Chemical and enzymic analyses of Pichia polymorpha cell walls

1980 ◽  
Vol 26 (2) ◽  
pp. 169-174 ◽  
Author(s):  
Tomas G. Villa ◽  
Vicente Notario ◽  
Julio R. Villanueva
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Amino Sugar ◽  
Stationary Growth Phase ◽  
Turnover Rates ◽  
Lytic Enzymes ◽  
Total Carbohydrate ◽  
Neutral Sugars ◽  
Early Stationary Growth Phase ◽  
Pichia Polymorpha

Conventional techniques of chemical analysis have shown that the cell walls of the yeast Pichia polymorpha, at early stationary growth phase, consisted of carbohydrate (about 85%), protein (8%), and lipid (7%). Glucose and mannose were the only neutral sugars and glucosamine the sole amino sugar present among the cell wall components. Paper and gas–liquid chromatographies of acid hydrolysates of purified cell walls and cell wall fractions proved that mannan and alkali-insoluble glucans, in that order, were the major polysaccharide components, accounting for 83% of the total carbohydrate content. The isolation of an alkali-soluble glucan, non-precipitable with Fehling's solution, has been achieved. Treatments of whole cell walls and their fractions with purified cell wall lytic enzymes have shown the presence of both 1, 3- and 1, 6-β-D-linkages in all the glucan fractions. 1, 3-α-Glucan was not detected. Mannan–glucan complexes have been found containing about 50% each of mannose and glucose. All polysaccharides exhibited different turnover rates when cells were grown in the presence of D-[U-14C]glucose. The morphogenetic implications of these results are discussed.

scholarly journals The composition of the cell wall of Fusicoccum amygdali

1971 ◽  
Vol 125 (2) ◽  
pp. 461-471 ◽  
Author(s):  
K. W. Buck ◽  
M. A. Obaidah
Keyword(s):  
Cell Wall ◽  
Sodium Hydroxide ◽  
Cell Walls ◽  
Digestive Enzymes ◽  
Helix Pomatia ◽  
Water Soluble ◽  
Extracellular Polysaccharides ◽  
Lytic Enzymes ◽  
Similar Nature ◽  
Dark Blue

1. The cell wall of Fusicoccum amygdali consisted of polysaccharides (85%), protein (4–6%), lipid (5%) and phosphorus (0.1%). 2. The main carbohydrate constituent was d-glucose; smaller amounts of d-glucosamine, d-galactose, d-mannose, l-rhamnose, xylose and arabinose were also identified, and 16 common amino acids were detected. 3. Chitin, which accounted for most of the cell-wall glucosamine, was isolated in an undegraded form by an enzymic method. Chitosan was not detected, but traces of glucosamine were found in alkali-soluble and water-soluble fractions. 4. Cell walls were stained dark blue by iodine and were attacked by α-amylase, with liberation of glucose, maltose and maltotriose, indicating the existence of chains of α-(1→4)-linked glucopyranose residues. 5. Glucose and gentiobiose were liberated from cell walls by the action of an exo-β-(1→3)-glucanase, giving evidence for both β-(1→3)- and β-(1→6)-glucopyranose linkages. 6. Incubation of cell walls with Helix pomatia digestive enzymes released glucose, N-acetyl-d-glucosamine and a non-diffusible fraction, containing most of the cell-wall galactose, mannose and rhamnose. Part of this fraction was released by incubating cell walls with Pronase; acid hydrolysis yielded galactose 6-phosphate and small amounts of mannose 6-phosphate and glucose 6-phosphate as well as other materials. Extracellular polysaccharides of a similar nature were isolated and may be formed by the action of lytic enzymes on the cell wall. 7. About 30% of the cell wall was resistant to the action of the H. pomatia digestive enzymes; the resistant fraction was shown to be a predominantly α-(1→3)-glucan. 8. Fractionation of the cell-wall complex with 1m-sodium hydroxide gave three principal glucan fractions: fraction BB had [α]D +236° (in 1m-sodium hydroxide) and showed two components on sedimentation analysis; fraction AA2 had [α]D −71° (in 1m-sodium hydroxide) and contained predominantly β-linkages; fraction AA1 had [α]D +40° (in 1m-sodium hydroxide) and may contain both α- and β-linkages.

scholarly journals Endo-β-1,3-glucanase (GH16 Family) from Trichoderma harzianum Participates in Cell Wall Biogenesis but Is Not Essential for Antagonism Against Plant Pathogens

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 781 ◽  
Author(s):  
Marcela Suriani Ribeiro ◽  
Renato Graciano de Paula ◽  
Aline Raquel Voltan ◽  
Raphaela Georg de Castro ◽  
Cláudia Batista Carraro ◽  
...  
Keyword(s):  
Cell Wall ◽  
Trichoderma Harzianum ◽  
Cell Walls ◽  
Plant Pathogens ◽  
Pcr Analysis ◽  
Glycosyl Hydrolases ◽  
Lytic Enzymes ◽  
Trichoderma Spp ◽  
Compensatory Response ◽  
Trichoderma Species

Trichoderma species are known for their ability to produce lytic enzymes, such as exoglucanases, endoglucanases, chitinases, and proteases, which play important roles in cell wall degradation of phytopathogens. β-glucanases play crucial roles in the morphogenetic-morphological process during the development and differentiation processes in Trichoderma species, which have β-glucans as the primary components of their cell walls. Despite the importance of glucanases in the mycoparasitism of Trichoderma spp., only a few functional analysis studies have been conducted on glucanases. In the present study, we used a functional genomics approach to investigate the functional role of the gluc31 gene, which encodes an endo-β-1,3-glucanase belonging to the GH16 family in Trichoderma harzianum ALL42. We demonstrated that the absence of the gluc31 gene did not affect the in vivo mycoparasitism ability of mutant T. harzianum ALL42; however, gluc31 evidently influenced cell wall organization. Polymer measurements and fluorescence microscopy analyses indicated that the lack of the gluc31 gene induced a compensatory response by increasing the production of chitin and glucan polymers on the cell walls of the mutant hyphae. The mutant strain became more resistant to the fungicide benomyl compared to the parental strain. Furthermore, qRT-PCR analysis showed that the absence of gluc31 in T. harzianum resulted in the differential expression of other glycosyl hydrolases belonging to the GH16 family, because of functional redundancy among the glucanases.

Compositional studies on the cell walls of the synnema and vegetative hyphae of Ceratocystis ulmi

1973 ◽  
Vol 51 (6) ◽  
pp. 1147-1153 ◽  
Author(s):  
James L. Harris ◽  
Willard A. Taber
Keyword(s):  
Electron Microscopy ◽  
Amino Acids ◽  
Cell Wall ◽  
Enzymatic Hydrolysis ◽  
Cell Walls ◽  
Amino Sugar ◽  
Fibrillar Structure ◽  
Dense Granules ◽  
Hyphal Wall ◽  
Hydrolysis Of

The composition of the cell walls of synnemal and vegetative hyphae of Ceratocystis ulmi was studied by fractionation and assay of released compounds. Residues after enzymatic hydrolyses were examined by electron microscopy. The synnemal wall was found to have 67% carbohydrate, 4.52% amino sugar, 5.02% protein, 1.6% lipid, and 0.59% ash, which accounted for 78.7% of the cell wall. The vegetative hyphal wall contained 56% carbohydrate, 3.44% amino sugar, 7.92% protein, 4.5% lipid, and 1.45% ash, which totaled 73.3% of the wall weight. Sugars identified were D-glucose, D-mannose, D-galactose, and L-rhamnose. Enzymatic hydrolysis of both wall types by cellulase and laminaranase indicated the presence of beta-1,3 and beta-1,4 linkages of glucose polymers. N-acetylglucosamine was liberated by chitinase. Most of the 16 amino acids detected in each wall type were at least twice as abundant in vegetative hyphal walls as in synnemal hyphal walls. Cellulase and laminaranase treatment of cell walls revealed a fibrillar structure. Chitinase-treated walls did not appear as fibrous, suggesting that the fibrous structure may be mostly chitinous. Synnemal cell walls are covered by electron-dense granules which may correspond to the pigment in the synnemal hyphae.

Melanin deposition in the hyphae of a species of Phomopsis

1975 ◽  
Vol 21 (4) ◽  
pp. 442-452 ◽  
Author(s):  
D. H. Ellis ◽  
D. A. Griffiths
Keyword(s):  
Cell Wall ◽  
Surface Analysis ◽  
Cell Walls ◽  
Lytic Enzymes ◽  
Fungal Cell ◽  
Ionizing Radiations ◽  
Dopa Melanin ◽  
Fungal Cell Walls ◽  
Cellular Organelles

Hyaline hyphae of Phomopsis become pigmented when exposed to short periods of light. Pigment was deposited in the form of melanin granules both within the cell wall and within mucilaginous excrescences that were developed irregularly over the hyphal surface. Analysis of the pigment showed it to have properties similar to that of "Dopa" melanin and to pigments previously isolated from fungal cell walls. Lysis of both hyaline and pigmented hyphal walls by means of lytic enzymes was minimal. It is suggested that the major role of melanin in this fungus is the protection of cellular organelles from harmful ionizing radiations.

Structure and composition of the cell wall of Choanephora cucurbitarum

1976 ◽  
Vol 22 (4) ◽  
pp. 486-494 ◽  
Author(s):  
D. R. Letourneau ◽  
J. M. Deven ◽  
M. S. Manocha
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Free Cell ◽  
Infrared Spectrophotometry ◽  
X Ray Diffraction ◽  
X Ray ◽  
Before And After ◽  
Neutral Sugars ◽  
Choanephora Cucurbitarum ◽  
Hyphal Wall

Mechanically isolated, cytoplasm-free cell walls of Choanephora cucurbitarum were analyzed qualitatively and quantitatively by use of microchemical methods, infrared spectrophotometry, and X-ray diffraction. Chemical analysis of cell wall revealed the presence of chitin (17%), chitosan (28.4%), neutral sugars (7.2%), uronic acid (2.4%), proteins (8.2%), and lipids (13.8%). The structure of hyphal wall, investigated by electron microscopy of shadowed replicas before and after alkali-acid hydrolysis, showed two distinct regions: microfibrillar and amorphous. The microfibrils, which were composed of mainly chitin, were organized into two distinct layers; an outer, thicker layer of randomly oriented microfibrils, and an inner, thin layer of parallel microfibrils. In its structure and chemical composition the cell wall of C. cucurbitarum resembles those of other zygomycetous fungi.

scholarly journals Structural lipids and carbohydrates of the deep mycelium of phoma-like micromycetes, potential mycoherbicides

2020 ◽  
Vol 23 ◽  
pp. 02011
Author(s):  
Sofia V. Sokornova ◽  
Galina M. Frolova ◽  
Evgeny A. Gusenkov ◽  
Daniel M. Malygin ◽  
Alexey L. Shavarda
Keyword(s):  
Growth Phase ◽  
Nutrient Medium ◽  
Biochemical Composition ◽  
Cell Walls ◽  
Stationary Growth Phase ◽  
Cirsium Arvense ◽  
Stationary Growth ◽  
Convolvulus Arvensis ◽  
Heracleum Sosnowskyi ◽  
Early Stationary Growth Phase

The work is devoted to the mycelium biochemical composition of Stagonospora cirsii C-211, Calophoma complanata 32.121, Didymella macrostoma 32.52. These phylogenetically distant species of phoma-like micromycetes are the potential mycoherbicides of Cirsium arvense, Heracleum sosnowskyi, and Convolvulus arvensis, respectively. The S. cirsii C-211, C. complanata 32.121, D. macrostoma 32.52 mycelium in the early stationary growth phase was obtained on sucrose-soybean nutrient medium. It was shown that the lipid and carbohydrate (polyols, sugars) profiles of these strains have much in common. We suppose that levels of arabitol and trehalose influence to the stress-resistant of phoma-like micromycetes. In particularly, these carbohydrates serve structural and protective roles in the cell walls during osmotic and temperatures stress. The ratio of phosphatidylcholine to phosphatidylethanolamine and the proportion of phosphatidylserine among structural lipids also determine the properties of mycelium, and can be used to assess its quality.

scholarly journals The ERECTA Receptor-Like Kinase Regulates Cell Wall–Mediated Resistance to Pathogens in Arabidopsis thaliana

2009 ◽  
Vol 22 (8) ◽  
pp. 953-963 ◽  
Author(s):  
Clara Sánchez-Rodríguez ◽  
José Manuel Estévez ◽  
Francisco Llorente ◽  
Camilo Hernández-Blanco ◽  
Lucía Jordá ◽  
...  
Keyword(s):  
Cell Wall ◽  
Disease Resistance ◽  
Cell Walls ◽  
Control Plant ◽  
Biological Processes ◽  
Uronic Acids ◽  
Wild Type ◽  
Receptor Like Kinase ◽  
Neutral Sugars ◽  
Plectosphaerella Cucumerina

Some receptor-like kinases (RLK) control plant development while others regulate immunity. The Arabidopsis ERECTA (ER) RLK regulates both biological processes. To discover specific components of ER-mediated immunity, a genetic screen was conducted to identify suppressors of erecta (ser) susceptibility to Plectosphaerella cucumerina fungus. The ser1 and ser2 mutations restored disease resistance to this pathogen to wild-type levels in the er-1 background but failed to suppress er-associated developmental phenotypes. The deposition of callose upon P. cucumerina inoculation, which was impaired in the er-1 plants, was also restored to near wild-type levels in the ser er-1 mutants. Analyses of er cell walls revealed that total neutral sugars were reduced and uronic acids increased relative to those of wild-type walls. Interestingly, in the ser er-1 walls, neutral sugars were elevated and uronic acids were reduced relative to both er-1 and wild-type plants. The cell-wall changes found in er-1 and the ser er-1 mutants are unlikely to contribute to their developmental alterations. However, they may influence disease resistance, as a positive correlation was found between uronic acids content and resistance to P. cucumerina. We propose a specific function for ER in regulating cell wall–mediated disease resistance that is distinct from its role in development.

Breakpoint: Cell Wall and Glycoproteins and their Crucial Role in the Phytopathogenic Fungi Infection

2020 ◽  
Vol 21 (3) ◽  
pp. 227-244 ◽  
Author(s):  
Verónica Plaza ◽  
Evelyn Silva-Moreno ◽  
Luis Castillo
Keyword(s):  
Cell Wall ◽  
Signaling Pathways ◽  
Crucial Role ◽  
Intracellular Signaling ◽  
Cell Walls ◽  
Wall Stress ◽  
Appressorium Formation ◽  
Cell Wall Degrading Enzymes ◽  
Lytic Enzymes ◽  
Fungal Cell

The cell wall that surrounds fungal cells is essential for their survival, provides protection against physical and chemical stresses, and plays relevant roles during infection. In general, the fungal cell wall is composed of an outer layer of glycoprotein and an inner skeletal layer of β-glucans or α- glucans and chitin. Chitin synthase genes have been shown to be important for septum formation, cell division and virulence. In the same way, chitin can act as a potent elicitor to activate defense response in several plant species; however, the fungi can convert chitin to chitosan during plant infection to evade plant defense mechanisms. Moreover, α-1,3-Glucan, a non-degradable polysaccharide in plants, represents a key feature in fungal cell walls formed in plants and plays a protective role for this fungus against plant lytic enzymes. A similar case is with β-1,3- and β-1,6-glucan which are essential for infection, structure rigidity and pathogenicity during fungal infection. Cell wall glycoproteins are also vital to fungi. They have been associated with conidial separation, the increase of chitin in conidial cell walls, germination, appressorium formation, as well as osmotic and cell wall stress and virulence; however, the specific roles of glycoproteins in filamentous fungi remain unknown. Fungi that can respond to environmental stimuli distinguish these signals and relay them through intracellular signaling pathways to change the cell wall composition. They play a crucial role in appressorium formation and penetration, and release cell wall degrading enzymes, which determine the outcome of the interaction with the host. In this review, we highlight the interaction of phypatophogen cell wall and signaling pathways with its host and their contribution to fungal pathogenesis.

Silicification of wood-cell walls

1994 ◽  
Vol 52 ◽  
pp. 428-429
Author(s):  
S. E. Keckler ◽  
D. M. Dabbs ◽  
N. Yao ◽  
I. A. Aksay
Keyword(s):  
Electron Microscopy ◽  
Cell Wall ◽  
Cell Walls ◽  
Lignin Content ◽  
Resin Composite ◽  
Middle Lamella ◽  
Cellulose Fibers ◽  
Complex Structures ◽  
Rich Region ◽  
Inorganic Precursors

Cellular organic structures such as wood can be used as scaffolds for the synthesis of complex structures of organic/ceramic nanocomposites. The wood cell is a fiber-reinforced resin composite of cellulose fibers in a lignin matrix. A single cell wall, containing several layers of different fiber orientations and lignin content, is separated from its neighboring wall by the middle lamella, a lignin-rich region. In order to achieve total mineralization, deposition on and in the cell wall must be achieved. Geological fossilization of wood occurs as permineralization (filling the void spaces with mineral) and petrifaction (mineralizing the cell wall as the organic component decays) through infiltration of wood with inorganics after growth. Conversely, living plants can incorporate inorganics into their cells and in some cases into the cell walls during growth. In a recent study, we mimicked geological fossilization by infiltrating inorganic precursors into wood cells in order to enhance the properties of wood. In the current work, we use electron microscopy to examine the structure of silica formed in the cell walls after infiltration of tetraethoxysilane (TEOS).

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