The Development and Behaviour of Mycelial Strands in Merulius lacrymans (Wulf.) Fr.: II. Hyphal Behaviour during Strand Formation

The system of hyphal branching by Merulius lacrymans was observedin mycelium which had grown from a wood food-base on to glassslides during incubation in sterile moist chambers. A hierarchyof branches and sub-branches arose from the region of clampconnexions, or nodes, of relatively wide main hyphae. Therewas evidence that the sequence of branches occurring at nodesin basipetal succession represented the time sequence of branchdevelopment at any one node. Later-formed branches at any nodewere smaller than earlier branches, but such earlier branchesusually became smaller towards the tip as growth continued.Mycelial strands were built up by growth and branching of thigmo-tropicallysensitive ‘tendril’ hyphae in association with thewide main hyphae. Tendril hyphae were characteristically narrow,thin-walled hyphae arising both as later-formed branches fromthe nodes of the main hyphae and as the narrowed tips of earlierbranches. Although this branching behaviour could be seen amongstaerial hyphae growing over agar media, hyphae growing in contactwith or within the agar behaved differently and did not formstrands.     

Purification of lectin from fruiting bodies of Lactarius rufus (Scop.: Fr.)Fr. and its carbohydrate specificity

A lectin from fruiting bodies of Lactarius rufus (Scop.: Fr.)Fr. has been purified by affinity chromatography on copolymer of polyvinyl alcohol with a blood group B specific substance. The lectin gives a single band at disk-electrophoresis in acidic (pH 4.3) and alkaline (pH 8.6) buffer systems. Under electrophoresis in 10-20% SDS-PAGE, the lectin consists of identical subunits with molecular weight 17 +/- 1 kDa. Molecular weight of the lectin is 98 kDa according to gel-chromatography on Tojopearl HW-55. It is supposed that the lectin contains six subunits. The lectin is quite enough stable in pH 4.0-10.0, its activity does not depend upon bivalent metal ions. When heating the lectin solution to 65 degrees C it lost more than 85% of its activity. The lectin agglutinates human etrythrocytes without any marked group specificity, it agglutinates 2-4 times worse rabbit erythrocytes, very weakly crucian erythrocytes and does not agglutinate sheep erythrocytes. Mono- and disaccharides are not inhibitors of the lectin activity, while alpha-phenyl-N-acethyl-D-glucosaminopyranosid (0.08 mM) and 4-nitrophenyl-beta-D-glucosamin are the best inhibitors of its activity. Among glycoproteins the best inhibitors of the lectin activity are: group-specific substances from human blood erythrocytes, asialosubmaxillary bovine mucin, human and bovine thyroglobulin and more weak inhibitors are fetuin, transferrin and human Ig G.     

The Distribution of Substances in the Sporangiophores of Peronospora parasitica (Pers. ex Fr.) Fr.

The distribution of nuclei, RNA, mitochondria, lipid material,protein, and insoluble carbohydrates in the developing sporangiophoresof Peronospora parasitica was demonstrated by cytochemical staining.Nuclei, mitochondria, and protein showed a more or less uniformdistribution throughout the young sporangiophores, but werelocated almost entirely within the mature spores when sporedispersal commenced. Lipid material had a similar distribution,but was absent from the sporangiophore apex and sporangiophorebranch tips during the early stages of development. RNA wasabundant in the sporangiophore apices during early development,but occurred only within the spores, in small quantities, atmaturation. Although insoluble carbohydrates were sparse, theyhad a similar distribution to the nuclei, mitochondria, andprotein. Glycogen was not detected. The major soluble carbohydrates, present in the mature sporesin about equal proportions, were identified by thin-layer chromatographyas trehalose and an aldo-hexose, either glucose or mannose.These sugars were present in about equal quantities in the immaturesporangiophores and spores, while in the mature sporangiophoresfrom which the spores had been removed, trehalose was the majorsugar present. Sugar alcohols were not detected.     

Rhizomorph Behaviour in Armillaria mellea (Fr.) Quel.: With one Figure in the text: Saprophytic Colonization of Woody Substrates in Soil

Rhizomorphs put out by Armillaria mellea from small woody inoculain glass tubes of moist soil were tested for their ability tocolonize segments of willow shoots, previously killed by autoclavingand then buried for various periods in soil before inoculationwith A. mellea. The degree of saprophytic colonization of thesesubstrate segments by A. mellea was assessed by reburying themin fresh tubes of moist soil, and recording weekly growth incrementsof rhizomorphs put out from them over a period of 5 weeks. Aperiod of previous burial in soil up to 3 weeks was found notto diminish availability of the substrate segments to A. mellea,but with longer periods substrate value for A. mellea progressivelydeclined. Segments of living, green shoots of willow provideda consistently better substrate for A. mellea than did deadsegments, and these living segments maintained their substratevalue over periods (up to 7 weeks) of previous burial in soil.These results are interpreted in terms of competition betweenA. mellea and other soil fungi. In the infection of living tissues,A. mellea benefits from the exclusion of competitors by hostresistance. In the saprophytic colonization of dead tissues,A. mellea has the advantage of its capacity to decompose celluloseand lignin, which are substrates restricted to a minority ofsoil fungi.     

Branched glucan from the fruiting bodies of Piptoporus betulinus (Bull.:Fr.) Karst

A new glucan, namely, piptoporane I, with a molecular mass of 270 kDa was isolated from fruiting bodies of Piptoporus betulinis (Bull.:Fr.) Karst. (Fomitopsidacaeae). Using a combination ofphysicochemical methods, it was established that piptoporane I was a branched glucan with a backbone consisting of alpha-( 1->3)-glucopyranose residues substituted at the C-6 position by single residues of beta-D-glucopyranose by 17.3%. A polysaccharide with such a structure was isolated for the first time from the fungus genus Piptoporus.     

Changes of the cytoplasmic ultrastructure during development of sclerotia in Claviceps purpurea (Fr.) Tul.

The development of sclerotia of Claviceps purpurea was investigated by light and electron microscopy. During the first days after infection sterigma and conidiospores are formed. The spores show a moderately developed vacuolar system, they are thick walled and contain about 20% lipid (related to the cell volume) embedded in glycogen. The sterigma are cylindrical unicellular hyphae with electron dense cytoplasm and isolated strongly contrasted lipid droplets. In maturing sclerotia the hyphae become septated with increasingly thick cell walls and a large lipid content. The lipid forms small droplets in young cells, while in the mature sclerotium it occurs in the form of very large drops, occupying the major part of the cell. Simultaneously the composition of the lipid is changed. The mature cells have several nuclei. They are partially connected by osmiophilic substances, forming a network of intercellular spaces.Abbreviations HEPES N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid - DMSO Dimethylsulfoxide     

Polysaccharides from the fruit bodies of the basidiomycete Laetiporus sulphureus (Bull.: Fr.) Murr

The two main polysaccharides from the basidiomycetous fungus Laetiporus sulphureus were isolated, purified and characterized. The structural assignments were carried out using (13)C, (1)H, and (1)H,(13) HSQC nuclear magnetic resonance spectroscopy, methylation analysis, and Smith degradation. One was a linear beta-glucan having a (1-->3)-linked main chain, namely laminaran. The other was a fucomannogalactan, which consisted of a main chain of (1-->6)-linked alpha-D-galactopyranosyl residues, a part of them being substituted at O-2 by 3-O-D-mannopyranosyl-L-fucopyranosyl, alpha-D-mannopyranosyl and in a minor proportion, alpha-L-fucopyranosyl groups. This heteropolysaccharide is related to those of other Basidiomycetes heterogalactans, although it differs distinctly in its side-chain structures. Whereas part of the single-unit L-fucopyranosyl and/or 3-O-alpha-mannopyranosyl-L-fucopyranosyl residues are present as side chains of the other heterogalactans, additional alpha-D-mannopyranosyl units are present in our fucomannogalactan of L. sulphureus.     

Structural and immunological studies of a major polysaccharide from spores of Ganoderma lucidum (Fr.) Karst

A polysaccharide isolated from spores of the fungus, Ganoderma lucidum, was found to be a complex glucan. On the basis of compositional and methylation analyses, periodate oxidation, Smith degradation, 1D and 2D NMR, and ESIMS experiments of the native polysaccharide and its degraded products, the polysaccharide was shown to have a backbone of beta-(1-->3)-linked D-glucopyranosyl residues, with branches of mono-, di- and oligosaccharide side chains substituting at the C-6 of the glucosyl residues in the main chain. Conformational analysis in aqueous solution and immunological activities of the native and degraded glucans were also investigated. The results suggested that the degree of substitution on the main chain and the length of side chains may be very important factors in determining the conformation and the biological activities of beta-(1-->3)-linked glucans.     

Effects of Chemical Growth Retardants on Growth and Development of Sweetpotato (Ipomoea batatas (L.) Lam.) in Vitro

Plant growth retardants were evaluated for their ability to reduce the growth rate of sweetpotato (Ipomoea batatas (L.) Lam.) in vitro. Nodal sections of cv. Jewel were cultured for 30 days on medium containing NDA, ancymidol, phosfon, TIBA, difenzoquat, chlormequat, ACC, mepiquat chloride, or daminozide at 0, 10⁻⁴, 10⁻⁵, 10⁻⁶, 10⁻⁷, or 10⁻⁸ m. Difenzoquat, NDA, phosfon, and TIBA, at 10⁻⁴ m, were lethal to axillary bud explants. A low concentration (10⁻⁸ m) of chlorflurenol or NDA stimulated shoot elongation. The effective concentration range for most growth retardants was 10⁻⁵ to 10⁻⁶ m. Small (2- to 4-mm diameter) storage root-like swellings were observed on roots in cultures containing TIBA or ancymidol. The growth-inhibiting effects of ancymidol and NDA were transitory and did not persist through a 180-day culture period. Shoots cultured on medium containing 10⁻⁵ m phosfon, TIBA, or difenzoquat were significantly shorter than control plants after a 180-day culture period. Culture on medium containing TIBA, NDA, ancymidol, or ACC resulted in abnormal leaf and stem development. Plants derived from nodal explants cultured on medium containing either phosfon or chlormequat were near normal in appearance but with some plants exhibiting interveinal chlorosis and reduced root system development. Received May 9, 1997; accepted August 14, 1997