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Microorganisms
Special Issues
Endophytic Fungus as Producers of New and/or Bioactive Substances
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Endophytic Fungus as Producers of New and/or Bioactive Substances

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Environmental Microbiology".

Deadline for manuscript submissions: 31 August 2025 | Viewed by 7773

Special Issue Editor

Prof. Dr. Cecilia Veronica Nunez

E-Mail Website
Guest Editor
Bioprospection and Biotechnology Laboratory, National Institute of Amazonian Research, Manaus 69067-375, Brazil
Interests: endophytic fungus; bioactive natural products; phytochemistry; in vitro plant cell culture

Special Issue Information

Dear Colleagues,

In recent years, fungi have attracted more attention from natural product chemists because of their great potential for producing new chemical entities, as well as bioactive substances. They can also produce substances similar to those produced by their host plant, which can be very useful in an industrial application. Sometimes, they may produce other molecules with very different chemical skeletons. Thus, this Special Issue will highlight fungi as producers of natural products (especially micromolecules, such as terpenes, flavonoids, alkaloids, etc.), though other biomolecules are also welcome. This Special Issue of Microorganisms will also provide a scientific platform for scientists performing chemical and biological studies related to endophytic fungus. Original studies and reviews are particularly welcome.

Prof. Dr. Cecilia Veronica Nunez
Guest Editor

Manuscript Submission Information

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Keywords

  • endophytic fungus
  • natural products
  • terpenes
  • alkaloids
  • flavonoids
  • peptides
  • nuclear magnetic resonance
  • bioactivity

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Published Papers (7 papers)

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Research

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13 pages, 456 KiB  
Article
5,6-Dihydro-5,6-Epoxymultiplolide A, Cytosporone C, and Uridine Production by Diaporthe hongkongensis, an Endophytic Fungus from Minquartia guianensis
by Andrei da Silva Alexandre, Luana Lopes Casas, David Ribeiro da Silva and Cecilia Veronica Nunez
Microorganisms 2025, 13(4), 792; https://doi.org/10.3390/microorganisms13040792 - 31 Mar 2025
Viewed by 291
Abstract
Endophytic fungi are valuable sources of bioactive secondary metabolites, with potential applications in pharmaceutical and agricultural fields. This study investigates the metabolic potential of Diaporthe hongkongensis, an endophytic fungus isolated from Minquartia guianensis. To date, no secondary metabolites have been identified [...] Read more.
Endophytic fungi are valuable sources of bioactive secondary metabolites, with potential applications in pharmaceutical and agricultural fields. This study investigates the metabolic potential of Diaporthe hongkongensis, an endophytic fungus isolated from Minquartia guianensis. To date, no secondary metabolites have been identified from this species, highlighting the novelty of this research and its contribution to understanding the chemical diversity of endophytic fungi. The fungus was cultivated on parboiled rice under static and dark conditions for 28 days, leading to the isolation of the following three compounds: 5,6-dihydro-5,6-epoxymultiplolide A (1), cytosporone C (2), and uridine (3). Structural identification was carried out using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. The results revealed the metabolic versatility of D. hongkongensis, as demonstrated by its ability to produce structurally diverse substances with biological relevance. Hence, it describes the first isolation of secondary metabolites from the endophytic fungus D. hongkongensis, marking a significant step in understanding its chemical profile. The identification of a known antifungal compound and a lactone derivative underscores the biosynthetic potential of this endophytic fungus, while the isolation of a nucleoside expands the chemical repertoire of fungal metabolites, suggesting possible roles in cellular metabolism and stress adaptation. These findings highlight the role of endophytic fungi as prolific sources of structurally diverse and potentially bioactive natural products, supporting further exploration of their biotechnological applications. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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17 pages, 3166 KiB  
Article
Exploring the Antimicrobial, Antioxidant and Extracellular Enzymatic Activities of Culturable Endophytic Fungi Isolated from the Leaves of Kirkia acuminata Oliv
by Sagwadi Kubayi, Raymond Tshepiso Makola and Khumiso Dithebe
Microorganisms 2025, 13(3), 692; https://doi.org/10.3390/microorganisms13030692 - 19 Mar 2025
Viewed by 375
Abstract
Fungal endophytes of medicinal plants produce diverse secondary metabolites and extracellular enzymes with therapeutic and biotechnological potential. However, the biological and biotechnological potential of fungal endophytes from South African medicinal plants remain relatively underexplored. In this study, the antimicrobial, antioxidant, anti-inflammatory and extracellular [...] Read more.
Fungal endophytes of medicinal plants produce diverse secondary metabolites and extracellular enzymes with therapeutic and biotechnological potential. However, the biological and biotechnological potential of fungal endophytes from South African medicinal plants remain relatively underexplored. In this study, the antimicrobial, antioxidant, anti-inflammatory and extracellular enzymatic capabilities of five fungal endophytes previously isolated from the leaves of Kirkia acuminata Oliv. were investigated. Sequencing of the internal transcribed spacer (ITS) regions revealed that the isolates belonged to the genera Setosphaeria, Diaporthe and Corynespora. The broth micro-dilution assay and the Folin–Ciocalteau reagent method were used to assess the antibacterial activity and the total phenolic content (TPC) of the fungal endophytes’ ethyl acetate crude extracts (CEs), respectively. The antioxidant activity was assessed using the ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assays. The influence of the CE of the Setosphaeria rostrata KaL-4 on the viability and LPS-induced interleukin-6 (IL-6) production in Raw 264.7 macrophages was assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and an ELISA, respectively. The ability of the isolates to produce extracellular proteases, laccases and peroxidases was also determined. The CEs displayed antimicrobial activity with MICs ranging from 0.63 to 1.25 mg/mL and reducing power and scavenging activity ranging from 40% to 18% and from 60% to 48%, respectively. The S. rostrata KaL-4 CE possessed the highest TPC and demonstrated dose-dependent cytotoxicity. The CE further demonstrated a significant reduction in IL-6 production at a concentration of 0.75 µg/mL. Only one isolate demonstrated the ability to produce proteases with an enzymatic index (EI) of 0.66, while laccases (EI range of 0.14 to 1.15) and peroxidases were produced by all of the isolates. These findings suggest that fungal endophytes from South African medicinal plants are promising sources of bioactive compounds and industry-significant extracellular enzymes. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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14 pages, 2524 KiB  
Article
Diversity of Endophytic Fungi Isolated from Prunus yedoensis and Their Antifungal Activity Against Wood Decay Fungi
by Misong Kim, Manh Ha Nguyen, Sanggon Lee, Wonjong Han, Minyoung Kim, Hyeongguk Jeon, Jinheung Lee, Sangtea Seo, Namkyu Kim and Keumchul Shin
Microorganisms 2025, 13(3), 617; https://doi.org/10.3390/microorganisms13030617 - 7 Mar 2025
Viewed by 505
Abstract
Endophytic fungi play a vital role in protecting and promoting the growth of their host plants. The diversity of fungal endophytes has been documented across different host plant species and varies depending on factors such as the species of the host, ecological conditions, [...] Read more.
Endophytic fungi play a vital role in protecting and promoting the growth of their host plants. The diversity of fungal endophytes has been documented across different host plant species and varies depending on factors such as the species of the host, ecological conditions, and the health status of the plant. We isolated endophytic fungi from Prunus yedoensis trees with different decay rates. A total of 31 species were isolated from decayed trees, while 33 species were obtained from healthy trees. The number of endophytic fungi exhibiting antifungal activities against wood decay fungi was higher in healthy trees, with 10 species showing activity compared to only 1 species from decayed trees. Endophytic fungus Fusarium acuminatum (BEN48) had the highest inhibition rates against Trametes versicolor, Ganoderma gibbosum, and Vanderbylia fraxinea. Heating conditions did not significantly affect the inhibitory ability of the culture filtrate from BEN48 on wood decay fungi. At 50% concentration, the inhibitory abilities of the culture filtrates against Trametes versicolor, Ganoderma gibbosum, and Vanderbylia fraxinea were 96.5%, 64.1%, and 92.7%, respectively. The inhibitory effects against Trametes versicolor decreased at concentrations of 30% and 10%, resulting in inhibition rates of 83.7% and 50.8%, respectively. For Ganoderma gibbosum, the inhibition rate reduced to 52.6% at 30% concentration and 24.5% at 10% concentration. For Vanderbylia fraxinea, there was no significant difference between the 30% and 10% concentrations, and the inhibition rates for both concentrations were high, measuring 89.9% and 88.8%, respectively. Hence, Fusarium acuminatum (BEN48) has promise as a biocontrol agent for managing wood decay fungi. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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17 pages, 2607 KiB  
Article
Identification and Pathogenicity of Fusarium Species from Herbaceous Plants on Grassland in Qiaojia County, China
by Yanzhu Gao, Zhixiao Zhang, Mei Ji, Sangzi Ze, Haodong Wang, Bin Yang, Lianrong Hu and Ning Zhao
Microorganisms 2025, 13(1), 113; https://doi.org/10.3390/microorganisms13010113 - 8 Jan 2025
Viewed by 1094
Abstract
The Fusarium species is an important plant pathogen that can cause plant diseases in grassland, leading to the degradation of grassland quality. However, the morphology of Fusarium is greatly affected by environmental factors, which makes it difficult to identify its species. In addition, [...] Read more.
The Fusarium species is an important plant pathogen that can cause plant diseases in grassland, leading to the degradation of grassland quality. However, the morphology of Fusarium is greatly affected by environmental factors, which makes it difficult to identify its species. In addition, the pathogenic ability of different Fusarium species in plants has not been fully studied. In this study, Fusarium isolates were obtained from grassland herbaceous plants via tissue separation. Through morphological means and based on ITS, RPB2, and TEF-1 gene sequences, we compared and constructed polygenic phylogenetic trees to classify and identify the Fusarium species. In addition, the pathogenicity of different Fusarium species was also analyzed. The results showed that a total of 24 Fusarium strains were successfully isolated from grassland, from which ten species were identified: F. flagelliforme, F. longifundum, F. clavum, F. scirpi, F. ipomoeae, F. oxysporum, etc. and were included in four complexes: Fusarium incarnatum-equiseti species complex (FIESC), Fusarium oxysporum species complex (FOSC), Fusarium tricinctum species complex (FTSC), and Fusarium sambucinum species complex (FSAMSC). Pathogenicity tests demonstrated that except for F. ipomoeae QJ5211, F. sambucinum QJ203, and F. acuminatum QJ1662, other Fusarium species had different degrees of pathogenic ability. This is the first study that discusses the effect of Fusarium on grassland disease control in this area. This study further provides clear pathogen information for the prevention and control of grassland diseases. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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24 pages, 1705 KiB  
Article
In Vitro Antioxidant and Antibacterial Activities of Ethyl Acetate Extracts of Ziziphus lotus Leaves and Five Associated Endophytic Fungi
by Amel Ghazi-Yaker, Bart Kraak, Jos Houbraken, El-hafid Nabti, Cristina Cruz, Noria Saadoun and Karim Houali
Microorganisms 2024, 12(12), 2671; https://doi.org/10.3390/microorganisms12122671 - 23 Dec 2024
Cited by 1 | Viewed by 1609
Abstract
The exploration of new pharmacological compounds from endophytic fungi offers infinite possibilities. The aim of this study was to evaluate the antibacterial and antioxidant activities of extracts from the leaves of Ziziphus lotus and five of its endophytic fungi and investigate the chemical [...] Read more.
The exploration of new pharmacological compounds from endophytic fungi offers infinite possibilities. The aim of this study was to evaluate the antibacterial and antioxidant activities of extracts from the leaves of Ziziphus lotus and five of its endophytic fungi and investigate the chemical diversity of the secondary metabolites produced. Isolated, purified, and molecularly identified endophytes and plant leaves were subjected to ethyl acetate extraction. The antibacterial potential of the extracts was assessed by the disc diffusion method against five bacterial strains: Staphylococcus aureus ATCC 25923; Staphylococcus aureus MU50; Enterococcus faecalis WDCM00009; Escherichia coli ATCC 25922; and Pseudomonas aeruginosa ATCC 27853. DPPH and reducing power tests were performed to assess antioxidant potential. GC–MS analysis was used to identify volatile compounds in extracts. Fungal endophytes were identified as Aspergillus cavernicola, Aspergillus persii, Alternaria alternata, Cladosporium asperlatum, and Fusarium incarnatum–equiseti complex, with respective accession numbers DTO 412-G6, DTO 412-I5, DTO 413-E7, DTO 412-G4, and DTO 414-I2. GC–MS analysis revealed a large number of bioactive compounds. All extracts showed antibacterial activity against at least two of the bacteria tested, and most showed antioxidant activity. The Aspergillus cavernicola extract stood out for its higher phenolic content and higher antioxidant and antibacterial activities in all tests. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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13 pages, 1249 KiB  
Article
In Vitro and In Vivo Antitumor Activity of Lophocereus marginatus (DC.) S. Arias & Terrazas Endophytic Aspergillus versicolor and Metarhizium anisopliae Extracts Against the Murine Lymphoma L5178Y-R
by Diana Laura Clark-Pérez, César Iván Romo-Sáenz, Jesica María Ramírez-Villalobos, Patricia Tamez-Guerra, Diana Caballero-Hernández, Ana Laura Delgado-Miranda, Andrés García, Joel Horacio Elizondo-Luevano, Cristina Rodríguez-Padilla and Ricardo Gomez-Flores
Microorganisms 2024, 12(11), 2310; https://doi.org/10.3390/microorganisms12112310 - 13 Nov 2024
Cited by 1 | Viewed by 1115
Abstract
Cancer belongs to a group of diseases characterized by uncontrolled cell growth. The search for new effective treatments for cancer has led to the discovery of different molecules from plants, bacteria, and fungi with pharmacological use. Plant endophytic fungi are large producers of [...] Read more.
Cancer belongs to a group of diseases characterized by uncontrolled cell growth. The search for new effective treatments for cancer has led to the discovery of different molecules from plants, bacteria, and fungi with pharmacological use. Plant endophytic fungi are large producers of metabolites with antitumor properties. We aimed to evaluate the in vitro and in vivo antitumor potential of extracts from Lophocereus marginatus endophytic fungi. We obtained ethyl acetate and hexane extracts from the L. marginatus endophytes Metarhizium anisopliae and Aspergillus versicolor and evaluated their antitumor activity against murine L5178Y-R lymphoma cells and human peripheral blood mononuclear cells, using the 3-(4,5-dimethylthiazol-2-yl)-2-diphenyltetrazolium bromide reduction colorimetric technique. M. anisopliae and A. versicolor ethyl acetate extracts showed IC50 values of 9.168 ± 1.21 μg/mL and 13.51 ± 1.62, respectively, and selectivity indices > 30. We also observed that the maximum tolerated dose (100 mg/kg) of ethyl acetate extracts and the vehicle in BALB/c mice did not cause hepatotoxicity. In addition, we evaluated the effects of ethyl acetate extracts on survival and tumor volume in the L5178Y-R lymphoma tumor model. An increase in survival (17 d) was observed in mice treated with A. versicolor extract. Furthermore, it did not increase tumor volume during 10 d, as compared with the control groups without treatment, vehicle, and M. anisopliae extract, which had a maximum survival of 10 d. A. versicolor ethyl acetate extract showed in vitro and in vivo antitumor activity against lymphoma L5178Y-R, increasing mice survival. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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Review

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22 pages, 1919 KiB  
Review
Endophytic Fungi Co-Culture: An Alternative Source of Antimicrobial Substances
by Lucas Silva Tironi, Lucilene Bento Carletto, Eliane Oliveira Silva, Jan Schripsema and Jaine Honorata Hortolan Luiz
Microorganisms 2024, 12(12), 2413; https://doi.org/10.3390/microorganisms12122413 - 25 Nov 2024
Cited by 4 | Viewed by 2063
Abstract
Antimicrobial resistance is becoming a critical issue due to the widespread and indiscriminate use of antibiotics and antifungals to treat common infections, leading to a growing shortage of effective drugs. Moreover, the increase in antimicrobial resistance is enhancing the pathogenicity and virulence of [...] Read more.
Antimicrobial resistance is becoming a critical issue due to the widespread and indiscriminate use of antibiotics and antifungals to treat common infections, leading to a growing shortage of effective drugs. Moreover, the increase in antimicrobial resistance is enhancing the pathogenicity and virulence of various pathogens. Microorganisms are key sources of chemically diverse specialized metabolites, which are produced in the final stages of their growth cycle. These metabolites hold significant value in chemical, pharmaceutical, and agrochemical industries. One of the major challenges researchers face in this field is the frequent isolation of already-known substances when classical protocols are used. To address this, several innovative strategies have been developed. The co-culture approach is a powerful tool for activating silent biosynthetic gene clusters, as it simulates natural microbial environments by creating artificial microbial communities. This method has shown promising results, with new compounds being isolated and the yields of target substances being improved. In this context, this review provides examples of antimicrobial compounds obtained from co-cultures of endophytic fungi, conducted in both liquid and solid media. Additionally, the review discusses the advantages and challenges of the co-culture technique. Significance and Impact of the Study: Microbial co-culture is a valuable strategy for discovering new natural products with antimicrobial activity, as well as for scaling up the production of target substances. This review aims to summarize important examples of endophyte co-cultures and highlights the potential of endophytic fungi co-culture for pharmacological applications. Full article
(This article belongs to the Special Issue Endophytic Fungus as Producers of New and/or Bioactive Substances)
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