracheids in the early stage of secondary cell wall formation, but not in CW tracheids because of the heavy accumulation of amorphous materials composed mainly of galactans and lignin. This result suggests that the ultrastructural deposition of cell wall components in the tracheid cell wall differ between NW and CW from the early stage of secondary cell wall formation. Delignified NW and CW tracheids showed similar structural changes during differentiating stages after xylanase or ß-mannanase treatment, whereas they exhibited clear differences in ultrastructure in mature stages. Although thin CMF bundles were exposed in both delignified mature NW and CW tracheids by xylanase treatment, ultrastructural changes following ß-mannanase treatment were only observed in CW tracheids. CW tracheids also showed different degradation patterns between xylanase and ß-mannanase. CMF bundles showed a smooth surface in delignified mature CW tracheids treated with xylanase, whereas they had an uneven surface in delignified mature CW tracheids treated with ß-mannanase, indicating that the uneven surface of CMF bundles was related to xylans. The present results suggest that ultrastructural deposition and organization of lignin and hemicelluloses in CW tracheids may differ from those of NW tracheids.
Assuntos
Cryptomeria/crescimento & desenvolvimento , Cryptomeria/ultraestrutura , Microscopia Eletrônica de Varredura/métodos , Madeira/crescimento & desenvolvimento , Madeira/ultraestrutura , Xilema/crescimento & desenvolvimento , Xilema/ultraestrutura , Cryptomeria/anatomia & histologia , Cryptomeria/efeitos dos fármacos , Endo-1,4-beta-Xilanases/farmacologia , Glicosídeo Hidrolases/farmacologia , Lignina/isolamento & purificação , Propriedades de Superfície/efeitos dos fármacos , Madeira/anatomia & histologia , Madeira/efeitos dos fármacos , Xilema/anatomia & histologia , Xilema/efeitos dos fármacosRESUMO
BACKGROUND AND AIMS: Although tension wood formation and the structure of gelatinous fibres (G-fibres) have been widely investigated, studies of the influence of the reaction phenomenon on phloem fibres have been few and incomplete in comparison with those of xylem wood fibres. This study was undertaken to clarify the influence of stem inclination on phloem fibres using several Japanese hardwood species that produce different G-fibre types in tension wood. METHODS: Eight hardwood species were inclined at 30-45° at the beginning of April. Specimens were collected in July and December. The cell-wall structure and lignin distribution of phloem fibres on both the tension and opposite sides were compared by light microscopy, ultraviolet microscopy, confocal laser scanning microscopy after staining with acriflavine, and transmission electron microscopy after staining with potassium permanganate. KEY RESULTS: Three types of changes were found in tension-side phloem fibres: (1) increases in the proportion of the syringyl unit in lignin in the S(1) and S(2) layers and compound middle lamella (Cercidiphyllum japonicum), (2) formation of unlignified gelatinous layers (Melia azedarach and Acer rufinerve) and (3) increases in the number of layers (n) in the multi-layered structure of S(1) + S(2) + n (G + L) (Mallotus japonicus). Other species showed no obvious change in cell-wall structure or lignin distribution. CONCLUSIONS: Phloem fibres of the tree species examined in our study showed three types of changes in lignin distribution and cell-wall structure. The reaction phenomenon may vary with tree species and may not be closely related to G-fibre type in tension wood.
Assuntos
Lignina/metabolismo , Magnoliopsida/anatomia & histologia , Floema/anatomia & histologia , Caules de Planta/anatomia & histologia , Xilema/anatomia & histologia , Parede Celular/metabolismo , Parede Celular/ultraestrutura , Magnoliopsida/metabolismo , Floema/metabolismo , Caules de Planta/metabolismo , Árvores/anatomia & histologia , Árvores/metabolismo , Madeira , Xilema/metabolismoRESUMO
Woody cells, such as tracheids, fibers, vessels, rays etc., have unique structural characteristics such as nano-scale ultrastructure represented by multilayers, microfibril angle (MFA), micro-scale anatomical properties and spatial arrangement. Simultaneous evaluation of the above indices is very important for their adequate quantification and extracting the effects of external stimuli from them. However, it is difficult in general to achieve the above only by traditional methodologies. To overcome the above point, a new methodological framework combining polarization optical microscopy, fluorescence microscopy, and image segmentation is proposed. The framework was tested to a model softwood species, Chamaecyparis obtusa for characterizing intra-annual transition of MFA and tracheid morphology in a radial file unit. According our result, this framework successfully traced the both characteristics tracheid by tracheid and revealed the high correlation (|r| > 0.5) between S2 microfibril angles and tracheidal morphology (lumen radial diameter, tangential wall thickness and cell wall occupancy). In addition, radial file based evaluation firstly revealed their complex transitional behavior in transition and latewood. The proposed framework has great potential as one of the unique tools to provide detailed insights into heterogeneity of intra and inter-cells in the wide field of view through the simultaneous evaluation of cells' ultrastructure and morphological properties.
Assuntos
Chamaecyparis , Microfibrilas , Microscopia , Madeira , Parede Celular/ultraestruturaRESUMO
Sorghum has been recognized as a promising energy crop. The composition and structure of lignin in the cell wall are important factors that affect the quality of plant biomass as a bioenergy feedstock. Silicon (Si) supply may affect the lignin content and structure, as both Si and lignin are possibly involved in plant mechanical strength. However, our understanding regarding the interaction between Si and lignin in sorghum is limited. Therefore, in this study, we analyzed the lignin in the cell walls of sorghum seedlings cultured hydroponically with or without Si supplementation. Limiting the Si supply significantly increased the thioglycolic acid lignin content and thioacidolysis-derived syringyl/guaiacyl monomer ratio. At least part of the modification may be attributable to the change in gene expression, as suggested by the upregulation of phenylpropanoid biosynthesis-related genes under -Si conditions. The cell walls of the -Si plants had a higher mechanical strength and calorific value than those of the +Si plants. These results provide some insights into the enhancement of the value of sorghum biomass as a feedstock for energy production by limiting Si uptake.
Assuntos
Sorghum , Biomassa , Parede Celular/metabolismo , Grão Comestível/metabolismo , Regulação da Expressão Gênica de Plantas , Lignina/metabolismo , Plântula/metabolismo , Silício/metabolismo , Sorghum/genéticaRESUMO
Wood is composed of various types of cells and each type of cell has different structural and functional properties. However, the temporal and spatial diversities of cell wall components in the cell wall between different cell types are rarely understood. To extend our understanding of distributional diversities of cell wall components among cells, we investigated the immunolabeling of mannans (O-acetyl-galactoglucomannans, GGMs) and xylans (arabino-4-O-methylglucuronoxylans, AGXs) in ray cells and pits. The labeling of GGMs and AGXs was temporally different in ray cells. GGM labeling began to be detected in ray cells at early stages of S(1) formation in tracheids, whereas AGX labeling began to be detected in ray cells at the S(2) formation stage in tracheids. The occurrence of GGM and AGX labeling in ray cells was also temporally different from that of tracheids. AGX labeling began to be detected much later in ray cells than in tracheids. GGM labeling also began to be detected in ray cells either slightly earlier or later than in tracheids. In pits, GGM labeling was detected in bordered and cross-field pit membranes at early stages of pit formation, but not observed in mature pits, indicating that enzymes capable of GGM degradation may be involved in pit membrane formation. In contrast to GGMs, AGXs were not detected in pit membranes during the entire developmental process of bordered and cross-field pits. AGXs showed structural and depositional variations in pit borders depending on the developmental stage of bordered and cross-field pits.
Assuntos
Diferenciação Celular , Cryptomeria/citologia , Cryptomeria/metabolismo , Mananas/metabolismo , Madeira/citologia , Madeira/metabolismo , Xilanos/metabolismo , Cryptomeria/ultraestrutura , Imunofluorescência , Fatores de Tempo , Madeira/ultraestruturaRESUMO
Compression wood (CW) tracheids have different cell wall components than normal wood (NW) tracheids. However, temporal and spatial information on cell wall components in CW tracheids is poorly understood. We investigated the distribution of arabino-4-O-methylglucuronoxylans (AGXs) and O-acetyl-galactoglucomannans (GGMs) in differentiating CW tracheids. AGX labeling began to be detected in the corner of the S(1) layer at the early S(1) formation stage. Subsequently, the cell corner middle lamella (ccML) showed strong AGX labeling when intercellular spaces were not fully formed. AGX labeling was uniformly distributed in the S(1) layer, but showed uneven distribution in the S(2) layer. AGX labeling was mainly detected in the inner S(2) layer after the beginning of the helical cavity formation. The outer S(2) layer showed almost no labeling of low substituted AGXs. Only a very small amount of high substituted AGXs was distributed in the outer S(2) layer. These patterns of AGX labeling in the S(2) layer opposed the lignin and ß-1-4-galactan distribution in CW tracheids. GGM labeling patterns were almost identical to AGX labeling in the early stages of CW tracheids, and GGM labeling was detected in the entire S(2) layer from the early S(2) formation stage of CW tracheids with some spatial differences in labeling density depending on developmental stage. Compared with NW tracheids, CW tracheids showed significantly different AGX distributions in the secondary cell wall but similar GGM labeling patterns. No significant differences were observed in labeling after delignification of CW tracheids.
Assuntos
Parede Celular/metabolismo , Cryptomeria/anatomia & histologia , Cryptomeria/metabolismo , Mananas/metabolismo , Organogênese , Madeira/anatomia & histologia , Xilanos/metabolismo , Anticorpos/imunologia , Cryptomeria/citologia , Cryptomeria/ultraestrutura , Imunofluorescência , Lignina/metabolismo , Mananas/ultraestrutura , Madeira/citologia , Madeira/metabolismo , Madeira/ultraestrutura , Xilanos/ultraestruturaRESUMO
This work demonstrates a unique approach of utilizing alkali lignin (AL), as smart additive to in situ BC fermentation in which it concurrently acts as promoter to microbial growth as well as reinforcing filler for fabrication of multifunctional composites. Traditionally, BC fermentation is accompanied by inhibitor formation with sudden drop in pH leading to low yield and biomass growth. AL due to its antioxidant nature prevents formation of gluconic acid as byproduct, at â¼0.25 wt.% AL based on inhibitory byproduct kinetics. Interestingly, AL self-assembles to form primary and secondary structures in BC pores, resulting in simultaneous improvement in thermal stability as well as toughness. The BC/AL films show strong UV-blocking capacity with prolonged radical scavenging activity and preventing browning of freshly cut apples making it suitable as food packaging. Therefore, present work opens up new avenues for fabrication of high-performance BC-based composites through selection of smart materials which can simultaneously improve BC bioprocessing.
RESUMO
We investigated the spatial and temporal distribution of xylans in the cell walls of differentiating earlywood tracheids of Cryptomeria japonica using two different types of monoclonal antibodies (LM10 and LM11) combined with immunomicroscopy. Xylans were first deposited in the corner of the S(1) layer in the early stages of S(1) formation in tracheids. Cell corner middle lamella also showed strong xylan labeling from the early stage of cell wall formation. During secondary cell wall formation, the innermost layer and the boundary between the S(1) and S(2) layers (S(1)/S(2) region) showed weaker labeling than other parts of the cell wall. However, mature tracheids had an almost uniform distribution of xylans throughout the entire cell wall. Xylan localization labeled with LM10 antibody was stronger in the outer S(2) layer than in the inner layer, whereas xylans labeled with LM11 antibody were almost uniformly distributed in the S(2) layer. In addition, the LM10 antibody showed almost no xylan labeling in the S(1)/S(2) region, whereas the LM11 antibody revealed strong xylan labeling in the S(1)/S(2) region. These findings suggest that structurally different types of xylans may be deposited in the tracheid cell wall depending on the developmental stage of, or location in, the cell wall. Our study also indicates that deposition of xylans in the early stages of tracheid cell wall formation may be spatially consistent with the early stage of lignin deposition in the tracheid cell wall.
Assuntos
Parede Celular/metabolismo , Cryptomeria/citologia , Cryptomeria/metabolismo , Xilanos/metabolismo , Imuno-Histoquímica , Microscopia de FluorescênciaRESUMO
We investigated the deposition of glucomannans (GMs) in differentiating earlywood tracheids of Cryptomeria japonica using immunocytochemical methods. GMs began to deposit at the corner of the cell wall at the early stages of S(1) formation and showed uneven distribution in the cell wall during S(1) formation. At the early stages of S(2) formation, limited GM labeling was observed in the S(2) layer, and then the labeling increased gradually. In mature tracheids, the boundary between the S(1) and S(2) layers and the innermost part of the cell wall showed stronger labeling than other parts of the cell wall. Deacetylation of GMs with mild alkali treatment led to a significant increase in GM labeling and a more uniform distribution of GMs in the cell wall than that observed before deacetylation, indicating that some GM epitopes may be masked by acetylation. However, the changes in GM labeling after deacetylation were not very pronounced until early stages of S(2) formation, indicating that GMs deposited in the cell wall at early stages of cell-wall formation may contain fewer acetyl groups than those deposited at later stages. Additionally, the density of GM labeling increased in the cell wall in both specimens before and after GM deacetylation, even after cell-wall formation was complete. This finding suggests that some acetyl groups may be removed from GMs after cell-wall formation is complete as part one of the tracheid cell aging processes.
Assuntos
Parede Celular/metabolismo , Cryptomeria/metabolismo , Mananas/metabolismo , Microscopia de FluorescênciaRESUMO
Compression wood (CW) contains higher quantities of beta-1-4-galactan than does normal wood (NW). However, the physiological roles and ultrastructural distribution of beta-1-4-galactan during CW formation are still not well understood. The present work investigated deposition of beta-1-4-galactan in differentiating tracheids of Cryptomeria japonica during CW formation using an immunological probe (LM5) combined with immunomicroscopy. Our immunolabeling studies clearly showed that differences in the distribution of beta-1-4-galactan between NW (and opposite wood, OW) and CW are initiated during the formation of the S(1) layer. At this stage, CW was strongly labeled in the S(1) layer, whereas no label was observed in the S(1) layer of NW and OW. Immunogold labeling showed that beta-1-4-galactan in the S(1) layer of CW tracheids significantly decreased during the formation of the S(2) layer. Most beta-1-4-galactan labeling was present in the outer S(2) region in mature CW tracheids, and was absent in the inner S(2) layer that contained helical cavities in the cell wall. In addition, delignified CW tracheids showed significantly more labeling of beta-1-4-galactan in the secondary cell wall, suggesting that lignin is likely to mask beta-1-4-galactan epitopes. The study clearly showed that beta-1-4-galactan in CW was mainly deposited in the outer portion of the secondary cell wall, indicating that its distribution may be spatially consistent with lignin distribution in CW tracheids of Cryptomeria japonica.
Assuntos
Cryptomeria/metabolismo , Galactanos/metabolismo , Lignina/metabolismo , Madeira , Microscopia/métodosRESUMO
This paper describes the design and synthesis of new trehalose-type diblock methylcellulose analogues with nonionic, cationic, and anionic cellobiosyl segments, namely 1-(tri-O-methyl-cellulosyl)-4-[ß-d-glucopyranosyl-(1â4)-ß-d-glucopyranosyloxymethyl]-1H-1,2,3-triazole (1), 1-(tri-O-methyl-cellulosyl)-4-[(6-amino-6-deoxy-ß-d-glucopyranosyl)-(1â4)- 6-amino-6-deoxy-ß-d-glucopyranosyloxymethyl]-1H-1,2,3-triazole (2), and 4-(tri-O-methyl-cellulosyloxymethyl)-1-[ß-d-glucopyranuronosyl-(1â4)-ß-d-glucopyranuronosyl]-1H-1,2,3-triazole (3), respectively. Aqueous solutions of all of the 1,2,3-triazole-linked diblock methylcellulose analogues possessed higher surface activities than that of industrially produced methylcellulose and exhibited lower critical solution temperatures, that allowed the formation of thermoresponsive supramolecular hydrogels at close to human body temperature. Supramolecular structures of thermo-reversible hydrogels based on compounds 1, 2, and 3 were investigated by means of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Detailed structure-property-function relationships of compounds 1, 2, and 3 were discussed. Not only nonionic hydrophilic segment but also ionic hydrophilic segments of diblock methylcellulose analogues were valid for the formation of thermo-reversible supramolecular hydrogels based on end-functionalized methylcellulose.
RESUMO
Important biological, nutritional and technological roles are attributed to cell wall polymers from cereal grains. The composition of cell walls in dry wheat grain has been well studied, however less is known about cell wall deposition and modification in the grain outer layers during grain development. In this study, the composition of cell walls in the outer layers of the wheat grain (Triticum aestivum Recital cultivar) was investigated during grain development, with a focus on cell wall phenolics. We discovered that lignification of outer layers begins earlier than previously reported and long before the grain reaches its final size. Cell wall feruloylation increased in development. However, in the late stages, the amount of ferulate releasable by mild alkaline hydrolysis was reduced as well as the yield of lignin-derived thioacidolysis monomers. These reductions indicate that new ferulate-mediated cross-linkages of cell wall polymers appeared as well as new resistant interunit bonds in lignins. The formation of these additional linkages more specifically occurred in the outer pericarp. Our results raised the possibility that stiffening of cell walls occur at late development stages in the outer pericarp and might contribute to the restriction of the grain radial growth.