PubMed · 14440969
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M SALES, VILLARD. [Progressive scleroderma (initial form)].. https://pubmed.ncbi.nlm.nih.gov/14440969/
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High-quality genomic DNA extraction from basidiomycete fungi remains challenging due to polysaccharide co-purification and nuclease-mediated DNA degradation. We systematically compared 22 experimental conditions combining five DNA extraction methods (one Dellaporta-based method, three CTAB-based variants, and one commercial kit) with five mycelium preparation techniques (scalpel scraping, fungal suspension, silica bead homogenization, liquid fermentation, and liquid nitrogen treatment) for Phanerodontia australis (BRM62389). DNA quality was assessed by 1% agarose gel electrophoresis, NanoDrop spectrophotometry, and Qubit fluorimetry. The optimal protocol combined liquid-fermentation mycelium with an adapted CTAB method, yielding Qubit-quantified concentrations of 34.2 and 62.6 ng/µL (samples S21 and S22), 260/280 ratios of 2.06 and 2.05, 260/230 ratios of 1.88 and 1.85, and no detectable DNA degradation by agarose gel electrophoresis. This protocol enabled whole-genome sequencing, yielding a 37.62 Mb assembly with 98.5% completeness as assessed by BUSCO v5 (basidiomycota_odb10 lineage dataset, n = 1,764). Liquid fermentation is associated with the production of younger, actively growing hyphae with reduced cell wall thickness, and the adapted CTAB method effectively removed residual polysaccharides. This protocol provides a reliable, cost-effective approach for obtaining sequencing-grade DNA from P. australis.
The wheat cultivar Kariega expresses complete adult plant resistance to stripe rust in South Africa. The aim of this investigation was to determine the extent and nature of variability in stripe rust resistance in a population of 150 doubled haploid lines generated from a cross between Kariega and the susceptible cultivar Avocet S. Analysis of field data for adult plant stripe rust resistance identified two major QTLs and two minor QTLs in the resistant cultivar Kariega. The two major QTLs were located on chromosomes 7D ( QYr.sgi-7D) and 2B ( QYr.sgi-2B.1), contributing 29% and 30% to the phenotypic variance, respectively. QYr.sgi-2B.1 is primarily associated with a chlorotic and/or necrotic response, unlike QYr.sgi-7D, which is believed to be the adult plant resistance gene Yr18. These two QTLs for adult plant resistance in Kariega appear to represent different forms of resistance, where QYr.sgi-7D may represent potentially more durable resistance than QYr.sgi-2B.1. Mixture model analysis of the field leaf infection scores suggested a genetic model involving two independent genes combining in a classical, epistatic manner. The results of the QTL analysis demonstrate its higher resolution power compared to the mixture model analysis by detecting the presence of minor QTLs.
The aim of the present work was to study the possibility of using the fatty acid content in the basidiospores as a taxonomic tool. Basidiospores of Armillaria borealis, Amanita muscaria, Agaricus sylvicola, Hypholoma capnoides, Cortinarius nemorensis and Russula delica were used. The content of fatty acids as well as other substances may vary to a certain degree depending on the part (pileus, stipe, lamella) or stage of development of the actual basidiocarp analysed. Moreover, substances from fungivorous invertebrates, parasitic fungi or bacteria may be found in the chemical analyses of the basidiocarps. Chemotaxonomic conclusions may, therefore, be burdened with serious uncertainties. On the other hand, the ripe basidiospores are terminated structures and belong to the most homogenous structures encountered from a basidiocarp. Their shape, size, colour and ornamentation are considerably homogenous within an actual species. Therefore, the basidiospores are often used as a reliable differentiating characteristic separating species as well as taxa of higher categories. From a practical point of view, ripe spores are easy to obtain in relatively large quantities with simple techniques, and they are not so prone to decay as the carpophore tissue. In the present study, gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS), after methanolysis of the fungus spores, were used to map essential fatty acids in basidiomycetes. Gas chromatography and gas chromatography-mass spectrometry revealed the presence of fatty acids of C12:0-C24:0 size in the basidiospores of these higher basidiomycetes. The major fatty acid in H. capnoides is C18:2, and the major fatty acid in the other species is C18:1. The basidiospores proved to be a good source of fatty acids for chemotaxonomic investigations of agarics.