STRUCTURE AND COMPOSITION OF ZOOSPORANGIAL DISCHARGE PAPILLAE IN THE FUNGUS ALLOMYCES

A study was made of the discharge papillae of zoosporangia (ZS), as well as of resistant sporangia and gametangia, of Allomyces (mainly A. × javanicus). It was found that they differ considerably. The discharge papillae of the resistant sporangia apparently arise by a blister-like modification of the inner wall, whereas those of the ZS and the gametangia consist of a pulley-shaped, refractile plug securely placed in a hole in the wall. This plug can be successfully removed from the ZS wall by treatment with sulfuric acid. Solubility and histochemical tests were used to identify the material of the ZS plugs and the results indicate that the material is a pectic substance, most likely protopectin. The plugs of the gametangia presumably have the same composition. A preliminary study was also made of the discharge papillae of representatives of other families of the aquatic Phycomycetes. The results indicate that such pulley-shaped plugs filling a pore in the wall are characteristic only of the ZS and the gametangia of members of the family Blastocladiaceae to which Allomyces belongs.     

The arrangement of microfibrils in sporangial walls of Allomyces

Summary The microfibrillar component of the walls of zoosporangia and resistant sporangia of the phycomycete Allomyces arbusculus has been studied in the electron microscope, after chemical removal of the amorphous wall materials. In the zoosporangium wall the microfibrils are randomly arranged, as in the outer layer of the hyphal walls, and the sporangial wall is of even thickness. In the resistant sporangia the microfibrillar layer of the wall is perforated by numerous pores 0.25 in diameter. The microfibrils are randomly arranged over much of the wall but tend to be concentrically arranged in the vicinity of the pores. On the inside of the wall the microfibrils form a thickened rim around the pore.     

THE CHEMICAL COMPOSITION OF THE HYPHAL WALLS OF THE FUNGUS ALLOMYCES

Aronson , Jerome M., and Leonard Machlis . (U. California, Berkeley.) The chemical composition of the hyphal walls of the fungus Allomyees. Amer. Jour. Bot. 46(4): 292–300. Illus. 1959.—The hyphal walls of Allomyces macrogynus were isloated by both alkaline digestion methods and by sonic oscillation. Both types of preparations showed the walls to consist of chitin, glucan, and ash. In addition, the mechanically isolated walls contained a protein fraction, the properties and significance of which were not determined. Hemicellulose-type polysaccharides, pectic substances, ether soluble lipids, and constituents giving rise to 3–0-α-earboxyethyl hexosamine were not found to be present in the walls. The walls of plants grown for 60–70 hr. under the prescribed conditions contain approximately 60% chitin, 15% glucan, 10% ash, and 10% protein intimately associated with the walls. The percentage of wall material in a mycelium, as well as the percentage of chitin in the walls, increases with the chronological age of the mycelium. These percentages were not, however, affected by variations in the composition of the nutrient medium. The chitin in the walls could be hydrolyzed in the presence of chitinase; lysozyme, however, had no detectable effect on the walls.     

Cladosporium carrionii and Hormoconis resinae (C. resinae): Cell Wall and Melanin Studies

Two phaeoid strains of the fungus Cladosporium carrionii (SR3 from a xerophyte species and PP8201 from a patient), and one strain of Hormoconis resinae (Cladosporium resinae), isolated from oil-impregnated soil, were analyzed for their cell wall composition by colorimetric methods, X-ray diffraction, infrared spectroscopy, and solid-state ¹³C-nuclear magnetic resonance. Results suggested that the cell walls were composed mainly of hexoses (34%–47%) as β-1,3-glucan (some galactose and mannose were also present) and melanin, chitin being absent. Electron microscopic observations suggested that melanin was found not only in the cell wall but also in intracellular bodies resembling melanosomes.     

Comparative studies onChlorella cell walls: Induction of protoplast formation

Among 12 strains ofChlorella ellipsoidea, C. vulgaris, andC. saccharophila tested, 4 strains (1,C. ellpsoidea; 2,C. vulgaris; 1,C. saccharophila) formed osmotically labile protoplasts after treatment with mixtures of polysaccharide degrading enzymes. The relationship between enzymatical digestibility and structure or composition ofChlorella cell walls were studied by electron microscopy and staining techniques with some specific dyes. The cell wall structures of the 12Chlorella strains were grouped into three types: (1) with a trilaminar outer layer, (2) with a thin outer monolayer, and (3) without an outer layer. Protoplasts were formed only from the strains with a cell wall of Type 2. In the strains with a cell wall of Type 1, the outer layer protected the inner major microfibrillar layer against enzymatic digestion. The cell wall of Type 3 was totally resistant to the enzymes; the chemical composition of the cell wall would be somewhat different from that of other types.     

BARINOPHYTON CITRULLIFORME (BARINOPHYTALES INCERTAE SEDIS,BARINOPHYTACEAE) FROM THE UPPER DEVONIAN OF PENNSYLVANIA

Barinophyton citrulliforme is definitely proven to be a tracheophyte. The vascular cylinder of the main axis is an exarch protostele composed of tracheids having a continuous secondary wall folded into protrusions into the cell lumen. These protrusions delineate the position of annular thickenings which were deposited earlier than the continuous secondary wall. Between successive protrusions, the later-deposited secondary wall is interrupted by minute pitlike structures. It is suggested that the secondary walls of the tracheids were laid down in a two-phase depositional sequence. The fertile system of B. citrulliforme consists of a main axis bearing spirally arranged strobili. The strobilus consists of an axis that bears two alternate rows of sporangiferous appendages. The sporangiferous appendages are borne laterally along the strobilar axis and recurve abaxially around the axis. The sporangia are attached along the inside curve of the appendages, one sporangium per appendage, each containing both microspores and megaspores. This species thus exhibits sporangial heterospory which is considered to be an adaptation to an aquatic habit and the sporangia are considered to be functional analogs of the sporocarps of Marsilea. The interpretation of the strobilus is morphologically identical to Ananiev's interpretation of the strobilus of Pectinophyton bipectinatum; consequently P. bipectinatum is here transferred to Barinophyton as B. robustius comb. nov.     

STRUCTURE AND GERMINATION OF BLASTOCLADIELLA EMERSONII RESISTANT SPORANGIA

Resistant sporangia of Blastocladiella emersonii were induced by the addition of bicarbonate, potassium chloride, sodium chloride, or ammonium chloride to the medium and by the exposure of the zoospores to ultraviolet irradiation. Mature resistant sporangia induced by all of these conditions exhibit similar areolate wall pitting. Under suitable conditions resistant sporangia in all cases examined germinated with the cracking of the outer sporangial wall, with the formation of exit tubes by the inner sporangial wall, and with the cleavage and release of zoospores through discharge papillae formed in the tips of the exit tubes.     

Comparative Chemical Characterization of Pigmented and Less Pigmented Cell Walls of Alternaria tenuissima

Alternaria tenuissima, the parasitic fungus, was obtained from the pruned upper-cut surfaces of mulberry stems. This fungus contains dark pigment because of the presence of melanin in the cell wall. To obtain less-pigmented cell walls, this fungus was grown under dark condition. When the pigmented and less-pigmented cell walls were chemically analyzed, no differences were observed in amino-acid composition, hexoses, or pentoses. However, in pigmented cell walls, higher contents of melanin (2.6%) were found than in less-pigmented cell walls (0.3%). Interestingly, a significant difference was observed in the relative fatty-acid compositions between these two types of cell walls. Among the major fatty acids, there were increased concentrations of tetradecanoic acid (C14:0), hexadecanoic acid (C16:0), 9-hexadecenoic acid (C16: 1,Δ9), and 9-octadecanoic acid (C18:1,Δ9) and a concomitant decrease in 9,12-octadecadienoic acid (C18:2,Δ9,12) in less-pigmented compared with pigmented cell walls. This difference in fatty-acid composition may be related to the higher percentage of melanin in the pigmented than the less-pigmented cell walls. Lesser amounts of 9,12-octadecadienoic acid in less-pigmented cell walls may have been caused by the growth of the fungus under environmental stress conditions. An interesting observation was the presence in pigmented cell walls only of methyl-substituted fatty acids with carbon numbers C14 to C17, but their occurrence could not be ascertained in the present study.     

Oligosaccharide side chains of wall molecules are essential for cell-wall lysis in Chlamydomonas reinhardtii

The glycoproteins of the cell walls of Chlamydomonas are lysed during the reproductive cycle by proteases (autolysins) which are specific for their substrates. The autolysin which digests the wall of sporangia to liberate the zoospore daughter cells in the vegetative life cycle is a collagenase-like enzyme which attacks only selected domains in its wall substrates containing (hydroxy)-proline clusters. Cell-wall fractions obtained by salt-extraction (NaClO₄) and oxidizing agents (NaClO₂) and the insoluble residue were tested as substrates. The most-crosslinked insoluble inner part of the wall is the best substrate for the sporangia autolysin. Oligosaccharides obtained from the insoluble cell-wall fraction of sporangia by hydrolysis with Ba(OH)₂ inhibit autolysin action. We conclude that the oligosaccharide side chains of wall substrates are essential for forming the reactive enzyme-substrate complex.Abbreviations CSW chlorite-soluble cell-wall fraction - ICW insoluble cell-wall fraction - PSW salt-soluble fraction - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Cytochemical evidence of a fungal cell wall alteration during infection of Eucalyptus roots by the ectomycorrhizal fungus Cenococcum geophilum

When the ectomycorrhizal fungus Cenococcum geophilum changes from a saprophytic to a symbiotic stage, its cell wall structure becomes simplified. The external hyphal wall layer which, in the saprophytic stage, is highly reactive to the Gomori-Swift test becomes poorly reactive and can no longer be distinguished from the internal wall layer in the Hartig net hyphae. The intensely stained external wall layer was also absent from pure cultures of Cenococcum geophilum grown on a medium with a low sugar content. This cell wall alteration could be due to a decrease in the amount of melanin or of melanin plus cystine-containing proteins. This change may be necessary for increased nutrient exchange between symbionts through hyphal walls.     

ACAULANGIUM GEN. N., A FERTILE MARATTIALEAN FROM THE UPPER PENNSYLVANIAN OF ILLINOIS

Material described by Graham as Cyathotrachus bulbaceus is believed to represent a new genus that is a common constituent of Upper Pennsylvanian coal balls. The sessile synangia of Acaulangium gen. n. are borne in a row on either side of the pinnule midrib and are composed of four to six short, tapering, laterally appressed sporangia. The sporangia have extended tips which curve over the inside of the synangium distally and delimit a small open area inside the synangium. The outer facing walls of the sporangia are two to three cells thick throughout while the inner facing walls are uniseriate. During dehiscence the sporangia separate laterally and spore release results from the rupture of a row of elongate cells along the inner sporangium midline. Among species of Scolecopteris the new genus resembles S. illinoensis and S. minor var. parvifolia but differs in its sessile synangial attachment. The additional parenchyma present between sporangial cavities in the synangia of Acaulangium, and the tendency toward bilateral symmetry suggests an early stage in the evolution of a bivalve synangium such as is present in Marattia.     

Changes in wall and internal structure of allomyces-resistant sporangia during germination

The electron microscope was used to examine changes which take place in wall, as well as in internal, structure during germination of mature resistant sporangia of Allomyces neo-moniliformis. When the resistant sporangia are first placed in water to initiate germination, nuclei, mitochondria, and endoplasmic reticulum are not evident, though after the sporangia have been in water for more than 30 min all of these structures become visible. At this time no cracks are evident in the resistant sporangial wall and the cell membrane appears highly convoluted. Within the next 30 min the outer wall splits and the inner wall expands considerably as the protoplast increases in volume. At the same time the cell membrane straightens out, apparently in response to the protoplasmic expansion. The “cementing substances” begin to dissolve about this time so that 1 1/2 hr after placement in water the outer wall is completely separated from the inner wall which now acts as the cell wall. Cleavage appears to be initiated by the invagination of the cell membrane and by the appearance of segments of endoplasmic reticulum with filled vesicles at one end. Between 2 1/2 and 3 hr after placement in water zoospores are released.     

Sporangila autolysis in Clostridium perfringens type A

The autolytic system functioning in the release of mature spores and enterotoxin from sporangia of Clostridium prefringens was partially characterized. After sporangial autolysis in buffer, the supernatant fluid of the suspension contained autolysin active against purified sporangial walls. The autolysin was most active at pH 8 and 37°C, in the presence of Co²⁺ (0.3 · 10⁻³ M CoCl₂) and trypsin (48 μg/ml). Sodium dodecyl sulfate-treated sporangial walls further extracted with trichloroacetic acid to remove teichoic acid were a better enzyme substrate than walls treated only with sodium dodecyl sulfate. N-Acetylmuramyl-l-alanine amidase activity which released N-terminal alanine, and endopeptidase activity which hydrolysed the d-alanyl-glycine linkage liberating N-terminal glycine and C-terminal alanine, were both functional at pH 8. It is not known if one or two enzyme are involved. Autolysin appeared in cells as early as 2 h after inoculation into sporulation medium. Two asporogenic Stage 0 mutants grown in sporulation medium also produced autolysin identical in mode of action to that of the sporogenic wild type. Although the active cellular autolysin concentration subsequently decreased as cells sporilated, the walls of 8-h-old sporangia containing refractile heat-resistant spores were more susceptible to digestion by autolysin, than those of 2-, 4-, or 6-h-old cells grown in sporulation medium or of 4- or 14-h vegetative cells from growth medium. The results suggest that a progressive change may occur in the structure of the sporangial wall during spore morphogenesis, thus increasing its susceptibility to autolysis.     

CELL WALL VARIABILITY IN THE GREEN SEAWEED CODIUM VERMILARA (BRYOPSIDALES CHLOROPHYTA) FROM THE ARGENTINE COAST1

Cell wall chemistry in the coencocytic green seaweed Codium vermilara (Olivi) Delle Chiaje (Bryopsidales, Chlorophyta) is well understood. These cell walls are composed of major amounts of neutral β‐(1→4)‐D‐ mannans (Mn), sulfated polysaccharides (SPs), which include pyranosic arabinan sulfates (ArpS), pyruvylated galactan sulfates (pGaS), and mannan sulfates (MnS); also minor amounts of O‐glycoproteins are present. In this study, cell wall samples of C. vermilara were investigated with regard to their monosaccharide composition and infrared spectra (using Fourier transform infrared spectroscopy coupled to principal component [FTIR‐PC] analysis). Samples from three different populations of C. vermilara from the Argentine coast showed: (i) an important variation in the relative arabinan content, which increases from north to south, and (ii) a measurable degree of cell wall variability in the sulfate distribution between the different sulfated polysaccharides, independent of the amount of each polysaccharide present and of total sulfate content. When cell wall composition was analyzed over three consecutive years in a single geographic location, the quantity of Mn and overall sulfate content on SPs remained constant, whereas the pGaS:ArpS molar ratio changed over the time. Besides, similar cell wall composition was found between actively growing and resting zones of the thallus, suggesting that cell wall composition is independent of growth stage and development. Overall, these results suggest that C. vermilara has developed a mechanism to adjust the total level of cell wall sulfation by modulating the ArpS:pGaS:MnS molar ratio and also by adjusting the sulfation level in each type of polymer, whereas nonsulfated Mn, as the main structural polysaccharide, did not change over the time or growing stage.     

Molecular structure of the resistant biopolymer in zygospore cell walls of Chlamydomonas monoica

The unicellular green alga Chlamydomonas monoica Strehlow is known to produce zygospores with a cell wall that is resistant against microbial and chemical attack. This resistance is thought to be due to the presence of a sporopollenin-like material. However, the resistant nature of sporopollenin-like materials seriously hampers their structural analysis. With complementary techniques such as ¹³C-nuclear magnetic resonance spectroscopy, Curie-point pyrolysis-gas chromatography/mass spectroscopy and RuO₄ chemical degradation, the chemical composition of resistant biopolymer in the isolated cell walls of C. monoica zygospores was determined. This material is composed of C₂₂–C₃₀ linear alcohols and carboxylic acids, intermolecularly linked via ester and ether-linkages similar to the resistant aliphatic biopolymers encountered in the walls of the vegetative cells of the algae Tetraedron minimum, Scenedesmus communis and Pediastrum boryanum. Received: 29 April 1998 / Accepted: 2 October 1998     

The fine structure of Micrococcus radiophilus and Micrococcus radioproteolyticus

The radiation resistant bacteria Micrococcus radiophilus and M. radioproteolyticus were studied by thin sectioning and freeze-etching techniques and the two species were found to be similar in the fine structure. The only significant difference was in the appearance of the surfaces of the cell walls in freeze-etched preparations.Since the two species, together with M. radiodurans, possess a unique cell wall structure and a cell wall peptidoglycan, which is different from that of other micrococci and Gram-positive cocci, it is recommended that they be reclassified into a new genus.     

THE COMPOSITION AND PHYLOGENETIC SIGNIFICANCE OF THE MOUGEOTIA (CHAROPHYCEAE) CELL WALL1

The two-layered, fibrillar cell wall of Mougeotia C. Agardh sp. consisted of 63.6% non-cellulosic carbohydrates and 13.4% cellulose. The orientation of cellulose microfibrils in the native cell wall agrees with the multinet growth hypothesis, which has been employed to explain the shift in microfibril orientation from transverse (inner wall) toward axial (outer wall). Monosaccharide analysis of isolated cell walls revealed the presence of ten sugars with glucose, xylose and galactose most abundant. Methylation analysis of the acid-modified, 1 N NaOH insoluble residue fraction showed that it was composed almost exclusively of 4-linked glucose, confirming the presence of cellulose. The major hemicellulosic carbohydrate was semi-purified by DEAE Sephacel (Cl^?) anion-exchange chromatography of the hot 1 N NaOH soluble fraction. This hemicellulose was a xylan consisting of a 4-xylosyl backbone and 2,4-xylosyl branch points. The major hot water soluble neutral polysaccharide was identified as a 3-linked galactan. Mougeotia cell wall composition is similar to that of (Charophyceae) and has homologies with vascular plant cell walls. Our observations support transtructural evidence which suggests that members of the Charophyceae represent the phylogenetic line that gave rise to vascular plants. Therefore, the primary cell walls of vascular plants many have evolved directly from structures typical of the filamentous green algal cell walls found in the Charophyceae.     

Cell wall composition of the yeast and mycelial forms of Yarrowia lipolytica

The cell walls of the yeast and mycelial forms of Yarrowia lipolytica were isolated and purified. Electron microscopy studies showed no differences between both types of cell walls. Chemical analysis revealed that the yeast cell wall contained 70% neutral carbohydrate, 7% amino sugars, 15% protein, 5% lipids and 0.8% phosphorus. Mycelial cell walls contained 70% carbohydrate, 14% aminosugars, 6% protein, 5% lipids and 0.6% phosphorus. Three polysaccharides: -glucan, mannan and chitin were detected. Proteins were solubilized from both cell wall fractions and separated by polyacrylamide gel electrophoresis. About 50 protein bands were detected, four of them corresponding to glycoproteins. The cell walls of the yeast and mycelial forms of Y. lipolytica were qualitatively similar and only quantitative differences were found.Abbreviations GlcNAc N-acetylglucosamine - FITC-WGA fluorescein isothiocyanate-wheat germ agglutinin - PAS periodic acid Schiff     

IDANOTHEKION GEN. N., A SYNANGIATE POLLEN ORGAN WITH SACCATE POLLEN FROM THE MIDDLE PENNSYLVANIAN OF ILLINOIS

Idanothekion glandulosum gen. et sp. n. is a synangiate pollen organ represented by approximately 30 specimens contained in coal balls from the middle Pennsylvanian of Illinois. Each synangium is composed of seven to nine elongate sporangia that are fused laterally for approximately four-fifths of their length, and are radially arranged about, and fused to, a short central column; the central column is restricted to the proximal one-third of the synangium. Distal to the column the sporangia surround a hollow central area. Dehiscence occurred by means of a longitudinal slit along the mid-line of the inner face of each sporangium. The outer walls of the sporangia have a complex histology involving an external epidermis, a middle presumably glandular layer containing scattered enlarged cells, and an inner layer made up of thin-walled parenchyma. Vascular tissue is present in the central column and outer walls of the sporangia. Each sporangium has a prominent, attenuate, multicellular tip. Large numbers of saccate pollen grains similar to those found in numerous fossil and extant coniferophytes as well as some Mesozoic pter-idosperms were produced in each sporangium. Idanolhekion resembles some synangia assignable to Paleozoic members of the Marattiales; however, the new genus compares most closely with pollen organs believed to have been produced by members of the Pteridospermales. It seems most likely that Idanothekion represents the pollen organ of some member of the Lyginopteridaceae that produced pollen of a type which up to now has not been known from Paleozoic seed ferns.     

Cell wall composition in the ascomycete sphaerostilbe repens at different developmental stages

Changes in the biochemical composition of isolated cell walls were analysed during the differentiation of coremia and rhizomorphs in Sphaerostilbe repens.Differentiation was accompanied by exclusively quantitative variations of the wall components: the content in carbohydrates, chitin and free amino sugars increased; on the contrary, amino acids, uronic acids, lipids and mineral substances decreased.Carbohydrates were composed of glucose, galactose and mannose; glucosamine was the main component of amino sugars. The predominant amino acid in the walls was cysteine the amount of which increased during hyphal aggregation, while quantities of the sixteen other determined amino acids decreased.Mineral matter was present in large quantities in the walls of the fungus, especially in vegetative mycelium. Iron, phosphorus and calcium were the most abundant elements.Possible relations between the variations in chemical composition of the wall and the capability of hyphae to aggregate are discussed.