Dynamic Changes in Distribution of Lignin and Hemicelluloses in Cell Walls During Differentiation of Secondary Xylem in Eucommia ulmoides

The dynamic changes in the distribution of lignin and hemicelluloses (xylans and xyloglucans) in cell walls during the differentiation of secondary xylem in Eucommia ulmoides Oliv. were studied by means of ultraviolet light microscopy and transmission electron microscopy combined with immunogold labelling. In the cambial zone and cell expansion zone, xyloglucans were localized both in the tangential and radial walls, but no xylans or lignin were found in these regions. With the formation of secondary wall S-1 layer, lignin occurred in the cell corners and middle lamella, while xylans appeared in S-1 layer, and xyloglucans were localized in the primary walls and middle lamella. In pace with the formation of secondary wall S-2 and S-3 layer, lignification extended to S-1, S-2 and S-3 layer in sequence, showing a patchy style of lignin deposition. Concurrently, xylans distributed in the whole secondary walls and xyloglucans, on the other hand, still localized in the primary walls and middle lamella. The results indicated that along with the formation and lignification of the secondary wall, great changes had taken place in the cell walls. Different parts of cell walls, such as cell corners, middle lamella, primary walls and various layers of secondary walls, had different kinds of hemicelluloses, which formed various cell wall architecture combined with lignin and other cell wall components.

Pectin methylesterase inhibitors GhPMEI53 and AtPMEI19 improve seed germination by modulating cell wall plasticity in cotton and Arabidopsis

The germination process of seeds is influenced by the interplay between two opposing factors,pectin methylesterase(PME)and pectin methylesterase inhibitor(PMEI),which collectively regulate patterns of pectin methylesterification.Despite the recognized importance of pectin methylesterification in seed germination,the specific mechanisms that govern this process remain unclear.In this study,we demonstrated that the overexpression of GhPMEI53is associated with a decrease in PME activity and an increase in pectin methylesterification.This leads to seed cell wall softening,which positively regulates cotton seed germination.AtPMEI19,the homologue in Arabidopsis thaliana,plays a similar role in seed germination to GhPMEI53,indicating a conserved function and mechanism of PMEI in seed germination regulation.Further studies revealed that GhPMEI53 and AtPMEI19 directly contribute to promoting radicle protrusion and seed germination by inducing cell wall softening and reducing mechanical strength.Additionally,the pathways of abscicic acid(ABA)and gibberellin(GA)in the transgenic materials showed significant changes,suggesting that GhPMEI53/AtPMEI19-mediated pectin methylesterification serves as a regulatory signal for the related phytohormones involved in seed germination.In summary,GhPMEI53 and its homologs alter the mechanical properties of cell walls,which influence the mechanical resistance of the endosperm or testa.Moreover,they impact cellular phytohormone pathways(e.g.,ABA and GA)to regulate seed germination.These findings enhance our understanding of pectin methylesterification in cellular morphological dynamics and signaling transduction,and contribute to a more comprehensive understanding of the PME/PMEI gene superfamily in plants.

Strigolactones modulate cotton fiber elongation and secondary cell wall thickening

Cotton is one of the most important economic crops in the world,and it is a major source of fiber in the textile industry.Strigolactones(SLs)are a class of carotenoid-derived plant hormones involved in many processes of plant growth and development,although the functions of SL in fiber development remain largely unknown.Here,we found that the endogenous SLs were significantly higher in fibers at 20 days post-anthesis(DPA).Exogenous SLs significantly increased fiber length and cell wall thickness.Furthermore,we cloned three key SL biosynthetic genes,namely GhD27,GhMAX3,and GhMAX4,which were highly expressed in fibers,and subcellular localization analyses revealed that GhD27,GhMAX3,and GhMAX4 were localized in the chloroplast.The exogenous expression of GhD27,GhMAX3,and GhMAX4 complemented the physiological phenotypes of d27,max3,and max4 mutations in Arabidopsis,respectively.Knockdown of GhD27,GhMAX3,and GhMAX4 in cotton resulted in increased numbers of axillary buds and leaves,reduced fiber length,and significantly reduced fiber thickness.These findings revealed that SLs participate in plant growth,fiber elongation,and secondary cell wall formation in cotton.These results provide new and effective genetic resources for improving cotton fiber yield and plant architecture.

Upregulation of the glycine-rich protein-encoding gene GhGRPL enhances plant tolerance to abiotic and biotic stressors by promoting secondary cell wall development

Abiotic and biotic stressors adversely affect plant survival,biomass generation,and crop yields.As the global availability of arable land declines and the impacts of global warming intensify,such stressors may have increasingly pronounced effects on agricultural productivity.Currently,researchers face the overarching challenge of comprehensively enhancing plant resilience to abiotic and biotic stressors.The secondary cell wall plays a crucial role in bolstering the stress resistance of plants.To increase plant resistance to stress through genetic manipulation of the secondary cell wall,we cloned a cell wall protein designated glycine-rich protein-like(GhGRPL)from cotton fibers,and found that it is specifically expressed during the period of secondary cell wall biosynthesis.Notably,this protein differs from its Arabidopsis homolog,AtGRP,since its glycine-rich domain is deficient in glycine residues.GhGRPL is involved in secondary cell wall deposition.Upregulation of GhGRPL enhances lignin accumulation and,consequently,the thickness of the secondary cell walls,thereby increasing the plant’s resistance to abiotic stressors,such as drought and salinity,and biotic threats,including Verticillium dahliae infection.Conversely,interference with GhGRPL expression in cotton reduces lignin accumulation and compromises that resistance.Taken together,our findings elucidate the role of GhGRPL in regulating secondary cell wall development through its influence on lignin deposition,which,in turn,reinforces cell wall robustness and impermeability.These findings highlight the promising near-future prospect of adopting GhGRPL as a viable,effective approach for enhancing plant resilience to abiotic and biotic stress factors.

Mechanical characterization of Pinus massoniana cell walls infected by blue-stain fungi using in situ nanoindentation

Characterizing the mechanical properties of wood cell walls will lead to better understanding and optimization of modifications made to wood infected by the blue-stain fungi.In this study,in situ nanoindentation was used to characterize the mechanical properties of the cell walls of Pinus massoniana infected by blue-stain fungi at the cellular level.The results show that in situ nanoindentation is an effective method for this purpose and that blue-stain fungi penetrate wood structures and degrade wood cell walls,significantly reducing the mechanical properties of the cell walls.The method can also be used to evaluate and improve the properties of other wood species infected by blue-stain fungi.

BRITTLE CULM16(BRITTLE NODE) is required for the formation of secondary cell walls in rice nodes

Plant cell walls constitute the skeletal structures of plant bodies,and thus confer lodging resistance for grain crops.While the basic cell wall synthesis machinery is relatively well established now,our understanding of how the process is regulated remains limited and fragmented.In this study,we report the identification and characterization of the novel rice（Oryza sativa L.）brittle culm16（brittle node;bc16）mutant.The brittle node phenotype of the bc16 mutant appears exclusively at nodes,and resembles the previously reported bc5 mutant.Combined histochemical staining and electron microscopy assays revealed that in the bc16 mutant,the secondary cell wall formation and thickening of node sclerenchyma tissues are seriously affected after heading.Furthermore,cell wall composition assays revealed that the bc16 mutation led to a significant reduction in cellulose and lignin contents.Using a map-based cloning approach,the bc16 locus is mapped to an approximately 1.7-Mb region of chromosome 4.Together,our findings strengthen evidence for discretely spatial differences in the secondary cell wall formation within plant bodies.

Identification of EDTA-Soluble Polysaccharides from Pea Epicotyl Cell Walls and Their Interaction with Xyloglucan

Nascent pectin and glucuronoxylan were prepared from membrane-bound enzymes obtained from pea epicotyls. They had previously been shown to exhibit a protein- and pH-dependent pattern of binding to cell wall ghosts and to xy-loglucan extracted from cell walls prepared from pea epicotyls;maximum binding required a pH of 3-4, and the pres-ence of cell wall proteins, namely assemblins. To determine whether wall polysaccharides deposited in cell walls be-have in the same manner as nascent polymers, radioactively labeled EDTA-soluble polymers were prepared from newly-deposited pea epicotyl cell walls. Different enzyme treatments followed by column chromatography, in addition to complete acid hydrolysis followed by paper and thin layer chromatography, indicated the presence of pectin, to-gether with smaller amounts of glucuronoxylan, in this EDTA-soluble extract. These radioactively labeled polysaccha-rides were found to bind to cell wall ghosts and to xyloglucan extracted from the second and third internodes of pea epicotyls cell walls in a pH-dependent manner, similar to the binding pattern obtained with nascent polymers. Maxi-mum binding occurred at pH 3-4, and also required the presence of protein.

Humans have intestinal bacteria that degrade the plant cell walls in herbivores

The cell walls of plants are mainly made of cellulose and contain a large number of calories.However,the main component,cellulose,is an indigestible plant fiber that is thought to be difficult for humans to use as energy.Herbivores acquire energy through the degradation of cell wall-derived dietary fiber by microorganisms in the digestive tract.Herbivores,especially horses,have a highly developed cecum and large intestine,and plants are fermented for their efficient use with the help of microorganisms.Humans also have an intestinal tract with a wide lumen on the proximal side of the large intestine,in which fermentation occurs.The digestive process of horses is similar to that of humans,and many of the intestinal bacteria found in horses that degrade plants are also found in humans.Therefore,it is thought that humans also obtain a certain amount of energy from cell wall-derived dietary fiber.However,the intake of dietary fiber by modern humans is low;thus,the amount of calories derived from indigestible plant fiber is considered to be very low.Cellulose in the plant cell wall is often accompanied by hemicellulose,pectin,lignin,suberin,and other materials.These materials are hard to degrade,and cellulose is therefore difficult for animals to utilize.If the cell wall can be degraded to some extent by cooking,it is thought that humans can obtain calories from cell wall-derived dietary fiber.If humans can use the calories from the cell wall for their diet,it may compensate for human food shortages.

Intratree Variation in Viscoelastic Properties of Cell Walls of Masson Pine (Pinus Massoniana Lamb)

In this study,Pinus massoniana Lamb at different heights,across the annual rings,and between earlywood and latewood was measured by X-ray diffraction and the chemical composition was analyzed by chemical treatment.Results indicated that the microfibril angle(MFA)decreased and the chemical composition changed little with the increase in height from 1 m to 9 m.In the radial direction,the MFA decreased and the chemical composition changed little with an increase in annual rings.The cellulose content of latewood was higher than that of early-wood.The viscoelastic changes of wood cell walls at different heights,across the annual rings by the method of quasi-static nanoindentation and dynamic modulus mapping techniques.Results indicated that the wood cell walls’elastic modulus increased,and the creep rate and creep compliance decreased with the increase in height;The elastic modulus and hardness increased with the annual rings.The cell walls’storage modulus increased and the loss modulus gradually decreased with an increase in height;the storage modulus gradually increased and the loss modulus decreased with the annual rings.

Observations of Wood Cell Walls with a Scanning Probe Microscope

Scanning Probe Microscopes (SPMs) observe specimen surfaces with probes by detecting the physical amount of a material between the cantilever and the surface. SPMs have a high resolution and can measure mechanical characteristics such as stiffness, adsorptive properties, and viscoelasticity. These features make it easy to identify the surface structure of complex materials;therefore, the use of SPMs has increased in recent years. Wood cell walls are primarily composed of cellulose, hemicellulose, and lignin. It is believed that hemicellulose and lignin surround the cellulose framework;however, their detailed formation remains unknown. Therefore, we observed wood cell walls via scanning probe microscopy to try to reveal the formation of the cellulose framework. We determined that the size of the cellulose microfibril bundle and hemicellulose lignin module composite was 18.48 nm based on topography and that the size of the cellulose microfibril bundle was 15.33 nm based on phase images. In the viscoelasticity image, we found that the viscoelasticities of each cell wall of the same cell were not the same. This is because the cellulose microfibrils in each cell wall lean in different directions. The angle between the leaning of the cellulose microfibril and the cantilever affects the viscoelasticity measurement.

Effects of cell wall property on compressive performance of aluminum foams

The effects of cell wall property on the compressive performance of high porosity, closed-cell aluminum foams prepared by gas injection method were investigated. The research was conducted both experimentally and numerically. Foam specimens prepared from conditioned melt were tested under uniaxial compressive loading condition. The cell wall microstructure and fracture were observed through optical microscope（OM） and scanning electron microscope（SEM）, which indicates that the cell wall property is impaired by the defects in cell walls and oxide films on the cell wall surface. Subsequently, finite element（FE） models based on three-dimensional thin shell Kelvin tetrakaidecahedron were developed based on the mechanical properties of the raw material and solid material that are determined by using experimental measurements. The simulation results show that the plateau stress of the nominal stress-strain curve exhibits a linear relationship with the yield strength of the cell wall material. The simulation plateau stress is higher than the experimental data, partly owing to the substitution of solid material for cell wall material in the process of the establishment of FE models.

Genetic analysis and QTL mapping of stalk cell wall components and digestibility in maize recombinant inbred lines from B73 × By804

The cell wall composition and structure of the maize stalk directly affects its digestibility and in turn its feed value.Previous studies of stem quality have focused mostly on common maize germplasm,and few studies have focused on high-oil cultivars with high grain and straw quality.Investigation of the genetic basis of cell wall composition and digestibility of maize stalk using high-oil maize is desirable for improving maize forage quality.In the present study,a high-oil inbred line(By804)was crossed as male parent with the maize inbred line B73 to construct a population of 188 recombinant inbred lines(RILs).The phenotypes of six cell-wall-related traits were recorded,and QTL analysis was performed with a genetic map constructed with SNP markers.All traits were significantly correlated with one another and showed high broad-sense heritability.Of 20 QTLs mapped,the QTL associated with each trait explained 10.0%–41.1%of phenotypic variation.Approximately half of the QTL each explained over 10%of the phenotypic variation.These results provide a theoretical basis for improving maize forage quality by marker-assisted selection.

QTL analysis of the developmental changes in cell wall components and forage digestibility in maize(Zea mays L.)

Cell wall architecture plays a key role in stalk strength and forage digestibility.Lignin,cellulose,and hemicellulose are the three main components of plant cell walls,and they can impact stalk quality by affecting the structure and strength of the cell wall.To explore cell wall development during secondary cell wall lignification in maize stalks,conventional and conditional genetic mapping were used to identify the dynamic quantitative trait loci(QTLs)of the cell wall components and digestibility traits during five growth stages after silking.Acid detergent lignin(ADL),cellulose(CEL),acid detergent fiber(ADF),neutral detergent fiber(NDF),and in vitro dry matter digestibility(IVDMD)were evaluated in a maize recombinant inbred line(RIL)population.ADL,CEL,ADF,and NDF gradually increased from 10 to 40 days after silking(DAS),and then they decreased.IVDMD initially decreased until 40 DAS,and then it increased slightly.Seventytwo QTLs were identified for the five traits,and each accounted for 3.48–24.04%of the phenotypic variation.Six QTL hotspots were found,and they were localized in the 1.08,2.04,2.07,7.03,8.05,and 9.03 bins of the maize genome.Within the interval of the pleiotropic QTL identified in bin 1.08 of the maize genome,six genes associated with cell wall component biosynthesis were identified as potential candidate genes for stalk strength as well as cell wall-related traits.In addition,26 conditional QTLs were detected in the five stages for all of the investigated traits.Twenty-two of the 26 conditional QTLs were found at 30 DAS conditioned using the values of 20 DAS,and at 50 DAS conditioned using the values of 40 DAS.These results indicated that cell wall-related traits are regulated by many genes,which are specifically expressed at different stages after silking.Simultaneous improvements in both forage digestibility and lodging resistance could be achieved by pyramiding multiple beneficial QTL alleles identified in this study.

Observation of the curative effect between Nocardia rubra cell wall skeleton-injection combined with the intrapleural chemotherapy and following the intrapleural chemotherapy

Objective:To observe and compare the curative effect between the intrapleural-chemotherapy combined Nocardia rubra cell wall skeleton-injection and the Nocardia rubra cell wall skeleton-injection after control of the malignant pleural effusion by chemotherapy.Methods:Every time after elimination of pleural effusion,we injected DDP and Nocardia rubra cell wall skeleton at the same time into pleural cavity;or after we emplaced an improved central vena catheter into pleural cavity then took out closed-drainage or eliminated the pleural effusion every day,then injected 5-FU 0.5 qd intrapleural-space.After the malignant pleural effusion had being controlled,800μg Nocardia rubra cell wall skeleton was injected intrapleural-space. (We call it as"improved following-therapy").Results:Using the improved following-therapy,the control rate of malignant pleural effusion increased:CR+PR,79.07%>65.79%(P<0.05),and the incidence rate of the encapsulated pleural effusion that would impact on patient’s respiratory faction decreased.Conclusion:Giving chemotherapy after closed-drainage or immediately after taking out pleural effusion everyday,then giving biotherapy after pleural effusion had been controlled,the curative effect would be enhanced and the side-reaction would be reduced.

Plant Cell Wall, a Challenge for Its Characterisation

The plant cell wall is a complex 3D network composed of polysaccharides, lignin and proteins. The knowledge of the structure and content of each cell wall polymer is a prerequisite to understand their functions during plant development and adaptation but also to optimise their industrial applications. The analysis of cell wall compounds is complicated by their multiple molecular interactions. In this review, we present numerous methods to purify, characterise and quantify proteins, polysaccharides and lignin from the wall. Two kinds of approaches are detailed: the first presents in vitro methods which involve the breakdown of the molecular linkages between polymers thanking to chemical, physical and/or enzymatic treatments. The second approach describes in situ methods that allow the cell wall polymer characterisation thanking to many analytical techniques coupled with microscopy. If microscopy is the common point of all of them, their development is associated with improvement of analytical techniques, increasing their power of resolution.

Brittle culm 25, which encodes an UDP-xylose synthase, affects cell wall properties in rice

Because plant mechanical strength influences plant growth and development,the regulatory mechanisms underlying cell-wall synthesis deserve investigation.Rice mutants are useful for such research.We have identified a novel brittle culm 25(bc25)mutant with reduced growth and partial sterility.BC25 encodes an UDP-glucuronic acid decarboxylase involved in cellulose synthesis and belongs to the UXS family.A single-nucleotide mutation in BC25 accounts for its altered cell morphology and cellwall composition.Transmission electron microscopy analysis showed that the thickness of the secondary cell wall was reduced in bc25.Monosaccharide analysis revealed significant increases in content of rhamnose and arabinose but not of other monosaccharides,indicating that BC25 was involved in xylose synthesis with some level of functional redundancy.Enzymatic assays suggested that BC25 functions with high activity to interconvert UDP-glucuronic acid(UDP-Glc A)and UDP-xylose.GUS staining showed that BC25 was ubiquitously expressed with higher expression in culm,root and sheath,in agreement with that shown by quantitative real-time(q RT)-PCR.RNA-seq further suggested that BC25 is involved in sugar metabolism.We conclude that BC25 strongly influences rice cell wall formation.

Comparison of cell wall changes of two different types of apple cultivars during fruit development and ripening

Fruit development and ripening is a complex procedure(Malus×domestica Borkh.)and can be caused by various factors such as cell structure,cell wall components,and cell wall hydrolytic enzymes.In our study,we focused on the variations in fruit firmness,cell wall morphology and components,the activity of cell wall hydrolytic enzymes and the expression patterns of associated genes during fruit development in two different types of apple cultivars,the hard-crisp cultivar and the loose-crisp cultivar.In this paper,the aim was to find out the causes of the texture variations between the different type cultivars.Cell wall materials(CWMs),hemicellulose and cellulose content were strongly associated with variations in fruit firmness during the fruit development.The content of water soluble pectin(WSP)and chelator soluble pectin(CSP)gradually increased,while the content of ionic soluble pectin(ISP)showed inconsistent trends in the four cultivars.The activities of polygalacturonase(PG),β-galactosidase(β-gal),cellulase(CEL),and pectate lyase(PL)gradually increased in four cultivars.And the activities of PG,β-gal,and CEL were higher in‘Fuji’and‘Honeycrisp’fruit with the fruit development,while the activity of PL of‘Fuji’and‘Honeycrisp’was lower than that of‘ENVY’and‘Modi’.Both four cultivars of fruit cells progressively became bigger as the fruit expanded,with looser cell arrangements and larger cell gaps.According to the qRT-PCR,the relative expression levels of MdACO and Mdβ-gal were notably enhanced.Our study showed that there were large differences in the content of ISP and hemicellulose,the activity of PL and the relative expression of Mdβ-gal between two different types of apple cultivars,and these differences might be responsible for the variations in the texture of the four cultivars.

Isolation of Mannooligosaccharides Corresponding to Antigenic Determinants of Pathogenic Yeast <i>Candida catenulata</i>Cell Wall Mannan

To investigate the chemical structure of cell wall mannan of pathogenic yeast, Candida catenulata IFO 0745 strain, which possess the epitopes of antigenic factors 1, 9, and 34 to genus Candida, we previously performed the two-dimensional nuclear magnetic resonance (NMR) analysis of this mannan, Fr. C, without the need for harsh procedures. In this study, three oligosaccharides, biose, triose, and tetraose, and mannose were isolated from Fr. C by acetolysis. The results of NMR analysis indicate that the chemical structures of these oligosaccharides were identified to Manα1-2Man, Manα1-2Manα1-2Man, and Manα1-3Manα1-2Manα1-2Man. The most of resultant mannose seems to be originated from the α-1,6-linked mannan backbone which is recognized by antiserum to factor 9. The inhibition assay of slide agglutination reaction between Fr. C and antigenic antibodies using three oligosaccharides indicate that the Manα1-2Manα1-2Man and Manα1-3Manα1-2Manα1-2Man possess domains corresponding to immunodominants of antigenic factors 1 and 34, respectively.

Impact of Expressing p-Coumaryl Transferase in Medicago sativa L. on Cell Wall Chemistry and Digestibility

The addition of p-coumaric acid (pCA) to lignin molecules is frequently found in members of the grass family. The role of this addition is not clearly understood, but is thought to potentially aid in the formation of syringyl-type lignin. This is because the incorporation is as a conjugate of pCA ester linked to sinapyl alcohol, a major component of lignin. The forage legume alfalfa (Medicago sativa L.) does not contain appreciable levels of pCA in its more heavily lignified stem tissues. The maize p-coumaryltransferase (pCAT) gene was used to transform alfalfa to determine its impact upon lignin composition and its potential to alter cell wall digestibility. A constitutive expression vector using the cassava vein mosaic virus (CsVMV) promoter was used to drive expression of maize pCAT in alfalfa. Expression of the pCAT transgene was detected in both leaves and stems. Though there was a range of pCAconcentration in transformed alfalfa stems (0.2 - 1.79 micrograms (μg)), this was a clear increase over bound pCA in control stems (0.15 - 0.2 mean = 0.17 micrograms (μg)). This did not lead to consistent responses concerning total lignin in the stem tissues. Leaf tissue, on the other hand, already has a relatively high level of pCA (0.85 - 1.2, mean = 0.99 micrograms (μg)) and those expressing pCAT gene showed on average a small increase, but there is a wide range of values among the transformants (0.38 - 1.55, mean = 1.06 micrograms (μg)). Lignin in leaves did not appear to be significantly impacted. However, incorporation of pCA into the wall appears to cause a shift in lignin composition. Testing the pCAT expressing stem cell walls for digestibility using a rumen in vitro system showed there was no change in the digestibility of the stem compared to empty vectors and control alfalfa stems. Although expression of pCAT gene in alfalfa changes the amount of wall bound pCA, it does not appear to change lignin levels or impact digestibility.

Asynchronous abdominal wall and sigmoid metastases in clear cell renal cell carcinoma: A case report and literature review

Sigmoid metastasis of renal cell carcinoma(RCC)is very rare.Herein we report a case of pathologically proven asynchronous abdominal wall and sigmoid metastases after a right nephrectomy.An 84-year-old man underwent right radical nephrectomy for clear cell renal cell carcinoma(ccRCC)13 years ago.Solitary contralateral abdominal wall metastasis was found for left abdominal mass 9 years after nephrectomy.The man experienced melena underwent resection of sigmoid colon tumor in February,2016.The postoperative pathological examinations revealed that the tumors were metastases of ccRCC.Recurrence more than 5 years after nephrectomy has been accepted as late recurrence by the majority of urologists now.Late recurrence is one of the specific biological behaviors of RCC.Asynchronous late recurrence of abdominal wall and sigmoid metastases in ccRCC has not been reported before.When patients have sigmoid mass after nephrectomy for RCC,doctors may consider the possibility of late recurrence.