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Characterization of circular mitochondrial plasmids in three Pythium species.

Four circular plasmids, with a monomer size ranging from 3.2 to 4.94 kb, have been identified in isolates of P. aphanidermatum (two different plasmids), P. torulosum, and an unidentified echinulate isolate. The mitochondrial location has been confirmed for three of the plasmids. Each fungal isolate contained a single plasmid, present in both monomeric and oligomeric forms; plasmid monomers were present as open circles and as supercoiled forms. Restriction maps of the plasmids were dissimilar. Hybridization studies using cloned plasmids revealed no DNA sequence similarity among the different plasmids or between the plasmids and the nuclear or mitochondrial genome of the isolates from which they were recovered. Hybridization of labeled plasmid DNA to Northern transfers of mitochondrial RNA for two isolates indicate that what appears to be the predominant RNA transcript is unit length in size. For three isolates, the plasmid was retained following subculturing and was present in all asexual and sexual single-spore progeny evaluated. For one isolate of P. aphanidermatum the plasmid was unstable and was lost during subculturing.

Blotting, Northern↗

Pythium aphanidermatum: culture, cell-wall composition, and isolation and structure of antitumour storage and solubilised cell-wall (1----3),(1----6)-beta-D-glucans.

Under optimal conditions for the culture of the fungus Phytium aphanidermatum, no polysaccharides were excreted into the medium. The mycelium contained up to 38% of a slightly branched, storage (1----3),(1----6)-beta-D-glucan with a MW of 20,000. The cell-wall polysaccharides of the mycelium comprised 18% of cellulose and 82% of (1----3),(1----6)-beta-D-glucans. Of the non-cellulosic glucans, approximately 33% could be solubilised by extraction with water at 121 degrees, and they had a MW of 10,000, were highly branched, and contained 6% of (1----6) linkages. Treatment of the cell wall with 0.1 M trifluoroacetic acid released approximately 50% of the non-cellulosic glucans. The acid-soluble cell-wall (1----3),-(1----6)-beta-D-glucans of lower MW (6000) were still highly branched and contained 14% of (1----6) and 8% of (1----4) linkages. The storage glucan and the hot-water-soluble cell-wall glucan exhibited strong activity against the Sarcoma 180 in CD-1 mice, whereas the acid-soluble cell-wall glucans were inactive. The hot-water-soluble cell-wall glucan was also active against the DBA/2-MC.SC-1 fibrosarcoma in DBA/2 mice.

Animals↗

Purification, crystallization and preliminary X-ray studies of sylvaticin, an elicitin-like protein from Pythium sylvaticum.

Sylvaticin belongs to the elicitin family. These 10 kDa oomycetous proteins induce a hypersensitive response in plants, including necrosis and cell death, but subsequently leading to a non-specific systemic acquired resistance (SAR) against other pathogens. Sylvaticin has been crystallized using PEG 2000 MME as a precipitant agent in the presence of nickel chloride. The crystals belong to space group C2, with unit-cell parameters a = 99.29, b = 25.67, c = 67.45 A, beta = 99.66 degrees. Diffraction data were recorded to 2.1 A resolution at a synchrotron-radiation source.

Algal Proteins↗