A Model Sheet Mineral System to Study Fungal Bioweathering of Mica

The general objective of this research was to examine fungal interactions with silicate minerals within the context of their roles in bioweathering. To achieve this, we used muscovite, a phyllosilicate mineral (KAl₂[(OH)₂|AlSi₃O₁₀]), in the form of a mineral sheet model system for ease of experimental manipulation and microscopic examination. It was found that test fungal species successfully colonized and degraded the surface of muscovite sheets in both laboratory and field experiments. After colonization by the common soil fungus Aspergillus niger, a network of hyphae covered the surface of the muscovite, and mineral dissolution or degradation was clearly evidenced by a network of fungal “footprints” that reflected coverage by the mycelium. For natural soil incubations, microorganisms associated with muscovite sheet material included biofilms of fungi and bacteria on the surface, while mineral encrustation or adhesion to microbial structures was also observed. Our results show that muscovite sheet is a good model mineral system for examination of microbial colonization and degradation, and this was demonstrated using laboratory and field systems, providing more evidence for the bioweathering significance of fungal activities in the context of silicate degradation and soil formation and development. The approach is also clearly applicable to other rock and mineral-based substrates and a variety of free-living and symbiotic microbial systems.     

Detection of metal-chelating compounds from wood-rotting fungi Trametes versicolorand Wolfiporia cocos

For many years, the wood decay process by fungi was associated almost exclusively with production of lignocellulolytic enzymes. However, recent studies by electron microscopy have shown that fungal enzymes are too large to penetrate into the cell wall at an early stage of decay. Thus, the hypothesis that low molecular mass agents may initiate the breakdown of both cellulose and lignin was proposed. The purpose of this work was to detect low molecular mass compounds, with metal-chelating capability, from liquid cultures of two wood-rot fungi. The brown-rot fungus Wolfiporia cocos produced the highest chrome azurol S (CAS) reaction, simultaneously reducing the pH of the malt extract medium. In contrast, the white-rot fungus Trametes versicolor did not react with CAS and the pH remained approximately constant during the culture period. The presence of hydroxamate derivatives and oxalic acid was detected in extracts of low molecular mass of both fungi. Moreover, in W. cocos extracts, catecholate derivatives were also detected. Accumulation of oxalic acid was greater in W. cocos than in T. versicolor at the end of the culture period, and this might be responsible for the strong response from W. cocos in the CAS reaction.     

草酸青霉对含钾矿物风化及钾溶出的影响

【目的】探讨含钾硅酸盐矿物在草酸青霉(Penicillium oxalicum)作用下的风化及钾溶出情况。【方法】利用等离子体发射光谱、X-射线能谱、X-射线光电子能谱分析了3种常见含钾硅酸盐矿物(钾长石、白云母和黑云母)在草酸青霉作用后浸出液和矿物表面元素含量的变化;通过X-射线衍射分析矿物晶相结构的变化;采用共聚焦激光扫描显微镜表征了草酸青霉在矿物表面形成的生物膜;测量了培养液中不同碳源与氮源组分对草酸青霉解钾的影响。【结果】草酸青霉对结构稳定的钾长石和白云母风化速率较低,相比而言黑云母容易被风化并释放可溶性钾元素;草酸青霉在矿物表面形成了网状结构的生物膜,有利于微环境的生成及有机酸在其内的富集,促进微环境内钾的释放,强化微生物对矿物的风化作用;草酸青霉对多种碳源及氮源都表现出较好的适用性。【结论】草酸青霉是一种能够促进多种含钾矿物风化和钾溶出的真菌,在堆肥和生物肥料领域具有广泛的应用前景。     

不同风化年限碳酸岩风化壳真菌群落结构特征

【目的】岩生真菌是促进碳酸岩生物风化的重要推动者,研究黔中典型喀斯特区不同风化年限碳酸岩风化壳真菌群落结构特征有利于了解真菌对岩石的风化作用。【方法】选取位于黔中花溪区南部的废弃碳酸岩墓碑为调查对象,对不同风化年限碳酸岩风化壳进行采样,运用宏基因组测序方法对风化壳样品进行基因测序,并利用统计学方法对真菌群落结构特征及功能特征进行分析。【结果】18个风化壳样品中共获得1087种真菌,分属于9个门、44纲、538属。不同样本之间真菌群落数量和组成差异较大,在碳酸岩风化过程中,子囊菌门(Ascomycota)始终为优势类群,平均相对丰度达到95%以上,随风化年限的增加呈现显著下降的趋势。Shannon指数和Simpson指数显示,碳酸岩风化壳真菌群落多样性随风化年限的增加呈现先减小后增加再减小的趋势。从所有样本中共检测出3 379 478个KEGG pathway level 3通路相关基因,主要与物质能量的代谢、运输等功能相关。与碳循环、氮循环和硫循环相关的主要微生物属于子囊菌门,随着风化年限的增加呈现下降的趋势。冗余分析(redundancyanalysis,RDA)结果表明,三氧化二铁...     

Biodegradation of aliphatic hydrocarbon by indigenous fungi isolated from used motor oil contaminated sites

Eighteen indigenous fungal isolates has been successfully isolated from samples of used motor oil, top five centimetres of soil and drainage water contaminated with used motor oil. All of the pure fungal isolates obtained were identified, characterized and subjected to preliminary screening by evaluating the average growth rate of each fungal isolates on minimal media containing 1% (v/v) used motor oil. Trichoderma asperellum strain TUB F-1067 (SA4), Trichoderma asperellum strain Tr48 (SA5), Trichoderma asperellum strain TUB F-756 (SA6), Penicillium species (P1), and Aspergillus species (P9) were further selected for their hydrocarbon biodegradation potential. Among these five fungal isolates selected, P1 strain presented a significant degree of degradation by degrading almost all of the n-alkanes (n-C-₁₅ to n-C-₂₃ range) present in the used motor oil, thus of greater potential in degrading the aliphatic hydrocarbon compounds of used motor oil. The authors would like to certify that the work have not been sent/considered to be published in other journals.     

Screening and production of erythritol by newly isolated osmophilic yeast-like fungi

Twenty-eight erythritol-producing strains were isolated from pollen, honey and high sugar food samples collected in Taiwan. Amongst these, six strains (166-2, 262-1, 278-3, 440, 441 and 442) were high erythritol-producers with a yield higher than 30% for 30% glucose. The erythritol productivity of these strains ranged from 90.9 to 116.4 g l⁻¹. ¹H- and ¹³C-NMR analyses confirmed that the fermentation product was erythritol. The results of morphological and physiological studies indicate that strains 166-2, 262-1, 278-3, 440, and 442 may be members of the genus Moniliella. More studies are required to determine the taxonomic position of strain 441. The use of a medium containing 30% glucose and 1% yeast extract gave the highest erythritol productivity. On batch fermentation in a 5-l fermentor using strain 166-2, a maximal erythritol productivity of 111.0 g l⁻¹ was obtained after cultivation for 144 h.     

Keratinophilic fungi isolated from antarctic soil

In the present study, 10 soil samples were collected aseptically from an equal number of areas of the Antarctic in the zone occupied by the 1986–1987 Italian expedition for research on keratinophilic fungi.Of particular interest was the isolation of a pathogenic fungus, Microsporum gypseum, from two sites in the base camp occupied by men and by skuas. Trichophyton terrestre was isolated from a site in which people worked and through which penguins and skuas passed.The most widespread fungal species were members of the genus Chrysosporium. Some of these species were isolated but not identified and this part of the study was still be completed.Another significant finding was the absence of fungi in one sample, while in another the widespread and abundant growth in all the seeded dishes of a single species of Chrysosporium.Other species in genera of general diffusion in many environments were also isolated : Aspergillus spp., Malbranchea sp., Mycelia sterilia spp., Paecilomyces sp., Penicillium spp. and Scopulariopsis spp.     

The determination of ferric iron in plants by HPLC using the microbial iron chelator desferrioxamine E

Common methods for plant iron determination are based on atomic absorption spectroscopy, radioactive measurements or extraction with subsequent spectrophotometry. However, accuracy is often a problem due to background, contamination and interfering compounds. We here describe a novel method for the easy determination of ferric iron in plants by chelation with a highly effective microbial siderophore and separation by high performance liquid chromatography (HPLC). After addition of colourless desferrioxamine E (DFE) to plant fluids, the soluble iron is trapped as a brown-red ferrioxamine E (FoxE) complex which is subsequently separated by HPLC on a reversed phase column. The formed FoxE complex can be identified due to its ligand-to-metal charge transfer band at 435 nm. Alternatively, elution of both, DFE and FoxE can be followed as separate peaks at 220 nm wavelength with characteristic retention times. The extraordinarily high stability constant of DFE with ferric iron of K=10³² enables extraction of iron from a variety of ferrous and ferric iron compounds and allows quantitation after separation by HPLC without interference by coloured by-products. Thus, iron bound to protein, amino acids, citrate and other organic acid ligands and even insoluble ferric hydroxides and phosphates can be solubilized in the presence desferrioxamine E. The “Ferrioxamine E method” can be applied to all kinds of plant fluids (apoplasmic, xylem, phloem, intracellular) either at physiological pH or even at acid pH values. The FoxE complex is stable down to pH 1 allowing protein removal by perchloric acid treatment and HPLC separation in the presence of trifluoroacetic acid containing eluents. Published online December 2004     

Detection of chrysotile asbestos by using a chrysotile-binding protein

In the current studies, we found that the DksA protein from Escherichia coli binds strongly to chrysotile, which is the most commonly used form of asbestos. We developed a convenient colorimetric assay for chrysotile using a fusion of DksA and alkaline phosphatase along with 5-bromo-4-chloro-3-indolyl-phosphate and nitro blue tetrazolium as substrates. Also, using a fusion of DksA and green-fluorescent protein, we were able to detect chrysotile by fluorescence microscopy.     

Degradation of keratin substrates by fungi isolated from sewage sludge

Four fungal species including two dermatophytes and two saprophytes were isolated from sewage sludge samples at Basrah (Iraq) they were tested for their degradative ability towards three types of keratin substrates (human hair, chicken feathers and wool). The rate of keratin degradation was expressed as weight loss over three weeks of incubation using a liquid culture medium. Human hair had the highest degradation rate by colonization of Chrysosporium pannicola and Microsporum gypseum at a rate of 62% and 4% respectively. Chicken feathers were highly degraded by Aspergillus flavus (32%) while wool degradation was highest by C. pannicola (45.5%) and Trichophyton mentagrophytes var. erinacei (38%). There was a significant difference (p < 0.00l) in keratin substrate degradation rates by the examined fungi. Keratinase activity was highest for C. pannicola and M. gypseum in the culture medium baited with human hair. Aspergillus flavus revealed the highest activity of this enzyme in cultures amended with chicken feathers while T. mentagrophytes var. erinacei showed highest keratinase activity in cultures with wool substrate. The amount of protein released into the culture medium varied among the tested fungi. The medium's alkalinity increased over incubation time from 6.5 to 7.8. Microscopic examination showed maceration of the keratin substrates by the fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

OTA-producing fungi isolated from stored cocoa beans

Aims:  The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates.
Methods and Results:  Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [−1 culture medium; LOD = 6  μ g kg⁻¹ culture medium].
Conclusions:  No correspondence was found between the OTA levels in cocoa beans and the presence of OTA-producing fungi. Nonetheless, some samples contained A. carbonarius with a high OTA-producing ability and, consequently, specific fungal controls should be set up during storage to avoid this toxin.
Significance and Impact of the Study:  Toxigenic fungi in cocoa beans are not well understood. This study attempted to identify these fungi and evaluate their OTA-producing ability.     

Growth of Frankia strains in leaf litter-amended soil and the rhizosphere of a nonactinorhizal plant

Verticillium leptobactrum , a rare fungal species, has repeatedly been isolated from serpentinic rocks in the Western Alps, thus suggesting that it adapts easily to this selective mineral substrate. The rRNA internal transcribed spacer region of several isolates has been sequenced to confirm their identity and taxonomic position within Verticillium , a recently revised polyphyletic entity. Isolates of V. leptobactrum have also been investigated to establish their ability to weather asbestos chrysotile, the most common mineral in the isolation sites. The results of solubilization assays on magnesium and silicon, as well as measurement of the Mg/Si ratio in the asbestos fibres after exposure to fungal mycelia, indicate a high bioweathering activity of V. leptobactrum towards chrysotile. Comparison with data on Fusarium oxysporum shows differences among species, with V. leptobactrum being more active than F. oxysporum in removing structural ions from chrysotile. Asbestos weathering by fungi could be envisaged as a bioremediation strategy for hazardous asbestos-rich soils (e.g. abandoned mines). Fungi that have adapted to live in serpentine sites could be good candidates for this purpose.     

Keratinophilic fungi isolated from children's sandpits in the Nablus area,West Bank of Jordan

The keratinophilic fungi of 29 sandpits from kindergarten schools and public parks in the city of Nablus was analysed to evaluate their role in the epidemiology of diseases caused by these fungi. Seventy two species were recovered 28 of which were common to both kindergartens and public parks sandpits. High percentage (57.4%) of fungal isolates found had been identified as the causes of various types of mycoses. Eight species of dermatophytes and closely related fungi were recovered, of which the followings were the most commonly found species in sandpits: Chrysosporium keratinophilum (20.7%), Microsporum gypseum (17.2%), Trichophyton mentagrophytes (6.9%), and C. evolceanui (6.9%).     

Molecular diversity of bamboo-associated fungi isolated from Japan

Bamboos are common and useful plants in Japan but little information is available about their endophytes. In this study, 257 fungal strains were isolated from bamboo tissues, and 71 representative strains were characterized by 18S rRNA gene and internal transcriber spacer region sequence analysis. Phylogenetic analysis showed that the fungal isolates were located in Sordariomycetes and Dothideomycetes. Xylariales was the dominant group within bamboos. Several rRNA gene sequences were not similar to any current sequence in the database and might be a novel species or genera that could represent sources of novel biological compounds. These findings reveal that bamboos are a huge reservoir of microorganisms that should be extensively investigated.     

Keratinophilic fungi isolated from soils of Mumbai,India

Eighty-seven soil samples were collected from various areas of Mumbai and its vicinity to determine the prevalence of keratinophilic fungi. From the 55 positive samples (63.21%), a total of four genera with nine species were isolated viz. Chrysosporium indicum (28.73%),C. lobatum (2.29 %), C. sp.I (1.14%), C. sp. II (1.14%),C. tropicum (1.14%),C. zonatum (2.29%),Ctenomyces serratus (2.29%) Malbranchea aurantiaca (2.29%) and Microsporum gypseum complex (21.83%). This revised version was published online in June 2006 with corrections to the Cover Date.     

Control of Bemisia tabaci by entomopathogenic fungi isolated from arid soils in Argentina

Entomopathogenic Hypocreales were isolated from arid soils in Argentina using Tenebrio molitor as bait and tested for their biological performance at 30°C and 45–65% RH. Conidial germination was tested in three vegetable oils (sunflower, olive and maize) at two concentrations (1% and 10%) to evaluate their compatibility for further liquid formulations. According to radial growth and germination results, we selected four isolates to test their pathogenicity against second instar B. tabaci nymphs with the selected oil formulations at 30°C. CEP381 and CEP401 showed the highest radial growth. Isolates CEP381, CEP401, CEP413 and CEP409 (Metarhizium spp.) had similar germination percentages as compared with water control when germinated on either sunflower, olive or maize oils at 10% v/v. The highest mortality of B. tabaci was observed for the isolates CEP381 in sunflower oil and CEP401 in olive oil. Molecular identification of isolates was performed using ITS4–5 primers. All isolates belong to the Metarhizium core group. Tested isolates could grow and infect B. tabaci nymphs at 30°C in some of the vegetable oils as carriers, providing new possibilities for integrated pest management of Bemisia tabaci.     

Fibrinolytic activity of some fungi isolated from self-heated composted fertilizer

Mesophilic fungi isolated from organic fertilizer compost samples accounted for 70.94% of the total fungal count, while thermophilic and thermotolerant fungi constituted 29.05% of that count. Eight mesophilic fungal species, namelyAspergillus niger, Monilia sitophila, Paecilomyces divaricata, Penicillium chrysogenum, P. fellutanum, Scopulariopsis brevicaulis, S. brumptii andZygorhynchus japonicus; two thermophilic fungiHumicola grisea andOidiodendron flavum and three thermotolerant speciesAspergillus fumigatus, Thermomyces lanuginosus andZygorhynchus vuilleminii were isolated during the study. Most of the tested fungi showed a proteolytic activity and liquified gelatin in the test tube method and in cup plates. The thermophilic fungusO. flavum was the most potent proteolytic fungus. The comparative fibrinolytic assay revealed the following sequence in the ability of the tested fungi to hydrolyse fibrin:O. fiavum>S. brevicaulis>H. grisea>A. fumigatus>T. lanuginosus.     

卷柏内生真菌多样性研究

为探究蕨类植物内生真菌的多样性,本研究采用可培养方法分离江西龙虎山和广东丹霞山的岩壁常见蕨类植物——卷柏的内生真菌,结合形态特征和ITS序列进行菌株鉴定、研究多样性。结果表明：从1 080个组织块中,共分离出234株菌株,卷柏内生真菌的总定殖率为18.98%,总分离率为21.67%。根据分离频率,毛壳菌属Chaetomium（26.50%）、青霉属Penicillium（18.38%）、棒束孢属Isaria（11.97%）、曲霉属Aspergillus（9.42%）为优势属。不同居群地春季卷柏的内生真菌多样性存在差异,龙虎山的分离率为34.72%,而丹霞山仅为6.39%,其优势属也不尽相同。对龙虎山不同季节的卷柏内生真菌多样性研究表明,春季的分离率为34.72%,高于冬季的23.89%,春、冬两季之间的真菌类群相似性系数仅为0.39,差异显著。研究结果将有助于进一步揭示丹霞地貌地区蕨类植物内生真菌物种组成及其影响因素。     

Survey of keratinophilic fungi isolated from city park soils of Pisa,Italy

A survey of geophilic dermatophytes and related keratinophilic fungi isolated from city park soils of Pisa is reported. Twenty-three (48%) soil samples out of 48 were positive by hair baiting. The following species were isolated: Microsporum gypseum (39%), Trichophyton ajelloi (31%), Chrysosporium keratinophilum (14%), T. terrestre (8%), M. fulvum, Ch. luteum, Ch. indicum (5% each) and M. cookei (2%). The presence of the different species is discussed in relation to the risk of fungal skin infections. This revised version was published online in June 2006 with corrections to the Cover Date.