Fungus Penicillium Research Articles

Novel Transcription Factor CXRD Regulates Cellulase and Xylanase Biosynthesis in Penicillium oxalicum under Solid-State Fermentation.

Penicillium oxalicum produces an integrated, extracellular cellulase and xylanase system, strictly regulated by several transcription factors. However, the understanding of the regulatory mechanism of cellulase and xylanase biosynthesis in P. oxalicum is limited, particularly under solid-state fermentation (SSF) conditions. In our study, deletion of a novel gene, cxrD (cellulolytic and xylanolytic regulator D), resulted in 49.3 to 2,230% enhanced production of cellulase and xylanase, except for 75.0% less xylanase at 2 days, compared with the P. oxalicum parental strain, when cultured on solid medium containing wheat bran plus rice straw for 2 to 4 days after transfer from glucose. In addition, the deletion of cxrD delayed conidiospore formation, leading to 45.1 to 81.8% reduced asexual spore production and altered mycelial accumulation to various extents. Comparative transcriptomics and real-time quantitative reverse transcription-PCR found that CXRD dynamically regulated the expression of major cellulase and xylanase genes and conidiation-regulatory gene brlA under SSF. In vitro electrophoretic mobility shift assays demonstrated that CXRD bound to the promoter regions of these genes. The core DNA sequence 5'-CYGTSW-3' was identified to be specifically bound by CXRD. These findings will contribute to understanding the molecular mechanism of negative regulation of fungal cellulase and xylanase biosynthesis under SSF. IMPORTANCE Application of plant cell wall-degrading enzymes (CWDEs) as catalysts in biorefining of lignocellulosic biomass into bioproducts and biofuels reduces both chemical waste production and carbon footprint. The filamentous fungus Penicillium oxalicum can secrete integrated CWDEs, with potential for industrial application. Solid-state fermentation (SSF), simulating the natural habitat of soil fungi, such as P. oxalicum, is used for CWDE production, but a limited understanding of CWDE biosynthesis hampers the improvement of CWDE yields through synthetic biology. Here, we identified a novel transcription factor CXRD, which negatively regulates the biosynthesis of cellulase and xylanase in P. oxalicum under SSF, providing a potential target for genetic engineering to improve CWDE production.

Action of the fungal compound citrinin, a bioherbicide candidate, on photosystem II.

Bioherbicides are becoming more attractive as safe weed control tools towards sustainable agriculture. Natural products constitute an important source chemicals and chemical leads for discovery and development of novel pesticide target sites. Citrinin is a bioactive compound produced by fungi of the genera Penicillium and Aspergillus. However, its physiological-biochemical mechanism as a phytotoxin remains unclear. Citrinin causes visible leaf lesions on Ageratina adenophora similar to those produced by the commercial herbicide bromoxynil. Phytotoxicity bioassay tests using 24 plant species confirmed that citrinin has a broad activity spectrum and therefore has potential as a bioherbicide. Based on chlorophyll fluorescence studies, citrinin mainly blocks PSII electron flow beyond plastoquinone QA at the acceptor side, resulting in the inactivation of PSII reaction centers. Furthermore, molecular modeling of citrinin docking to the A. adenophora D1 protein suggests that it binds to the plastoquinone QB site by a hydrogen bond between the O1 hydroxy oxygen atom of citrinin and the histidine 215 of the D1 protein, the same way as classical phenolic PSII herbicides do. Finally, 32 new citrinin derivatives were designed and sorted according to free energies on the basis of the molecular model of an interaction between the citrinin molecule and the D1 protein. Five of the modeled compounds had much higher ligand binding affinity within the D1 protein compared with lead compound citrinin. Citrinin is a novel natural PSII inhibitor that has the potential to be developed into a bioherbicide or utilized as a lead compound for discovery of new derivatives with high herbicidal potency. © 2023 Society of Chemical Industry.

Addition of Organic Matter to Pine Plantations on Agricultural Land Positively Alters the Mycobiome of Agricultural Soils

Afforestation of former agricultural land poses a real challenge for foresters because soil life is often severely limited by the loss of natural soil fungal diversity. In addition, former agricultural soils have low levels of fungal species typical of forest soils, which have a unique microbiome that plays a protective role (antagonists, plant growth promoters, mycorrhizal fungi, etc.). This study aimed to determine the effect of using organic material in the form of bark compost, wood waste, and sawdust to improve the soil mycobiome of soils that have been damaged by their agricultural use. This study used experimental plots established 20 years ago, and we compared the biodiversity of the treated soils with that of the control soils by analysing soil samples with powerful molecular methods. Next-generation sequencing analysis of DNA extracted from soil samples and subsequent analysis of their species composition and biodiversity showed that the mycobiome of soil fungi has been altered by the addition of various forms of organic material. The proportion of fungi belonging to the Ascomycota decreased in favour of species from the Basidiomycota and Mucoromycota. The dominant fungal groups in the soil of the control area were Sagenomella, Wilcoxina, Oidiodendron, Meliniomyces, and Penicillium. Enrichment with organic matter by adding bark compost under the roots led to an increase in Penicillium, Inocybe, and Amphinema. The application of bark compost on the surface led to an increase in the dominance of Inocybe fungi in the soil. The mycobiome of the plant to which woody debris was applied was characterised by a marked dominance of fungi of the genera Russula, Oidiodendron, and Penicillium. Similar ratios were found in the plant to which sawdust was applied, where the fungi Meliniomyces, Penicillium, Oidiodendron, and Russula dominated. A comparative analysis of fungal diversity with the Shannon diversity index showed that the most diverse fungal communities were found in the sawdust plant (6.56), while the control sample (a soil sample from an agricultural area where no organic material was applied) had an index of 5.71. After the treatments, more potential antagonists against pine pathogens and mycorrhizal fungi were found to form beneficial symbiotic relationships with them. In our opinion, the results of this study show that it is worthwhile to introduce different forms of organic matter to post-agricultural land to improve soil biodiversity and mycorrhizal associations of pine roots with fungi to ensure the sustainability of the first generation of forests created.

Effects of mushroom-tobacco rotation on microbial community structure in continuous cropping tobacco soil.

Rotation is an effective strategy for controlling crop diseases and improving plant health. However, the effect of a mushroom-tobacco rotation on the composition and structure of microbial communities in continuous cropping soil is unclear. This study analysed the structure and function of soil bacterial and fungal communities using Illumina MiSeq high-throughput 16S rRNA gene sequencing. The results showed that the physicochemical properties (organic matter, available nitrogen, available phosphorus, and available potassium) and enzymatic activity (phosphatase, catalase, urease and invertase activity) in the rotation treatments (Y1, M1, Y2 and M2) were significantly higher than those in the control (continuous cropping) treatment (CK) and reached peak values in the M2 treatment. PCA showed that the soil microbial community structure in each rotation treatment was different from that in the control. The dominant bacterial phyla of the different soil treatments were Proteobacteria and Actinobacteriota, and the dominant fungal phyla of the different soil treatments were Ascomycota and Basidiomycota. The M2 rotation significantly reduced the relative abundance of harmful fungi (Penicillium and Gibberella) compared to the other treatments. RDA showed that the most abundant bacterial taxa were negatively correlated with pH and positively correlated with physicochemical properties. However, the most abundant fungal taxa were positively correlated with pH and negatively correlated with physicochemical properties. The mushroom-tobacco rotation can effectively maintain the ecological balance of the substrate microbial environment, and provide a more effective way to prevent the continuous cropping of tobacco.

TLR/MyD88-Mediated Inflammation Induced in Porcine Intestinal Epithelial Cells by Ochratoxin A Affects Intestinal Barrier Function.

The intestinal epithelium performs vital functions such as nutrient absorption and acting as an intestinal barrier to maintain the host's homeostasis. Mycotoxin, which affects the processing and storage of animal feedstuff, is a problematic pollutant in farming products. Ochratoxin A generated by Aspergillus and Penicillium fungi causes inflammation, intestinal dysfunction, decline in growth, and reduced intake in porcine and other livestock. Despite these ongoing problems, OTA-related studies in intestinal epithelium are lacking. This study aimed to demonstrate that OTA regulates TLR/MyD88 signaling in IPEC-J2 cells and induces barrier function impairment through tight junction reduction. We measured expression of TLR/MyD88 signaling-related mRNAs and proteins. The indicator of intestinal barrier integrity was confirmed through immunofluorescence and transepithelial electrical resistance. Additionally, we confirmed whether inflammatory cytokines and barrier function were affected by MyD88 inhibition. MyD88 inhibition alleviated inflammatory cytokine levels, tight junction reduction, and damage to barrier function due to OTA. These results indicate that OTA induces TLR/MyD88 signaling-related genes and impairs tight junctions and intestinal barrier function in IPEC-J2 cells. MyD88 regulation in OTA-treated IPEC-J2 cells mitigates the tight junction and intestinal barrier function impairments. Our findings provide a molecular understanding of OTA toxicity in porcine intestinal epithelial cells.

Nine undescribed oxidized ergosterols from the endophytic fungus Penicillium herquei and their cytotoxic activity

A chemical investigation of the EtOAc extract of the endophytic fungus Penicillium herquei led to the isolation of nine undescribed oxidized ergosterols, penicisterols A–I (1–9), along with ten known analogs (10–19). Their structures and absolute configurations were elucidated by a combination of spectroscopic data analysis, quantum-chemical electronic circular dichroism (ECD) calculations and comparisons, [Rh2(OCOCF3)4]-induced ECD experiments, DFT-calculated 13C chemical shifts and DP4+ probability analysis. Compound 1 was a rare example of ergosterol in which the bond between C-8 and C-9 was cleaved to form an enol ether. Moreover, compound 2 possessed a rare (2,5-dioxo-4-imidazolidinyl)-carbamic acid ester group substituted at C-3. All undescribed oxidized ergosterols (1–9) were evaluated for their cytotoxic activity against five cancer cell lines including 4T1 (mouse breast carcinoma), A549 (human pulmonary carcinoma), HCT-116 (human colorectal carcinoma), HeLa (human cervical carcinoma) and Hepg2 (human hepatoma carcinoma) cells. Compounds 2 and 3 displayed moderate cytotoxic activity against 4T1, A549 and HeLa cells, with IC50 values ranging from 17.22 to 31.35 μM.

Temporal and Habitat Dynamics of Soil Fungal Diversity in Gravel-Sand Mulching Watermelon Fields in the Semi-Arid Loess Plateau of China.

Mulching is an important agricultural management tool for increasing watermelon productivity and land-use efficiency because it helps improve water use efficiency and reduce soil erosion. However, there is relatively little available information regarding the effects of long-term continuous monoculture farming on soil fungal communities and related fungal pathogens in arid and semiarid regions. In this study, we characterized the fungal communities of four treatment groups, including gravel-sand-mulched farmland, gravel-sand-mulched grassland, fallow gravel-sand-mulched grassland, and native grassland, using amplicon sequencing. Our results revealed that the soil fungal communities differed significantly between mulched farmland and mulched grassland as well as the fallow mulched grassland. Gravel-sand mulch significantly impaired the diversity and composition of soil fungal communities. Soil fungal communities were more sensitive to gravel-sand mulch in grassland than in other habitats. Long-term continuous monoculture (more than 10 years) led to decreased abundance of Fusarium species, which contains include agronomically important plant pathogens. In the gravel-mulched cropland, some Penicillium and Mortierella fungi were significantly enriched with increasing mulch duration, suggesting potential beneficial properties of those fungi that could be applied to disease control. We also found that long-term gravel mulching in continuous monoculture farming could potentially form disease-suppressive soils and alter soil microbial biodiversity and fertility. Our study provides insights into the exploration of novel agricultural management strategies along with continuous monoculture practice to control watermelon wilt disease by maintaining a more sustainable and healthier soil environment. IMPORTANCE Gravel-sand mulching is a traditional agricultural practice in arid and semiarid regions, providing a surface barrier for soil and water conservation. However, application of such practice in monocropping systems may lead to outbreaks of several devastating plant diseases, such as watermelon Fusarium wilt. Our results with amplicon sequencing suggest that soil fungal communities differ significantly between mulched farmland and mulched grassland and are more sensitive to gravel-sand mulch in grassland. Under continuous monoculture regimens, long-term gravel mulch is not necessarily detrimental and may result in decreased Fusarium abundance. However, some known beneficial soil fungi may be enriched in the gravel-mulch cropland as mulch duration increases. A possible explanation for the reduction in Fusarium abundance may be the formation of disease-suppressive soils. This study provides insight into the need to explore alternative strategies using beneficial microbes for sustainable watermelon wilt control in continuous monocropping system.

Isolation and characterization of three pairs of verrucosidin epimers from the marine sediment-derived fungus Penicillium cyclopium and configuration revision of penicyrone A and related analogues

Verrucosidins, a methylated α-pyrone class of polyketides rarely reported upon, have been implicated in one or more neurological diseases. Despite the significance of verrucosidins as neurotoxins, the absolute configurations of most of the derivatives have not been accurately characterized yet. In this study, three pairs of C-9 epimeric verrucosidin derivatives, including the known compounds penicyrones A and B (1a/1b) and 9-O-methylpenicyrones A and B (2a/2b), the new compounds 9-O-ethylpenicyrones A and B (3a/3b), together with the related known derivative verrucosidin (4), were isolated and identified from the culture extract of Penicillium cyclopium SD-413, which was obtained from the marine sediment collected from the East China sea. Their structures were established based on an in-depth analysis of nuclear magnetic resonances (NMR) and mass spectroscopic data. Determination of the absolute configurations of these compounds was accomplished by Mosher’s method and time-dependent density functional theory (TDDFT) calculations of electronic circular dichroism (ECD) and optical rotation (OR). The configurational assignment of penicyrone A demonstrated that the previously reported C-6 absolute configuration of verrucosidin derivatives needs to be revised from (6S) to (6R). The 9R/9S epimers of compounds 1–3 were found to exhibit growth inhibition against some pathogenic bacteria, indicating that they have potential as lead compounds for the creation of antimicrobial agents.

Kinase POGSK-3β modulates fungal plant polysaccharide-degrading enzyme production and development

The filamentous fungus Penicillium oxalicum secretes integrative plant polysaccharide-degrading enzymes (PPDEs) applicable to biotechnology. Glycogen synthase kinase-3β (GSK-3β) mediates various cellular processes in eukaryotic cells, but the regulatory mechanisms of PPDE biosynthesis in filamentous fungi remain poorly understood. In this study, POGSK-3β (POX_c04478), a homolog of GSK-3β in P. oxalicum, was characterised using biochemical, microbiological and omics approaches. Knockdown of POGSK-3β in P. oxalicum using a copper-responsive promoter replacement system led to 53.5 - 63.6%, 79.0 - 92.8% and 76.8 - 94.7% decreases in the production of filter paper cellulase, soluble starch-degrading enzyme and raw starch-degrading enzyme, respectively, compared with the parental strain ΔKu70. POGSK-3β promoted mycelial growth and conidiation. Transcriptomic profiling and real-time quantitative reverse transcription PCR analyses revealed that POGSK-3β dynamically regulated the expression of genes encoding major PPDEs, as well as fungal development-associated genes. The results broadened our understanding of the regulatory functions of GKS-3β and provided a promising target for genetic engineering to improve PPDE production in filamentous fungi. KEY POINTS: • The roles of glycogen synthase kinase-3β were investigated in P. oxalicum. • POGSK-3β regulated PPDE production, mycelial growth and conidiation. • POGSK-3β controlled the expression of major PPDE genes and regulatory genes.

Anti-inflammatory potential of Penicillium brefeldianum endophytic fungus supported with phytochemical profiling

Various factors contribute to the development of the acute inflammation process, like the pro-inflammatory cytokines, certain enzymes as well as oxidative stress mediators. The anti-inflammatory potential of the endophytic fungus Penicillium brefeldianum was explored in carrageenan-induced inflammation in rats. After isolation of the fungus from Acalypha hispida leaves, it was identified by 18S rRNA gene sequencing. Then, its phytochemical profile was elucidated using LC–ESI–MS/MS technique. There was a remarkable decrease in the edema weight in the endophytic fungi-treated group (200 mg/kg). Also, this group had few inflammatory cells and thickened epidermis with underlying moderate collagenosis when stained with haematoxylin and eosin. Besides, immunostaining with monoclonal antibodies of cyclooxygenase-2 and tumor necrosis factor alpha showed a decrease in the positive immune cells in the endophytic fungi treated group (200 mg/kg) in relation to the positive control. Interestingly, the levels of the inflammatory as well as oxidative stress markers, including prostaglandin E2, nitric oxide, and malondialdehyde, which are hallmarks of the inflammatory process, considerably diminished (p < 0.05) in this group. qRT-PCR was utilised to elucidate the impact of the endophytic fungi treatment on the expression of interleukins (IL-1β and IL-6) genes, which decreased in comparison with the positive control group. Consequently, we can deduce that P. brefeldianum endophytic fungus has a promising anti-inflammatory potential and should be extensively studied on a broader range in the near future.Graphical

Fungal communities, nutritional, physiological and sensory characteristics of sweet potato under three Chinese representative storages

Changes in fungi communities, nutritional, physiological and sensory characteristics of sweet potato tuber roots (TRs) under Chinese representative storages (under-ground, U; semi-underground, S; above-ground, A) were investigated. Diversity of fungal communities of sweet potato TRs under different storages was decreased with storage extension and rotting process. A storage showed the slowest decrease of starch and the highest umami taste in sweet potato TRs than U and S storages, indicating its better edible quality. In addition, communities and quality correspondence analysis indicated that sucrose and total phenol content (TPC) was positively correlated with DPPH and Phenylalanine ammonia-lyase, but negatively correlated with malondialdehyde (MDA). MDA had a negative correlation with beneficial fungi Penicillium, but a positive correlation with pathogen fungi Fusarium. This study revealed changes in fungal communities and quality of sweet potato TRs during storage, providing a basis for effective storage ways development.

Anti-Diabetic Activity of a Novel Exopolysaccharide Produced by the Mangrove Endophytic Fungus Penicillium janthinellum N29

Marine microorganisms often produce exopolysaccharides with novel structures and diverse biological activities due to their specific marine environment. The novel active exopolysaccharides from marine microorganisms have become an important research area in new drug discovery, and show enormous development prospects. In the present study, a homogeneous exopolysaccharide from the fermented broth of the mangrove endophytic fungus Penicillium janthinellum N29, designated as PJ1-1, was obtained. The results of chemical and spectroscopic analyses showed that PJ1-1 was a novel galactomannan with a molecular weight of about 10.24 kDa. The backbone of PJ1-1 was composed of →2)-α-d-Manp-(1→, →4)-α-d-Manp-(1→, →3)-β-d-Galf-(1→ and →2)-β-d-Galf-(1→ units with partial glycosylation at C-3 of →2)-β-d-Galf-(1→ unit. PJ1-1 had a strong hypoglycemic activity in vitro, evaluated using the assay of α-glucosidase inhibition. The anti-diabetic effect of PJ1-1 in vivo was further investigated using mice with type 2 diabetes mellitus induced by a high-fat diet and streptozotocin. The results indicated that PJ1-1 markedly reduced blood glucose level and improved glucose tolerance. Notably, PJ1-1 increased insulin sensitivity and ameliorated insulin resistance. Moreover, PJ1-1 significantly decreased the levels of serum total cholesterol, triglyceride and low-density lipoprotein cholesterol, enhanced the level of serum high-density lipoprotein cholesterol and alleviated dyslipidemia. These results revealed that PJ1-1 could be a potential source of anti-diabetic agent.

DNA metabarcoding analysis of fungal community on surface of four root herbs

ObjectiveAngelicae Sinensis Radix (ASR, Danggui in Chinese), Cistanches Herba (CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma (PG, Renshen in Chinese), and Panacis Quinquefolii Radix (PQ, Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding. MethodsA total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region (ITS2). ResultsAll the 12 samples were detected with fungal contamination. Rhizopus (13.04%−74.03%), Aspergillus (1.76%−23.92%), and Fusarium (0.26%−15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi (Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi (Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups, while fungal community in PG and PQ groups was relatively similar. ConclusionDNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Pegriseofamines A−E: Five cyclopiazonic acid related indole alkaloids from the fungus Penicillium griseofulvum

Five new cyclopiazonic acid related indole alkaloids, pegriseofamines A−E (1–5), were isolated from the fungus Penicillium griseofulvum. Their structures and absolute configurations were determined by NMR, HRESIMS, quantum-chemical calculation, and X-ray diffraction experiments. Among them, pegriseofamine A (1) possesses an undescribed 6/5/6/7 tetracyclic ring system generated by the fusion of an azepine and an indole unit via a cyclohexane, and the postulated biosynthetic origin of 1 was discussed. Compound 4 could relieve liver injury and prevent hepatocyte apoptosis in ConA-induced autoimmune liver disease.

Two new secondary metabolites isolated from the fungus Penicillium virgatum T49-A

The extensively chemical investigation of the EtOAc extract of the soil fungus Penicillium virgatum T49-A has successfully led to the isolation of two undescribed secondary metabolites penivirtone A (1) and peniviramide B (2) together with six known compounds. Their chemical structures including the absolute configurations of the two new compounds were comprehensively established by extensive analyses of NMR and HRESIMS spectra as well as ECD powered by theoretical calculations. Moreover, the cytotoxic and antibacterial activities of compounds 1–2 were also evaluated, whereas the two novel compounds showed no notable cytotoxic and antibacterial activities.

Identification by DNA barcode of culturable, airborne, environmental fungi isolated from the Shepherd University campus

Diverse fungi are abundant within soil communities and commonly found in association with many plant and tree species. This study investigates the culturable, airborne fungi found in the outside environment of the Shepherd University campus. Viable fungi were captured in two different locations on two different culture media, using the open plate method. Captured fungi were isolated and putative species identified by sequencing the internal transcribed spacer region (ITS) of the ribosomal RNA (rRNA) gene cassette. Due to insufficient sequence variability within the ITS region, most isolates could not be identified to species. 15 fungal isolates were identified to order, family, genus, or species complex: Dothideales (1 isolate), Didymellaceae (4 isolates), Aureobasidium (1 isolate), Cladosporium (1 isolate), Mucor (3 isolates), Nigrospora (1 isolate), Pestalotiopsis (2 isolates), and Penicillium (2 isolates), including Penicillium thiersii (1 isolate). These isolates are similar to those isolated from inside a teaching laboratory on the Shepherd University campus; Didymellaceae, Cladosporiaceae, and Penicillium fungi were isolated multiple times within the teaching laboratory. This research was supported by the Shepherd University Biology Department, a student fellowship from NASA-WVSGC (J. Kellison), and a research enhancement award from NASA-WVSGC (L. Robertson).

An efficient Agrobacterium-mediated system based on the pyrG auxotrophic marker for recombinant expression in the filamentous fungus Penicillium rubens

This work aimed to construct a versatile, effective, and food-grade Agrobacterium tumefaciens-mediated transformation (ATMT) system for recombinant expression in the filamentous fungus Penicillium rubens (also known as Pencillium chrysogenum). In this study, the wild-type P. chrysogenum VTCC 31172 strain was re-classified as P. rubens by a multilocus sequencing analysis. Further, the pyrG gene required for uridine/uracil biosynthesis was successfully deleted in the VTCC 31172 strain by homologous recombination to generate a stable uridine/uracil auxotrophic mutant (ΔpyrG). The growth of the P. rubens ΔpyrG strain could be restored by uridine/uracil supplementation, and a new ATMT system based on the uridine/uracil auxotrophic mechanism was established for this strain. The optimal ATMT efficiency could reach 1750 transformants for 106 spores (equivalent to 0.18%). In addition, supplementation of uridine/uracil at the concentrations of 0.005-0.02% during the co-cultivation process significantly promoted transformation efficiency. Especially, we demonstrated that the pyrG marker and the amyB promoter from the koji mold Aspergillus oryzae were fully functional in P. rubens ΔpyrG. Expression of the DsRed reporter gene under the regulation of the A. oryzae amyB promoter lighted up the mycelium of P. rubens with a robust red signal under fluorescence microscopy. Furthermore, genomic integration of multiple copies of the Aspergillus fumigatus phyA gene under the control of the amyB promoter significantly enhanced phytase activity in P. rubens. The ATMT system developed in our work provides a safe genetic platform for producing recombinant products in P. rubens without using drug resistance markers.

Three new α-pyrone derivatives from the soil-derived fungus Penicillium herquei MA-370

Three new α-pyrone derivatives, annularins L–N (1–3), were isolated from the EtOAc extract of Penicillium herquei MA-370, a fungus obtained from the rhizospheric soil of the mangrove plant Rhizophora mucronata. The planar structures of compounds 1–3 were determined based on comprehensive spectral interpretation of the NMR and MS data. The absolute configuration of 1 was determined by X-ray crystallographic data and that of 2 was assigned by TDDFT calculations of its ECD spectrum and cotton effects comparison with those of 1. The antimicrobial activity of compounds 1–3 was evaluated.

Nature-Inspired Chemistry of Complex Alkaloids: Combining Targeted Molecular Networking Approach and Semisynthetic Strategy to Access Rare Communesins in a Marine-Derived Penicillium expansum.

Communesins are rare alkaloids isolated from fungi of the genus Penicillium. In this work, the extract of a marine-derived Penicillium expansum strain was studied using targeted molecular networking approach allowing to detect 65 communesins including 55 new ones. A fragmentation pattern for dimethylvinyl communesins was established and a script was implemented allowing to predict the structure and map all communesins in a global molecular network. A semisynthetic strategy was carried out to obtain some minor congeners from the two isolated communesins A and B. Nine communesins were then synthetised: two of them were already described as produced by the studied strain; four are new natural products which occurrence in the extracts was confirmed; three are new semi-synthetic analogues never described so far. These communesins were evaluated for their cytotoxicity on two human cancer cell lines KB and MCF-7 leading to a preliminary study of their structure-activity relationships.

Antibiofilm activity of LAE (ethyl lauroyl arginate) against food-borne fungi and its application in polystyrene surface coating

Several filamentous fungi species as Fusarium oxysporum or Cladosporium sp. can form biofilms by themselves or by participating in polymicrobial biofilms with bacteria. However, despite the high impact of biofilm on the food industry and the high efforts done to control biofilm produced by bacteria in the food area, there has been little study of strategies to control fungal biofilm in this area. In this study, the antibiofilm activity of the safe antimicrobial compound ethyl lauroyl arginate (LAE) was investigated against food spoilage fungi (Cladosporium cladosporioides, Aspergillus ochraceus, Penicillium italicum, Botrytis cynerea and Fusarium oxyspoum). Finally, the efficacy of a varnish-based coating incorporating LAE and coated onto polystyrene microtiter plates has been evaluated as a strategy to reduce fungal biofilm formation. The results of the 2,3-bis-(2-metoxi-4-nitro-5-sulfofenil)-2H-tetrazoilo-5-carboxanilida (XTT) assay, which measure the biofilm metabolic activity of moulds, demonstrated that LAE reduced significantly the formation of fungal biofilm at concentrations from 6 to 25 mg/L. This reduction was confirmed by the micrographs obtained by scanning electronic microscopy (SEM). In addition, LAE also showed antifungal activity against established biofilms. Particularly, it reduced their metabolic activity and viability at concentrations from 6 to 25 mg/L according to results obtained in the XTT assay and observations made by confocal laser scanning microscopy (CLSM). Finally, active coating incorporating from 2% of LAE proved to reduce significantly the biofilm formation in C. cladosporioides, B. cynerea and F. oxyspoum according to the results obtained in the XTT assay. However, the released studies indicated that the retention of LAE in the coating should be improved to prolong their activity.