-
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 Plant Science 2022Gravity is known as an important environmental factor involved in the regulation of plant architecture. To identify genes related to the gravitropism of Tartary...
Gravity is known as an important environmental factor involved in the regulation of plant architecture. To identify genes related to the gravitropism of Tartary buckwheat, a creeping line was obtained and designated as from the mutant bank by Co-γ ray radiation. Genetic analysis indicated that the creeping phenotype of was attributed to a single recessive locus. As revealed by the horizontal and inverted suspension tests, was completely lacking in shoot negative gravitropism. The creeping growth of occurred at the early seedling stage, which could not be recovered by exogenous heteroauxin, hormodin, α-rhodofix, or gibberellin. Different from the well-organized and equivalent cell elongation of wild type (WT), exhibited dilated, distorted, and abnormally arranged cells in the bending stem. However, no statistical difference of indole-3-acetic acid (IAA) levels was found between the far- and near-ground bending sides in , which suggests that the asymmetric cell elongation of was not induced by auxin gradient. Whereas, showed up-expressed gibberellin-regulated genes by quantitative real-time PCR (qRT-PCR) as well as significantly higher levels of gibberellin, suggesting that gibberellin might be partly involved in the regulation of creeping growth in . RNA sequencing (RNA-seq) identified a number of differentially expressed genes (DEGs) related to gravitropism at stages I (before bending), II (bending), and III (after bending) between WT and . Venn diagram indicated that only was down-expressed at stages I [Log fold change (LogFC): -3.20], II (LogFC: -4.97), and III (LogFC: -1.23) in , compared with WT. Gene sequencing revealed that a fragment deletion occurred in the coding region of , which induced the destruction of a pbH domain in of . qRT-PCR indicated that was extremely down-expressed in at stage II (0.02-fold of WT). Meanwhile, showed the affected expression of lignin- and cellulose-related genes and cumulatively abnormal levels of pectin, lignin, and cellulose. These results demonstrate the possibility that functions as the key gene that could mediate primary cell wall metabolism and get involved in the asymmetric cell elongation regulation of .
PubMed: 35574111
DOI: 10.3389/fpls.2022.815131 -
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 -
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 -
Scientific Reports Dec 2020Contemplating the exemplary benefits of pectin on human health, we precisely characterized and evaluated the antibacterial and anticancer activities from purified...
Contemplating the exemplary benefits of pectin on human health, we precisely characterized and evaluated the antibacterial and anticancer activities from purified Mulberry Fruit Pectins (MFP). Here, we tested BR-2 and S-13 varieties of mulberry fruit pectins against six bacterial strains and two human cancer cell lines (HT-29 and Hep G-2), using MIC and an in vitro cell-based assay respectively. The BR-2 mulberry fruit pectin performs superior to S-13 by inhibiting strong bacterial growth (MIC = 500-1000 μg/mL) against tested bacterial strains and cytotoxic activities at the lowest concentration (10 µg/ml) against the Hep G-2 cell line. However, both tested drugs failed to exhibit cytotoxicity on the human colon cancer cell line (HT-29). Based on molecular interaction through docking, pectin binds effectively with the receptors (1e3g, 3t0c, 5czz, 6j7l, 6v40, 5ibs, 5zsy, and 6ggb) and proven to be a promising antimicrobial and anti-cancer agents. The pursuit of unexploited drugs from mulberry fruit pectin will potentially combat against bacterial and cancer diseases. Finally, future perspectives of MFP for the treatment of many chronic diseases will help immensely due to their therapeutic properties.
Topics: Anti-Bacterial Agents; Cytotoxins; Drug Screening Assays, Antitumor; Fruit; HT29 Cells; Hep G2 Cells; Humans; Molecular Docking Simulation; Morus; Pectins
PubMed: 33311512
DOI: 10.1038/s41598-020-78086-8 -
Bio-medical Materials and Engineering 2018Novel pectin-honey hydrogels have been developed and characterized as medical device. Ideally, a wound dressing should maintain optimal fluid affinity, permit moisture...
BACKGROUND
Novel pectin-honey hydrogels have been developed and characterized as medical device. Ideally, a wound dressing should maintain optimal fluid affinity, permit moisture evaporation, protect the wound from microbes, and have shape-conformability, biocompatibility, and antibacterial activity.
OBJECTIVE
A novel, simple and fast method to produce pectin-honey wound dressings is described.
METHODS
The properties of these pectin-honey hydrogels were investigated, including swelling ability, water vapour transmission rate, hydrogen peroxide production, methylglyoxal content and antibacterial activity. Biocompatibility was assessed by proliferation assays using cultured fibroblast cells and by in vivo study with subcutaneous and intraperitoneal implantation in rats.
RESULTS
Hydrogel showed a good water vapour transmission rate, fluid uptake and were not cytotoxic for fibroblasts. The hydrogel demonstrated good antibacterial activity toward clinically relevant pathogens, including S. aureus and E. coli. Biocompatibility was confirmed by the measurement of plasma levels of interleukin (IL)1 beta, IL-6, tumour necrosis factor (TNF) alpha, and prostaglandin (PG)E2. No histological changes were observed.
CONCLUSIONS
The presence of a natural active component, conformability, and complete resorbability are the main characteristics of this new biocompatible biomaterial that is well tolerated by the body, possibly improves healing, may be used for surgical complications prevention, with a simple and inexpensive production process.
Topics: Animals; Anti-Bacterial Agents; Bandages; Biocompatible Materials; Cell Line; Escherichia coli; Escherichia coli Infections; Fibroblasts; Honey; Hydrogels; Male; Materials Testing; Mice; Pectins; Rats, Sprague-Dawley; Staphylococcal Infections; Staphylococcus aureus; Wound Healing
PubMed: 29578463
DOI: 10.3233/BME-181730 -
Antioxidants (Basel, Switzerland) Jan 2020This study aimed to identify and quantify the chemical composition and polyphenolic profile of 19 cultivars of Chaenomeles × superba, a, and by liquid chromatography...
ABTS On-Line Antioxidant, α-Amylase, α-Glucosidase, Pancreatic Lipase, Acetyl- and Butyrylcholinesterase Inhibition Activity of Fruits Determined by Polyphenols and other Chemical Compounds.
This study aimed to identify and quantify the chemical composition and polyphenolic profile of 19 cultivars of Chaenomeles × superba, a, and by liquid chromatography coupled with photodiode array detector and quadrupole time-of-flight electrospray ionization mass spectrometry (LC-PDA-QTOF-ESI-MS). Antioxidant (ABTS on-line, ABTS, FRAP, and ORAC), as well as in vitro biological activities, i.e., the ability to inhibit α-amylase, α-glucosidase, pancreatic lipase, acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and 15-lipoxygenase (15-LOX) were determined. Most of the species and cultivars analyzed in this study have not been examined in this respect until now. Fruits contained 30.26 to 195.05 mg of vitamin C, 0.65 to 1.69 g of pectin, 0.32 to 0.64 g of ash, 0.60 to 3.98 g of sugars, and 41.64 to 110.31 g of organic acids in 100 g fresh weight. The lowest content of total polyphenols showed 'Rubra' (57.84 g/kg dry weight, dw) while × 'Nicoline' (170.38 g/kg dw) exhibited the highest concentration of those compounds. In the phenolic compounds, polymeric procyanidin fraction predominated (65%) with procyanidin B2, C1, and (-)-epicatechin the most abundant. The antioxidant capacity measured by ABTS assay was mainly formed by polymeric procyanidins and flavan-3-ols, which was confirmed by ABTS on-line profiling. Chaenomeles fruits showed high potential for inhibition of α-glucosidase and pancreatic lipase. The analyzed cultivars displayed greater potential for acetylcholinesterase (AChE) inhibition than for butyrylcholinesterase (BuChE). The data indicate that Chaenomeles fruits could be regarded as a promising source of bioactive functional food.
PubMed: 31936619
DOI: 10.3390/antiox9010060 -
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 -
Precision Clinical Medicine Jun 2020Increasing resistance to antibiotics has ledthat molecular testing is appropriate as a sub to adoption of seven different bismuth quadruple therapies (BQT) in China...
Increasing resistance to antibiotics has ledthat molecular testing is appropriate as a sub to adoption of seven different bismuth quadruple therapies (BQT) in China without differentiation of first-line or second-line regimens. The objective of this study was to evaluate the efficacy of susceptibility-guided BQT for patients who had experienced previous treatment failures. A total of 133 patients was included and was successfully cultured from 101 patients (75.9%) for subsequent antimicrobial susceptibility testing (AST). Based on the AST results, 88 patients completed one of five AST-guided 14-day BQT regimens: esomeprazole and bismuth colloidal pectin, along with either, amoxicillin and clarithromycin (EBAC), amoxicillin and levofloxacin (EBAL), amoxicillin and furazolidone (EBAF), amoxicillin and tetracycline (EBAT), or tetracycline and furazolidone (EBTF). eradication rates were 100% for EBAC (5/5), EBAL (13/13), EBAF (14/14), and EBTF (43/43), but 76.9% for EBAT (10/13). The three patients that failed the EBAT regimen were all cured after subsequent treatment with the EBTF regimen. Our study demonstrates the excellent efficacy of the AST-guided BQT for referred patients, and that the current EBAT regimen, used in clinics, needs to be optimized. In addition, 57 of the isolates were subjected to whole-genome sequencing. Analysis of the sequences revealed that point mutations in 23S rRNA correlated well with the phenotypic clarithromycin resistance with a concordance of 91.2%, while the concordance between phenotypic levofloxacin resistance and point mutations was 82.3%. This suggests that molecular testing is appropriate as a substitute for AST as a more rapid and cost-effective method for determining clarithromycin and levofloxacin resistance in Chinese patients.
PubMed: 35692608
DOI: 10.1093/pcmedi/pbaa010 -
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