Involvement of an Extracellular Glucan Sheath during Degradation of Populus Wood by Phanerochaete chrysosporium

Observations by transmission electron microscopy of wood samples of Populus tremula inoculated with the white rot fungus Phanerochaete chrysosporium showed that, at certain stages of their growth cycle, hyphae were encapsulated by a sheath which seems to play an active role in the wood cell wall degradation. Chemical and immunochemical techniques and C nuclear magnetic resonance spectroscopy were applied to demonstrate the beta-1,3-1,6-d-glucan nature of the sheath. Double-staining methods revealed the interaction between the extracellular peroxidases involved in lignin degradation and the glucan mucilage. The glucan was also shown to establish a material junction between the fungus and the wood cell wall. It was concluded that, by means of these interactions, the sheath provides a transient junction between the hyphae and the wood, thus establishing a point of attachment to the site of the degradation. The association of peroxidases to the glucan matrix is in favor of the role of the sheath as a supporting structure. Furthermore, that the sheath was hydrolyzed during the attack demonstrated its active role both in providing the H(2)O(2) necessary to the action of peroxidases and in providing a mode of transport of the fungal enzymes to their substrates at the surface of the wood cell wall.     

Callus formation and induction of a cell suspension culture from Acacia senegal

Cell proliferation has been induced from the cambial zone of a branch of Acacia senegal, kept on the basal Knop and Ball medium in the presence of auxin. Transferred on the more complete nutrient medium of Schenk and Hildebrandt, the colonies gave rise to several cell lines. One of the friable lines, consisting of aggregates of parenchymatous cells, gave a cell suspension culture in an agitated liquid medium which is maintained as a strain of illimited growth. The heterogeneous suspension did not undergo noticeable changes after eight transfers. Aggregates collected on a 1000-m nylon seive were able to grow on a solid medium and gave back a friable callus similar to the initial colonies.     

Mode of action of a xylanase and its significance for the structural investigation of the branched l-arabino-d-glucurono-d-xylan from redwood (Sequoia sempervirens)

The substitution pattern of the water-soluble l-arabino-(4-O-methyl-d-glucurono)-d-xylan from redwood (Sequoia sempervirens) has been studied by enzymic degradation. Exhaustive hydrolysis by an endo-xylanase (EC 3.2.1.8) from a Basidiomycete Sporotrichum dimorphosporum left a residue accounting for 20% of the original d-xylan. In the dialyzable material, oligosaccharides having arabinose or 4-O-methylglucuronic acid residues attached to the non-reducing d-xylosyl end-group of xylobiose or xylotriose, respectively, were the smallest branched oligomers released. Action of the xylanase appears to involve a region of the polysaccharide backbone having three xylosyl residues. A mode of action is proposed that requires unsubstituted hydroxyl groups at C-2, C-3, and C-2′ of a xylobiosyl residue. The binding site seems to correspond to a shallow cavity. The composition and structure of the final residue of attack shows that the enzyme has no action when the xylosyl residues branched through O-2 are separated by only one, unsubstituted xylose residue. This pattern of action, the nature of the dialyzable products, and the production of a final residue in which the substituents are accumulated, suggest that the arabinosyl and glucosyl-uronic groups are irregularly distributed on the main chain of the xylan from redwood and that in some regions they are in close vicinity when not actually on adjacent xylosyl residues.     

The use of bacteriophage depolymerization in the structural investigation of the capsular polysaccharide from Klebsiella serotype K3

The structure of the repeating unit of the capsular polysaccharide from Klebsiella serotype K3 has been established from the results of n.m.r. (1H and 13C) spectroscopy and methylation analysis of P1, the pyruvic acetal-bearing pentasaccharide obtained on depolymerization of the polysaccharide with a bacteriophage-borne endogalactosidase, reduced deacetalated P1, and the native polysaccharide. The data permit the assignment of the following structure to the repeating unit: (formula see text)     

Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Modification of hemicellulose content by antisense down-regulation of UDP-glucuronate decarboxylase in tobacco and its consequences for cellulose extractability

Extractability and recovery of cellulose from cell walls influences many industrial processes and also the utilisation of biomass for energy purposes. The utility of genetic manipulation of lignin has proven potential for optimising such processes and is also advantageous for the environment. Hemicelluloses, particularly secondary wall xylans, also influence the extractability of cellulose. UDP-glucuronate decarboxylase produces UDP-xylose, the precursor for xylans and the effect of its down-regulation on cell wall structure and cellulose extractability in transgenic tobacco has been investigated. Since there are a number of potential UDP-glucuronate decarboxylase genes, a 490bp sequence of high similarity between members of the family, was chosen for general alteration of the expression of the gene family. Sense and antisense transgenic lines were analysed for enzyme activity using a modified and optimised electrophoretic assay, for enzyme levels by western blotting and for secondary cell wall composition. Some of the down-regulated antisense plants showed high glucose to xylose ratios in xylem walls due to less xylose-containing polymers, while arabinose and uronic acid contents, which could also have been affected by any change in UDP-xylose provision, were unchanged. The overall morphology and stem lignin content of the modified lines remained little changed compared with wild-type. However, there were some changes in vascular organisation and reduction of xylans in the secondary walls was confirmed by immunocytochemistry. Pulping analysis showed a decreased pulp yield and a higher Kappa number in some lines compared with controls, indicating that they were less delignified, although the level of residual alkali was reduced. Such traits probably indicate that lignin was less available for removal in a reduced background of xylans. However, the viscosity was higher in most antisense lines, meaning that the cellulose was less broken-down during the pulping process. This is one of the first studies of a directed manipulation of hemicellulose content on cellulose extractability and shows both positive and negative outcomes.