Soil Filamentous Fungi Research Articles

Resistance and Activity of Phytopathogenic and Potential Antagonistic Soil Filamentous Fungi under the Action of Heavy Metals

Resistance and Activity of Phytopathogenic and Potential Antagonistic Soil Filamentous Fungi under the Action of Heavy Metals

Mutagenic effects of a commercial glyphosate-based herbicide formulation on the soil filamentous fungus Aspergillus nidulans depending on the mode of exposure.

Glyphosate-based herbicides (GBH) are the most used pesticides worldwide. This widespread dissemination raises the question of non-target effects on a wide range of organisms, including soil micro-organisms. Despite a large body of scientific studies reporting the harmful effects of GBHs, the health and environmental safety of glyphosate and its commercial formulations remains controversial. In particular, contradictory results have been obtained on the possible genotoxicity of these herbicides depending on the organisms or biological systems tested, the modes and durations of exposure and the sensitivity of the detection technique used. We previously showed that the well-characterized soil filamentous fungus Aspergillus nidulans was highly affected by a commercial GBH formulation containing 450g/L of glyphosate (R450), even when used at doses far below the agricultural application rate. In the present study, we analysed the possible mutagenicity of R450 in A. nidulans by screening for specific mutants after different modes of exposure to the herbicide. R450 was found to exert a mutagenic effect only after repeated exposure during growth on agar-medium, and depending on the metabolic status of the tested strain. The nature of some mutants and their ability to tolerate the herbicide better than did the wild-type strain suggested that their emergence may reflect an adaptive response of the fungus to offset the herbicide effects. The use of a non-selective molecular approach, the quantitative random amplified polymorphic DNA (RAPD-qPCR), showed that R450 could also exert a mutagenic effect after a one-shot overnight exposure during growth in liquid culture. However, this effect was subtle and no longer detectable when the fungus had previously been repeatedly exposed to the herbicide on a solid medium. This indicated an elevation of the sensitivity threshold of A. nidulans to the R450 mutagenicity, and thus confirmed the adaptive capacity of the fungus to the herbicide.

Review on Performance of Aspergillus and Penicillium Species in Biodegradation of Organochlorine and Organophosphorus Pesticides.

The use of pesticides in agricultural practices raises concerns considering the toxic effects they generate in the environment; thus, their sustainable application in crop production remains a challenge. One of the frequently addressed issues regarding their application includes the development of a sustainable and ecofriendly approach for their degradation. Since the filamentous fungi can bioremediate various xenobiotics owing to their efficient and versatile enzymatic machinery, this review has addressed their performance in the biodegradation of organochlorine and organophosphorus pesticides. It is focused particularly on fungal strains belonging to the genera Aspergillus and Penicillium, since both are ubiquitous in the environment, and often abundant in soils contaminated with xenobiotics. Most of the recent reviews on microbial biodegradation of pesticides focus primarily on bacteria, and the soil filamentous fungi are mentioned only marginally there. Therefore, in this review, we have attempted to demonstrate and highlight the exceptional potential of aspergilli and penicillia in degrading the organochlorine and organophosphorus pesticides (e.g., endosulfan, lindane, chlorpyrifos, and methyl parathion). These biologically active xenobiotics have been degraded by fungi into various metabolites efficaciously, or these are completely mineralized within a few days. Since they have demonstrated high rates of degradation activity, as well as high tolerance to pesticides, most of the Aspergillus and Penicillium species strains listed in this review are excellent candidates for the remediation of pesticide-contaminated soils.

Interaction of different pigmented micromycetes with humic substances and stability of soil biomes: spectral characterization

Humic substances (HSs) support the sustainable functioning of soils. Filamentous fungi are involved in HSs turnover, demonstrating the ability to degrade and simultaneously synthesize HSs. The aim of the work was to study the transformation of HS samples by soil filamentous fungi using a set of spectral indices and to connect those findings to stability of soil biomes. We used three fungal strains: melanin containing Alternaria alternatа, non-pigmented Fusarium solani and Trichoderma harzianum. Absorption and fluorescence spectra were measured after cultivating fungi in various concentrations of HSs (200 mg/L for HPs and 100 mg/L for HAs) and sucrose (3 and 0.3 g/L). The transformations of HSs under cultivation of A alternatа led to increase of absorbance ratio A250/A365, shortening of the fluorescence emission wavelength, and growth in fluorescence quantum yield QY355. Under cultivation of F. solani and T. harzianum we did not observe these spectral changes. This indicates that among three strains studied, A. alternata degrades HSs the most actively. The data we obtained and principal component analysis showed that phytopathogenic A. alternata, F. solani and antagonistically active against alternaria and fusaria T. harzianum have different patterns of interaction with HSs; this should be considered when humic-based biostimulants are used.

Antagonistic Potentiality of Some Soil Fungi Against Six Fungal Pathogens Isolated From Cotton (Gossypium Hirsutum L.) Seeds

The antagonistic potentials of four soil filamentous fungi viz., Aspergillus flavus Link, A. fumigatus Fresenius, A. niger van Tieghem and Trichoderma viride Pers. against six pathogens isolated from 14 varieties of cotton (Gossypium hirsutum L.) were evaluated by "dual culture colony interaction”, volatile and non-volatile metabolites. In dual culture colony interaction, out of four soil fungi, T. viride was found be the most effective to control the growth of the cotton pathogens. T. viride showed the highest growth inhibition against Curvularia lunata, Fusarium moniliforme, Mucor sp. and Rhizoctonia solani. On the other hand A. niger showed the highest growth inhibition against Fusarium nivale and A. fumigatus showed the maximum growth inhibition against C. gloeosporioides. The highest inhibition of radial growth of C. lunata, F. nivale and F. moniliforme was observed might be due to the secretion of volatile metabolites of T. viride whereas, the maximum inhibition of radial growth of C. gloeosporioides was observed because of the volatile metabolites of A. flavus. Mucor sp. and R. solani were inhibited due to the volatile metabolites of A. fumigatus. The greatest radial growth inhibition of C. lunata and F. moniliforme were noticed in case of T. viride owing to the effect of non-volatile metabolites. On the other hand, the radial growth of C. gloeosporioides and F. nivale were inhibited highest amount for the effect of non-volatile metabolites of A. niger, whereas highest growth inhibition of Mucor sp. and R. solani was observed due to the non-volatile effect of A. fumigatus. The present investigation suggests that the isolates of Aspergillus and Trichoderma may be further exploited as potential biocontrol agents against the fungal pathogens of cotton in field trial.
 Asiat. Soc. Bangladesh, Sci. 46(2): 143-153, December 2020

Growth rate trades off with enzymatic investment in soil filamentous fungi

Saprobic soil fungi drive many important ecosystem processes, including decomposition, and many of their effects are related to growth rate and enzymatic ability. In mycology, there has long been the implicit assumption of a trade-off between growth and enzymatic investment, which we test here using a set of filamentous fungi from the same soil. For these fungi we measured growth rate (as colony radial extension) and enzymatic repertoire (activities of four enzymes: laccase, cellobiohydrolase, leucine aminopeptidase and acid phosphatase), and explored the interaction between the traits based on phylogenetically corrected methods. Our results support the existence of a trade-off, however only for the enzymes presumably representing a larger metabolic cost (laccase and cellobiohydrolase). Our study offers new insights into potential functional complementarity within the soil fungal community in ecosystem processes, and experimentally supports an enzymatic investment/growth rate trade-off underpinning phenomena including substrate succession.

Diversity of Soil Filamentous Fungi Influenced by Marine Environment in São Luís, Maranhão, Brazil.

Introduction In recent decades, there has been an intensification of environmental problems, which are becoming increasingly critical and frequent due to population growth. Microorganisms, including soilborne fungi, play an essential role in maintaining and balancing the environment. One of the most impacted ecosystems in São Luís, Maranhão, Brazil, is the Jansen Lagoon State Park, an important tourist spot, which has suffered anthropogenic actions such as the dumping of household waste (sewage) in its body of water. As a consequence, these pollutants can accumulate in the adjacent soil, since the body of water is near this substrate. The objectives were to isolate and identify filamentous fungi from the soil of the Jansen Lagoon State Park. Methods Monthly soil samples were collected and later processed using the modified suspension technique according to Clark (1965). Results The isolated genera were Aspergillus, Penicillium, Trichoderma, Absidia, and Fusarium. Aspergillus is the fungal genus of greater dominance in the soil of the Jansen Lagoon State Park. Aspergillus niger was the dominant species (37%), followed by A. tamarii (21.6%). Conclusion The main isolated fungi from the Jansen Lagoon State Park were Aspergillus niger and Aspergillus tamrii. These fungi can be used as biological markers of pollution and as biodegraders and/or bioremediators to improve the area studied.

Effect of Exogenic Humic Substances on Various Growth Endpoints of Alternaria alternata and Trichoderma harzianum in the Experimental Conditions

Industrial humic products (HPs) manufactured from various organic matter resources, including fossils, peat, lake-bottom sediments and organic waste materials, have found multiple applications in soil environments. Fungi play a crucial role in the turnover of humic substances (HSs). However, the data on the biological effects of HSs on soil fungi are contradictory. Revealing mechanisms of how HSs interact with soil filamentous fungi is essential for understanding a sustainable soil system. Here we study the responses of phytopathogenic Alternaria alternata and the potential antagonist Trichoderma harzianum, to exogenic HPs (from leonardite and lignosulfonate) at two concentrations (0.1 and 0.02%) in growth media with varying sucrose concentrations (0, 3 and 30 g/L). We assessed the fungal growth endpoints including the mycelium biomass accumulation, fungal colony size and conidia production. Our results demonstrate the significant relationships between the extent of inhibitory/stimulation effect of HPs on the investigated fungi and the carbon status of the growth medium, some features of the fungal species, as well as the type and concentration of the introduced HPs. The responses of plant pathogens correlated specifically with various types of HPs. HP from lignosulfonate exhibited a more stimulating effect on A. alternata compared to the effect of HP from leonardite.

Transcriptome profiling of the fungus Aspergillus nidulans exposed to a commercial glyphosate-based herbicide under conditions of apparent herbicide tolerance

Glyphosate-based herbicides, such as Roundup®, are the most widely used non-selective, broad-spectrum herbicides. The release of these compounds in large amounts into the environment is susceptible to affect soil quality and health, especially because of the non-target effects on a large range of organisms including soil microorganisms. The soil filamentous fungus Aspergillus nidulans, a well-characterized experimental model organism that can be used as a bio-indicator for agricultural soil health, has been previously shown to be highly affected by Roundup GT Plus (R450: 450 g/L of glyphosate) at concentrations far below recommended agricultural application rate, including at a dose that does not cause any macroscopic effect. In this study, we determined alterations in the transcriptome of A. nidulans when exposed to R450 at a dose corresponding to the no-observed-adverse-effect level (NOAEL) for macroscopic parameters. A total of 1816 distinct genes had their expression altered. The most affected biological functions were protein synthesis, amino acids and secondary metabolisms, stress response, as well as detoxification pathways through cytochromes P450, glutathione-S-transferases, and ABC transporters. These results partly explain the molecular mechanisms underlying alterations in growth parameters detected at higher concentrations for this ascomycete fungus. In conclusion, our results highlight molecular disturbances in a soil fungus under conditions of apparent tolerance to the herbicide, and thus confirm the need to question the principle of “substantial equivalence” when applied to plants made tolerant to herbicides.

Biodegradation of humic substances by microscopic filamentous fungi: chromatographic and spectroscopic proxies

The study of interactions between humic substances (HSs) and soil filamentous fungi is the key to understanding the sustainable soil functioning. The present work aims to examine the decomposition of HSs by filamentous dark-pigmented fungus Alternaria alternatа under the laboratory conditions and to determine the effect of easily assimilable organic carbon on this process. Analyzing such polydisperse substances like HSs by a complex integrated methodology makes it possible to explore the data on their decomposition by microorganisms. To achieve the aforementioned goals, we used chromatographic and spectroscopic approaches: low-pressure size-exclusion and hydrophobic interaction chromatography accompanied by absorption and fluorescence spectroscopy. To determine the effect cometabolism conditions produced on HS decomposition, two types of carbon substrates were added to the nutrient media: easily assimilable organic carbon (standard 0.3% or reduced 0.03% sucrose content) and hardly assimilable organic carbon (HSs), as well as their combinations. Five HS samples of different organic matter origin have been inspected: potassium humates (HPs) and humic acids (HAs) from coal, peat, and lignosulfonate. Correlation matrix and principal component analysis (PCA) were calculated for comprehensive data analysis. Transformations of the investigated HSs under fungal cultivation lead to the increase in the low molecular weight fraction, rise of hydrophilic fraction, enlargement of absorbance ratio A250/A365, shortening of the emission wavelength of the humic-type fluorescence, and growth in the fluorescence quantum yield measured with excitation at 355 nm. A positive correlation was observed between the accumulation of fungal biomass and the degree of HS decomposition. PCA analysis confirms that the difference in the results of HS decomposition largely depends on the sucrose content and the nature of HSs. We divided all the HS samples into four groups according to the degree of HS decomposition: original HS solutions, HPs altered using fungal cultivation at 0.03% sucrose, HAs after fungal cultivation at 0.03% sucrose, and finally, HSs (both HPs and HAs) after fungal cultivation at 0.3% sucrose. In the laboratory experiments, we showed that (1) the isolated HAs were more effectively degraded than the parent HPs, and this process was more pronounced at a reduced sucrose content, and (2) the decomposition of stable organic compounds (HSs) was activated by the easily assimilable carbon sources (especially 0.3% sucrose) being present. We assume that it is the easily assimilable organic carbon that most likely triggers the HS degradation working as the priming effect in natural environments.

Enzyme production by soil filamentous fungi under solid-state fermentation in banana stalk

The adequate disposal of agricultural waste is one of the major concerns of public officials and a research challenge to obtain sustainable solutions to the problem. In this sense, the objective of this study was to evaluate the banana leaf stalk use as substrate in solid-state fermentation (SSF) for production of amylolytic and cellulolytic enzymes by Penicillium spp. LEMI A11 strain grown under different substrate concentrations, pH and temperature. Effects of different pH conditions (5.0 and 6.0), temperature (30 and 35 °C) and substrate concentration 70 and 90% (in relation to the final volume) of the fermentation were evaluated over 120 hours of fermentation. The results indicated that Penicillium spp. LEMI A11 was able to use the banana stalk as substrate under SSF. The maximum activities for amylase dextraining, amylase saccharifying and CMCase were 0.18; 0.13 and 04 U.g-1, respectively. The effect of environmental factors related to the substrate concentration was significant for saccharifying amylase and CMCase activity only. The interaction between the environmental factors tested was significant for the dextrinizing amylase activity only. It was verified enzyme activity reduction after 96 hours of fermentation for all enzymes. It concluded that banana stalk is an alternative carbon source to be used in SSF for enzyme production by of Penicillium spp. LEMI A11.

Proteomic analysis of the soil filamentous fungus Aspergillus nidulans exposed to a Roundup formulation at a dose causing no macroscopic effect: a functional study.

Roundup® is a glyphosate-based herbicide (GBH) used worldwide both in agriculture and private gardens. Thus, it constitutes a substantial source of environmental contaminations, especially for water and soil, and may impact a number of non-target organisms essential for ecosystem balance. The soil filamentous fungus Aspergillus nidulans has been shown to be highly affected by a commercial formulation of Roundup® (R450), containing 450g/L of glyphosate (GLY), at doses far below recommended agricultural application rate. In the present study, we used two-dimensional gel electrophoresis combined to mass spectrometry to analyze proteomic pattern changes in A. nidulans exposed to R450 at a dose corresponding to the no-observed-adverse-effect level (NOAEL) for macroscopic parameters (31.5mg/L GLY among adjuvants). Comparative analysis revealed a total of 82 differentially expressed proteins between control and R450-treated samples, and 85% of them (70) were unambiguously identified. Their molecular functions were mainly assigned to cell detoxification and stress response (16%), protein synthesis (14%), amino acid metabolism (13%), glycolysis/gluconeogenesis/glycerol metabolism/pentose phosphate pathway (13%) and Krebs TCA cycle/acetyl-CoA synthesis/ATP metabolism (10%). These results bring new insights into the understanding of the toxicity induced by higher doses of this herbicide in the soil model organism A. nidulans. To our knowledge, this study represents the first evidence of protein expression modulation and, thus, possible metabolic disturbance, in response to an herbicide treatment at a dose that does not cause any visible effect. These data are likely to challenge the concept of "substantial equivalence" when applied to herbicide-tolerant plants.

Resilience of Fungal Communities to Elevated CO2.

Soil filamentous fungi play a prominent role in regulating ecosystem functioning in terrestrial ecosystems. This necessitates understanding their responses to climate change drivers in order to predict how nutrient cycling and ecosystem services will be influenced in the future. Here, we provide a quantitative synthesis of ten studies on soil fungal community responses to elevated CO2. Many of these studies reported contradictory diversity responses. We identify the duration of the study as an influential parameter that determines the outcome of experimentation. Our analysis reconciles the existing globally distributed experiments on fungal community responses to elevated CO2 and provides a framework for comparing results of future CO2 enrichment studies.

Multiple effects of a commercial Roundup® formulation on the soil filamentous fungus Aspergillus nidulans at low doses: evidence of an unexpected impact on energetic metabolism.

Soil microorganisms are highly exposed to glyphosate-based herbicides (GBH), especially to Roundup® which is widely used worldwide. However, studies on the effects of GBH formulations on specific non-rhizosphere soil microbial species are scarce. We evaluated the toxicity of a commercial formulation of Roundup® (R450), containing 450g/L of glyphosate (GLY), on the soil filamentous fungus Aspergillus nidulans, an experimental model microorganism. The median lethal dose (LD50) on solid media was between 90 and 112mg/L GLY (among adjuvants, which are also included in the Roundup® formulation), which corresponds to a dilution percentage about 100 times lower than that used in agriculture. The LOAEL and NOAEL (lowest- and no-observed-adverse-effect levels) associated to morphology and growth were 33.75 and 31.5mg/L GLY among adjuvants, respectively. The formulation R450 proved to be much more active than technical GLY. At the LD50 and lower concentrations, R450 impaired growth, cellular polarity, endocytosis, and mitochondria (average number, total volume and metabolism). In contrast with the depletion of mitochondrial activities reported in animal studies, R450 caused a stimulation of mitochondrial enzyme activities, thus revealing a different mode of action of Roundup® on energetic metabolism. These mitochondrial disruptions were also evident at a low dose corresponding to the NOAEL for macroscopic parameters, indicating that these mitochondrial biomarkers are more sensitive than those for growth and morphological ones. Altogether, our data indicate that GBH toxic effects on soil filamentous fungi, and thus potential impairment of soil ecosystems, may occur at doses far below recommended agricultural application rate.

Temperature priming and memory in soil filamentous fungi

Abstract Most soil fungi experience a constantly fluctuating environment, and coping with resulting biotic and abiotic stressors can come at a considerable metabolic cost. We know that organisms respond better to severe stresses (triggering stress) when they have experienced a similar milder stress (priming stress) before. Asking how long organisms can remember a priming event is a compelling question. We here studied priming by temperature stress in filamentous fungi isolated from the same grassland soil. We hypothesized that filamentous fungi can show priming responses, and that their memory-spans correlate with their growth rates. Fungal colonies of 19 different filamentous fungi were first primed at 35 °C for 5 h (as priming stress) and after 0, 6, 12, 24 or 48 h they were exposed to 40 °C for 10 h (as triggering stress). The variable lag time between the stress applications allowed us to assess memory. Our main response variable was growth rate. Of the 19 fungal isolates tested, eight showed temperature priming ability. The Mucoromycotina isolates (Mortierellales) showed a mean growth increase following triggering stress that was 2.75-fold higher than in unprimed colonies (log-response-ratio). Mucoromycotina isolates had a memory half-life span (power-law-relationships) of 5.65 h. Considering fungal traits like growth rate to predict priming responses, we found a positive relationship between priming response (with 12 h memory phase) and growth rate. The differential ability to be primed in co-occurring isolates may have direct consequences for fungal communities and coexistence in soil.

Potential of Microscopic Fungi Isolated from Mercury Contaminated Soils to Accumulate and Volatilize Mercury(II)

This article evaluates mercury-resistant autochthonous filamentous fungi’s role in mercury transformation and mobility with respect to mercury biovolatilization. Aspergillus, Cladosporium, Trichoderma, and Alternaria genera isolated from mercury contaminated and non-contaminated soils were cultivated under static conditions in 8.2–32.7 mg L−1 mercury(II) concentration range to evaluate and compare mercury bioaccumulation, fungal strain resistance, and biovolatilization efficiency. Results indicate the enormous fungal capacity for mercury removal and volatilization, especially by Aspergillus niger and Cladosporium isolates, which volatilized almost 80 % of initial mercury content during 7-day static cultivation in the dark. We presume that the mercury detoxification mechanism changed above our 8.2 mg L−1 initial media concentration with significant domination of mercury volatilization. We suggest that mercury biovolatilization, rather than its deposition or efflux in non-volatile forms, is the major filamentous fungal detoxification mechanism. Our results highlight that the soil filamentous fungi’s contribution on mercury biogeochemical cycle may be considered significant, if the conditions for fungal growth are sufficient. Still, when contaminated soil is treated with respect to enhanced fungal growth, biovolatilization may be beneficiary for natural remediation processes applied for mercury contaminated soils.

A fast and efficient method for total DNA extraction from soil filamentous fungi

espanolLa extraccion de ADN de alta calidad de hongos puede ser afectada debido a la presencia de una pared celular compleja, alto contenido de polisacaridos o metabolitos secundarios. En esta investigacion se adapto un simple metodo de extraccion y purificacion de ADN de hongos filamentosos aislados del suelo, para aplicaciones tipo Polymerase Chain Reaction (PCR). La metodologia descrita es rapida y de bajo costo. EnglishExtraction of high quality of DNA from fungus can be affected because the presence of the complex cell wall, high content of polysaccharides or secondary metabolites. In this research, we adapt a simple method of DNA extraction and purification of from fungus isolated from soil for Polymerase Chain Reaction (PCR) applications. The methodology described is both rapid and cost effective.

Identification of biological secondary metabolites in three Penicillium species, P. goditanum, P. moldavicum, and P. corylophilum

Microorganisms are important components of soil. Some soil filamentous fungi such asPenicilium produce many bioactive small molecules, or secondary metabolites, that range frombeneficial bioactive compounds to harmful toxins. In this study, the metabolites of threepenicillium species (P. goditanum, P. moldavicum and P. corylophilum) were extracted byadding ethyl acetate to liquid cultures. The metabolites were determined using gaschromatography and mass spectrometry. The results obtained from the GC-MS analysis showedthat Penicillium species are sources of bioactive compounds. We have identified differentgroups of compounds, such as alkaloids, alkenes, sesquiterpens, fatty acids, and essential oils.Among them, 1,3,8-p-Menthatriene, 2-methylenecyclohexane, anthracene, neoisolongifolene,[14] annulene and thioxanthene in P. goditanum, isocyclocitral, coumarin-6-ol and 2, 4, 6-Trimethoxystyrene in P. moldavicum as well as asarone in P. corylophilum were majorcompounds. Moreover, several compounds, such as oxalic acid, dibenzothiophene,hexadecanoic acid, and alkane hydrocarbons, were identified in all species.

Fermentative production of nervonic acid by Mortierella capitata RD000969.

A high-nervonic acid (cis-15-tetracosenoic acid, C24:1, n-9)-producing filamentous fungus of the Mortierella species was discovered among soil filamentous fungi. The filamentous fungal strain -RD000969- was isolated from soil collected in Kanagawa Prefecture (Japan) and was found to accumulate nervonic acid at a rate of 6.94% of the total cellular fatty acids. The base sequences of 28S rDNA D1/D2 and ITS 5.8S rDNA showed 100% homology with Mortierella capitata CBS 293.96. In addition to nervonic acid, strain RD000969 produced a large amount of long-chain monounsaturated fatty acids (C20:1, 12.22%; C22:1, 4.07%; C26:1, 5.91%) and a small amount of ultra-long-chain fatty acids (C28:1, 0.44%; C30:1, 0.06%; C32:1, trace). In the fungal cells, 98.87% of nervonic acid was localized at the sn-1,3 position of triacylglycerol. Nervonic acid production was maximum (186.3 mg·L(-1)) when the fungus was cultured in potato dextrose (PD) medium containing yeast extract, CaCl2, and MgSO4·7H2O.

Energy-dependent uptake of benzo[a]pyrene and its cytoskeleton-dependent intracellular transport by the telluric fungus Fusarium solani

In screening indigenous soil filamentous fungi for polycyclic aromatic hydrocarbons (PAHs) degradation, an isolate of the Fusarium solani was found to incorporate benzo[a]pyrene (BaP) into fungal hyphae before degradation and mineralization. The mechanisms involved in BaP uptake and intracellular transport remain unresolved. To address this, the incorporation of two PAHs, BaP, and phenanthrene (PHE) were studied in this fungus. The fungus incorporated more BaP into cells than PHE, despite the 400-fold higher aqueous solubility of PHE compared with BaP, indicating that PAH incorporation is not based on a simple diffusion mechanism. To identify the mechanism of BaP incorporation and transport, microscopic studies were undertaken with the fluorescence probes Congo Red, BODIPY®493/503, and FM®4-64, targeting different cell compartments respectively fungal cell walls, lipids, and endocytosis. The metabolic inhibitor sodium azide at 100 mM totally blocked BaP incorporation into fungal cells indicating an energy-requirement for PAH uptake into the mycelium. Cytochalasins also inhibited BaP uptake by the fungus and probably its intracellular transport into fungal hyphae. The perfect co-localization of BaP and BODIPY reveals that lipid bodies constitute the intracellular storage sites of BaP in F. solani. Our results demonstrate an energy-dependent uptake of BaP and its cytoskeleton-dependent intracellular transport by F. solani.