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Cell Wall Composition of Neurospora crassa Under Conditions of Copper Toxicity.

The mycelia of Neurospora crassa grown in the presence of high concentrations of copper were blue in color, but only on a medium containing inorganic nitrate and phosphate as the nitrogen and phosphate sources, respectively. The cell wall isolate of the blue mycelia contained large amounts (12%) of copper and higher amounts of chitosan, phosphate, and amino groups, with a 42% decrease in the chitin content. Although all the glucosamine of the cell wall of control cultures could be released within 6 h of hydrolysis with acid, that of the blue mycelium required prolonged hydrolysis for 24 h. On removal of copper, the cell wall of the blue mycelium could quantitatively bind again to copper as well as to zinc. Although zinc binding was fivefold greater, copper alone was preferentially bound from a mixture of the two metal ions. Supplementation of iron along with copper in the culture medium resulted in the disappearance of the blue color of the mycelium and restoration of normal growth and composition of the cell wall, probably by limiting the uptake of copper from the medium. The possibility of the cell wall being a specific site of lesion in copper toxicity in the mold is discussed.

Journal Article↗

Internode structure and cell wall composition in maturing tillers of switchgrass (Panicum virgatum. L).

This work examined cell composition gradients in maturing tillers of switchgrass (Panicum virgatum L.) with the aim of developing baseline information on this important forage and biomass crop. Flowering tillers were collected from plants raised from seeds in a greenhouse and field, harvested at soil level and separated into internodes beginning with the node subtending the peduncle. Internodes were analyzed using microscopy, by fiber digestion, high-performance liquid chromatography and by gas chromatography-mass spectrometry to obtain anatomical and compositional data. Microscopy demonstrated the development and maturation of cortical fibers which eventually became confluent with the fiber sheath surrounding vascular bundles in the lower internodes. Detergent fiber analysis indicated increasing cellulose and lignin contents and decreases in cell solubles and hemicelluloses with increasing distance of the internodes from the top of the plant. Soluble phenolics were greatest in amounts and complexity in top internodes. The lower internodes contained greater levels of wall-bound phenolic acids, principally as 4-coumarate and ferulate.

Biomass↗

Cell wall composition and deoxyribonucleic acid similarities among the anaerobic coryneforms, classical propionibacteria, and strains of Arachnia propionica.

Eighty strains of anaerobic coryneforms were compared with 29 strains of classical propionibacteria and 8 strains of Arachnia propionica by cell wall analysis, deoxyribonucleic acid (DNA) base compositions, and nucleotide sequence similarities. The anaerobic coryneforms have DNA base compositions in the range of 58 to 64% guanine + cytosine (GC) and show at least three homology groups. The largest group corresponds to organisms identified as Propionibacterium acnes and shows about 50% homology to strains in the P. avidum homology group. The third group, P. granulosum, shows low levels of similarities to the other two. All strains of anaerobic coryneforms have some combination of galactose, glucose, or mannose as cell wall sugars, and most have alanine (ala), glutamic acid (glu), glycine (gly), and l-alpha-epsilon-diaminopimelic acid (l-DAP) as amino acids of peptidoglycan. However, a few strains in the P. acnes and P. avidum homology groups have meso-DAP and minimal amounts of glycine. Two serological types, based on cell wall antigens, were found in the P. acnes homology group. One type had galactose, glucose, and mannose as cell wall sugars, the other glucose and mannose only. The classical propionibacteria have DNA base compositions in the range of 65 to 68% GC and show four homology groups which correspond closely to van Niel's classification as given in the 7th edition of Bergey's Manual. The P. jensenii group showed about 50% homology to the P. thoenii group and about 30 to 35% to the P. acidi-propionici group. The P. freudenreichii strains showed a rather lower level of similarity (8 to 25%) to the other homology groups. Most of the strains of classical propionibacteria also have some combination of galactose, glucose, or mannose as cell wall sugars and ala, glu, gly, and l-DAP as peptidoglycan amino acids, but P. shermanii and P. freudenreichii strains, which form a single homology group, have galactose, mannose, and rhamnose as cell wall sugars and ala, glu, and meso-DAP in their peptidoglycan. There is a rather low level of DNA homology (10 to 20%) between the anaerobic coryneforms and classical propionibacteria. However, the strains of A. propionica which have a GC content of 64 to 65% and form a single homology group, show no homology to either of the other two major groups.

Actinomycetales↗

Mutations in PMR5 result in powdery mildew resistance and altered cell wall composition.

Powdery mildews and other obligate biotrophic pathogens are highly adapted to their hosts and often show limited host ranges. One facet of such host specialization is likely to be penetration of the host cell wall, a major barrier to infection. A mutation in the pmr5 gene rendered Arabidopsis resistant to the powdery mildew species Erysiphe cichoracearum and Erysiphe orontii, but not to the unrelated pathogens Pseudomonas syringae or Peronospora parasitica. PMR5 belongs to a large family of plant-specific genes of unknown function. pmr5-mediated resistance did not require signaling through either the salicylic acid or jasmonic acid/ethylene defense pathways, suggesting resistance in this mutant may be due either to the loss of a susceptibility factor or to the activation of a novel form of defense. Based on Fourier transform infrared analysis, the pmr5 cell walls were enriched in pectin and exhibited a reduced degree of pectin modification relative to wild-type cell walls. In addition, the mutant had smaller cells, suggesting a defect in cell expansion. A double mutant with pmr6 (defective in a glycosylphosphatidylinositol-anchored pectate lyase-like gene) exhibited a strong increase in total uronic acid content and a more severe reduction in size, relative to the single mutants, suggesting that the two genes affect pectin composition, either directly or indirectly, via different mechanisms. These two mutants highlight the importance of the host cell wall in plant-microbe interactions.

Arabidopsis↗

Resistance to the plant PR-5 protein osmotin in the model fungus Saccharomyces cerevisiae is mediated by the regulatory effects of SSD1 on cell wall composition.

The capacity of plants to counter the challenge of pathogenic fungal attack depends in part on the ability of plant defense proteins to overcome fungal resistance by being able to recognize and eradicate the invading fungi. Fungal genes that control resistance to plant defense proteins are therefore important determinants that define the range of fungi from which an induced defense protein can protect the plant. Resistance of the model fungus Saccharomyces cerevisiae to osmotin, a plant defense PR-5 protein, is strongly dependent on the natural polymorphism of the SSD1 gene. Expression of the SSD1-v allele afforded resistance to the antifungal protein. Conversely, yeast strains carrying the SSD1-d allele or a null ssd1Delta mutation displayed high sensitivity to osmotin. The SSD1-v protein mediates osmotin resistance in a cell wall-dependent manner. Deletion of SSD1-v or SSD1-d impeded sorting of the PIR proteins (osmotin-resistance factors) to the cell wall without affecting mRNA levels, indicating that SSD1 functions in post-transcriptional regulation of gene expression. The sensitivity of ssd1Delta cells to osmotin was only partially suppressed by over-accumulation of PIR proteins in the cell wall, suggesting an additional function for SSD1 in cell wall-mediated resistance. Accordingly, cells carrying a null ssd1 mutation also displayed aberrant cell-wall morphology and lower levels of alkali-insoluble cell-wall glucans. Therefore SSD1 is an important regulator of fungal cell-wall biogenesis and composition, including the deposition of PIR proteins which block the action of plant antifungal PR-5 proteins.

Alleles↗

Cell-wall composition and polysaccharide synthase activity changes following photoinduction in Trichoderma viride.

The differentiation and metabolism in the soil-borne saprophytic deuteromycete Trichoderma viride are subject to control by light. We have investigated the effect of illumination of mycelia on the activities of cell-wall synthesizing enzymes beta-1,3-glucan synthase and chitin synthase and on the composition of cell alls. After 10 min illumination of dark-grown mycelia with white fluorescent light at 600 1x, a gradual rise in specific activity of membrane-bound enzymes beta-1,3-glucan synthase by about 130% and a decrease in specific activity of chitin synthase by about 50% in relation to the dark control were observed. The changes in enzyme activities were caused by de novo synthesis of corresponding polysaccharide synthases(s) and/or their regulatory components since they were not observed when protein synthesis was blocked with 50 microg/ml cycloheximide. The content of beta-1,3-glucan in the cell walls of illuminated mycelia has increased by 35-50% in comparison to the dark control while the content of chitin remained practically unchanged.

Cell Wall↗

Changes in cell wall composition of three Fragaria x ananassa cultivars with different softening rate during ripening.

Fleshy fruit soften during ripening mainly as a consequence of solubilization and depolymerization of cell wall components. We have performed a comparative study of the polysaccharide content of fruit cell walls during final steps of development and ripening of three strawberry (Fragaria x ananassa Duch.) cultivars with different softening rates. The three chosen varieties showed very different firmness; Camarosa was the firmest, Toyonaka the softest, and Pajaro intermediate between them. Cell walls were extracted, quantified and fractioned by sequential extraction to obtain particular subclasses of cell wall polymers. Cell wall content diminished during the process in the three cultivars. Differences among cultivar cell wall contents were detected only in immature stages. The amount of water soluble polymers (WSP) increased in all cultivars from small green (SG) to white (W) stage, although from the W to 100% red (100%R) stage the WSP remained constant in Camarosa and Pajaro and decreased in Toyonaka. On the contrary, the hydrochloric acid-soluble pectins (HSP) decreased during ripening of all the cultivars analyzed. Camarosa had the largest amount of HSP, but there were no differences between Pajaro and Toyonaka. The amount of hemicellulosic polysaccharides and cellulose also decreased in the three cultivars. Camarosa had the highest amounts of both polysaccharides while Toyonaka had the lowest at immature stages, but there were no differences among cultivars at 100%R stage. WSP showed depolymerization only in Toyonaka cultivar, while HSP showed depolymerization in Pajaro and Toyonaka cultivars. A slight depolymerization was observed in hemicelluloses extracted from any of three cultivars.

Cell Wall↗