Inter- and intra-species heterogeneity in germination of Aspergillus conidia

Aspergilli are among the most abundant fungi worldwide. They degrade organic material and can be pathogens of plants and animals. Aspergilli spread by forming high numbers of conidia. Germination of these stress resistant asexual spores is characterized by a swelling and a germ tube stage. Here, we show that conidia of Aspergillusniger,Aspergillusoryzae,Aspergillusclavatus, Aspergillusnidulans and Aspergillusterreus show different swelling and germ tube formation dynamics in pure water or in water supplemented with (in)organic nutrients. Apart from inter-species heterogeneity, intra-species heterogeneity was observed within spore populations of the aspergilli except for A.terreus. Sub-populations of conidia differing in size and/or contrast showed different swelling and germ tube formation dynamics. Together, data imply that aspergilli differ in their competitive potential depending on the substrate. Moreover, results suggest that intra-species heterogeneity provides a bet hedging mechanism to optimize survival of aspergilli

Heat resistance acquirement of the spoilage yeast Saccharomyces diastaticus during heat exposure

The main fungal cause of spoilage of carbonated fermented beverages in the brewing industry is the amylolytic budding yeast Saccharomyces cerevisiae subsp. diastaticus (Saccharomyces diastaticus). Heat treatment is used to avoid microbial spoilage of the fermented beverages. Therefore, the spoilage capacity of S. diastaticus may be linked to its relative high heat resistance. Here, we assessed whether S. diastaticus can acquire heat resistance when exposed to heat stress. To this end, ascospores of S. diastaticus strain MB523 were treated at 60°C for 10 min followed by growing the surviving spores on a glucose‐containing medium. The resulting vegetative cells were then allowed to sporulate again in sporulation medium. This cycle of heat treatment, vegetative growth, and sporulation was performed eight times in three independent lineages. After these eight cycles, the sporulation rate was similar to the start (∼75%) but the resulting ascospores were more heat resistant. The time needed to kill 90% of the population at 60°C (i.e. the D60‐value) increased from 6.5 to 9.0 min (p = 0.005). The vegetative cells also showed a trend to increased heat resistance with an increase in the D52‐value from 9.2 to 16.2 min (p = 0.1). In contrast, heat resistance of the vegetative cells that had not been exposed to heat during the eight cycles had been reduced with a D52‐value of 4.2 min (p = 0.003). Together, these data show that S. diastaticus MB523 can easily acquire heat resistance by inbreeding while subjected to heat stress. Conversely, heat resistance can be easily lost in the absence of this stress condition, indicative of a trade‐off for heat resistance

The role of the Flb protein family in the life cycle of Aspergillus niger

Genes flbA-E are involved in sporulation and vegetative growth in Aspergillus nidulans. Inactivation of either of these genes results in a fluffy phenotype with delayed or even abolished sporulation. Previously, a non-sporulating phenotype was obtained by inactivating flbA in Aspergillus niger, which was accompanied by lysis, thinner cell walls, and an increased secretome complexity. Here, we further studied the role of the flb genes of A. niger. Strains ΔflbA, ΔflbB and ΔflbE showed increased biomass formation, while inactivation of flbA-D reduced, or even abolished, formation of conidia. Strain ΔflbA was more sensitive to H2O2, DTT, and the cell wall integrity stress compounds SDS and Congo Red (CR). Also, ΔflbC was more sensitive to SDS, while ΔflbB, ΔflbD, and ΔflbE were more sensitive to CR. On the other hand, inactivation of flbE increased resistance to H2O2. Enzyme secretion was impacted when the Δflb strains were grown on xylose. Strain ΔflbE showed reduced xylanase, cellulase and amylase secretion. On the other hand, amylase secretion at the periphery of the ΔflbA colony was reduced but not in its center, while secretion of this enzyme was increased in the center of the ΔflbB colony but not at its periphery. Inactivation of flbC and flbD also impacted zonal cellulase and amylase activity. Together, the Flb protein family of A. niger function in biomass formation, sporulation, stress response, and protein secretion

SapB and the rodlins are required for development of Streptomyces coelicolor in high osmolarity media

Streptomyces coelicolor produces spore-forming aerial hyphae after a period of vegetative growth. These aerial structures are decorated with a hydrophobic coating of rodlets consisting of chaplins and rodlins. Here, we show that rodlins and the surface-active peptide SapB are essential for development during growth in a medium with high osmolarity. To this end, both vegetative and aerial hyphae secrete SapB, whereas rodlins are only secreted by the spore-forming aerial hyphae.

Discoloration of textile dyes by spent mushroom substrate of Agaricus bisporus

The textile industry discharges up to 5 % of their dyes in aqueous effluents. Here, use of spent mushroom substrate (SMS) of commercial white button mushroom production and its aqueous extract, SMS tea, was assessed to remove textile dyes from water. A total of 30–90 % and 5–85 % of the dyes was removed after a 24 h incubation in SMS and SMS tea, respectively. Removal of malachite green and remazol brilliant blue R was similar in SMS and its tea. In contrast, removal of crystal violet, orange G, and rose bengal was higher in SMS, explained by sorption to SMS and by the role of non-water-extractable SMS components in discoloration. Heat-treating SMS and its tea, thereby inactivating enzymes, reduced dye removal to 8–58 % and 0–31 %, respectively, indicating that dyes are removed by both enzymatic and non-enzymatic activities. Together, SMS of white button mushroom production has high potential to treat textile-dye-polluted aqueous effluents.</p

Binding of micro-nutrients to the cell wall of the fungus Schizophyllum commune

The cell wall fulfils several functions in the biology of fungi. For instance, it provides mechanical strength, interacts with the (a)biotic environment, and acts as a molecular sieve. Recently, it was shown that proteins and β-glucans in the cell wall of Schizophyllum commune bind Cu2+. We here show that the cell wall of this mushroom forming fungus also binds other (micro-)nutrients. Ca2+, Mg2+, Mn2+, NO3–, PO43-, and SO42- bound at levels > 1 mg per gram dry weight cell wall, while binding of BO3-, Cu2+, Zn2+ and MoO42- was lower. Sorption of Ca2+, Mn2+, Zn2+ and PO43- was promoted at alkaline pH. These compounds as well as BO33-, Cu2+, Mg2+, NO3–, and SO42- that had bound at pH 4, 6, or 8 could be released from the cell wall at pH 4 with a maximum efficiency of 46–93 %. Solid-state NMR spectroscopy showed that the metals had the same binding sites as Cu2+ when a low concentration of this ion is used. Moreover, data indicate that anions bind to the cell wall as well as to the metal ions. Together, it is shown that the cell wall of S. commune binds various (micro-)nutrients and that this binding is higher than the uptake by hyphae. The binding to the cell wall may be used as a storage mechanism or may reduce availability of these molecules to competitors or prevent toxic influx in the cytoplasm

Enzymatic and non-enzymatic removal of organic micropollutants with spent mushroom substrate of Agaricus bisporus

Water bodies are increasingly contaminated with a diversity of organic micropollutants (OMPs). This impacts the quality of ecosystems due to their recalcitrant nature. In this study, we assessed the removal of OMPs by spent mushroom substrate (SMS) of the white button mushroom (Agaricus bisporus) and by its aqueous tea extract. Removal of acesulfame K, antipyrine, bentazon, caffeine, carbamazepine, chloridazon, clofibric acid, and N, N-diethyl-meta-toluamide (DEET) by SMS and its tea was between 10 and 90% and 0–26%, respectively, in a 7-day period. Sorption to SMS particles was between 0 and 29%, which can thus not explain the removal difference between SMS and its tea, the latter lacking these particles. Carbamazepine was removed most efficiently by both SMS and its tea. Removal of OMPs (except caffeine) by SMS tea was not affected by heat treatment. By contrast, heat-treatment of SMS reduced OMP removal to &lt; 10% except for carbamazepine with a removal of 90%. These results indicate that OMP removal by SMS and its tea is mediated by both enzymatic and non-enzymatic activities. The presence of copper, manganese, and iron (0.03, 0.88, and 0.33 µg L -1, respectively) as well as H 2O 2 (1.5 µM) in SMS tea indicated that the Fenton reaction represents (part of) the non-enzymatic activity. Indeed, the in vitro reconstituted Fenton reaction removed OMPs &gt; 50% better than the teas. From these data it is concluded that spent mushroom substrate of the white button mushroom, which is widely available as a waste-stream, can be used to purify water from OMPs.</p