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Aniline agar: A simple medium useful in characterizing white-rot higher fungi in culture
Abstract
Species of fungi causing a white wood-rot comprise about 90 % of the wood-rotters. In the absence of a fruitbody it is possible, once isolated from the attacked wood, to determine the species responsible for the rot by using cultural features. We hereby propose a new test useful in fungal taxonomy which allows the separation of those fungi able to produce extracellular enzymes which react with aniline in an acid medium. The test consists in inoculating a small portion of mycelium in an acid aniline-containing medium. After 7 days it is possible to observe a brown halo around the inoculum in those species giving a positive reaction. A screening test was carried out with 80 strains of species belonging to the orders Agaricales, Aphyllophorales and Auriculariales. All strains of Aphyllophorales causing brown-rot gave a negative reaction whereas two thirds of the test species causing white-rot gave positive results. The aniline test allows the separation, within the white-rotters, of those having extracellular enzymes capable of reacting with aniline. This test, added to the series of Nobles and Stalpers may be an additional useful element in taxonomy based on cultural characters.
... To classify the axenic cultures as a white or brown-rot fungus, aniline agar medium was used according to Alberto and Wright [24]. To prepare this medium, two solutions were employed: solution A (a mixture of 20 mL of aniline, 5 mL of concentrated nitric acid, and 500 mL of deionized water) and solution B (a mixture of 20 g of agar with 500 mL of deionized water). ...
This study explores the biotechnological potential of lignocellulolytic fungi collected in an oak forest. Fungal collections were obtained from natural reserves located in Boyacá-Colombia, ranging from 2700 to 3000 m.a.s.l. Twenty-three strains were isolated on malt agar, molecular characterization was performed, and ligninolytic and cellulolytic enzymatic activities were screened. Several white-rot fungi of biotechnological importance were identified as follows: Trametes sp., Trametes versicolor, Trametes villosa, Pycnoporus sanguineus, Bjerkandera adjusta, Lentinula boryana, Panus conchatus, Antrodia neotropica, Brunneoporus malicola, Laetiporus gilbertsonii, Stereum sp., Ganoderma sp., and Dichomitus sp. The strains T. versicolor 0554 and 0583, T. villosa 0562, and B. adusta 0556 showed the highest response in the qualitative enzymatic assays. These strains were used to determine their ability to decolorate the dyes aniline blue and Congo red, and it was found that T. villosa 0562 reached a level of decolorization close to 90% after 48 h of submerged culture. The fungal strains obtained here could offer alternatives to develop a process to accomplish sustainable development objectives.
... Production of oxidases and phenoloxidases was evaluated by growing the strain on tannic and gallic media (Nobles, 1965). Tyrosine, p-cresol and guaiacol Spots tests (Boidin, 1954), and the aniline test (Albertó & Wright, 1997) reactions were measured by the intensity ofthe colour halo produced. All the information thus obtained was codified in a "species code" according to Nobles (1965). ...
Using a strain determined by Singer 37 years ago as Pleurotus lindquistii, fruitbodies were obtained using traditional methods for edible mushroom culture. The new basidiomata allowed us to redescribe the species and to transfer it to Lentinus. Mating studies between monosporic cultures revealed a tetrapolar mating system. Descriptions of cultural characters and Nobles' code are given.
... In this work 26 native Argentinean strains of white rot fungi belonging to the orders Agaricales, Aphyllophorales and Auriculariales (Albert o and Wright, 1997) were examined for the decolorization of six industrial dyes (azo, triphenylmethane, anthraquinonic, heterocyclic and polymeric dyes). Their decolorizing abilities were compared with strains of P. chrysosporium, recognized as very efficient dye-degrading microorganisms (Cripps et al., 1990;Spadaro et al., 1992;Rodriguez et al., 1999;Podgornik et al., 2001). ...
The decolorizing capacity of 26 white rot fungi from Argentina was investigated. Extracellular production of ligninolytic enzymes by mycelium growing on solid malt extract/glucose medium supplemented with different dyes (Malachite Green, Azure B, Poly R-478, Anthraquinone Blue, Congo Red and Xylidine), dye decolorization and the relationship between these two processes were studied. Only ten strains decolorized all the dyes, all ten strains produced laccase, lignin peroxidase and manganese peroxidase on solid medium. However, six of the strains could not decolorize any of the dyes; all six strains tested negative for lignin peroxidase, and produced less than 0.05 U/g agar of manganese peroxidase. Comparing the isolates with the well-known dye-degrader Phanerochaete chrysosporium, a new fungus was identified: Coriolus versicolor f. antarcticus, potentially a candidate for use in biodecoloration processes. Eighteen day-old cultures of this fungus were able to decolorize in an hour 28%, 30%, 43%, 88% and 98% of Xylidine (24 mg/l), Poly R-478 (75 mg/l), Remazol Brilliant Blue R (9 mg/l), Malachite Green (6 mg/l) and Indigo Carmine (23 mg/l), respectively. Laccase activity was 0.13 U/ml, but neither lignin peroxidase nor manganese peroxidase were detected in the extracellular fluids for that day of incubation.
... Different qualitative and quantitative methodologies to detect extracellular oxidases and peroxidases have already been used by several investigators [4,7,11]. Exist previous works [13][14] on enzymatic characterization of different white-rot Basidiomycetes using several aromatic compounds in spot-tests. However, there is no information available as regards the production of extracellular oxidorreductases by autochthonous fungal strains belonging to different ecophysiological and taxonomic groups. ...
The screening for extracellular oxidases and peroxidases from autochthonous filamentous fungi isolated from different substrates is an important step towards the detection of extracellular fungal oxidative systems. Thirty-one autochthonous fungal strains from Argentina, belonging to different ecophysiological and taxonomic groups, were plate-screened for their ability to produce extracellular oxidoreductases. Modified Kirk solid medium containing the chromogen 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) was used to determine the presence of this extracellular activity. The fungi tested were grouped according to the colour intensity of the modified Kirk medium in: a) species without extracellular ABTS-oxidizing activity; b) species with low extracellular ABTS-oxidizing activity; c) species with moderate extracellular ABTS-oxidizing activity; d) species with high extracellular ABTS-oxidizing activity. The assay revealed extracellular ABTS-oxidizing activity in 90% of the strains tested. All species of Basidiomycetes used exhibited ABTS-oxidizing activity, except Laeticorticium roseum. Aspergillus terreus and Epicoccum purpurascens (Deuteromycetes) did not show extracellular oxidative activity on ABTS. Agrocybe aegerita, Amauroderma boleticeum, Cladosporium cladosporioides, Coriolopsis rigida, Grammothele subargentea, Graphium putredinis, Hexagona hydnoides, Hexagona papyraceae, Loweporus lividus, Peniophora albobadia, Phellinus everhartii, Phellinus gilvus; Phellinus linteus; Pleurotus laciniatocrenatus, Pycnoporus sanguineus, Rigidoporus ulmarius, Steccherinum rawakense, Talaromyces helicus, Trametes elegans, Trametes pavonia, Trametes villosa and Trichaptum sector are reported here for the first time as species capable of producing ABTS-oxidizing extracellular oxidorreductases.
The decolorizing capacity of 26 white rot fungi from Argentina was investigated. Extracellular production of ligninolytic enzymes by mycelium growing on solid malt extract/glucose medium supplemented with different dyes (Malachite Green, Azure B, Poly R-478, Anthraquinone Blue, Congo Red and Xylidine), dye decolorization and the relationship between these two processes were studied. Only ten strains decolorized all the dyes, all ten strains produced laccase, lignin peroxidase and manganese peroxidase on solid medium. However, six of the strains could not decolorize any of the dyes; all six strains tested negative for lignin peroxidase, and produced less than 0.05 U/g agar of manganese peroxidase. Comparing the isolates with the well-known dye-degrader Phanerochaete chrysosporium, a new fungus was identified: Coriolus versicolor f. antarcticus, potentially a candidate for use in biodecoloration processes. Eighteen day-old cultures of this fungus were able to decolorize in an hour 28%, 30%, 43%, 88% and 98% of Xylidine (24 mg/l), Poly R-478 (75 mg/l), Remazol Brilliant Blue R (9 mg/l), Malachite Green (6 mg/l) and Indigo Carmine (23 mg/l), respectively. Laccase activity was 0.13 U/ml, but neither lignin peroxidase nor manganese peroxidase were detected in the extracellular fluids for that day of incubation.
This listing covers the period May 1, 1997 through to June 30, 1997. Citations are arranged in groups which roughly correspond with the British Mycological Society's Special Interest Committees. All correspondence about this item should be addressed to the Executive Editor. Reprints of this feature will not be available.
This listing covers the period September 1, 1997 through to October 31, 1997. Citations are arranged in groups which roughly correspond with the British Mycological Society's Special Interest Committees. All correspondence about this item should be addressed to the Executive Editor. Reprints of this feature will not be available.
Diazonium blue B reagent was used to test staining reactions of mycelia of 96 species of ectomycorrhizal and ectomycorrhizal-like fungi grown for five weeks in liquid culture. Red to violet staining was seen in harvested mycelia of basidiomycetes treated in cold KOH while negative reactions indicated fungi with ascomycetous affinities. Non-treated mycelia gave a variety of different color responses which were generally species-specific. Mycelia of Cortinarius species and most boletes did not stain while most species of Hebeloma, Hygrophorus, Lacearia and Tricholoma stained yellow. A purely red staining reaction was restricted mainly to a few species within the Boletaceae. Results reveal potential value of diazonium blue B staining as a taxonomic character for identification of mycelia isolated from field-collected ectomycorrhizae.
An oxygenase enzyme was isolated from the basidiomycete fungus Trametes versicolor, that is capable of attacking lignin and a large number of di- and tri-substituted benzene rings containing at least one hydroxy group. This enzyme system was produced late in the growth cycle without the requirement for any inducer. This non-selective enzyme system is thermophilic and operates at pH 3–5 in the presence of air or oxygen. The action of this enzyme system caused the loss of UV absorption in ferulic acid solution, the formation of hydroxy muconic semialdehyde from catechol, and transformation with the production of CO2 from a number of hydroxy aromatics as well as lignin.
Roaring Billy Forest Walk, leg. M. Rajchenberg, 8-V-89 (strain 0582)
- Haast Park
- Pass
Park, Haast Pass, Roaring Billy Forest Walk, leg. M. Rajchenberg, 8-V-89 (strain
0582). Isol. & Det. M. Rajchenberg.
Bjerkandera adusta: ARG., Tucurnan, Horco Molle, leg. D. Job & A.
082; Neuquen, Nat'l Park Lanin
- Leg M Avellaneda
- Pose
Avellaneda, Leg. M. Pose, 24-IV-93
(strain 2105), Isol. & Det. A. Gottlieb BAFC 33.082; Neuquen, Nat'l Park Lanin, Lo.
Loloy, leg. P. Wielong & M. Rajchenberg, 13-IV-92 (strain 2563), Isol. & Det. M.
Rajchenberg, BAFC 33.056.
Ganoderma resinaceum s. lato: ARG., Entre Rios, Concordia, Parque