-
Genomic deletions in Aureobasidium pullulans by an AMA1 plasmid for gRNA and CRISPR/Cas9 expression.Fungal Biology and Biotechnology Jun 2024Aureobasidium pullulans is a generalist polyextremotolerant black yeast fungus. It tolerates temperatures below 0 °C or salt concentrations up to 18%, among other...
BACKGROUND
Aureobasidium pullulans is a generalist polyextremotolerant black yeast fungus. It tolerates temperatures below 0 °C or salt concentrations up to 18%, among other stresses. A. pullulans genome sequencing revealed a high potential for producing bioactive metabolites. Only few molecular tools exist to edit the genome of A. pullulans, hence it is important to make full use of its potential. Two CRISPR/Cas9 methods have been proposed for the protoplast-based transformation of A. pullulans. These methods require the integration of a marker gene into the locus of the gene to be deleted, when the deletion of this gene does not yield a selectable phenotype. We present the adaptation of a plasmid-based CRISPR/Cas9 system developed in Aspergillus niger for A. pullulans to create deletion strains.
RESULTS
The A. niger CRISPR/Cas9 plasmid led to efficient genomic deletions in A. pullulans. In this study, strains with deletions ranging from 30 to 862 bp were obtained by using an AMA1 plasmid-based genome editing strategy.
CONCLUSION
The CRISPR/Cas9 transformation system presented in this study provides new opportunities for strain engineering of A. pullulans. This system allows expression of Cas9 and antibiotic resistance while being easy to adapt. This strategy could open the path to intensive genomic engineering in A. pullulans.
PubMed: 38824542
DOI: 10.1186/s40694-024-00175-4 -
Ecotoxicology and Environmental Safety Jul 2024Abrus cantoniensis Hance (ACH) is an ancient Chinese medicine herb known for its therapeutic effects. This study investigated the potential protective effect of ACH...
Abrus cantoniensis Hance (ACH) is an ancient Chinese medicine herb known for its therapeutic effects. This study investigated the potential protective effect of ACH against carbon tetrachloride (CCl)-induced liver damage in mice. Fifty (n= 50) ICR mice were grouped into five groups. CCl was intraperitoneally injected into different mice groups: AM (CCl induced), AD (ACH-treated with 25 mg/kg), AZ (ACH-treated with 50 mg/kg), and AG (ACH-treated with100mg/kg) after every three days for a total of 31 days. The control group was denoted as AC. Additionally, groups AD, AZ, and AG received daily doses of ACH via gavage throughout the study period. According to our findings, ACH administration prominently mitigated liver pathological lesions and the increased liver index induced by CCl in mice (p < 0.05). Treatment with ACH resulted in a dose-dependent recovery of GSH-px, SOD, and CAT activities (p < 0.001). Moreover, the levels of TNF-α, MDA, and ALT showed significanlty decreasing trends with various doses of ACH (p < 0.001). Furthermore, 16 S rRNA gene sequencing demonstrated that ACH increased the abundance of beneficial genera of Comoclathris, Aureobasidium, and Kazachstania while decreased the presence of pathogenic genera such as Sporobolomyces and Filobasidium. Additionally, ACH treatment ameliorated the changes in liver metabolism due to CCl and enhanced the beneficial liver metabolites. In conclusion, ACH shows potential in protecting the liver against oxidative stress and inflammation caused by CCl exposure, possibly through its effects on gut microbiota and liver metabolism. Therefore, the use of ACH may offer an effective approach for alleviating CCl-induced liver injury.
Topics: Animals; Mice, Inbred ICR; Gastrointestinal Microbiome; Liver; Mice; Chemical and Drug Induced Liver Injury; Male; Carbon Tetrachloride; Abrus; Protective Agents; Drugs, Chinese Herbal; Oxidative Stress
PubMed: 38820876
DOI: 10.1016/j.ecoenv.2024.116495 -
Medical Mycology Case Reports Jun 2024Spontaneous mycosis caused by is documented in roach in a cyprinid-prevalent water reservoir in Czechia. Gross pathological lesions included pale gills and...
Spontaneous mycosis caused by is documented in roach in a cyprinid-prevalent water reservoir in Czechia. Gross pathological lesions included pale gills and splenomegaly, as revealed during necropsy examination. Histological examination showed extensive foci with fungal elements in the kidney. The isolated fungus was identified through phenotypic and molecular characterization, including phylogeny. This report represents the first case of infection in fish and cold-blooded vertebrates, to the best of our knowledge.
PubMed: 38799503
DOI: 10.1016/j.mmcr.2024.100652 -
Foods (Basel, Switzerland) May 2024The composition and continuous succession of natural microbial communities during grape growth play important roles in grape health and flavor quality as well as in...
The composition and continuous succession of natural microbial communities during grape growth play important roles in grape health and flavor quality as well as in characterizing the regional wine terroir. This study explored the diversity and dynamics of fruit epidermal microbes at each growth and developmental stage of Ecolly grapes under an extremely simplified eco-cultivation model, analyzed microbial interactions and associations of weather parameters to specific communities, and emphasized metabolic functional characteristics of microecology. The results indicated that the natural microbial community changed significantly during the grape growth phase. The dominant fungal genera mainly included , , , , , , and , and the dominant bacterial genera mainly contained , , , , , , and . In summary, filamentous fungi gradually shifted to basidiomycetous yeasts along with fruit ripening, with a decline in the number of Gram-negative bacteria and a relative increase in Gram-positive bacteria. The community assembly process reflects the fact that microbial ecology may be influenced by a variety of factors, but the fungal community was more stable, and the bacterial community fluctuated more from year to year, which may reflect their response to weather conditions over the years. Overall, our study helps to comprehensively profile the ecological characteristics of the grape microbial system, highlights the natural ecological viticulture concept, and promotes the sustainable development of the grape and wine industry.
PubMed: 38790880
DOI: 10.3390/foods13101580 -
Frontiers in Microbiology 2024Grape-associated microbial community is influenced by a combination of viticultural, climatic, pedological and anthropological factors, collectively known as ....
Grape-associated microbial community is influenced by a combination of viticultural, climatic, pedological and anthropological factors, collectively known as . Therefore, grapes of the same cultivar grown in different areas can be appreciated for their distinctive biogeographic characteristics. In our previous study, we showed that the phenotypic response of Aglianico and Cabernet grapevines from Molise and Sicily regions is significantly influenced by the prevailing pedoclimatic conditions, particularly soil physical properties. However, the scale at which microbial communities differ could be important in clarifying the concept of , including whether it is linked to the grape variety present in a particular vineyard. To explore this further, in the research presented here, a comparative study on the fungal communities inhabiting the berry surfaces of Cabernet and Aglianico cultivars was conducted on different vineyards located in Southern Italy (Molise, Sicily and Campania regions, the first two of which had been involved in our previous study) by using high-throughput sequencing (HTS) and multivariate data analysis. The descriptive approach through relative abundance analysis showed the most abundant (Ascomycota, Basidiomycota, and Chytridiomycota), families (, , , , , , , , and ) and genera (, , , and ) detected on grape berries. The multivariate data analysis performed by using different packages (phyloseq, Vegan, mixOmics, microbiomeMarker and ggplot2) highlighted that the variable "vineyard location" significantly affect the fungal community, while the variable "grape variety" has no significant effect. Thus, some taxa are found to be part of specific vineyard ecosystems rather than specific grape varieties, giving additional information on the microbial contribution to wine quality, thanks to the presence of fermentative yeasts or, conversely, to the involvement in negative or detrimental roles, due to the presence of grape-deriving fungi implied in the spoilage of wine or in grapevine pathogenesis. In this connection, the main functions of core taxa fungi, whose role in the vineyard environment is still poorly understood, are also described.
PubMed: 38725687
DOI: 10.3389/fmicb.2024.1399968 -
Bioresources and Bioprocessing Aug 2023Aureobasidium pullulans (A. pullulans) has a wide range of applications. Ultraviolet (UV) rays from the sun can cause skin photoaging. In order to explore the protective...
BACKGROUND
Aureobasidium pullulans (A. pullulans) has a wide range of applications. Ultraviolet (UV) rays from the sun can cause skin photoaging. In order to explore the protective effect and application potential of A. pullulans lysate on UV-damaged human skin fibroblasts (HSF) and HaCaT Cells, this study investigates the anti-aging and anti-inflammatory effects of A. pullulans lysate as well as the mechanism of anti-oxidative stress at the cellular and molecular levels through cytotoxicity experiments, enzyme-linked immunosorbent assays (ELISA), and real-time quantitative PCR (RT-qPCR).
RESULTS
The experimental results have shown that the A. pullulans lysate can effectively reduce the loss of extracellular matrix components (EMC), such as collagen and hyaluronic acid (HA). It is also capable of scavenging excess reactive oxygen species (ROS) from the body, thereby increasing the activity of catalase, decreasing the overexpression of intracellular matrix metalloproteinases (MMPs), enhancing the gene expression of metalloproteinase inhibitors (TIMPs), and decreasing the level of inflammatory factors, reducing UV-induced apoptosis of HaCaT cells. Meanwhile, oxidative stress homeostasis is also regulated through the Nrf2/Keap1 and MAPK signaling pathways.
CONCLUSIONS
This study shows that the A. pullulans lysate has the potential to resist photoaging.
PubMed: 38647892
DOI: 10.1186/s40643-023-00678-9 -
Microbiology Resource Announcements Jun 2024
PubMed: 38647292
DOI: 10.1128/mra.00323-24 -
International Journal of Food... Jun 2024This study delved into the evolution of fungal population during the fermentation of Spanish-style green table olives (Manzanilla cultivar), determining the influence of...
This study delved into the evolution of fungal population during the fermentation of Spanish-style green table olives (Manzanilla cultivar), determining the influence of different factors such as fermentation matrix (brine or fruit) or the use of a lactic acid bacteria inoculum, on its distribution. The samples (n = 24) were directly obtained from industrial fermentation vessels with approximately 10.000 kg of fruits and 6.000 L of brines. Our findings showcased a synchronized uptick in lactic acid bacteria counts alongside fungi proliferation. Metataxonomic analysis of the Internal Transcribed Spacer (ITS) region unearthed noteworthy disparities across different fermentation time points (0, 24, and 83 days). Statistical analysis pinpointed two Amplicon Sequence Variants (ASV), Candida and Aureobasidium, as accountable for the observed variances among the different fermentation time samples. Notably, Candida exhibited a marked increase during 83 days of fermentation, opposite to Aureobasidium, which demonstrated a decline. Fungal biodiversity was slightly higher in brines than in fruits, whilst no effect of inoculation was noticed. At the onset of fermentation, prominently detected genera were also Mycosphaerella (19.82 %) and Apohysomyces (16.31 %), hitherto unreported in the context of table olive processing. However, their prevalence dwindled to nearly negligible levels from 24th day fermentation onwards (<2 %). On the contrary, they were replaced by the fermentative yeasts Saccharomyces and Isstachenkia. Results obtained in this work will be useful for designing new strategies for better control of table olive fermentations.
Topics: Olea; Fermentation; Biodiversity; Lactobacillales; Food Microbiology; Fungi; Spain; Fruit; Salts
PubMed: 38621325
DOI: 10.1016/j.ijfoodmicro.2024.110689 -
International Journal of Food... Jun 2024Taggiasca table olives are typical of Liguria, a Northwestern Italian region, produced with a spontaneous fermentation carried out by placing the raw drupes directly...
Monitoring the yeasts ecology and volatiles profile throughout the spontaneous fermentation of Taggiasca cv. table olives through culture-dependent and independent methods.
Taggiasca table olives are typical of Liguria, a Northwestern Italian region, produced with a spontaneous fermentation carried out by placing the raw drupes directly into brine with a salt concentration of 8-12 % w/v. Such concentrations limit the development of unwanted microbes and favor the growth of yeasts. This process usually lasts up to 8 months. Yeasts are found throughout the entire fermentation process and they are mainly involved in the production of volatile organic compounds, which strongly impact the quality of the final product. The aim of this study was to evaluate the dynamics of autochthonous yeasts in brines and olives in a spontaneous process with no lye pre-treatment or addition of acids in the fermenting brine with 10 % NaCl (w/v) in two batches during 2021 harvest. Three hundred seventy-three yeast colonies were isolated, characterized by rep-PCR and identified by the D1/D2 region of the 26S rRNA gene sequencing. Mycobiota was also studied by 26S rRNA gene metataxonomics, while metabolome was assessed through GC-MS analysis. Traditional culture-dependent methods showed the dominance of Candida diddensiae, Wickerhamomyces anomalus, Pichia membranifaciens and Aureobasidium pullulans, with differences in species distribution between batches, sampling time and type of sample (olives/brines). Amplicon-based sequencing confirmed the dominance of W. anomalus in batch 1 throughout the entire fermentation, while Cyteromyces nyonsensis and Aureobasidium spp. were most abundant in the fermentation in batch 2. Volatilome results were analyzed and correlated to the mycobiota data, confirming differences between fermentation stages. Given the high appreciation for this traditional food, this study helps elucidate the mycobiota associated to Taggiasca cv. table olives and its relationship with the quality of the final product.
Topics: Fermentation; Olea; Volatile Organic Compounds; Yeasts; Food Microbiology; Italy; Salts
PubMed: 38615425
DOI: 10.1016/j.ijfoodmicro.2024.110688 -
Persoonia Jun 2023Novel species of fungi described in this study include those from various countries as follows: , on whitefly, on bark of , from soil under , on leaf spot of , and...
Novel species of fungi described in this study include those from various countries as follows: , on whitefly, on bark of , from soil under , on leaf spot of , and on leaf spot of . , on fully submersed siliceous schist in high-mountain streams, and on the lower part and apothecial discs of on a twig. , on decaying wood, from moist soil with leaf litter, on a trunk of a living unknown hardwood tree species, and on dead twigs of unidentified plant. , on sandy soil in a plantation of . , on dead bark of , and on dead bark of . , on fruit lesion of . , on corticioid , on sp. , on calcareous soils in dry forests and park habitats. , on sandy soil under , and on leaves of . , on decaying bark of logs, on unidentified woody substrate, from soil, on the trunk of , and on elephant dung. , on infected leaves of . , (incl. gen. nov.) from . , on acidic soil. , on dead leaf of , and on dead leaves of . , on dead culms of , (incl. gen. nov.) on culms of , (incl. gen. nov.) on branch of , on dead standing culms of , on culms of , and on dead bamboo sticks. , half-buried and moss-covered pieces of rotting wood in grass-grown path. , on soil. , (incl. gen. nov.) from resin of ssp. , from sooty mould community on , and from a gallery of on . , on mossy areas of laurel forest areas planted with , and from a biofilm covering a biodeteriorated limestone wall. , from hypersaline sea water, and from water sample collected from hypersaline lagoon. , on culm of , on , (incl. gen. nov.) on culms of , on nest of cases of bag worm moths () on , on leaves of , on stems of , from the roots of × , and (incl. gen. nov.) on leaf of . , on decaying leaves of sp. from pond. , on the bark of fallen trees of , from surface-sterilised, asymptomatic roots of , on soil in mixed forest, on calcareous soil in mixed forest, on acidic soils, from roots of × , on leaves of sp., and from soil. , on calcareous soil. , on pupa, buried in soil, on larva, buried in soil, and on pupa, buried in soil. , on dead leaf of . , on clay loamy soils. , (incl. gen. nov.) on leaves of . , on recently dead stem of , (incl. gen. nov.) from water, and from swab of coil surface. Morphological and culture characteristics for these new taxa are supported by DNA barcodes. : Crous PW, Osieck ER, Shivas RG, et al. 2023. Fungal Planet description sheets: 1478-1549. Persoonia 50: 158- 310. https://doi.org/10.3767/persoonia.2023.50.05.
PubMed: 38567263
DOI: 10.3767/persoonia.2023.50.05