Analysis of cell wall hardening and cell wall enzymes of salt-stressed maize (Zea mays) leaves

2001 ◽  
Vol 28 (2) ◽  
pp. 101 ◽  
Author(s):  
Grant R. Cramer ◽  
Connie L. Schmidt ◽  
Chad Bidart
Keyword(s):  
Cell Wall ◽  
Zea Mays ◽  
Cell Walls ◽  
Leaf Growth ◽  
Physical Property ◽  
Physical Structure ◽  
In Vitro Assays ◽  
Xyloglucan Endotransglycosylase ◽  
Yield Threshold

It has been indicated that salinity inhibits maize (Zea mays L.) leaf growth and leaf cell expansion by increasing the apparent yield threshold of the cell wall. We tested whether this increase in the apparent yield threshold was a physical property of cell walls, using in vitro creep-type assays. Salinity had no significant effects on cell wall structural properties based upon several different in vitro assays. In support of these results, there were no differences between control and salt-stressed plants in their total apoplastic concentration of cell wall proteins, in the activity of apoplastic peroxidases or xyloglucan endotransglycosylase. We conclude that short-term salinity does not appear to inhibit maize leaf elongation by hardening the physical structure of the cell walls of the growing zone.

scholarly journals Confirmation of Cage Effect and Prebiotic Production Potential of a β-Mannanase, with SBM as Substrate Using Microscopy and Wet Chemistry

2021 ◽  
Vol 13 (2) ◽  
pp. 23
Author(s):  
L. M. Gomez-Osorio ◽  
Hwa Gyun Oh ◽  
Jung Jin Lee
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Degree Of Polymerization ◽  
In Vitro Assays ◽  
Wall Structure ◽  
Specific Substrate ◽  
Mannan Oligosaccharides

In vitro assays were carried out to investigate the solubilization of cell walls and generation of mannan oligosaccharides of a b-mannanase-containing commercial product on SBM. Using commercial dosages of the b-mannanase (500 g per ton of feed) cell wall degradation of mannan in SBM cell walls was visualized and an increase in reducing ends (0.12±0.02 mg/mL) and the generation of mannan oligosaccharides of degree of polymerization 2 and 4 (22.9±3.2 mg/L and 398.8±25.4 mg/L) were also measured using HPLC. Mannan, which is H-bonded to cellulose and xyloglucan, was solubilized using a single monocomponent enzyme, allowing for visualization of the disintegration of the entire SBM cell wall structure. This work is the first of its kind using strictly commercial dosage levels of enzyme for evaluating efficacy of the same microscopically. These data confirm the hypothesis that there most likely is a need for only a single relevant NSP enzyme targeting its specific substrate, independent of the concentration of the latter within the complex polysaccharide matrix in the plant cell wall to experience the beneficial effects of the enzyme both in vitro and in vivo. An analogy to compare our data would be destruction of the foundation (mannan) of a building or a bridge (soybean cell wall) which would inevitably lead to dismantling or demolition the entire building or bridge.

Napropamide Binding in Corn (Zea mays) Root Tissue

Weed Science ◽  
1983 ◽  
Vol 31 (5) ◽  
pp. 712-719 ◽  
Author(s):  
Michael Barrett ◽  
Floyd M. Ashton
Keyword(s):  
Cell Wall ◽  
Zea Mays ◽  
Cell Walls ◽  
Wall Material ◽  
Cell Wall Material ◽  
Protease Treatment ◽  
Wall Components ◽  
Soluble Components ◽  
Soluble Fractions

Napropamide [2-(α-naphthoxy)-N,N-diethylpropionamide]-binding in excised root segments of corn (Zea maysL. ‘NC + 59′) was confined to cell wall fractions (residue and 500gpellet) remaining after homogenization and to components of the 100 000gsupernatant. Binding increased in both the cell wall and soluble fractions with continued exposure to napropamide. Microautoradiographs revealed that the napropamide bound in the cell walls was located in epidermal, cortical, and stelar tissue. Various proteins were capable of binding napropamide in vitro; however, protease treatment did not liberate the radioactivity bound in the cell wall fragments. Carbohydrate release from the cell wall material with cellulase was not correlated with the solubilization of bound radioactivity and wall carbohydrate monomers did not appear to bind to napropamide in vitro. A portion of the radioactivity found in the soluble components (at 100 000g) was associated with a molecule of MW > 600. The continued influx of napropamide was due to binding to cell wall components and molecules within the cell.

scholarly journals Co-operative action by endo- and exo-β-(1→3)-glucanases from parasitic fungi in the degradation of cell-wall glucans of Sclerotinia sclerotiorum (Lib.) de Bary

1974 ◽  
Vol 140 (1) ◽  
pp. 47-55 ◽  
Author(s):  
David Jones ◽  
Alex. H. Gordon ◽  
John S. D. Bacon
Keyword(s):  
Cell Wall ◽  
Sclerotinia Sclerotiorum ◽  
Culture Filtrate ◽  
Cell Walls ◽  
Trichoderma Viride ◽  
Major Constituent ◽  
Coniothyrium Minitans ◽  
Parasitic Fungi ◽  
Complete Breakdown

1. Two fungi, Coniothyrium minitans Campbell and Trichoderma viride Pers. ex Fr., were grown on autoclaved crushed sclerotia of the species Sclerotinia sclerotiorum, which they parasitize. 2. in vitro the crude culture filtrates would lyse walls isolated from hyphal cells or the inner pseudoparenchymatous cells of the sclerotia, in which a branched β-(1→3)-β-(1→6)-glucan, sclerotan, is a major constituent. 3. Chromatographic fractionation of the enzymes in each culture filtrate revealed the presence of several laminarinases, the most active being an exo-β-(1→3)-glucanase, known from previous studies to attack sclerotan. Acting alone this brought about a limited degradation of the glucan, but the addition of fractions containing an endo-β-(1→3)-glucanase led to almost complete breakdown. A similar synergism between the two enzymes was found in their lytic action on cell walls. 4. When acting alone the endo-β-(1→3)-glucanase had a restricted action, the products including a trisaccharide, tentatively identified as 62-β-glucosyl-laminaribiose. 5. These results are discussed in relation to the structure of the cell walls and of their glucan constituents.

scholarly journals A genomic and proteomic analysis of surface proteins in bovine-adapted lineages of Staphylococcus aureus

2020 ◽  
Vol 2 (7A) ◽  
Author(s):  
Shauna D. Drumm ◽  
Rebecca Owens ◽  
Jennifer Mitchell ◽  
Orla M. Keane
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Surface Proteins ◽  
Host Cells ◽  
Mass Spectrometry Analysis ◽  
In Vitro Assays ◽  
Immune Recognition ◽  
Independent Manner ◽  
Genes Encoding

In Ireland, Staphylococcus aureus is the most common cause of intramammary infection (IMI) in cattle with the bovine-adapted lineages CC151 and CC97 most commonly found. Surface proteins play a major role in establishing and maintaining the infection. A previous study revealed that a strain from the CC151 lineage showed significant decay in genes encoding predicted surface proteins. Twenty-three S. aureus strains, twelve belonging to CC151 and eleven belonging to CC97, isolated from clinical IMI, were sequenced and genes encoding cell wall anchored (CWA) proteins predicted. Analysis showed that a minority of genes encoding putative CWA proteins were intact in the CC151 strains compared to CC97. Of the 26 known CWA proteins in S. aureus, the CC151 strains only encoded 10 intact genes while CC97 encoded on average 18 genes. Also within the CC97 lineage, the repertoire of genes varied depending on individual strains, with strains encoding between 17-20 intact genes. Although CC151 is reported to internalize within bovine host cells, it does so in a fibronectin-binding protein (FnBPA and FnBPB) independent manner. In-vitro assays were performed and results showed that strains from CC151, and surprisingly also CC97, weakly bound bovine fibronectin and that the FnBPs were poorly expressed in both these lineages. Mass spectrometry analysis of cell wall extracts revealed that SdrE and AdsA were the most highly expressed CWA proteins in both lineages. These results demonstrate significant differences between CC151 and CC97 in their repertoire of genes encoding CWA proteins, which may impact immune recognition of these strains and their interactions with host cells.

scholarly journals Melanin Externalization in Candida albicans Depends on Cell Wall Chitin Structures

2010 ◽  
Vol 9 (9) ◽  
pp. 1329-1342 ◽  
Author(s):  
Claire A. Walker ◽  
Beatriz L. Gómez ◽  
Héctor M. Mora-Montes ◽  
Kevin S. Mackenzie ◽  
Carol A. Munro ◽  
...  
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Fungal Pathogen ◽  
Cell Walls ◽  
Chitin Synthesis ◽  
Melanin Production ◽  
Content Type ◽  
Encoding Gene ◽  
Chitinase Genes

ABSTRACT The fungal pathogen Candida albicans produces dark-pigmented melanin after 3 to 4 days of incubation in medium containing l-3,4-dihydroxyphenylalanine (l-DOPA) as a substrate. Expression profiling of C. albicans revealed very few genes significantly up- or downregulated by growth in l-DOPA. We were unable to determine a possible role for melanin in the virulence of C. albicans. However, we showed that melanin was externalized from the fungal cells in the form of electron-dense melanosomes that were free or often loosely bound to the cell wall exterior. Melanin production was boosted by the addition of N-acetylglucosamine to the medium, indicating a possible association between melanin production and chitin synthesis. Melanin externalization was blocked in a mutant specifically disrupted in the chitin synthase-encoding gene CHS2. Melanosomes remained within the outermost cell wall layers in chs3Δ and chs2Δ chs3Δ mutants but were fully externalized in chs8Δ and chs2Δ chs8Δ mutants. All the CHS mutants synthesized dark pigment at equivalent rates from mixed membrane fractions in vitro, suggesting it was the form of chitin structure produced by the enzymes, not the enzymes themselves, that was involved in the melanin externalization process. Mutants with single and double disruptions of the chitinase genes CHT2 and CHT3 and the chitin pathway regulator ECM33 also showed impaired melanin externalization. We hypothesize that the chitin product of Chs3 forms a scaffold essential for normal externalization of melanosomes, while the Chs8 chitin product, probably produced in cell walls in greater quantity in the absence of CHS2, impedes externalization.

In vitro digestibility, breakdown when eaten and physical structure of stovers and straws compared with lucerne hay and sweet potato haulm

1999 ◽  
Vol 132 (4) ◽  
pp. 491-498 ◽  
Author(s):  
D. WILMAN ◽  
YILUN JI ◽  
E. J. MTENGETI ◽  
NAZIR AHMAD
Keyword(s):  
Cell Wall ◽  
Sweet Potato ◽  
Rice Straw ◽  
Physical Structure ◽  
Vascular Bundles ◽  
Milled Material ◽  
Maize Leaves ◽  
Stem Internode ◽  
The Mean

In order to learn more about the digestibility, breakdown when eaten and physical structure of stovers and straws, seven diets were compared in one experiment and eight in another. The diets in the first experiment were: the upper and lower parts of lucerne (Medicago sativa) hay, the leaves, upper stem and lower stem of maize (Zea mays) stover and the leaves and stem of sorghum (Sorghum vulgare) stover. The diets in the second experiment were: the upper and lower parts of lucerne hay, the leaves, stems and whole stover of millet (Setaria italica), wheat (Triticum aestivum) straw, rice (Oryza sativa) straw and sweet potato (Ipomoea batatas) haulm. The diets were examined for in vitro digestibility in three forms: milled, chopped (1 cm lengths) and chewed (by cattle). The lengths and widths of the chewed particles were recorded in both experiments. Aspects of the morphology and vascular structure of plant parts of the diets and of chewed particles were recorded in the second experiment.The in vitro digestibility of chopped or chewed plant material was lower (by 11 and 12 percentage units, respectively, on average) than that of milled material. The difference between chewed and milled material in in vitro digestibility was smaller with leaves (5 percentage units, on average) than with stems (18 percentage units, on average) in maize, sorghum and millet. Digestibility of chewed material was similar to or lower than that of chopped material, except with the leaves of maize, sorghum and millet. Millet stover had higher in vitro digestibility than wheat straw or rice straw, whether the diet was milled, chopped or chewed.The thinnest stem internode vascular bundles recorded (0·14 mm) were in wheat. The percentage of stem internode cross-sectional area occupied by vascular bundles was in the range 6–8 for lucerne, millet, wheat and rice. The mean numbers of veins or vascular bundles per chewed particle were in the range 6–23. The mean length of the chewed particles varied from 10 mm with the upper stem of maize to 18 mm with sorghum stem and maize leaves. The mean width of the chewed particles varied from 1·8 mm with rice straw to 5·4 mm with maize leaves.It is suggested that the low digestibility of mature plant tissue such as stovers and straws is not particularly due to thick vascular strands or to a high proportion of vascular tissue, but rather to a high proportion of cell wall and incomplete and delayed access by rumen microorganisms to much of the cell wall.

scholarly journals Evaluation of Antifungal Activity and Mode of Action of New Coumarin Derivative, 7-Hydroxy-6-nitro-2H-1-benzopyran-2-one, againstAspergillusspp.

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Felipe Queiroga Sarmento Guerra ◽  
Rodrigo Santos Aquino de Araújo ◽  
Janiere Pereira de Sousa ◽  
Fillipe de Oliveira Pereira ◽  
Francisco J. B. Mendonça-Junior ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mode Of Action ◽  
Cell Walls ◽  
Coumarin Derivative ◽  
Antifungal Drugs ◽  
Azole Resistance ◽  
Subinhibitory Concentration ◽  
Fungal Cell ◽  
Mycelia Growth

Aspergillusspp. produce a wide variety of diseases. For the treatment of such infections, the azoles and Amphotericin B are used in various formulations. The treatment of fungal diseases is often ineffective, because of increases in azole resistance and their several associated adverse effects. To overcome these problems, natural products and their derivatives are interesting alternatives. The aim of this study was to examine the effects of coumarin derivative, 7-hydroxy-6-nitro-2H-1-benzopyran-2-one (Cou-NO2), both alone and with antifungal drugs. Its mode of action againstAspergillusspp. Cou-NO2was tested to evaluate its effects on mycelia growth and germination of fungal conidia ofAspergillusspp. We also investigated possible Cou-NO2action on cell walls (0.8 M sorbitol) and on Cou-NO2to ergosterol binding in the cell membrane. The study shows that Cou-NO2is capable of inhibiting both the mycelia growth and germination of conidia for the species tested, and that its action affects the structure of the fungal cell wall. At subinhibitory concentration, Cou-NO2enhanced thein vitroeffects of azoles. Moreover, in combination with azoles (voriconazole and itraconazole) Cou-NO2displays an additive effect. Thus, our study supports the use of coumarin derivative 7-hydroxy-6-nitro-2H-1-benzopyran-2-one as an antifungal agent againstAspergillusspecies.

scholarly journals Putative Role of β-1,3 Glucans in Candida albicans Biofilm Resistance

2006 ◽  
Vol 51 (2) ◽  
pp. 510-520 ◽  
Author(s):  
Jeniel Nett ◽  
Leslie Lincoln ◽  
Karen Marchillo ◽  
Randall Massey ◽  
Kathleen Holoyda ◽  
...  
Keyword(s):  
Cell Wall ◽  
Candida Albicans ◽  
Cell Viability ◽  
Amphotericin B ◽  
Cell Walls ◽  
Positive Impact ◽  
Indirect Evidence ◽  
Phenotypic Properties

ABSTRACT Biofilms are microbial communities, embedded in a polymeric matrix, growing attached to a surface. Nearly all device-associated infections involve growth in the biofilm life style. Biofilm communities have characteristic architecture and distinct phenotypic properties. The most clinically important phenotype involves extraordinary resistance to antimicrobial therapy, making biofilm infections very difficulty to cure without device removal. The current studies examine drug resistance in Candida albicans biofilms. Similar to previous reports, we observed marked fluconazole and amphotericin B resistance in a C. albicans biofilm both in vitro and in vivo. We identified biofilm-associated cell wall architectural changes and increased β-1,3 glucan content in C. albicans cell walls from a biofilm compared to planktonic organisms. Elevated β-1,3 glucan levels were also found in the surrounding biofilm milieu and as part of the matrix both from in vitro and in vivo biofilm models. We thus investigated the possible contribution of β-glucans to antimicrobial resistance in Candida albicans biofilms. Initial studies examined the ability of cell wall and cell supernatant from biofilm and planktonic C. albicans to bind fluconazole. The cell walls from both environmental conditions bound fluconazole; however, four- to fivefold more compound was bound to the biofilm cell walls. Culture supernatant from the biofilm, but not planktonic cells, bound a measurable amount of this antifungal agent. We next investigated the effect of enzymatic modification of β-1,3 glucans on biofilm cell viability and the susceptibility of biofilm cells to fluconazole and amphotericin B. We observed a dose-dependent killing of in vitro biofilm cells in the presence of three different β-glucanase preparations. These same concentrations had no impact on planktonic cell viability. β-1,3 Glucanase markedly enhanced the activity of both fluconazole and amphotericin B. These observations were corroborated with an in vivo biofilm model. Exogenous biofilm matrix and commercial β-1,3 glucan reduced the activity of fluconazole against planktonic C. albicans in vitro. In sum, the current investigation identified glucan changes associated with C. albicans biofilm cells, demonstrated preferential binding of these biofilm cell components to antifungals, and showed a positive impact of the modification of biofilm β-1,3 glucans on drug susceptibility. These results provide indirect evidence suggesting a role for glucans in biofilm resistance and present a strong rationale for further molecular dissection of this resistance mechanism to identify new drug targets to treat biofilm infections.

Combined effects of hormones and light during growth promotion in primary leaves of Phaseolus vulgaris

1993 ◽  
Vol 71 (3) ◽  
pp. 501-505 ◽  
Author(s):  
Thomas G. Brock
Keyword(s):  
Cell Wall ◽  
Phaseolus Vulgaris ◽  
Gibberellic Acid ◽  
White Light ◽  
Osmotic Potential ◽  
Leaf Growth ◽  
Growth Potential ◽  
Combined Effects ◽  
Osmotic Concentration ◽  
Yield Threshold

Cell enlargement in primary leaves of bean is promoted by bright white light, gibberellic acid, or the cytokinin N6-benzyladenine. I examine the combined effects of light and hormones on growth, cell wall properties, and osmotic parameters during growth over 24 h. Applied alone, benzyladenine (10 μM), gibberellic acid (10 μM), and white light produced similar increases in the length and fresh weight of excised leaf strips over 24 h. The combined effects of hormones and light on growth were much less than additive. Individually, all three treatments significantly increased cell wall plastic extensibility over 24 h. However, benzyladenine combined with white light were additive in effect on plastic extensibility, and gibberellic acid combined with white light were synergistic. The differences in effects of hormones in white light on growth versus plastic extensibility indicate a decrease in growth potential, which is attributable in part to hormonal effects on osmotic concentration. Although white light alone increased osmotic concentration, both benzyladenine and gibberellic acid greatly decreased it, with or without white light. Furthermore, because growth potential is a function of both osmotic potential and wall yield threshold, it appears that yield threshold does not decline in parallel with osmotic potential in hormone-treated bean leaf strips. Finally, both benzyladenine and gibberellic acid inhibit the increase in osmotic solutes normally produced by white light. This effect, coupled with water uptake during cell expansion, would produce the observed decreases in osmotic concentration in hormone-treated strips. Hence, both benzyladenine and gibberellic acid interfere with light-induced growth, primarily through effects on the apparent ability of light to direct solute accumulation. Key words: Phaseolus vulgaris, leaf growth, cytokinin, gibberellic acid, light.

Isolation of anatomically defined cell walls from fodder kale, and their contributions to determining the in vitro cellulase digestibility of the whole plant

1984 ◽  
Vol 103 (2) ◽  
pp. 347-352 ◽  
Author(s):  
J. G. McCluskey ◽  
M. J. Allison ◽  
H. J. Duncan ◽  
M. C. Jarvis
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Stem Anatomy ◽  
Vascular Cell ◽  
Cellulase Digestion ◽  
Whole Plant ◽  
Digestible Organic Matter ◽  
Component Cell ◽  
Whole Plants

SUMMARYVascular and non-vascular cell walls were isolated separately from leaves, upper stems and lower stems of 12 kale (Brassica oleracea L.) cultivars, by a sieving technique. The digestible organic matter in the dry matter (DOMD) of the cell walls and of the whole plants was determined by pepsin-cellulase digestion. The measured whole-plant DOMD correlated closely with the DOMD predicted by adding together the amounts of non-digested material derived from all the plant's component cell-wall fractions. Differences in DOMD between cultivars were determined primarily by the amount of vascular cell walls in the stems, particularly the lower stems; that is, by the stem anatomy. The vascular cell walls of the upper stems had a wider range of DOMD values and a higher mean DOMD than the vascular cell walls of the lower stems. Thus cell-wall composition made some contribution to determining the whole-plant DOMD, although it contributed less than the anatomy of the stem.

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