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White root rot disease (WRD), caused by Rigidoporus microporus, poses a serious threat to various tropical trees, resulting in substantial crop damage and even plant death. However, there is still a lack of information regarding WRD in tea plants. Due to soil-borne in nature, synthetic fungicide is not recommended to control WRD. Therefore, biocontrol using natural microorganisms found in the tea’s rhizosphere has been explored as a potential solution. This study aimed to identify rhizospheric fungi of tea plants with antagonistic properties that can be used as a biocontrol agent against R. microporus. The study involved a field survey and laboratory works had been conducted in a tea plantation at Rancabali, West Java and the Phytopathology Laboratory, Faculty of Agriculture, Universitas Padjadjaran from March to October 2021. The disease incidence and severity were assessed in three blocks of land representing different topographic conditions of the plantation while antagonism tests were conducted using the completely randomized design and dual culture method. The results showed that the highest occurrence of WRD was in the central part of the hill with the disease incidence and severity of 70 and 50%, respectively. Thirteen fungal isolates were obtained from the rhizosphere of tea plants, all of which were found to inhibit the growth of R. microporus. These isolates belonged to the genera of Trichoderma, Gliocladium, Paecilomyces, Penicillium, Aureobasidium, Acremonium and Rhizopus. The considerable inhibition of R. microporus colony growth was performed by the isolates of Aureobasidium sp. (51.23%), Rhizopus sp. (50%), and Trichoderma sp. (43.96%). The results suggested that rhizospheric fungi of tea plants have the potential as biocontrol agents against R. microporus. [ABSTRACT FROM AUTHOR]

Aureobasidium is omnipresent and can be isolated from air, water bodies, soil, wood, and other plant materials, as well as inorganic materials such as rocks and marble. A total of 32 species of this fungal genus have been identified at the level of DNA, of which Aureobasidium pullulans is best known. Aureobasidium is of interest for a sustainable economy because it can be used to produce a wide variety of compounds, including enzymes, polysaccharides, and biosurfactants. Moreover, it can be used to promote plant growth and protect wood and crops. To this end, Aureobasidium cells adhere to wood or plants by producing extracellular polysaccharides, thereby forming a biofilm. This biofilm provides a sustainable alternative to petrol-based coatings and toxic chemicals. This and the fact that Aureobasidium biofilms have the potential of self-repair make them a potential engineered living material avant la lettre. Key points: •Aureobasidium produces products of interest to the industry •Aureobasidium can stimulate plant growth and protect crops •Biofinish of A. pullulans is a sustainable alternative to petrol-based coatings •Aureobasidium biofilms have the potential to function as engineered living materials [ABSTRACT FROM AUTHOR]

Aureobasidium pullulans is an extremotolerant, cosmopolitan yeast-like fungus that successfully colonises vastly different ecological niches. The species is widely used in biotechnology and successfully applied as a commercial biocontrol agent against postharvest diseases and fireblight. However, the exact mechanisms that are responsible for its antagonistic activity against diverse plant pathogens are not known at the molecular level. Thus, it is difficult to optimise and improve the biocontrol applications of this species. As a foundation for elucidating biocontrol mechanisms, we have de novo assembled a high-quality reference genome of a strongly antagonistic A. pullulans strain, performed dual RNA-seq experiments, and analysed proteins secreted during the interaction with the plant pathogen Fusarium oxysporum. Based on the genome annotation, potential biocontrol genes were predicted to encode secreted hydrolases or to be part of secondary metabolite clusters (e.g., NRPS-like, NRPS, T1PKS, terpene, and β-lactone clusters). Transcriptome and secretome analyses defined a subset of 79 A. pullulans genes (among the 10,925 annotated genes) that were transcriptionally upregulated or exclusively detected at the protein level during the competition with F. oxysporum. These potential biocontrol genes comprised predicted secreted hydrolases such as glycosylases, esterases, and proteases, as well as genes encoding enzymes, which are predicted to be involved in the synthesis of secondary metabolites. This study highlights the value of a sequential approach starting with genome mining and consecutive transcriptome and secretome analyses in order to identify a limited number of potential target genes for detailed, functional analyses.

Gracilaria verrucosa is a red seaweed that has been widely utilized in the food and pharmaceutical industries due to its biological properties. The utilization of biological agents in obtaining certain bioactive compounds would confront unavoidable issues, particularly its bioactive sustainability. Hence, microbial fermentation has been reported as a practical approach to maintaining bioactive production and boosting its properties. Our study aimed to evaluate the potential of marine yeast Aureobasidium melanogenum MTGK.31 as a fermenting agent for G. verrucosa and characterize the seaweed metabolite profile and antioxidant activity after fermentation. The seaweed was fermented using A. melanogenum MTGK.31 in a medium consisting of yeast extract, peptone, and glucose. The fermentation was done for 24, 48, and 72 hours. Total plate count and pH were measured after each fermentation period. The primary and secondary metabolites of G. verrucosa in each fermentation were observed. Antioxidant assay using the 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) method was conducted, followed by total phenolic content using the Folin-Ciocalteu method. It was highlighted that yeast colony increased during the fermentation, while the pH level was decreasing. We found that the fermentation not only boosted some elements in primary metabolites, but also increased G. verrucosa bioactive groups. After 72 hours of fermentation, the G. verrucosa percent radical scavenging activity (%RSA) increased more than two times compared to the fresh G. verrucosa with a %RSA value of 16.09±6.57. Nevertheless, the highest total phenolic content of 5.62±0.00028 mg GAE/g extract was shown by G. verrucosa after 48 hours of fermentation.

Background: This work studied the occurrence, density, and diversity of yeast species associated with the leaves and fruits of Sabicea brasiliensis and Anacardium humile. Methods: Fragments of leaves and fruits were subjected to maceration and placed under agitation in peptone water. One hundred µL aliquots of each of the decimal dilutions were plated on YM chloramphenicol agar medium. The different morphotypes were identified through sequence analysis of the 26S rDNA. Results: A total of 83 yeast isolates were recovered from the organs sampled, 40 from fruits and 19 from the leaves of S. brasiliensis, while 14 were recovered from fruits and 10 from the leaves of A. humile. The isolates were identified as belonging to two phyla, six classes, eight orders, and eight families. Most isolates, at 71 (84.3%), belong to Ascomycota, with Saccharomycetes dominant in fruits and Dothideomycetes in leaves. The remaining 12 (15.7%) isolates belong to the phylum Basidiomycota. The most frequent genera were Aureobasidium, Candida, Rhodotorula, and Wickerhamiella. The Wickerhamiella isolates made possible the subsequent description of a new species within this genus. Conclusions: Our data reveal the yeast species that inhabit different Cerrado plant organs and indicate that these hosts can be considered an important habitat for yeast diversity, including new and/or previously unreported species in this biome. [ABSTRACT FROM AUTHOR]

In Poland's pine forests, mistletoe (Viscum album subsp. austriacum) abundance has increased over the past 20 years. This mistletoe infestation has decreased the growth and vigour of Scots pine trees. In this study, we surveyed the culturable fungi from healthy and diseased V. album subsp. austriacum leaves from two stands in Poland. In total, 63 distinct species were identified, 99.8% of which belonged to the phylum Ascomycota. The community compositions of fungi in mistletoe leaves were similar in healthy and diseased leaves as Alternaria alternata, Aureobasidium pullulans, Botryosphaeria visci, Fusarium paeoniae and Microsphaeropsis olivacea were consistently found in leaves of all symptom types. The most frequently isolated fungus from asymptomatic leaves was M. olivacea, followed by A. alternata, A. pullulans and Hypoxylon rubiginosum. In comparison, the most frequently isolated fungi from leaves with rusty‐brown necrotic spots were (in decreasing order) M. olivacea, B. visci, F. paeoniae and A. alternata, while the most frequently isolated fungi from leaves with black or dark brown spots were M. olivacea, A. alternata, A. pullulans, Epicoccum layuense and F. paeoniae. This study was the first comprehensive report showing that certain fungal species may be pathogens of V. album subsp. austriacum in Poland. This study was also the first report of F. paeoniae, F. juglandicola, Diaporthe vacuae and Heterotruncatella spartii from V. album, and the first report of D. vacuae and H. spartii in Poland. [ABSTRACT FROM AUTHOR]

Aureobasidium pullulans is a yeast‐like fungus with remarkable phenotypic plasticity widely studied for its importance for the pharmaceutical and food industries. So far, genomic studies with strains from all over the world suggest they constitute a genetically unstructured population, with no association by habitat. However, the mechanisms by which this genome supports so many phenotypic permutations are still poorly understood. Recent works have shown the importance of sequencing yeast genomes from extreme environments to increase the repertoire of phenotypic diversity of unconventional yeasts. In this study, we present the genomic draft of A. pullulans strain from a Patagonian yeast diversity hotspot, re‐evaluate its taxonomic classification based on taxogenomic approaches, and annotate its genome with high‐depth transcriptomic data. Our analysis suggests this isolate could be considered a novel variant at an early stage of the speciation process. The discovery of divergent strains in a genomically homogeneous group, such as A. pullulans, can be valuable in understanding the evolution of the species. The identification and characterization of new variants will not only allow finding unique traits of biotechnological importance, but also optimize the choice of strains whose phenotypes will be characterized, providing new elements to explore questions about plasticity and adaptation. Take away: New high‐quality omics resources of Aureobasidium pullulans were developed.A battery of genes encoding antioxidant and photoprotective enzymes was found.Taxogenomics approach unveils intriguing intraspecific variability in Patagonia.RPB2 marker and genomic indexes are good proxies to detect divergent strains.A. pullulans CRUB 1823 shows evidence of belonging to a distinct lineage. [ABSTRACT FROM AUTHOR]

The role of melanin in Aureobasidium pullulans ATCC 15233 and Zalaria obscura LS31012019, under simulated osmotic, oxidative, and high temperature stress conditions, on the susceptibility to essential oils (EOs) or antifungals and on the resistance to UV-C radiation was investigated. 93.6% of melanized A. pullulans and 92% of Z. obscura survived to 40 °C for 1 h compared to 77% and 76% of the non-melanized ones, while both yeasts tolerated a high concentration of NaCl (up to 30%) and H 2 O 2 (up to 400 mM) regardless of melanin production. Higher EOs antifungal efficacy was observed in non-melanized cells (growth inhibition zone >30 mm) compared to the melanized ones (25 mm). Similarly, the lowest Minimum Inhibitory Concentrations (MIC) and Minimum Fungicidal Concentration (MFC) values were evidenced for Fluconazole, Clotrimazole, Bifonazole and Amphotericin in the non-melanized fungi. Increasing UV-C intensity (up to 2004.5 J/m2) caused total death in the non-melanized strains compared to about 30% growth reduction in the melanized ones. The results of this investigation, the first focused on the biological role of melanin in "black-fungi" , are novel and encourage a better understanding of the biochemical features of melanin in the environmental adaptive ability of the new species Z. obscura. • We investigated the role of melanin in black yeasts A. pullulans and Z. obscura. • Melanized and non-melanized cells have different behaviour under environmental stress. • Melanized cells resulted more resistant to heat treatment, antimicrobials and UV-C [ABSTRACT FROM AUTHOR]

The ability to acquire three-dimensional (3D) information of cellular structures without the need for fluorescent tags or staining makes holotomographic imaging a powerful tool in cellular biology. It provides valuable insights by measuring the refractive index (RI), an optical parameter describing the phase delay of light that passes through the living cell. Here, we demonstrate holotomographic imaging on industrial relevant ascomycete fungi and study their development and morphogenesis. This includes conidial germination, subcellular dynamics, and cytoplasmic flow during hyphal growth in Aspergillus niger. In addition, growth and budding of Aureobasidium pullulans cells are captured using holotomographic microscopy. Coupled to fluorescence imaging, lipid droplets, vacuoles, the mitochondrial network, and nuclei are targeted and analyzed in the 3D RI reconstructed images. While lipid droplets and vacuoles can be assigned to a specific RI pattern, mitochondria and nuclei were not pronounced. We show, that the lower sensitivity of RI measurements derives from the fungal cell wall that acts as an additional barrier for the illumination light of the microscope. After cell wall digest of hyphae and protoplast formation of A. niger expressing GFP-tagged histone H2A, location of nuclei could be determined by non-invasive RI measurements. Furthermore, we used coupled fluorescence microscopy to observe migration of nuclei in unperturbed hyphal segments and duplication during growth on a single-cell level. Detailed micromorphological studies in Saccharomyces cerevisiae and Trichoderma reesei are challenging due to cell size restrictions. Overall, holotomography opens up new avenues for exploring dynamic cellular processes in real time and enables the visualization of fungi from a new perspective. • Exploring applications and limitations of holotomographic imaging for ascomycete fungi based on refractive index (RI) measurements. • Time-lapse imaging shows subcellular structures and cytoplasmatic streaming in A. niger and A. pullulans. • Cell wall degradation enables visualization of nuclei in the hyphae of A. niger. • Coupled with fluorescence imaging, nuclear migration and division are observed during polarized growth of A. niger. [ABSTRACT FROM AUTHOR]

Background: This work studied the occurrence, density, and diversity of yeast species associated with the leaves and fruits of Sabicea brasiliensis and Anacardium humile. Methods: Fragments of leaves and fruits were subjected to maceration and placed under agitation in peptone water. One hundred µL aliquots of each of the decimal dilutions were plated on YM chloramphenicol agar medium. The different morphotypes were identified through sequence analysis of the 26S rDNA. Results: A total of 83 yeast isolates were recovered from the organs sampled, 40 from fruits and 19 from the leaves of S. brasiliensis, while 14 were recovered from fruits and 10 from the leaves of A. humile. The isolates were identified as belonging to two phyla, six classes, eight orders, and eight families. Most isolates, at 71 (84.3%), belong to Ascomycota, with Saccharomycetes dominant in fruits and Dothideomycetes in leaves. The remaining 12 (15.7%) isolates belong to the phylum Basidiomycota. The most frequent genera were Aureobasidium, Candida, Rhodotorula, and Wickerhamiella. The Wickerhamiella isolates made possible the subsequent description of a new species within this genus. Conclusions: Our data reveal the yeast species that inhabit different Cerrado plant organs and indicate that these hosts can be considered an important habitat for yeast diversity, including new and/or previously unreported species in this biome.

Microbial biofouling on polymer surfaces can lead to their biodeterioration. This may result in deterioration of the surface, leading to cracking and fracturing. Fungal spores from Aspergillus niger 1957, Aspergillus niger 1988 and Aureobasidium pullulans were tested to determine their strength of attachment on three surfaces, p(γ-MPS-co-MMA), p(γ-MPS-co-LMA) and spin-coated poly(methyl methacrylate) (PMMAsc), using lateral force measurements. The results demonstrate that A. niger 1957 and A. niger 1988 spores were most easily removed from the p(γ-MPS-co-MMA) surface, which was the surface with the highest Ra value. The A. niger 1957 and A. pullulans spores were most difficult to remove from the PMMAsc surface, which was the hardest surface. A. niger 1988 spores were the most difficult to remove from p(γ-MPS-co-LMA), the most hydrophobic surface. The results with A. pullulans were difficult to elucidate since the spores bound to all three surfaces and were removed with similar rates of force. The lateral force results demonstrate that spore attachment to a surface is a multi-factorial process, and independent surface and microbial factors influence spore binding. Thus, each environmental scenario needs to be considered on an individual basis, since a solution to one biofouling issue will probably not translate across to other systems. This article is part of the theme issue 'Nanocracks in nature and industry'. [ABSTRACT FROM AUTHOR]

Three fungi not previously reported on desiccated human remains were identified on cadavers at the Forensic Investigation Research Station (FIRS) in Whitewater, Colorado. The location of the FIRS provides the unique opportunity to observe the stages of decomposition in a high desert environment. The two cadavers used in the study were in the late stages of decomposition (PMI of approximately 1520 and 1820 days) to the point of desiccation and had developed an extensive black crust on the skin that remained. Skin samples of the two cadavers were taken and plated onto potato dextrose agar to determine whether fungi were present on the desiccated tissues. Three different fungi consistently dominated cultures grown from numerous samples taken from each cadaver. Based on morphological observations, nuclear rDNA sequence data, and phylogenetic analyses, two fungi were identified to species (Aureobasidium melanogenum and Didymella glomerata) and one fungus was identified to the genus level (Alternaria). These results will contribute to the understanding of the role that fungi might play in late‐stage decomposition and the extended postmortem period. [ABSTRACT FROM AUTHOR]

Species of the genus Aureobasidium are ubiquitous, polyextremotolerant, "yeast-like" ascomycetes used for the industrial production of pullulan and other products and as biocontrol agents in agriculture. Their application potential and wide-spread occurrence make Aureobasidium spp. interesting study objects. The availability of a fast and efficient genome editing method is an obvious advantage for future basic and applied research on Aureobasidium. In this study, we describe the development of a CRISPR/Cas9-based genome editing method using ribonucleoproteins (RNPs) in A. pullulans and A. melanogenum. We demonstrate that this method can be used for single and multiplex genome editing using only RNPs by targeting URA3 (encoding for orotidine-5′-phosphate decarboxylase), ADE2 (encoding for phosphoribosylaminoimidazole carboxylase) and ARG4 (encoding for argininosuccinate lyase). We demonstrate the applicability of Trichoderma reesei pyr4 and Aspergillus fumigatus pyrG to complement the URA3 deficiency. Further, we show that using RNPs improves the homologous recombination rate and 20 bp long homologous flanks are sufficient. Therefore, the repair cassettes can be constructed by a single PCR, abolishing the need for laborious and time-consuming cloning, which is necessary for previously described methods for CRISPR-mediated genome editing in these fungi. The here presented method allows fast and efficient genome editing for gene deletions, modifications, and insertions in Auresobasidium with a minimized risk of off-target effects. • Cas9 ribonucleoproteins allow single- and multiplex genome editing of Aureobasidium pullulans and A. melanogenum. • Advantages are enhanced speed, easier handling, lower risk of off-target changes, multiplex genome editing. • Cas9 ribonucleoproteins can be used to boost the homologous integration rate even with flanks as short as 20 bp. • Improvements of the transformation and delivery method will allow genome editing in difficult strains. • Uridine, arginine and adenine auxotrophic strains were constructed. [ABSTRACT FROM AUTHOR]

Abstract Fructooligosaccharides (FOS) are a type of important prebiotics and produced by transfructosylating enzymes. In this study, sugarcane molasses was used as the substrate for production of transfructosylating enzymes by Aureobasidium pullulans FRR 5284. NaNO3 was a superior nitrogen source to yeast extract for production of transfructosylating enzymes by A. pullulans FRR 5284 and decreasing the ratio of NaNO3 to yeast extract nitrogen from 1:0 to 1:1 resulted in the reduction of the total transfructosylating activity from 109.8 U/mL to 82.5 U/mL. The addition of only 4.4 g/L NaNO3 into molasses-based medium containing 100 g/L mono- and di-saccharides resulted in total transfructosylating activity of 123.8 U/mL. Scale-up of the A. pullulans FRR 5284 transfructosylating enzyme production process from shake flasks to 1 L bioreactors improved the enzyme activity and productivity to 171.7 U/mL and 3.58 U/mL/h, 39% and 108% higher than those achieved from shake flasks, respectively. Sucrose (500 g/L) was used as a substrate for extracellular, intracellular, and total A. pullulans FRR 5284 transfructosylating enzymes, with a maximum yield of 61%. Intracellular, extracellular, and total A. pullulans FRR 5284 transfructosylating enzymes from different production systems resulted in different FOS profiles, indicating that FOS profiles can be controlled by adjusting intracellular and extracellular enzyme ratios and, hence prebiotic activity.

Aureobasidium pullulans is an extremotolerant, cosmopolitan yeast-like fungus that successfully colonises vastly different ecological niches. The species is widely used in biotechnology and successfully applied as a commercial biocontrol agent against postharvest diseases and fireblight. However, the exact mechanisms that are responsible for its antagonistic activity against diverse plant pathogens are not known at the molecular level. Thus, it is difficult to optimise and improve the biocontrol applications of this species. As a foundation for elucidating biocontrol mechanisms, we have de novo assembled a high-quality reference genome of a strongly antagonistic A. pullulans strain, performed dual RNA-seq experiments, and analysed proteins secreted during the interaction with the plant pathogen Fusarium oxysporum. Based on the genome annotation, potential biocontrol genes were predicted to encode secreted hydrolases or to be part of secondary metabolite clusters (e.g., NRPS-like, NRPS, T1PKS, terpene, and β-lactone clusters). Transcriptome and secretome analyses defined a subset of 79 A. pullulans genes (among the 10,925 annotated genes) that were transcriptionally upregulated or exclusively detected at the protein level during the competition with F. oxysporum. These potential biocontrol genes comprised predicted secreted hydrolases such as glycosylases, esterases, and proteases, as well as genes encoding enzymes, which are predicted to be involved in the synthesis of secondary metabolites. This study highlights the value of a sequential approach starting with genome mining and consecutive transcriptome and secretome analyses in order to identify a limited number of potential target genes for detailed, functional analyses.

Black yeasts have been isolated from acidic, low water activity, and thermally processed foods as well as from surfaces in food manufacturing plants. The genomic basis for their relative tolerance to food-relevant environmental stresses has not been well defined. In this study, we performed whole genome sequencing (WGS) on seven black yeast strains including Aureobasidium (n=5) and Exophiala (n=2) which were isolated from food or food production environments. These strains were previously characterized for their tolerance to heat, hyperosmotic pressure, high pressure processing, hypochlorite sanitizers, and ultraviolet light. Based on the WGS data, three of the strains previously identified as A. pullulans were reassigned as A. melanogenum. Both haploid and diploid A. melanogenum strains were identified in this collection. Single-locus phylogenies based on beta tubulin, RNA polymerase II, or translation elongation factor protein sequences were compared to the phylogeny produced through SNP analysis, revealing that duplication of the fungal genome in diploid strains complicates the use of single-locus phylogenetics. There was not a strong association between phylogeny and either environmental source or stress tolerance phenotype, nor were trends in the copy numbers of stress-related genes associated with extremotolerance within this collection. While there were obvious differences between the genera, the heterogenous distribution of stress tolerance phenotypes and genotypes suggests that food-relevant black yeasts may be ubiquitous rather than specialists associated with particular ecological niches. However, further evaluation of additional strains and the potential impact of gene sequence modification is necessary to confirm these findings.

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Nanoemulsions (NEs) have been made for improving the delivery and disperse of bioactive compounds. In this study, it was found that the best ingredients for the stable Massoia lactone-loaded and food-grade NEs making were 560.0 µL of Tween-80, 240.0 µL of Span-80 and 200.0 µL of Massoia lactone. Then, 9.0 mL of distilled water was titrated into the mixture under continuous magnetic stirring (750 rotations min−1) with about 2 drops per second for 20 min. Finally, the system was treated by ultrasonication using an ultrasonic generator (180 W and 22 KHz) for 5 min. All the prepared particles with a mean droplet diameter of 43 nm were spherical, had uniform size distribution and were equally distributed in the Massoia lactone-loaded NEs. The obtained Massoia lactone-loaded nanoemulsions (NEs) were very stable without changes of the mean droplet diameter and polydispersity indexes (PDI) for over two months under different conditions. As with free Massoia lactone, Massoia lactone loaded in the NEs had high anti-fungal activity against Metschnikowia bicuspidate LIAO, a pathogenic yeast causing milky disease in the Chinese mitten crab by damaging its cell membrane and causing cellular necrosis. Massoia lactone loaded in the NEs also had the DPPH radical scavenging activity and the hydroxyl radical scavenging activity. [ABSTRACT FROM AUTHOR]

Plants can ‘catch’ and mitigate airborne pollutants and are assisted by fungi inhabiting their leaves. The structure and function of the fungal communities inhabiting the phyllosphere of hornbeam trees growing in two chronically polluted areas, the oilfield of Bóbrka and the city center of Warsaw, were compared to the ones growing in one nature reserve, the Białowieża National Park. Fungi were isolated and characterized both phylogenetically and functionally for their potential role in air pollution mitigation. Both culture-dependent (e.g., enzyme assays and tolerance tests) and culture-independent methods (e.g., ITS and shotgun sequencings) were used. Furthermore, the degradation potential of the fungi was assessed by gas chromatography mass spectrometry (GC-MS). Shotgun sequencing showed that the phyllosphere fungal communities were dominated by fungi belonging to the phylum Ascomycota. Aureobasidium was the only genus detected at the three locations with a relative abundance ≥ 1.0%. Among the cultivated epiphytic fungi from Bóbrka, Fusarium sporotrichioides AT11, Phoma herbarum AT15, and Lophiostoma sp. AT37 showed in vitro aromatic hydrocarbon degradation potential with laccase activities of 1.24, 3.62, and 7.2 μU L-1, respectively, and peroxidase enzymes with activities of 3.46, 2.28, and 7.49 μU L-1, respectively. Furthermore, Fusarium sporotrichioides AT11 and Phoma herbarum AT15 tolerated exposure to airborne naphthalene and benzene. Lophiostoma sp. AT37 was the most tolerant to exposure to these pollutants, in line with being the best potential aromatic hydrocarbon degrader isolated in this study. [ABSTRACT FROM AUTHOR]

Abstract Background Yeasts, which are ubiquitous in agroecosystems, are known to degrade various xenobiotics. The aim of this study was to analyze the effect of fungicides on the abundance of natural yeast communities colonizing winter wheat leaves, to evaluate the sensitivity of yeast isolates to fungicides in vivo, and to select yeasts that degrade propiconazole. Results Fungicides applied during the growing season generally did not affect the counts of endophytic yeasts colonizing wheat leaves. Propiconazole and a commercial mixture of flusilazole and carbendazim decreased the counts of epiphytic yeasts, but the size of the yeast community was restored after 10 days. Epoxiconazole and a commercial mixture of fluoxastrobin and prothioconazole clearly stimulated epiphyte growth. The predominant species isolated from leaves were Aureobasidium pullulans and Rhodotorula glutinis. In the disk diffusion test, 14 out of 75 yeast isolates were not sensitive to any of the tested fungicides. After 48 h of incubation in an aqueous solution of propiconazole, the Rhodotorula glutinis Rg 55 isolate degraded the fungicide in 75%. Isolates Rh. glutinis Rg 92 and Rg 55 minimized the phytotoxic effects of propiconazole under greenhouse conditions. The first isolate contributed to an increase in the dry matter content of wheat seedlings, whereas the other reduced the severity of chlorosis. Conclusion Not sensitivity of many yeast colonizing wheat leaves on the fungicides and the potential of isolate Rhodotorula glutinis Rg 55 to degrade of propiconazole was established. Yeast may partially eliminate the ecologically negative effect of fungicides.

Poly(methyl methacrylate) (PMMA) surfaces, (commercial PMMA (PMMAc), spin coated PMMA (PMMAsc) and a 90% methylmethacrylate/10% 3-methacryloxypropyltrimethoxysilane random copolymer (P(MMA-co-gMPS)) were used to determine the effect of surface properties on conidia biofouling. The contact angles of the substrates demonstrated that the PMMAsc and the P(MMA-co-gMPS) polymer (62.8°) were more wettable than the PMMAc surface (71.0°). The PMMAsc had the greatest roughness value (32.0 nm) followed by the PMMAc (3.0 nm), then P(MMA-co-gMPS) (1 nm). Aspergillus niger 1957 conidia were spherical, smooth and hydrophobic (12.1%). Aspergillus niger 1988 conidia were spherical with spikes and hydrophobic (17.1%). Aureobasidium pullulans was elliptical with longitudinal ridges and hydrophilic (79.9%). Following attachment assays, cPMMA attached the greatest numbers of conidia. Following the adhesion and retention assays (washing step included in the protocol), A. niger 1957 and A. niger 1988 were least adhered to the P(MMA-co-gMPS) surface, whilst A. pulluans was least adhered to the PMMAsc surface. This work demonstrated that in the absence of a washing step, only the surface properties influenced the conidia attachment, whilst in the presence of a washing step, both the properties of the surfaces and the conidia affected conidia adhesion and retention. Hence, the methodology used (with or without a washing step) should reflect the environment in which the surface is to be applied.

Aureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all three proteins showed proton pump activity with maximal activity in green light. Among them, ApOps2 exhibited the most pronounced proton pump activity with current amplitudes occasionally extending 10 pA/pF at 0 mV. Proton pump activity was further supported in the presence of extracellular weak organic acids. Furthermore, we used site-directed mutagenesis to reshape protein functions and thereby implemented light-gated proton channels. We discuss the difference to other well-known proton pumps and the potential of these rhodopsins for optogenetic applications.

Volatile organic compounds (VOCs) produced by yeasts can positively affect crops, acting as antifungals or biostimulants. In this study, Aureobasidium pullulans and Metschnikowia pulcherrima were evaluated as potential antagonists of Trichoderma spp., common fungal pathogen in mushroom cultivation. To assess the biocontrol ability and biostimulant properties of the selected yeast species, in vitro co-culture and VOCs exposure assays were conducted. In both assays, VOCs produced by Aureobasidium spp. showed the stronger antifungal activity with a growth inhibition up to 30 %. This result was further confirmed by the higher volatilome alcohol content revealed by solid phase microextraction-gas chromatography mass spectrometry (SPME/GC-MS). Overall, Aureobasidium strains can be potentially used as biocontrol agent in Pleorotus ostreatus and Cyclocybe cylindracea mycelial growth, without affecting their development as demonstrated by VOCs exposure assay and Fourier-transform infrared spectroscopy (FT-IR). Conversely, M. pulcherrima was characterized by a lower or absent antifungal properties and by a volatilome composition rich in isobutyl acetate, an ester often recognized as plant growth promoter. As confirmed by FT-IR, Lentinula mycelia exposed to M. pulcherrima VOCs showed a higher content of proteins and lipids, suggesting an improvement of some biochemical properties. Our study emphasizes that VOCs produced by specific yeast strains are potentially powerful alternative to synthetic fungicide in the vegetative growth of mushroom-forming fungi and also able to modify their biochemical composition., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 British Mycological Society. All rights reserved.)

A high-yielding microbial polysaccharide-producing strain, named RM1603, was isolated from rhizosphere soil and identified by morphological and phylogenetic analysis. The extracellular polysaccharides (EPS) were identified by thin-layer chromatography and infrared spectroscopy. The fermentation conditions were optimized by single factor experiments in shake flasks and a 5-L fermentor. The results of morphological and phylogenetic tree analysis showed that RM1603 was a strain of Aureobasidium pullulans. Its microbial polysaccharide was identified as pullulan, and the EPS production capacity reached 33.07 ± 1.03 g L -1 in shake flasks. The fermentation conditions were optimized in a 5-L fermentor, and were found to encompass an initial pH of 6.5, aeration rate of 2 vvm, rotor speed of 600 rpm, and inoculum size of 2 %. Under these conditions, the pullulan yield of RM1603 reached 62.52 ± 0.24 g L -1 . Thus, this study contributes RM1603 as a new isolation with high-yielding pullulan and potential application value in biotechnology., (Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The CAZy auxiliary activity family 3 (AA3) comprises FAD-dependent enzymes belonging to the superfamily of glucose-methanol-choline (GMC) oxidoreductases. Glucose oxidase (GOx; EC 1.1.3.4) and glucose dehydrogenase (GDH; EC 1.1.5.9) are part of subfamily AA3_2 and catalyze the oxidation of β-D-glucose at its anomeric carbon to D-glucono-1,5-lactone. Recent phylogenetic analysis showed that AA3_2 glucose oxidoreductases can be grouped into four major clades, GOx I and GDH I–III, and in minor clades such as GOx II or distinct subclades. This wide sequence space of AA3_2 glucose oxidoreductases has, however, not been studied in detail, with mainly members of GOx I and GDH I studied biochemically or structurally. Here, we report the biochemical characterization of four fungal glucose oxidoreductases from distinct, hitherto unexplored clades or subclades. The enzyme from Aureobasidium subglaciale, belonging to the minor GOx II clade, showed a typical preference for oxygen and glucose, confirming the correct annotation of this clade. The other three enzymes exhibited strict dehydrogenase activity with different substrate specificities. GDH II from Trichoderma virens showed an almost six-fold higher catalytic efficiency for maltose compared to glucose. The preferred substrate for the two GDH III enzymes from Rhizoctonia solani and Ustilago maydis was gentiobiose, a β(1→6) disaccharide, as judged from the catalytic efficiency. Overall, the newly studied AA3_2 glucose oxidoreductases showed a much broader substrate spectrum than the archetypal GOx from Aspergillus niger, which belongs to clade GOx I. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the percentage of fungal contamination in wheat seeds in the stores of seed production companies in Nineveh Governorate, Iraq for the year 2019, the results of the study showed that all samples of wheat seeds were contaminated with fungi in different percentage and the sample of wheat seeds was Baraka/certified 2/ Mabain-Al-Nahrain Company is the most contaminated wheat Varieties with percentage 72%. More than 811 different fungi were isolated and identification from the studied wheat seed samples, Aureobasidium spp. appetence in high frequency 100% and was present in all the studied wheat seed samples, followed by Rhizopus stolonifer, which appeared at a frequency 76.92% and it was present in most samples, and the isolation results showed the appearance of Penicillium sp. With a frequency of 61%, then Alternaria alternata appeared at a frequency of 53.81%. The isolation results showed five species of Aspergillus. They are A. flavus, A. ochraceuos, A. paraciticus, A. fumigatus, A. terrus and A. niger with a frequency of 46.16%, 38.46%, 38.46%, 23.07%, 15.38% and 7.69%, respectively. Rhizoctonia solani isolated with a frequency 23.08%. Finally, Drechslera spp. Was detected. With a frequency 15.38% in the studied wheat seed samples. The results showed a difference in the sensitivity of the studied varieties to fungi Aureobasidium spp., R. solani, A. alternata and Dreshslera spp. The results refer to all Varieties treated with Aureobasidium spp. recorded good growth represented by a high percentage germination percentage and the variety was Abu Gharaib3/Certified/Iraqia Company the most affected. [ABSTRACT FROM AUTHOR]

The black yeast-like fungus Aureobasidium melanogenum is an opportunistic human pathogen frequently found indoors. Its traits, potentially linked to pathogenesis, have never been systematically studied. Here, we examine 49 A. melanogenum strains for growth at 37 °C, siderophore production, hemolytic activity, and assimilation of hydrocarbons and human neurotransmitters and report within-species variability. All but one strain grew at 37 °C. All strains produced siderophores and showed some hemolytic activity. The largest differences between strains were observed in the assimilation of hydrocarbons and human neurotransmitters. We show for the first time that fungi from the order Dothideales can assimilate aromatic hydrocarbons. To explain the background, we sequenced the genomes of all 49 strains and identified genes putatively involved in siderophore production and hemolysis. Genomic analysis revealed a fairly structured population of A. melanogenum, raising the possibility that some phylogenetic lineages have higher virulence potential than others. Population genomics indicated that the species is strictly clonal, although more than half of the genomes were diploid. The existence of relatively heterozygous diploids in an otherwise clonal species is described for only the second time in fungi. The genomic and phenotypic data from this study should help to resolve the non-trivial taxonomy of the genus Aureobasidium and reduce the medical hazards of exploiting the biotechnological potential of other, non-pathogenic species of this genus. [ABSTRACT FROM AUTHOR]

Aureobasidium melanogenum has been used as an animal feed additive for improving thehealth of pets, however, it has not yet been applied in honey bees. Here, a fungal strain CK-CsC isolated from bee bread pollen, was identified as A. melanogenum. Following characterizing CK-CsC fermentation broth, the 4-days fermentation broth (SYM medium or bee pollen) of the CK-CsC was used to feed newly emerged adult honey bees in cages under laboratory-controlled conditions for analysis of survival, gene expression of nutrient and antibacterial peptide, and gut microbiota of honey bees. It was found that the CK–CsC fermentation broth (SYM medium or bee pollen) is nontoxic to honey bees, and can regularly increase nutrient gene expression of honey bees. However, significant mortality of bees was observed after bees were fed on the supernatant liquid of the fermentation broth. Notably, this mortality can be lowered by the simultaneous consumption of bee pollen. The honey bees that were fed bee pollen exhibited more γ-Proteobacteria, Bacteriodetes, and Actinobacteria in their gut flora than did the honey bees fed only crude supernatant liquid extract. These findings indicate that A. melanogenum CK–CsC has high potential as a bee probiotic when it was fermented with bee pollen. [ABSTRACT FROM AUTHOR]

Fructooligosaccharides (FOS) are a type of important prebiotics and produced by transfructosylating enzymes. In this study, sugarcane molasses was used as the substrate for production of transfructosylating enzymes by Aureobasidium pullulans FRR 5284. NaNO3 was a superior nitrogen source to yeast extract for production of transfructosylating enzymes by A. pullulans FRR 5284 and decreasing the ratio of NaNO3 to yeast extract nitrogen from 1:0 to 1:1 resulted in the reduction of the total transfructosylating activity from 109.8 U/mL to 82.5 U/mL. The addition of only 4.4 g/L NaNO3 into molasses-based medium containing 100 g/L mono- and di-saccharides resulted in total transfructosylating activity of 123.8 U/mL. Scale-up of the A. pullulans FRR 5284 transfructosylating enzyme production process from shake flasks to 1 L bioreactors improved the enzyme activity and productivity to 171.7 U/mL and 3.58 U/mL/h, 39% and 108% higher than those achieved from shake flasks, respectively. Sucrose (500 g/L) was used as a substrate for extracellular, intracellular, and total A. pullulans FRR 5284 transfructosylating enzymes, with a maximum yield of 61%. Intracellular, extracellular, and total A. pullulans FRR 5284 transfructosylating enzymes from different production systems resulted in different FOS profiles, indicating that FOS profiles can be controlled by adjusting intracellular and extracellular enzyme ratios and, hence prebiotic activity. [ABSTRACT FROM AUTHOR]

Foliar fungi, especially endophytic fungi, constitute an important part of the microbiome of plants. Yet little is known about the composition of these communities. In this study, we isolated fungi from leaf tissues of the desert plant Welwitschia mirabilis to determine the culturable diversity of the foliar fungal community. The isolated fungal taxa, which grouped into 17 distinct lineages, were identified by sequencing elongation factor 1 alpha, beta-tubulin 1, beta-tubulin 2 and the internal transcribed spacer region. The culturable community was mainly composed of cosmopolitan fungal genera despite the unique taxonomic position of the plant and its geographic isolation. To test for endemism in two of the common fungal genera, Alternaria and Aureobasidium, we built haplotype networks using a global data set. Even this broad data set showed little evidence for specialisation within this unique host or its geographical location. The data suggest that the culturable members of communities of leaf-associated fungi in habitats with little plant coverage, such as the Namib Desert, are mainly established by long-distance aerially distributed fungal inocula and few of these taxa co-evolve with the host within the habitat. Significance: • The culturable members of fungal communities associated with an ecological and evolutionary isolated plant have not co-speciated with their hosts, but to a large extent are composed of globally distributed fungal species. • Harsh environmental conditions and the geographic isolation of host plants seem to favour ubiquitous fungal species over more specialist fungal species. Open data sets*: https://www.ncbi.nlm.nih.gov/genbank *see accession numbers in text

Foliar fungi, especially endophytic fungi, constitute an important part of the microbiome of plants. Yet little is known about the composition of these communities. In this study, we isolated fungi from leaf tissues of the desert plant Welwitschia mirabilis to determine the culturable diversity of the foliar fungal community. The isolated fungal taxa, which grouped into 17 distinct lineages, were identified by sequencing elongation factor 1 alpha, beta-tubulin 1, beta-tubulin 2 and the internal transcribed spacer region. The culturable community was mainly composed of cosmopolitan fungal genera despite the unique taxonomic position of the plant and its geographic isolation. To test for endemism in two of the common fungal genera, Alternaria and Aureobasidium, we built haplotype networks using a global data set. Even this broad data set showed little evidence for specialisation within this unique host or its geographical location. The data suggest that the culturable members of communities of leaf-associated fungi in habitats with little plant coverage, such as the Namib Desert, are mainly established by long-distance aerially distributed fungal inocula and few of these taxa co-evolve with the host within the habitat. [ABSTRACT FROM AUTHOR]

Nanoemulsions (NEs) have been made for improving the delivery and disperse of bioactive compounds. In this study, it was found that the best ingredients for the stable Massoia lactone-loaded and food-grade NEs making were 560.0 µL of Tween-80, 240.0 µL of Span-80 and 200.0 µL of Massoia lactone. Then, 9.0 mL of distilled water was titrated into the mixture under continuous magnetic stirring (750 rotations min−1) with about 2 drops per second for 20 min. Finally, the system was treated by ultrasonication using an ultrasonic generator (180 W and 22 KHz) for 5 min. All the prepared particles with a mean droplet diameter of 43 nm were spherical, had uniform size distribution and were equally distributed in the Massoia lactone-loaded NEs. The obtained Massoia lactone-loaded nanoemulsions (NEs) were very stable without changes of the mean droplet diameter and polydispersity indexes (PDI) for over two months under different conditions. As with free Massoia lactone, Massoia lactone loaded in the NEs had high anti-fungal activity against Metschnikowia bicuspidate LIAO, a pathogenic yeast causing milky disease in the Chinese mitten crab by damaging its cell membrane and causing cellular necrosis. Massoia lactone loaded in the NEs also had the DPPH radical scavenging activity and the hydroxyl radical scavenging activity.

Asparagine is one of the precursors of acrylamide and toxic fungal secondary metabolites, both carcinogenic compounds. In the present study, the optimal conditions to deplete asparagine by Aureobasidium pullulans (L1 and L8) from potato and wheat flour matrices were investigated. Through a colorimetric plate-assay with phenol red as indicator dye, both strains demonstrated to be able to produce L-asparaginase from 20 ∘C to 30 ∘C for L1 and only at 20 ∘C for L8 strain starting from 48 h of incubation. The ability of both yeasts to reduce asparagine content in potato and wheat flour was studied by in vitro spectrophotometric assay. Both strains showed a great ability to totally reduce asparagine at 20 ∘C after 15 min of incubation in potato homogenate, conversely in wheat flour, the highest reduction was detected after a longer exposition time (60 min). As known, L1 and L8 diamine asparaginase to aspartic acid. For this reason, both amino acids were tested to verify the antifungal effect against Rhizoctonia solani (Rs1) and Fusarium graminearum (F3) mycelial growth. Asparagine (120 mg/L) increased Rs1 and F3 mycelial growth respectively by 4.4% and 18.9%; conversely, aspartic acid significantly inhibited both respectively by 8.2% and 12.0%.

Two Castanea plant species, C. henryi and C. mollissima, are cultivated in China to produce chestnut crops. Leaf spot diseases commonly occur in Castanea plantations, however, little is known about the fungal species associated with chestnut leaf spots. In this study, leaf samples of C. henryi and C. mollissima were collected from Beijing, Guizhou, Hunan, Sichuan and Yunnan Provinces, and leaf-inhabiting fungi were identified based on morphology and phylogeny. As a result, twenty-six fungal species were confirmed, including one new family, one new genus, and five new species. The new taxa are Pyrisporaceae fam. nov., Pyrispora gen. nov., Aureobasidium castaneae sp. nov., Discosia castaneae sp. nov., Monochaetia castaneae sp. nov., Neopestalotiopsis sichuanensis sp. nov. and Pyrispora castaneae sp. nov. [ABSTRACT FROM AUTHOR]

Curcumin is a multitargeting nutraceutical with numerous health benefits, however, its efficacy is limited due to poor aqueous solubility and reduced bioavailability. While nano-formulation has emerged as an alternative to encounter such issues, it often involves use of toxic solvents. Microbial synthesis may be an innovative solution to address this lacuna. Present study, for the first time, reports exploitation of Aureobasidium pullulans RBF4A3 for production of nano-curcumin. For this purpose, Aureobasidium pullulans RBF4A3 was inoculated in YPD media along with curcumin (0.1 mg/mL) and incubated for 24 h, 48 h, and 72 h. Subsequently, residual sugar, biomass, EPS concentration, curcumin concentration, and curcumin nanoparticle size were measured. As a result, nano-curcumin with an average particle size of 31.63 nm and enhanced aqueous solubility was obtained after 72 h. Further, investigations suggested that pullulan, a reducing polysaccharide, played a significant role in curcumin nano-formulation. Pullulan-mediated nano-curcumin formulation, with an average particle size of 24 nm was achieved with conversion rate of around 59.19 %, suggesting improved aqueous solubility. Additionally, the anti-oxidant assay of the resulting nano-curcumin was around 53.7 % per μg. Moreover, kinetics and thermodynamic studies of pullulan-based nano-curcumin revealed that it followed first-order kinetics and was favored by elevated temperature for efficient bio-conversion. Also, various physico-chemical investigations like FT-IR, NMR, and XRD reveal that pullulan backbone remains intact while forming curcumin nanoparticle. This study may open up new avenues for synthesizing nano-polyphenols through a completely green and solvent free process with plausible diverse applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Aureobasidium pullulans is a ubiquitous fungus with a wide variety of morphologies and growth modes including "typical" single-budding yeast, and interestingly, larger multinucleate yeast than can make multiple buds in a single cell cycle. The study of A. pullulans promises to uncover novel cell biology, but currently tools are lacking to achieve this goal. Here, we describe initial components of a cell biology toolkit for A. pullulans , which is used to express and image fluorescent probes for nuclei as well as components of the cytoskeleton. These tools allowed live-cell imaging of the multinucleate and multibudding cycles, revealing highly synchronous mitoses in multinucleate yeast that occur in a semiopen manner with an intact but permeable nuclear envelope. These findings open the door to using this ubiquitous polyextremotolerant fungus as a model for evolutionary cell biology.

Different preventive strategies are needed to minimize the intake risks of mycotoxins, including zearalenone (ZEN). The aim of this study was to determine the ZEN adsorption ability of an autolyzed biomass preparation of polymorphic yeast Aureobasidium pullulans A.p.-3. The evaluation of the antitoxic properties of the preparation was also performed in relation to Saccharomyces cerevisiae yeast (ATCC 2366, ATCC 7090 and ATCC 9763) used as a model cell exposed to a toxic ZEN dose. The preparation at a dose of 5 mg/mL showed the adsorption of ZEN present in model systems at concentrations between 1 μg/mL to 100 μg/mL. The highest degree of adsorption was established for ZEN concentrations of 1 μg/mL and 5 μg/mL, becoming limited at higher doses of the toxin. Based on the Langmuir model of adsorption isotherms, the predicted maximum ZEN adsorption was approx. 190 µg/mL, regardless of pH. The growth of three strains of S. cerevisiae yeast cells in the medium with ZEN at concentrations within the range of 1.56 μg/mL-100 μg/mL was analyzed to determine the minimum inhibitory concentration. The growth of all tested strains was especially limited by high doses of ZEN, i.e., 50 and 100 μg/mL. The protective effect of the tested preparation was noted in relation to yeast cells exposed to toxic 100 μg/mL ZEN doses. The highest yeast cell growth (app. 36% percentage) was noted for a S. cerevisiae ATCC 9763 strain compared to the medium with ZEN but without preparation. More detailed tests determining the antitoxic mechanisms of the A. pullulans preparation are planned in the future, including cell culture bioassays and animal digestive tract models.

Co-fermentation of multiple substrates has emerged as the most effective method to improve the yield of bioproducts. Herein, sustainable rubberwood enzymatic hydrolysates (RWH) were co-fermented by Aureobasidium pullulans to produce poly(β-L-malic acid) (PMA), and RWH + glucose/xylose was also investigated as co-substrates. Owing to low inhibitor concentration and abundant natural nitrogen source content of RWH, a high PMA yield of 0.45 g/g and a productivity of 0.32 g/L/h were obtained by RWH substrate fermentation. After optimization, PMA yields following the fermentation of RWH + glucose and RWH + xylose reached 59.92 g/L and 53.71 g/L, respectively, which were 52 % and 36 % higher than that after the fermentation of RWH. RWH + glucose more significantly affected the correlation between PMA yield and substrate concentration than RWH + xylose. The results demonstrated that the co-fermentation of RWH co-substrate is a promising method for the synthesis of bioproducts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Natural melanin is a biopolymer with wide application prospects in medicine, food, cosmetics, environmental protection, agriculture, and so on. Microbial fermentation is an important and effective way to produce melanin. In this study, Aureobasidium melanogenum, known as black yeast with cellular pleomorphism, was used for the production of melanin. Based on the characteristic of A. melanogenum secreting melanin under oligotrophic stress, a simple medium containing only glucose, MgSO 4 ·7H 2 O, and KCl was constructed for the production of melanin. The melanin titer of 6.64 ± 0.22 g/L was obtained after 20 days of fermentation without pH control. The cell morphological changes of A. melanogenum during the production of melanin were recorded, and the results showed that chlamydospore might be the most favorable cell morphology for melanin synthesis. Then, different fermentation strategies with cell morphology analysis were developed to further improve the production of melanin in a 5-L fermenter. Results showed that the maximum titer of melanin reached 18.50 g/L by using the fermentation strategy integrating pH control, ammonium salt addition, and H 2 O 2 stimulation, which increased by 178.6% than that of the strategy without pH control. Furthermore, the melanin obtained from the fermentation broth was characterized as eumelanin containing an indole structure. This study provided a potentially feasible fermentation strategy for the industrial production of melanin., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Fusarium head blight (FHB) is the most dangerous disease in all regions where bread wheat and durum wheat are grown, mostly due to grain contamination with trichothecenes produced by fungi of the genus Fusarium. Biological control of FHB with the use of yeast suspensions could pose a viable alternative to fungicides. The aim of this study was to perform in vitro selection of yeast isolates capable of inhibiting the development of FHB and reducing the concentrations of type A and type B trichothecenes in grain. In a field experiment, the inoculation of durum wheat spikes with Fusarium culmorum decreased grain yield by 9.13% and led to the highest accumulation of deoxynivalenol (DON) in grain at 11.704 mg kg-1. Isolates Candida sake Cs58, Rhodotorula glutinis Rg64 and Debaryomyces hansenii Dh53 most effectively inhibited the decline in durum wheat yield. Biological treatments induced a 10-fold decrease in the DON content of inoculated grain. Spike inoculation with F. culmorum decreased bread wheat yield by 19.88%. The isolate Aureobasidium pullulans Ap24 was most effective in decreasing FHB symptoms, whereas the isolate D. hansenii Dh53 reduced DON concentration by 11.33-fold in inoculated grain and decreased nivalenol concentration by 18.12-fold in the grain of noninoculated bread wheat. [ABSTRACT FROM AUTHOR]

The identification of yeasts or yeasts-like fungi and verify their diversity are principal aspect for bioindustry and ecosystem sustainability. Taxonomic approach provides identification tool to ensure the taxonomic position of yeasts and yeasts-like fungi which definitely set to utilization concerns. The aim of this study is to understanding the taxonomic position of yeasts and yeasts-like fungi from the distinctive of its sequences relationship. Yeasts and yeasts-like fungi strains were isolated through various culture dependent methods from natural resources samples of Karimun Besar Island, Province of Riau Islands, Indonesia. The identification process was performed through amplifying the accurate DNA-based in D1/D2 region of large subunit (26S) ribosomal DNA. As the result, a total of 85 isolates of yeasts and yeasts-like fungi were obtained with 16 closest related taxa through phylogenetic tree construction. Ascomycetous was the predominating group representing 91% of the total isolates sequences followed by Basidiomycetous (8%) and Zygomycetous (1%). The black yeasts (yeasts-like) known as Aureobasidium melanogenum was predominant species with represent to 54% of total isolates and present in particular habitat. Taxonomically, there are six isolates are represent to be novel taxa candidates which pretend to enhance genetic resources of yeasts and yeasts-like fungi especially from Indonesia. In addition, this information provides specific technique to reach specific yeasts or yeasts-like fungi species in nature by managing the sample collection and culture methods.

Aureobasidium pullulans is the most abundant and ubiquitous species within the genus and is also considered a core component of the grape juice microflora. So far, a small number of other Aureobasidium species have been reported, that in contrast to A. pullulans , appear far more constrained to specific habitats. It is unknown whether grape juice is a reservoir of novel Aureobasidium species, overlooked in the course of conventional morphological and meta-barcoding analyses. In this study, eight isolates from grape juice taxonomically classified as Aureobasidium through ITS sequencing were subjected to whole-genome phylogenetic, synteny and nucleotide identity analyses, which revealed three isolates to likely represent newly discovered Aureobasidium species. Analyses of ITS and metagenomic sequencing datasets show that these species can be present in grape juice samples from different locations and vintages. Functional annotation revealed the Aureobasidium isolates possess the genetic potential to support growth on the surface of plants and grapes. However, the loss of several genes associated with tolerance to diverse environmental stresses suggest a more constrained ecological range than A. pullulans. [ABSTRACT FROM AUTHOR]

Data on the number and diversity of psychrotolerant yeast isolated from high-altitude and surface samples of atmospheric aerosol from southwestern Siberia are derived from a microbiological study. According to certain phenotypic and genomic characteristics, the yeast isolates are referred to the Saccharomyces, Candida, Sporidiobolus, Aureobasidium, Sporobolomyces, Cryptococcus, Rhodotorula, and some other genera. Both saprophytic and pathogenic microorganisms are detected. Aureobasidium yeast strains are revealed, which actively produce exopolysaccharides and melanin and are promising for biotechnological developments. [ABSTRACT FROM AUTHOR]

Background: Yeasts, which are ubiquitous in agroecosystems, are known to degrade various xenobiotics. The aim of this study was to analyze the effect of fungicides on the abundance of natural yeast communities colonizing winter wheat leaves, to evaluate the sensitivity of yeast isolates to fungicides in vivo, and to select yeasts that degrade propiconazole. Results: Fungicides applied during the growing season generally did not affect the counts of endophytic yeasts colonizing wheat leaves. Propiconazole and a commercial mixture of flusilazole and carbendazim decreased the counts of epiphytic yeasts, but the size of the yeast community was restored after 10 days. Epoxiconazole and a commercial mixture of fluoxastrobin and prothioconazole clearly stimulated epiphyte growth. The predominant species isolated from leaves were Aureobasidium pullulans and Rhodotorula glutinis. In the disk diffusion test, 14 out of 75 yeast isolates were not sensitive to any of the tested fungicides. After 48 h of incubation in an aqueous solution of propiconazole, the Rhodotorula glutinis Rg 55 isolate degraded the fungicide in 75%. Isolates Rh. glutinis Rg 92 and Rg 55 minimized the phytotoxic effects of propiconazole under greenhouse conditions. The first isolate contributed to an increase in the dry matter content of wheat seedlings, whereas the other reduced the severity of chlorosis. Conclusion: Not sensitivity of many yeast colonizing wheat leaves on the fungicides and the potential of isolate Rhodotorula glutinis Rg 55 to degrade of propiconazole was established. Yeast may partially eliminate the ecologically negative effect of fungicides. [ABSTRACT FROM AUTHOR]

It has been well known that poly(β- l -malic acid)(PMA) has many potential applications. However, it is still completely unknown how PMA is biosynthesized in Aureobasidium spp. In this study, it was found that malic acid from TCA cycle was the main source for PMA biosynthesis. Especially, the novel PMA synthetase encoded by the PMAs gene, a non-ribosomal peptide synthetase (NRPS) containing A like domain, T domain and C like domain was the key enzyme for polymerization of malate into PMA. Therefore, abolishment of the PMAs gene encoding the novel PMA synthetase rendered the mutant Δ PMAs - 3 totally to lose the ability to synthesize any PMA and complementation of the PMAs gene partially restored PMA biosynthesis, but the mutant could grow normally on the YPD plate and in the PMA medium with CaCO 3. The transcriptional activator Crz1 in the Ca2+-signal pathway controlled expression of the PMAs gene and PMA biosynthesis. The complete elucidation of the PMA biosynthesis pathway and its regulation was of significant for a deeper understanding of detailed yeast-like fungal PMA synthesis, metabolic engineering and molecular editing for modifying PMA biosynthesis and its physicochemical properties. • The PMA synthetase was the key enzyme for polymerization of malate into PMA. • The multi-domain enzyme has A like domain, T domain and C like domain. • The expression of the PAMs gene was regulated by the Crz1 in the Ca2+ signaling pathway. [ABSTRACT FROM AUTHOR]

Poly(methyl methacrylate) (PMMA) surfaces, (commercial PMMA (PMMAc), spin coated PMMA (PMMAsc) and a 90% methylmethacrylate/10% 3-methacryloxypropyltrimethoxysilane random copolymer (P(MMA-co-gMPS)) were used to determine the effect of surface properties on conidia biofouling. The contact angles of the substrates demonstrated that the PMMAsc and the P(MMA-co-gMPS) polymer (62.8°) were more wettable than the PMMAc surface (71.0°). The PMMAsc had the greatest roughness value (32.0 nm) followed by the PMMAc (3.0 nm), then P(MMA-co-gMPS) (1 nm). Aspergillus niger 1957 conidia were spherical, smooth and hydrophobic (12.1%). Aspergillus niger 1988 conidia were spherical with spikes and hydrophobic (17.1%). Aureobasidium pullulans was elliptical with longitudinal ridges and hydrophilic (79.9%). Following attachment assays, cPMMA attached the greatest numbers of conidia. Following the adhesion and retention assays (washing step included in the protocol), A. niger 1957 and A. niger 1988 were least adhered to the P(MMA-co-gMPS) surface, whilst A. pulluans was least adhered to the PMMAsc surface. This work demonstrated that in the absence of a washing step, only the surface properties influenced the conidia attachment, whilst in the presence of a washing step, both the properties of the surfaces and the conidia affected conidia adhesion and retention. Hence, the methodology used (with or without a washing step) should reflect the environment in which the surface is to be applied. [ABSTRACT FROM AUTHOR]

The biodiversity and evolution of fungal communities were monitored over a period of 3 vintages in a new winery. Samples were collected before grape receipt and 3 months after fermentation from 3 different wine related environments (WRE): floor, walls and equipment and analyzed using Illumina Mi-Seq. Genera of mold and filamentous fungi (294), non-enological (10) and wine-associated yeasts (25) were detected on all WREs before the arrival of the first harvest. Among them, genera like Alternaria and Aureobasidium persisted during two vintages. Therefore, these genera are not specific to winery environment and appear to be adapted to natural or anthropic environments due to their ubiquitous character. Some genera like Candida were also detected before the first harvest but only on one WREs, whereas, on the other WREs they were found after the harvest. The ubiquitous character and phenotypic traits of these fungal genera can explain their dynamics. After the first harvest and during 3 vintages the initial consortium was enriched by oenological genera like Starmerella introduced either by harvest or by potential transfers between the different WREs. However, these establishing genera, including Saccharomyces, do not appear to persist due to their low adaptation to the stressful conditions of winery environment. [ABSTRACT FROM AUTHOR]

Xylose, the main component of xylan, is the second most abundant sugar in nature after glucose. Consequently, xylose represents an attractive feedstock for the production of value-added compounds such as biosurfactants (BSs), which are produced by various bacteria and yeasts. In this study, we screened and isolated yeast strains that synthesize BSs using xylose as the sole carbon source. We applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to screen for BS-producing yeasts and isolated eight strains as the liamocin producers. Two of the eight strains, AS37 and SK25, were identified as Aureobasidium melanogenum , which is known as black yeasts, by based on 26S ribosomal RNA gene sequences. Both strains produced a wide variety of liamocin structures from not only xylose but also glucose and sucrose. According to the MALDI-TOF MS analysis, signals corresponding to sodium ion adducts of di-, tri-, tetra-, penta- and hexa-acylated C6-liamocins and di-, tri- and tetra-acylated C5-liamocins were detected. In addition, their mono-acetylated form was also detected. The dominant sugar component of liamocins produced by strains AS37 and SK25 is mannitol as estimated by HPLC analysis. This is the first report to describe the screening of liamocins-producing yeasts using xylose as the sole carbon source. [ABSTRACT FROM AUTHOR]