Fungal Strains Research Articles

Bioprospecting Endophytic Fungi of Forest Plants for Bioactive Metabolites with Antibacterial, Antifungal, and Antioxidant Potentials.

The growing emergence of multi-drug resistant microbial strains has kept the scientific world searching for novel bioactive compounds with specific chemical characteristics. Accordingly, researchers have started exploring the understudied metabolites from endophytes as a new source of bioactive compounds. In this context, the current study was designed to evaluate the bioactive properties of endophytic fungi from the Mokrzański forest in Wrocław, Poland that have not yet been fully researched. Forty-three endophytic fungi were isolated from twelve distinct plants. Following their cultivation, fungal extracts were separately prepared from biomass and cell-free filtrates, and their antibacterial, antifungal (against human and plant pathogens), and antioxidant properties were examined. Five promising fungi after screening were identified to possess all of these activities. These strains and their respective plant hosts were Trichoderma harzianum BUK-T (Fagus sylvatica), Aspergillus ochraceus ROB-L1 (Robinia pseudoacacia), Chaetomium cochliodes KLON-L1, Fusarium tricinctum KLON-L2 (Acer platanoides), and Penicillium chrysogenum SOS-B2 (Pinus sylvestris). Moreover, gamma irradiation at several doses (Gy) was separately applied to the fungal cultures to study their effects on the recorded activities. Finally, compounds after preparative thin-layer chromatography fractionation of the five fungal strains were identified by GC-MS. These findings suggest that the isolated endophytic fungi could serve as novel sources of bioactive metabolites with antibacterial, antifungal, and antioxidant properties, potentially paving the way for future research and the development of new bioactive compounds.

Exploration of three native strains of entomopathogenic fungi against rice yellow hairy caterpillar, <i>Psalis pennatula</i> (Lepidoptera: Lymantriidae): An emerging pest of rainfed rice in Assam

Rice yellow hairy caterpillar Psalis pennatula Fabricius (Lepidoptera: Lymantriidae) is an emerging pest of rice and an increased incidence is expected in different rice-growing areas of Assam in the coming years. The caterpillar is a voracious leaf feeder and can consume the whole leaves within a day. Keeping this in view, an attempt was made to evaluate the bio-efficacy of three important native isolates of entomopathogenic fungal strains viz. Beauveria bassiana (KR855715), Cordyceps javanica (OM321438) and Isaria fumosorosea (MH414514) against P. pennatula larvae under laboratory conditions during the Sali season of rice. All three EPF strains at different conidial dilutions (1x103, 1x105,1x107, 1x108 and1x109 conidia/ml) were found to be pathogenic to the 3rd instar larvae of P. pennatula. Among the tested fungal strains, the strain Isaria fumosorosea (MH414514) showed the highest mortality (100%) at 1x107 conidia/ml and Cordyceps javanica (OM321438) showed the lowest mortality (51%) at 1x107conidia/ml at 10th day of post-treatment. The tested fungal isolates could play a vital role as microbial biopesticides in suppressing the P. pennatula population in Assam and could be incorporated into IPM strategies.

A Preliminary Experimental Study on Biodegradation of 3D-Printed Samples from Biomass–Fungi Composite Materials

Products made from petroleum-derived plastic materials are linked to many environmental problems, such as greenhouse gas emissions and plastic pollution. It is desirable to manufacture products from environmentally friendly materials instead of petroleum-based plastic materials. Products made from biomass–fungi composite materials are biodegradable and can be utilized for packaging, construction, and furniture. In biomass–fungi composite materials, biomass particles (derived from agricultural wastes) serve as the substrate, and the fungal hyphae network binds the biomass particles together. There are many reported studies on the 3D printing of biomass–fungi composite materials. However, there are no reported studies on the biodegradation of 3D-printed samples from biomass–fungi composite materials. In this study, two types of biomass materials were used to prepare printable mixture hemp hurd and beechwood sawdust. The fungi strain used was Trametes versicolor. Extrusion based 3D printing was used to print samples. 3D-printed samples were left for five days to allow fungi to grow. The samples were then dried in an oven for 4 h at 120 °C to kill all the fungi in the samples. The samples were buried in the soil using a mesh bag and kept in an environmental chamber at 25 °C with a relative humidity of 48%. The weight of these samples was measured every week over a period of three months. During the testing period, the hemp hurd test samples lost about 33% of their original weight, whereas the beechwood sawdust samples lost about 30% of their original weight. The SEM (scanning electron microscope) micrographs showed the presence of zygospores in the test samples, providing evidence of biodegradation of the test samples in the soils. Additionally, the difference in peak intensity between the control samples and test samples (for both hemp hurd and beechwood sawdust) showed additional evidence of biodegradation of the test samples in the soils.

Green Synthesis and Characterization of Apple Peel‐Derived Selenium Nanoparticles for Anti‐Fungal Activity and Effects of MexA Gene Expression on Efflux Pumps in Acinetobacter baumannii

ABSTRACTSelenium nanoparticles (Se‐NPs) were produced adopting an environmentally benign green synthetic approach from the waste biomaterial obtained from the apple peel extract (APE). The produced NPs were characterized through UV–Visible spectroscopy (UV–Vis), atomic force microscopy (AFM), Fourier‐transform infrared spectroscopy (FT‐IR), X‐ray diffraction (XRD), field emission‐scanning electron microscopy (FE‐SEM), and transmission electron microscopy (TEM). The UV–Visible spectra exhibited the absorption peak λmax at 295 nm, confirming the formation of Se‐NPs. The AFM examination revealed the roughness and distribution of the nanoparticles, and FT‐IR ascertained the surface‐contained functional groups presence. The XRD confirmed the crystalline nature of the sample, and FE‐SEM validated the surface morphology and the size range of 52 to 84 nm. The TEM demonstrated the spherical shapes of the Se‐NPs with a face‐centered cubic crystal nature, whereas the size distribution analysis showed mean size of 70 nm. The minimum inhibitory concentration (MIC) at 500 μg mL−1 against Candida spp., namely, C. albicans, C. guilliermondii, and C. ciferrii, confirmed their anti‐fungal activity with the in control and minor statistical difference (p < 0.05) in the dissemination mean zones of inhibition for the Se‐NPs treatment against these fungi. The Se‐NPs effects on MexA gene expression demonstrated the efflux pump role in the anti‐fungal activity and gene's down regulation for the treated fungal strains. The Se‐NPs exhibited their anti‐fungal activity against the multi‐drug resistant microbes.

Formulation Strategies to Overcome Amphotericin B Induced Toxicity.

Fungal infection poses a major global threat to public health because of its wide prevalence, severe mortality rate, challenges involved in diagnosis and treatment, and the emergence of drug-resistant fungal strains. Millions of people are getting affected by fungal infection, and around 3.8 million people face death per year due to fungal infection, as per the latest report. The polyene antibiotic AmB has an extensive record of use as a therapeutic moiety against systemic fungal infection and leishmaniasis since 1960. AmB has broad-spectrum fungistatic and fungicidal activity. AmB exerts its therapeutic activity at the cellular level by binding to fungal sterol and forming hydrophilic pores, releasing essential cellular components and ions into the extracellular fluid, leading to cell death. Despite using AmB as an antifungal and antileishmanial at a broad scale, its clinical use is limited due to drug-induced nephrotoxicity resulting from binding the aggregated form of the drug to mammalian sterol. To mitigate AmB-induced toxicity and to get better anti-fungal therapeutic outcomes, researchers have developed nanoformulations, self-assembled formulations, prodrugs, cholesterol- and albumin-based AmB formulations, AmB-mAb combination therapy, and AmB cochleates. These formulations have helped to reduce toxicity to a certain extent by controlling the aggregation state of AmB, providing sustained drug release, and altering the physicochemical and pharmacokinetic parameters of AmB. Although the preclinical outcome of AmB formulations is quite satisfactory, its parallel result at the clinical level is insignificant. However, the safety and efficacy of AmB therapy can be improved at the clinical stage by continuous investigation and collaboration among researchers, clinicians, and pharmaceutical companies.

A Taxonomic and Phylogenetic Study of Anamorphic Strains of Daldinia (Hypoxylaceae, Xylariales) in Southern China.

In an extensive fungal investigation conducted in southern China, a large number of fungal strains were isolated by collecting and treating diseased and decayed leaves. Using internal transcribed spacer regions (ITSs) sequence data for a BLAST search to screen for suspected strains of Daldinia, followed by phylogenetic analysis using internal transcribed spacer regions, partial sequences of the large subunit of the rDNA (LSU), RNA polymerase II (rpb2), and beta tubulin (tub2) sequence data, combined with morphological characteristics of anamorphic species, ninety-four strains of Daldinia were identified. Furthermore, their geographical distribution and host specificity of the genus were thoroughly analyzed and summarized. Additionally, seven new anamorphic species of the genus Daldinia were also detected, Daldinia ehretiae sp. nov., D. jianfengensis sp. nov., D. ledongensis sp. nov., D. menghaiensis sp. nov., D. rhododendri sp. nov., D. spatholobi sp. nov., and D. thunbergiae sp. nov.

Dothistroma septosporum and Dothistroma pini, the causal agents of Dothistroma needle blight, are infected by multiple viruses

Dothistroma septosporum and Dothistroma pini are severe foliar pathogens of conifers. They infect a broad spectrum of hosts (mainly Pinus spp.), causing chlorosis, defoliation of needles, and eventually the death of pine trees in extreme cases. Mycoviruses represent a novel and innovative avenue for controlling pathogens. To search for possible viruses hosted by Dothistroma spp. we screened a subset of isolates (20 strains of D. septosporum and one D. pini) originating from the Czech Republic, Slovenia, Italy, Austria and Ireland for viral dsRNA segments. Only five of them showed the presence of dsRNA segments. A total of 21 fungal isolates were prepared for total RNA extractions. RNA samples were pooled, and two separate RNA libraries were constructed for stranded total RNA sequencing. RNA-Seq data processing, pairwise sequence comparisons (PASC) and phylogenetic analyses revealed the presence of thirteen novel putative viruses with varying genome types: seven negative-sense single-stranded RNA viruses, including six bunya-like viruses and one new member of the order Mononegavirales; three positive-sense single-stranded RNA viruses, two of which are similar to those of the family Narnaviridae, while the genome of the third correspond to those of the family Gammaflexiviridae; and three double-stranded RNA viruses, comprising two novel members of the family Chrysoviridae and a potentially new species of gammapartitivirus. The results were confirmed with RT-PCR screening that the fungal pathogens hosted all the viruses and showed that particular fungal strains harbour multiple virus infections and that they are transmitted vertically. In this study, we described the narnavirus infecting D. pini. To our knowledge, this is the first virus discovered in D. pini.

Effects of phosphate-solubilizing bacteria Aspergillus flavus AF-LRH1 on promoting phosphorus solubilization, wheat growth and soil heavy metal remediation

Increasing nutrient supply level and eliminating pollution risk are essential to promote farmland soil use. In the soil-plant nutrition cycle, phosphorus is a limiting nutrient, toxic metals such as Cd and Pb are stress threats. Utilizing microorganisms as a bio-amendment could be a viable approach to enhance the cycling of soil phosphorus nutrients and immobilize Cd and Pb in the soil. This study involved the isolation of Aspergillus flavus AF-LRH1, a phosphate solubilizing fungal strain from soil. The strain exhibited the capacity to solubilize calcium phosphate, attaining a phosphorus concentration of 220.73 mg L−1 in the medium. High performance liquid chromatography analysis showed that AF-LRH1 produced a combination of organic acids (tartaric acid 260.7 mg L−1, fumaric acid 0.8 mg L−1, citric acid 911.9 mg L−1) to dissolve Ca3(PO4)2 by providing H+ ions. AF-LRH1 also utilizes NH4+ for the biosynthesis of amino acids and releases protons, which facilitate the dissolution of insoluble Ca3(PO4)2 and calcium phytate. Through these processes, calcium phosphate is partly converted into hydroxyapatite because of biological solubilization. AF-LRH1 strain showed the capability to recover phosphorus from various types of sludge-based solid residue by increasing the solubility of insoluble phosphate compounds for plant nutrient supply. In addition, the combining addition of AF-LRH1 strain and phosphate-containing amendments (calcium phosphate, sludge ash and biochar) benefited the decontamination of Pb and Cd contaminated soil. This finding confirmed the AF-LRH1 could be a promising environmental-friendly biofertilizer suitable for agricultural applications and bioremediation of heavy metal contaminated soil.

Enhancing the biotransformation of progesterone to the anticancer compound testololactone by Penicillium chrysogenum Ras3009: kinetic modelling and efficiency maximization

BackgroundBiotransformation of steroid compounds into therapeutic products using microorganisms offers an eco-friendly and economically sustainable approach to the pharmaceutical industry rather than a chemical synthesis way. The biotransformation efficiency of progesterone into the anticancer compound testololactone using Penicillium chrysogenum Ras3009 has been investigated. Besides, maximization of testololactone formation was achieved by studying the kinetic modelling and impact of some fermentation conditions on the biotransformation process.ResultsThe fungal strain Ras3009 was selected among twelve fungal strains as the most runner for the transformation of 81.18% of progesterone into testololactone. Ras3009 was identified phenotypically and genotypically as Penicillium chrysogenum, its 18 S rRNA nucleotide sequence was deposited in the GenBank database by the accession number OR480104. Studying the impact of fermentation conditions on biotransformation efficiency indicated a positive correlation between substrate concentration and testololactone formation until reaching the maximum velocity vmax. Kinetic studies revealed that vmax was \\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\:0.0482$$\\end{document} gL− 1hr− 1 with high accuracy, giving R2 of 0.977. The progesterone transformation efficiency generally increased with time, reaching a maximum of 100% at 42 h with testololactone yield (Ypt/s) 0.8700 mg/mg. Moreover, the study indicated that the enzymatic conversion by P. chrysogenum Ras3009 showed high affinity to the substrate, intracellularly expressed, and released during cell disruption, leading to higher efficiency when using whole microbial cell extract.ConclusionsFungi can be promising biocatalysts for steroid transformation into valuable chemicals and pharmaceutical compounds. The study revealed that the new fungal isolate P. chrysogenum Ras3009 possesses a great catalytic ability to convert progesterone into testololactone. Kinetic modelling analysis and optimization of the fermentation conditions lead to higher transformation efficiency and provide a better understanding of the transformation processes.Graphical abstract

Evaluation of Phytochemical Composition and Antifungal Potential of Millingtonia hortensis Leaf Extract

Millingtonia hortensis, commonly known as the Indian cork tree, has been traditionally used in Southern Asian medicine for various therapeutic purposes. This study aimed to investigate the antifungal properties of M. hortensis leaf extract and evaluate its potential as a natural alternative to synthetic antifungal agents. Leaf samples were collected, authenticated, and subjected to aqueous and methanolic extraction. Phytochemical screening revealed the presence of alkaloids, flavonoids, cardiac glycosides, terpenoids, tannins, phenolic compounds, and proteins. The antioxidant activity of the extracts was assessed using the DPPH free radical scavenging assay, while the antifungal potential was evaluated through the agar-well diffusion method against selected fungal strains. The aqueous extract yielded 4.4% w/w, while the methanolic extract yielded 2.9% w/w. Both extracts demonstrated significant antioxidant activity, with the methanolic extract showing slightly higher potency. The antifungal assay revealed dose-dependent inhibition of fungal growth, with the highest concentration (300 μg/ml) exhibiting the maximum zone of inhibition. Ketoconazole served as a positive control, while 10% DMSO was used as a negative control. The results suggest that M. hortensis leaf extract possesses promising antifungal properties, likely attributed to its rich phytochemical composition. This study provides a foundation for further research into the development of novel, plant-based antifungal therapies and highlights the potential of M. hortensis as a source of bioactive compounds for pharmaceutical applications

Bioremediation petroleum wastewater and oil-polluted soils by the non-toxigenic indigenous fungi.

Soil and wastewater samples contaminated by petroleum-related industries were collected from various locations in Saudi Arabia, a country known for its vast oil reserves. The samples were analyzed for their physicochemical properties, including the presence of metals, petroleum hydrocarbons, and aromatic compounds. A total of 264 fungal isolates were analyzed and categorized into eight groups of Aspergillus (194 isolates) and four groups of Penicillium (70 isolates). The potential of these fungal groups to grow in oil or its derivatives was investigated. Two isolates, Aspergillus tubingensis FA-KSU5 and A. niger FU-KSU69, were utilized in two remediation experiments-one targeting wastewater and the other focusing on polluted soil. The FA-KSU5 strain demonstrated complete removal of Fe3+, As3+, Cr6+, Zn2+, Mn2+, Cu2+ and Cd2+, with bioremediation efficiency for petroleum hydrocarbons in the wastewater from these sites ranging between 90.80 and 98.58%. Additionally, the FU-KSU69 strain achieved up to 100% reduction of Co2+, Ba2+, B3+, V+, Ni2+, Pb2+ and Hg2+, with removal efficiency ranging from 93.17 to 96.02% for aromatic hydrocarbons after 180min of wastewater treatment. After 21days of soil incubation with Aspergillus tubingensis FA-KSU5, there was a 93.15% to 98.48% reduction in total petroleum hydrocarbons (TPHs) and an 88.11% to 97.31% decrease in polycyclic aromatic hydrocarbons (PAHs). This strain exhibited the highest removal rates for Cd2+ and As3+ followed by Fe3+, Zn2+, Cr6+, Se4+ and Cu2+. Aspergillus niger FU-KSU69 achieved a 90.37% to 94.90% reduction in TPHs and a 95.13% to 98.15% decrease in PAHs, with significant removal of Ni2+, Pb2+ and Hg2+, followed by Co2+, V+, Ba2+ and B3+. The enzymatic activity in the treated soils increased by 1.54- to 3.57-fold compared to the polluted soil. Although the mixture of wastewater and polluted soil exhibited high cytotoxicity against normal human cell lines, following mycoremediation, all treated soils and effluents with the dead fungal biomass showed no toxicity against normal human cell lines at concentrations up to 500 µL/mL, with IC50 values ≥ 1000 µL/mL. SEM and IR analysis revealed morphological and biochemical alterations in the biomass of A. tubingensis FA-KSU5 and A. niger FA-KSU69 when exposed to petroleum effluents. This study successfully introduces non-toxigenic and environmentally friendly fungal strains play a crucial role in the bioremediation of contaminated environments. Both strains serve as low-cost and effective adsorbents for bio-remediating petroleum wastewater and oil-contaminated soil. Heavy metals and hydrocarbons, the primary pollutants, were either completely removed or reduced to permissible levels according to international guidelines using the dead biomass of FA-KSU5 and FA-KSU69 fungi. Consequently, the environments associated with this globally significant industry are rendered biologically safe, particularly for humans, as evidenced by the absence of cytotoxicity in samples treated with A. tubingensis FA-KSU5 and A. niger FA-KSU69 on various human cell types.

Fungal Lung Disease.

Fungal lung disease encompasses a wide spectrum of organisms and associated clinical conditions presenting a significant global health challenge with type and severity determined by underlying host immunity and infecting fungal strain. The most common group of diseases are associated with the filamentous fungus Aspergillus spp. and include allergic bronchopulmonary aspergillosis (ABPA), sensitization, aspergilloma, and chronic and invasive pulmonary aspergillosis. Fungal lung disease remains epidemiologically heterogenous and is influenced by geography, environment, and host comorbidities. Diagnostic modalities continue to evolve and now include novel molecular assays and biomarkers, however, persisting challenges include achieving rapid and accurate diagnosis, particularly in resource-limited settings and in differentiating fungal infection from other pulmonary conditions. Treatment strategies for fungal lung diseases rely mainly on antifungal agents, however, the emergence of drug-resistant strains poses a substantial global threat and adds complexity to existing therapeutic challenges. Emerging antifungal agents and increasing insight into lung mycobiome via may offer fresh and personalized approaches to diagnosis and treatment, while innovative methodologies are required to mitigate drug resistance and adverse effects of treatment. This state-of-the-art review describes the current landscape of fungal lung disease, highlighting key clinical insights, current challenges, and emerging approaches for its diagnosis and treatment.

A-273 Rapid, Portable Invasive Fungal Infection Diagnostic and Candida Antimicrobial Susceptibility Testing

Abstract Background Traumatic wounds are associated with a higher risk of invasive fungal infections (IFI), which lead to a significant increase in amputation or death. Actionable treatment decisions are hampered because true infections can be difficult to differentiate from external wound contaminates, and tissue biopsy and quantification of fungal load is required to determine if treatment is necessary, which currently takes several days. GeneCapture is developing the CAPTURE 60-minute, portable infection identification platform to detect infectious levels of common fungi including the genera Aspergillus, Fusarium, and Scedosporium, the family Herpotrichiellaceae, and the order Mucorales. The company is also pursuing rapid identification and antimicrobial susceptibility testing for Candida species from multiple infection types. Methods This study seeks to determine the Limit of Detection (LoD), sensitivity and specificity of the CAPTURE ID assay for fungal pathogens. Since tissue biopsy samples must be used to assess the presence of IFI in wounds, GeneCapture has developed a manual process for the crude homogenization of muscle tissue to allow this matrix to be used as the starting sample for the ID assay. Mock samples were tested for inclusivity and exclusivity on a series of IFI ID panels against multiple strains of the included fungi, against fungi not yet specifically detected on the panel, and against bacteria that commonly infect wounds. While the LoD for yeasts may be determined in CFU/mL, there is no similar, easy-to-define metric for molds. GeneCapture has developed a quantitative PCR method to detect the single copy beta-tubulin gene to roughly correlate fungal dry mass and nuclei number to assay signal for molds cultured in Potato Dextrose Broth. Determining beta-tubulin copy numbers for extracts from mock samples then allowed detection of the progression of IFI over the course of infection. Lastly, antimicrobial susceptibility testing (AST) was performed on two species of Candida to validate utilizing GeneCapture’s rapid phenotypic AST method for fungal infections. Results To date, the CAPTURE ID panel has been tested against 27 included or excluded strains of fungi or bacteria with 97% sensitivity and 100% specificity. The current LoD for mold organisms varies from approximately 1,000 nuclei for Aspergillus species, to 100,000 nuclei for Fusarium species. Additional work to understand this range is underway. Lastly, we have correctly determined the susceptibility of Candida species using our novel approach within 4-6 hours. Conclusions GeneCapture’s ID and AST technology offer a unique solution to address the increasing prevalence of fungal infections. Our platform can identify the most common fungal components in a wound within 60 minutes of receiving a tissue biopsy sample. Candida infections occur across multiple matrices and rapidly identifying the species of Candida and then determining its phenotypic susceptibility profile will have a large impact on treating these difficult pathogens.

First Report of Leaf Rot on Catalpa bungei caused by Fusarium verticillioides in China.

Catalpa bungei originates from China. Because of its well-developed root system and strong resistance to wind and soil, it is one of the top ten recommended species of ecological management (Jian et al., 2022). In September 2023, a severe leaf rot of C. bungei was observed at Lanlake farm (500 acres) in Nanyang (33°3'23" N, 112°28'50" E), China. The incidence rate of leaf rot reached 45% (n = 100). The pale-yellow spots initially appeared on the adaxial surface of leaf margins, subsequently enlarging to form irregular black rot lesions, with the yellow halos around the necrotic area of the lesion, ultimately causing the entire leaves to wither. Diseased leaves (20) were collected, cut into pieces, sterilized, and then placed on potato dextrose agar (PDA). A total of 25 purified fungal strains were isolated, and three strains (QS2-1, QS2-2, QS2-3) from distinct areas were selected for further analysis. Each strain produced abundant aerial mycelium, initially white, which later developed purple pigments. The aerial conidiophores were sparsely branched, ending with verticillate phialides. The strains generally produced many more microconidia than macroconidia on PDA media. Microconidia were clavate and measured 3.9 to 6.6 × 1.1 to 2.4 μm (n = 50). To produce macroconidia, we used YPG liquid medium (0.3% yeast extract, 1% peptone, and 2% glucose) with shaking (200 r.p.m.) for 5 days. Macroconidia were slender, straight, and measured 19.5 to 27.1 × 1.9 to 3.5 μm, with 3 to 5 septa (n = 50). The morphological characteristics matched the species description of Fusarium verticillioides (Sacc.) Nirenberg 1976 (Leslie and Summerell, 2006). The rDNA internal transcribed spacer (ITS), β-tubulin gene (tub2), translation elongation factor 1-alpha gene (tef1), calmodulin (cmdA), RNA polymerase II largest subunit (rpb1), and RNA polymerase II second largest subunit (rpb2) were amplified for molecular identification (O'Donnell et al., 2022). The sequences were submitted to GenBank with accession numbers OR741762, OR741763, OR741765 (ITS), OR762222, OR762223, OR939807 (tub2), OR939799, OR939800, PQ035927 (tef1), OR778611, OR939808, OR939809 (cmdA), PQ035921, PQ035922, PQ035923 (rpb1), and PQ035924, PQ035925, PQ035926 (rpb2). BLASTn analysis of QS2-1 sequences exhibited 99% similarity with F. verticillioides sequences (strains CBS 576.78) MT010888 of cmdA (711/713, 99%), MT010956 of rpb1 (1790/1791, 99%), and MT010972 of rpb2 (868/870, 99%). A phylogenetic tree was constructed using concatenated sequences along with the sequences of the type strains employing the neighbor-joining method, showing the three strains formed a clade with the type strain CBS 576.78. Pathogenicity was tested on 10 healthy potted seedlings by spraying them with a conidial suspension (106 conidia ml-1), while 5 seedlings were sprayed with sterilized water as a control. The plants were placed in climate incubators. Ten days after inoculation, typical lesions were observed on the treated plants, but not on the control group. The reisolated strains were identified as F. verticillioides through morphological characterization, thus fulfilling Koch's postulates. F. verticillioides is known to cause Fusarium ear rot on maize, and other plants including Brassica rapa (Akram et al., 2020) and Schizonepeta tenuifolia (Li et al., 2024). This is the first documented instance of F. verticillioides causing leaf rot on C. bungei globally. Identifying the pathogen is critical to implementing effective disease management strategies, especially in choosing proper pesticide agents and screening disease-resistant varieties.

Isolation and identification of endophytic fungi from Alhagi sparsifolia Shap. and their antibacterial activity

In order to explore the endophytic resources of Alhagi sparsifolia Shap. and identified novel antibacterial substances. Thirty endophytic fungal strains were isolated from the stems and roots of A. sparsifolia Shap. Morphological and molecular biology methods were used to identify ten strains of fungi, including four strains of Aspergillus niger, three strains of Alternaria alternata, two strains of Aspergillus flavus, and one strain of Fusarium incarnatum. All these strains were isolated from A. sparsifolia Shap. for the first time, and of these Aspergillus was the dominant genus. Antibacterial activity of the ten strains against Escherichia coli, Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa were evaluated using the disc diffusion method. The results demonstrated that the metabolites from all the strains had inhibitory effects on at least one indicator bacterium. Notably, the endophytic fungi AFJ3 and AFG6 demonstrated strong broad-spectrum antibacterial activity, particularly against E. coli, with inhibition zones measuring 32.0 ± 0.3 and 31.3 ± 0.3 mm, respectively. The three endophytic fungi (AFG1, AFG2, and AFG3) isolated from the roots demonstrated significant antibacterial activity against P. aeruginosa forming an inhibition zone of diameter 31.3 ± 0.1, 25.6 ± 0.2, and 25.6 ± 0.2 mm, respectively. However, the strains of endophytic fungi demonstrated no significant inhibitory effects on C. albicans. Ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry/mass spectrometry (UPLC-QTOF-MS/MS) analysis depicted that the ethyl acetate phase of AFJ3 and AFG6 fermentation broth predominantly contained organic acids, phenolic acids, flavonoids, and fatty acids. These secondary metabolites often exhibited good antibacterial activity. This study broadens our understanding of endophytic fungi in A. sparsifolia Shap. The antibacterial activity of some strains of endophytic fungi was significant, making it worthy of further research on their active material.

Fungal degradation of phenylacetate focusing on CRISPR/Cas9-assisted characterization of two oxidative enzyme genes of Akanthomyces muscarius AM1091

The degradation of phenylacetate (PA) was investigated as a model to explore aromatic compound breakdown in the fungal system. Fungal strains capable of utilizing PA as their sole carbon source were isolated using a minimal solid medium supplemented with 0.5 % PA. Subsequent cultivation in minimum liquid medium revealed that selected fungal strains, including Trametes versicolor TV0876 and TV3295, Paecilomyces hepiali PH4477, and Akanthomyces muscarius AM1091, efficiently removed PA within 24 h. HPLC analysis of culture supernatants from various fungal strains revealed a time-dependent accumulation of 2-hydroxyphenylacetate (2-HPA) and 4-hydroxyphenylacetate (4-HPA), two key major metabolic products primarily found in ascomycetes and basidiomycetes, respectively. This suggests that the first hydroxylation of PA is catalyzed by two distinct hydroxylases, one for each fungal group. Furthermore, fungal species that make 4-HPA also produce phenylethanol (PE), indicating a distinct catabolic mechanism to remove PA by direct reduction of PA to PE. A. muscarius AM1091, identified as the most efficient PA degrader in this study, was studied further to determine the biochemical pathway of PA degradation. RNA-Seq and RT-PCR analyses of AM1091 revealed two oxidative enzyme genes, CYP1 and DIO4, upregulated in the presence of PA. Targeted disruption utilizing preassembled Cas9-gRNA ribonucleoprotein complexes and homologous DNAs harboring the URA3 gene as an auxotrophic marker resulted in the cyp1 and dio4 mutant strains. The cyp1 mutant was incapable of converting PA to 2-HPA, indicating its involvement in the C2 hydroxylation, whereas the dio4 mutant was unable to degrade 2,5-dihydroxyphenylacetate (2,5-DHPA), resulting in the accumulation of 2,5-DHPA. Our findings indicate that A. muscarius AM1091 degrades PA through the activities of CYP1 and DIO4 for the C2 hydroxylation and subsequent ring-opening reactions, respectively.

Diastereomeric pure pyrazolyl-indolyl dihydrofurans: Unveiling isomeric selectivity in antibacterial action via in vitro and in silico insights

Developing pure diastereoisomeric molecular hybrids for the selective inhibition of bacterial growth opened new avenues for combating the ever-increasing microbial resistance. Considering this, a series of diastereoisomeric pure pyrazolyl-dihydrofurans (7a-7y) were synthesized and characterized using NMR, LCMS, and X-ray crystallography. DFT based method was used to explore the configurational stability of cis over trans isomeric form. Considering 7a and 8a as representative isomeric forms with same structural framework, the difference in their bio-efficacy against bacterial and fungal strains was assessed using serial dilution method. The relatively high inhibition of bacterial growth by the cis isomeric form (7a) (MIC = 1.562 µg/mL), amoxicillin (MIC = 3.125 µg/mL) inspired us to broaden the substrate scope for synthesizing a series of pure diastereoisomeric cis forms as selective anti-bacterial agents. However, both the isomers displayed antifungal activity less than the standard drug (Fluconazole) employed in the study. All the reactions proceeded smoothly and yielded a diverse array of dihydrofuran derivatives. The developed synthetics were found to be non-cytotoxic against mouse fibroblast cells and didn’t affect the seed germination of Brassica nigra seeds when treated at a concentration of 1 mg/mL. The experimentally determined in vitro results were further validated using in silico molecular docking and dynamics studies.

Novel Co(II), Cu(II) and Zn(II) complexes derived from thiazole containing Schiff base ligands: Green synthesis, antimicrobial and anti-cancer studies

Six novel metal complexes of Co(II), Cu(II), and Zn(II) were synthesized using a microwave-assisted method. These complexes were derived from Schiff base ligands prepared through the condensation of 4-(p-fluorophenyl)-5-methyl-2-aminothiazole with 3-methoxysalicyldehyde (L1) and 2-hydroxynaphthaldehyde (L2). Characterization techniques, including UV–vis, IR, NMR, and HR-MS confirm the tridentate nature of Co(II), Zn(II), and bidentate nature of Cu(II) complexes. Elemental and thermogravimetric analysis showed 1:2 (M: L) stoichiometry in metal complexes. Meanwhile, magnetic susceptibility and powder X-ray diffraction express the geometry and crystal systems of complexes. The antimicrobial analysis of the synthesized compounds indicates Cu(II) complexes have greater potency against gram-positive bacterial and fungal strains while Zn(II) complexes show a greater zone of inhibition against gram-negative bacterial strains. Moreover, MIC values of the most potent complexes [Cu(L2)2] and [Zn(L2)2] range from 190-375 µg/mL. Furthermore, ligands and their complexes have been tested for anticancer potential against MDA-MB 231 and A549 cell lines by MTT assay. [Zn(L1)2] and [Zn(L2)2] complexes were found to have potent activity with IC50 values of 16.7 µM, 24.1 µM, and 8.7 µM, 13.4 µM respectively on MDA-MB-231 and A549 cell lines compared to other complexes and parent ligands. Molecular docking study performed using the PyRx 0.8 (with RF-Score V2 scoring function) program against EGFR (PDB: 5XGM) showed strong binding interactions which supports the experimental results.

Synthesis, spectroscopic characterization, antimicrobial activity, and computational studies of five and/or six heterocyclic nitrogen rings linked to thienopyrazole moiety

A new series of thienopyrazoles linked to five and/or six heterocyclic nitrogen rings such as pyrazole, triazole-3-thione, oxadiazole, thiazole-2-thione and/or pyrimidone moieties was synthesized starting from 5-amino-1,3-diphenyl-1H-thieno[3,2-c]pyrazole-6-carboxylic acid hydrazide as a key precursor. For the synthesis of new thienopyrazoles 2-13, the carboxylic acid hydrazide derivative 1 served as a crucial intermediary. These novel heterocyclic structures were confirmed by analytical and spectroscopic methods (m.p., IR, 1H-NMR, 13C-NMR, and MS). The new compounds 2-13 were evaluated for their antimicrobial activity to identify new drug candidates to combat diseases caused by microbial strains. Against numerous bacterial and fungal strains, certain derivatives of the produced compounds 9 and 11 demonstrated impressive antimicrobial activity. Besides, molecular docking of the newly compounds was conducted against methicillin-resistant Staphylococcus aureus (MRSA) (2 × 3f.pdb). Among the series, compounds 9 and 11 exhibited the best binding affinity towards the target enzyme -10.1 and -10.5 kcal/mol, respectively. The optimized structures and molecular orbitals of the best docked compounds were predicted using DFT (Density Functional Theory) analysis. Our findings represented that both derivatives 9 and 11 could be utilized for development antimicrobial drug candidates.

Chemical Composition, Antioxidant and Antimicrobial Activities of Anacyclus maroccanus Ball. and Anacyclus radiatus Loisel. Essential Oils Against Multidrug Resistant Microbial Pathogens

Objective: The present study aimed to characterize the phytochemical composition, the antioxidant and antimicrobial potencies of two essential oils obtained from the Moroccan ecotypes A. maroccanus and A. radiatus. Methods: The chemical composition of EOs was analysed by means of Gas Chromatography-Mass Spectroscopy analysis (GC-MS); moreover, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assays ware made to evaluate the antioxidant activity. In addition, disk diffusion method, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were employed to evaluate the antimicrobial activities against some resistant and pathogenic microorganism strains. Results: Results showed that the Anacyclus maroccanus and Anacyclus radiatus EOs were characterized by high amounts of oxygenated monoterpenes and sesquiterpenes, along with radical scavenging and reducing properties. Furthermore, EOs exhibited marked antimicrobial effects against all the tested bacterial and fungal strains. Conclusion: The current findings suggest that EOs from Anacyclus maroccanus and Anacyclus radiatus Moroccan ecotypes are a promising source of natural antioxidants that could be used in the pharmaceutical industry, particularly against drug-resistant microbial strains.