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Biology Jul 2022Improved cellulose biosynthesis and plant biomass represent important economic targets for several biotechnological applications including bioenergy and biofuel...
Improved cellulose biosynthesis and plant biomass represent important economic targets for several biotechnological applications including bioenergy and biofuel production. The attempts to increase the biosynthesis of cellulose by overexpressing CesAs proteins, components of the cellulose synthase complex, has not always produced consistent results. Analyses of morphological and molecular data and of the chemical composition of cell walls showed that tobacco plants (F1 line), stably expressing the CesA6 fused to GFP, exhibits a "giant" phenotype with no apparent other morphological aberrations. In the F1 line, all evaluated growth parameters, such as stem and root length, leaf size, and lignified secondary xylem, were significantly higher than in wt. Furthermore, F1 line exhibited increased flower and seed number, and an advance of about 20 days in the anthesis. In the leaves of F1 seedlings, the expression of primary (, , and ) was enhanced, as well as of proteins involved in the biosynthesis of non-cellulosic polysaccharides (xyloglucans and galacturonans, , ), cell wall remodeling ( and XTHs), and cell expansion ( and ). While in leaves the expression level of all secondary cell wall (, , and ) did not change significantly, both primary and secondary were differentially expressed in the stem. The amount of cellulose and matrix polysaccharides significantly increased in the F1 seedlings with no differences in pectin and hemicellulose glycosyl composition. Our results highlight the potentiality to overexpress primary in tobacco plants to enhance cellulose synthesis and biomass production.
PubMed: 36009766
DOI: 10.3390/biology11081139 -
Frontiers in Pharmacology 2021infection (CDI) is a leading cause of antibiotic-associated diarrhoea. Adhesion of this Gram-positive pathogen to the intestinal epithelium is a crucial step in CDI,...
infection (CDI) is a leading cause of antibiotic-associated diarrhoea. Adhesion of this Gram-positive pathogen to the intestinal epithelium is a crucial step in CDI, with recurrence and relapse of disease dependent on epithelial interaction of its endospores. Close proximity, or adhesion of, hypervirulent strains to the intestinal mucosa are also likely to be necessary for the release of toxins, which when internalized, result in intestinal epithelial cell rounding, damage, inflammation, loss of barrier function and diarrhoea. Interrupting these -epithelium interactions could therefore represent a promising therapeutic strategy to prevent and treat CDI. Intake of dietary fibre is widely recognised as being beneficial for intestinal health, and we have previously shown that soluble non-starch polysaccharides (NSP) from plantain banana ( spp.), can block epithelial adhesion and invasion of a number of gut pathogens, such as and Salmonellae. Here, we assessed the action of plantain NSP, and a range of alternative soluble plant fibres, for inhibitory action on epithelial interactions of clinical isolates, purified endospore preparations and toxins. We found that plantain NSP possessed ability to disrupt epithelial adhesion of vegetative cells and spores, with inhibitory activity against found within the acidic (pectin-rich) polysaccharide component, through interaction with the intestinal epithelium. Similar activity was found with NSP purified from broccoli and leek, although seen to be less potent than NSP from plantain. Whilst plantain NSP could not block the interaction and intracellular action of purified toxins, it significantly diminished the epithelial impact of , reducing both bacteria and toxin induced inflammation, activation of caspase 3/7 and cytotoxicity in human intestinal cell-line and murine intestinal organoid cultures. Dietary supplementation with soluble NSP from plantain may therefore confer a protective effect in CDI patients by preventing adhesion of to the mucosa, i.e. a "contrabiotic" effect, and diminishing its epithelial impact. This suggests that plantain soluble dietary fibre may be a therapeutically effective nutritional product for use in the prevention or treatment of CDI and antibiotic-associated diarrhoea.
PubMed: 34955836
DOI: 10.3389/fphar.2021.766293 -
Horticulture Research 2020Glyoxalase I (Gly I) is the first enzyme in the glutathionine-dependent glyoxalase pathway for detoxification of methylglyoxal (MG) under stress conditions. Transgenic...
Glyoxalase I (Gly I) is the first enzyme in the glutathionine-dependent glyoxalase pathway for detoxification of methylglyoxal (MG) under stress conditions. Transgenic tomato 'Money Maker' plants overexpressing tomato gene (tomato unigene accession SGN-U582631/Solyc09g082120.3.1) were generated and homozygous lines were obtained after four generations of self-pollination. In this study, overepxressing line (GlyI), wild type (WT, negative control) and plants transformed with empty vector (ECtr, positive control), were subjected to Al-treatment by growing in Magnavaca's nutrient solution (pH 4.5) supplemented with 20 µM Al ion activity. After 30 days of treatments, the fresh and dry weight of shoots and roots of plants from Al-treated conditions decreased significantly compared to the non-treated conditions for all the three lines. When compared across the three lines, root fresh and dry weight of GlyI was significant higher than WT and ECtr, whereas there was no difference in shoot tissues. The basal 5 mm root-tips of GlyI plants expressed a significantly higher level of glyoxalase activity under both non-Al-treated and Al-treated conditions compared to the two control lines. Under Al-treated condition, there was a significant increase in MG content in ECtr and WT lines, but not in GlyI line. Quantitative proteomics analysis using tandem mass tags mass spectrometry identified 4080 quantifiable proteins and 201 Al-induced differentially expressed proteins (DEPs) in root-tip tissues from GlyI, and 4273 proteins and 230 DEPs from ECtr. The Al-down-regulated DEPs were classified into molecular pathways of gene transcription, RNA splicing and protein biosynthesis in both GlyI and ECtr lines. The Al-induced DEPs in GlyI associated with tolerance to Al and MG toxicity are involved in callose degradation, cell wall components (xylan acetylation and pectin degradation), oxidative stress (antioxidants) and turnover of Al-damaged epidermal cells, repair of damaged DNA, epigenetics, gene transcription, and protein translation. A protein-protein association network was constructed to aid the selection of proteins in the same pathway but differentially regulated in GlyI or ECtr lines. Proteomics data are available via ProteomeXchange with identifiers PXD009456 under project title '25Dec2017_Suping_XSexp2_ITAG3.2' for overexpressing tomato plants and PXD009848 under project title '25Dec2017_Suping_XSexp3_ITAG3.2' for positive control ECtr line transformed with empty vector.
PubMed: 32257229
DOI: 10.1038/s41438-020-0264-x -
International Journal of Molecular... Sep 2023Common smut caused by is one of the dominant fungal diseases in plants. The resistance mechanism to infection involving alterations in the cell wall is poorly studied....
Common smut caused by is one of the dominant fungal diseases in plants. The resistance mechanism to infection involving alterations in the cell wall is poorly studied. In this study, the resistant single segment substitution line (SSSL) R445 and its susceptible recurrent parent line Ye478 of maize were infected with , and the changes in cell wall components and structure were studied at 0, 2, 4, 8, and 12 days postinfection. In R445 and Ye478, the contents of cellulose, hemicellulose, pectin, and lignin increased by varying degrees, and pectin methylesterase (PME) activity increased. The changes in hemicellulose and pectin in the cell wall after infection were analyzed via immunolabeling using monoclonal antibodies against hemicellulsic xylans and high/low-methylated pectin. infection altered methyl esterification of pectin, and the degree of methyl esterification was correlated with the resistance of maize to . Furthermore, the relationship between methyl esterification of pectin and host resistance was validated using 15 maize inbred lines with different resistance levels. The results revealed that cell wall components, particularly pectin, were important factors affecting the colonization and propagation of in maize, and methyl esterification of pectin played a role in the resistance of maize to infection.
Topics: Plant Diseases; Esterification; Zea mays; Pectins; Ustilago; Cell Wall
PubMed: 37834187
DOI: 10.3390/ijms241914737 -
Protective, Antioxidant and Antiproliferative Activity of Grapefruit IntegroPectin on SH-SY5Y Cells.International Journal of Molecular... Aug 2021Tested in vitro on SH-SY5Y neuroblastoma cells, grapefruit IntegroPectin is a powerful protective, antioxidant and antiproliferative agent. The strong antioxidant...
Tested in vitro on SH-SY5Y neuroblastoma cells, grapefruit IntegroPectin is a powerful protective, antioxidant and antiproliferative agent. The strong antioxidant properties of this new citrus pectin, and its ability to preserve mitochondrial membrane potential and morphology, severely impaired in neurodegenerative disorders, make it an attractive therapeutic and preventive agent for the treatment of oxidative stress-associated brain disorders. Similarly, the ability of this pectic polymer rich in RG-I regions, as well as in naringin, linalool, linalool oxide and limonene adsorbed at the outer surface, to inhibit cell proliferation or even kill, at high doses, neoplastic cells may have opened up new therapeutic strategies in cancer research. In order to take full advantage of its vast therapeutic and preventive potential, detailed studies of the molecular mechanism involved in the antiproliferative and neuroprotective of this IntegroPectin are urgently needed.
Topics: Antioxidants; Cell Line, Tumor; Cell Proliferation; Cell Survival; Citrus paradisi; Humans; Membrane Potential, Mitochondrial; Neuroblastoma; Neuroprotective Agents; Pectins; X-Ray Diffraction
PubMed: 34502276
DOI: 10.3390/ijms22179368 -
Carbohydrate Polymers Sep 2022Roots of Aconitum carmichaelii are used in Asian countries due to its content of bioactive alkaloids. In the production of root preparations, tons of leaves are usually...
Roots of Aconitum carmichaelii are used in Asian countries due to its content of bioactive alkaloids. In the production of root preparations, tons of leaves are usually discarded, leading to a huge waste of herbal material. The aim of this study is to investigate the polysaccharides in these unutilized leaves. A neutral polysaccharide (AL-N) appeared to be a mixture of heteromannans, and two purified acidic polysaccharides (AL-I-I and AL-I-II) were shown to be pectins containing a homogalacturonan backbone substituted with terminal β-Xylp-units. AL-I-I consisted of a type-I rhamnogalacturonan core, with arabinan and type-II arabinogalactan domains while AL-I-II was less branched. AL-N and AL-I-I were able to modulate the complement system, while AL-I-II was inactive. Interestingly, AL-N, AL-I-I and AL-I-II were shown to exert anti-inflammatory effects on porcine enterocyte IPEC-J2 cells. AL-I-I and AL-I-II were able to down-regulate the expression of toll-like receptor 4 (TLR4) and nucleotide-binding oligomerization domain 1 (NOD1).
Topics: Aconitum; Alkaloids; Animals; Anti-Inflammatory Agents; Cell Line; Enterocytes; Plant Leaves; Polysaccharides; Swine
PubMed: 35698356
DOI: 10.1016/j.carbpol.2022.119655 -
Polymers May 2023In recent years, natural polysaccharides have been widely used in the preparation of drug delivery systems. In this paper, novel polysaccharide-based nanoparticles were...
In recent years, natural polysaccharides have been widely used in the preparation of drug delivery systems. In this paper, novel polysaccharide-based nanoparticles were prepared by layer-by-layer assembly technology using silica as a template. The layers of nanoparticles were constructed based on the electrostatic interaction between a new pectin named NPGP and chitosan (CS). The targeting ability of nanoparticles was formed by grafting the RGD peptide, a tri-peptide motif containing arginine, glycine, and aspartic acid with high affinity to integrin receptors. The layer-by-layer assembly nanoparticles (RGD-(NPGP/CS)NPGP) exhibited a high encapsulation efficiency (83.23 ± 6.12%), loading capacity (76.51 ± 1.24%), and pH-sensitive release property for doxorubicin. The RGD-(NPGP/CS)NPGP nanoparticles showed better targeting to HCT-116 cells, the integrin αvβ3 high expression human colonic epithelial tumor cell line with higher uptake efficiency than MCF7 cells, the human breast carcinoma cell line with normal integrin expression. In vitro antitumor activity tests showed that the doxorubicin-loaded nanoparticles could effectively inhibit the proliferation of the HCT-116 cells. In conclusion, RGD-(NPGP/CS)NPGP nanoparticles have potential as novel anticancer drug carriers because of their good targeting and drug-carrying activity.
PubMed: 37299309
DOI: 10.3390/polym15112510 -
Frontiers in Plant Science 2021Flesh firmness of watermelon is an important quality trait for commercial fruit values, including fruit storability, transportability, and shelf life. To date, knowledge...
Flesh firmness of watermelon is an important quality trait for commercial fruit values, including fruit storability, transportability, and shelf life. To date, knowledge of the gene networks underlying this trait is still limited. Herein, we used weighted genes co-expression network analysis (WGCNA) based on correlation and the association of phenotypic data (cell wall contents) with significantly differentially expressed genes between two materials, a near isogeneic line "HWF" (with high average flesh firmness) and inbred line "203Z" (with low average flesh firmness), to identify the gene networks responsible for changes in fruit flesh firmness. We identified three gene modules harboring 354 genes; these gene modules demonstrated significant correlation with water-soluble pectin, cellulose, hemicellulose, and protopectin. Based on intramodular significance, eight genes involved in cell wall biosynthesis and ethylene pathway are identified as hub genes within these modules. Among these genes, two genes, (Cellulose synthase) and (Pectinesterase), were significantly correlated with cellulose ( = 0.83) and protopectin ( = 0.81); three genes, (ERF1), (Cellulose synthase), and (Galactosyltransferase), had a significant correlation with water-soluble pectin ( = 0.91), cellulose ( = 0.9), and protopectin ( = 0.92); and three genes, (ERF2a), (probable glycosyltransferase), and (Xyloglucan endotransglucosylase/hydrolase), were correlated with hemicellulose ( = 0.85), cellulose ( = 0.8), and protopectin ( = 0.8). This study generated important insights of biosynthesis of a cell wall structure and ethylene signaling transduction pathway, the mechanism controlling the flesh firmness changes in watermelon, which provide a significant source to accelerate future functional analysis in watermelon to facilitate crop improvement.
PubMed: 34239519
DOI: 10.3389/fpls.2021.630243 -
A Guide to Elucidate the Hidden Multicomponent Layered Structure of Plant Cuticles by Raman Imaging.Frontiers in Plant Science 2021The cuticle covers almost all plant organs as the outermost layer and serves as a transpiration barrier, sunscreen, and first line of defense against pathogens. Waxes,...
The cuticle covers almost all plant organs as the outermost layer and serves as a transpiration barrier, sunscreen, and first line of defense against pathogens. Waxes, fatty acids, and aromatic components build chemically and structurally diverse layers with different functionality. So far, electron microscopy has elucidated structure, while isolation, extraction, and analysis procedures have revealed chemistry. With this method paper, we close the missing link by demonstrating how Raman microscopy gives detailed information about chemistry and structure of the native cuticle on the microscale. We introduce an optimized experimental workflow, covering the whole process of sample preparation, Raman imaging experiment, data analysis, and interpretation and show the versatility of the approach on cuticles of a spruce needle, a tomato peel, and an Arabidopsis stem. We include laser polarization experiments to deduce the orientation of molecules and multivariate data analysis to separate cuticle layers and verify their molecular composition. Based on the three investigated cuticles, we discuss the chemical and structural diversity and validate our findings by comparing models based on our spectroscopic data with the current view of the cuticle. We amend the model by adding the distribution of cinnamic acids and flavonoids within the cuticle layers and their transition to the epidermal layer. Raman imaging proves as a non-destructive and fast approach to assess the chemical and structural variability in space and time. It might become a valuable tool to tackle knowledge gaps in plant cuticle research.
PubMed: 34975980
DOI: 10.3389/fpls.2021.793330 -
Frontiers in Nutrition 2021The purpose of this study is to quantify the effect of adding sodium alginate and pectin to a carbohydrate (CHO) beverage on exogenous glucose (ExGluc) oxidation rate...
PURPOSE
The purpose of this study is to quantify the effect of adding sodium alginate and pectin to a carbohydrate (CHO) beverage on exogenous glucose (ExGluc) oxidation rate compared with an isocaloric CHO beverage.
METHODS
Following familiarization, eight well-trained endurance athletes performed four bouts of prolonged running (105 min; 71 ± 4% of VOmax) while ingesting 175 mL of one of the experimental beverages every 15 min. In randomized order, participants consumed either 70 gh of maltodextrin and fructose (10% CHO; NORM), 70 gh of maltodextrin, fructose, sodium alginate, and pectin (10% CHO; ENCAP), 180 gh of maltodextrin, fructose, sodium alginate, and pectin (26% CHO; HiENCAP), or water (WAT). All CHO beverages had a maltodextrin:fructose ratio of 1:0.7 and contained 1.5 gL of sodium chloride. Total substrate oxidation, ExGluc oxidation rate, blood glucose, blood lactate, serum non-esterified fatty acid (NEFA) concentration, and RPE were measured for every 15 min. Every 30 min participants provided information regarding their gastrointestinal discomfort (GID).
RESULTS
There was no significant difference in peak ExGluc oxidation between NORM and ENCAP (0.63 ± 0.07 and 0.64 ± 0.11 gmin, respectively; > 0.5), both of which were significantly lower than HiENCAP (1.13 ± 0.13 gmin, < 0.01). Both NORM and HiENCAP demonstrated higher total CHO oxidation than WAT from 60 and 75 min, respectively, until the end of exercise, with no differences between CHO trials. During the first 60 min, blood glucose was significantly lower in WAT compared with NORM and HiENCAP, but no differences were found between CHO beverages. Both ENCAP and HiENCAP demonstrated a higher blood glucose concentration from 60-105 min than WAT, and ENCAP was significantly higher than HiENCAP. There were no significant differences in reported GID symptoms between the trials.
CONCLUSIONS
At moderate ingestion rates (i.e., 70 gh), the addition of sodium alginate and pectin did not influence the ExGluc oxidation rate compared with an isocaloric CHO beverage. At very high ingestion rates (i.e., 180 gh), high rates of ExGluc oxidation were achieved in line with the literature.
PubMed: 35127792
DOI: 10.3389/fnut.2021.810041