Diaporthe species associated with the maritime grass Festuca rubra subsp. pruinosa.

Festuca rubra subsp. pruinosa is a perennial grass growing in sea cliffs where plants are highly exposed to salinity and marine winds, and often grow in rock fissures where soil is absent. Diaporthe species are one of the most abundant components of the root microbiome of this grass and several Diaporthe isolates have been found to produce beneficial effects in their host and other plant species of agronomic importance. In this study, 22 strains of Diaporthe isolated as endophytes from roots of Festuca rubra subsp. pruinosa were characterized by molecular, morphological, and biochemical analyses. Sequences of the nuclear ribosomal internal transcribed spacers (ITS), translation elongation factor 1-α (TEF1), beta-tubulin (TUB), histone-3 (HIS), and calmodulin (CAL) genes were analyzed to identify the isolates. A multi-locus phylogenetic analysis of the combined five gene regions led to the identification of two new species named Diaporthe atlantica and Diaporthe iberica. Diaporthe atlantica is the most abundant Diaporthe species in its host plant, and Diaporthe iberica was also isolated from Celtica gigantea, another grass species growing in semiarid inland habitats. An in vitro biochemical characterization showed that all cultures of D. atlantica produced indole-3-acetic acid and ammonium, and the strains of D. iberica produced indole 3-acetic acid, ammonium, siderophores, and cellulase. Diaporthe atlantica is closely related to D. sclerotioides, a pathogen of cucurbits, and caused a growth reduction when inoculated in cucumber, melon, and watermelon.

Serial Exhaustive Extraction Revealed Antimicrobial and Antioxidant Properties of Platycerium stemaria (Beauv) Desv.

Microbial infections are increasing worldwide, and the widespread emergence of antibiotic-resistant pathogens poses a severe threat to public health. Medicinal plants are well-known sources of bioactive ingredients. This study was designed to determine the antimicrobial and antioxidant activities of extracts from Platycerium stemaria. The serial exhaustive extraction method using a solvent of increasing polarity from nonpolar (hexane) to polar (water) was designed to prepare crude extracts; liquid-liquid partition was used to fractionate of active extracts. The extracts and fractions were screened for antimicrobial activity on bacteria and yeasts using the microdilution method. The antioxidant activity was done using DPPH and FRAP assays. Out of the sixteen extracts screened, four (PsHex, PsH2O(H), PsMeOH(EA), and PsMeOH) exhibited potency with minimal inhibitory concentration (MIC) values ranging from 31.25 to 500 µg/mL. Out of the four extracts, two, including PsMeOH and PsMeOH(EA), exhibited DPPH radical scavenging activity with the antiradical power of 8.94 × 10-5 and 47.96 × 10-5, respectively, and ferric reducing antioxidant power values ranging from 0.34 to 61.53 µg equivalent Vit C/g of extract. The phytochemical screening of the promising crude extracts revealed flavonoids, glycosides, phenols, tannins, terpenoids, saponins, and anthraquinones. This study reports the antimicrobial and antioxidant activities of P. stemaria for the first time. The results showed that the serial exhaustive extraction approach used in this study allowed capturing the antimicrobial and antioxidant metabolites beyond the single extraction, indicating the need for a rigorous choice of an appropriate solvent and method for extracting P. stemaria. Further investigation is needed to characterize the active ingredients present in the promising extracts.

Antibacterial and Mode of Action of Extracts from Endophytic Fungi Derived from Terminalia mantaly, Terminalia catappa, and Cananga odorata.

Emerging drug-resistant bacteria creates an urgent need to search for antibiotics drugs with novel mechanisms of action. Endophytes have established a reputation as a source of structurally novel secondary metabolites with a wide range of biological activities. In the present study, we explore the antibacterial potential of endophytic fungi isolated from different tissues of Terminalia mantaly, Terminalia catappa, and Cananga odorata. The crude ethyl acetate extracts of 56 different endophytic fungi were screened against seven bacterial strains using the broth microdilution method. The antibacterial modes of action of the most active extracts (04) were evaluated using E. coli ATCC 25922 and H. influenzae ATCC 49247 strains. Both the DPPH and FRAP assays were used to investigate their antioxidant activity, and their cytotoxicity against the Vero cell line was evaluated using the MTT assay. Out of the 56 crude extracts tested, about 13% were considered very active, 66% partially active, and 21% nonactive against all tested bacterial strains with MIC values ranging from 0.32 µg/mL to 25 µg/mL. The four more potent extracts (MIC <5 µg/mL) (from Aspergillus sp. N454, Aspergillus sp. N13, Curvularia sp. N101, and Aspergillus sp. N18) significantly lysed the bacteria cells, increased outer membrane permeability, reduced salt tolerance, and inhibited bacterial catalase activity. They exhibited a DPPH free radical scavenging activity with IC50 ranging from 150.71 to 936.08 µg/mL. Three of the four potent extracts were noncytotoxic against the Vero cells line (CC50 > 100 µg/mL). Results from this investigation demonstrated that endophytes from Cameroonian medicinal plants might content potent antibacterial metabolites. The bioguided fractionation of these potent extracts is ongoing to isolate and characterise potential active ingredients.

Bioprospecting endophytic fungi from Fusarium genus as sources of bioactive metabolites.

Endophytic fungi became an attractive source for the discovery of new leads, because of the complexity and the structural diversity of their secondary metabolites. The genus Fusarium comprising about 70 species is extremely variable in terms of genetics, biology, ecology, and consequently, secondary metabolism and have been isolated from countless plants genera from diverse habitats. These endophytic microbes may provide protection and survival strategies in their host plants with production of a repertoire of chemically diverse and structurally unprecedented secondary metabolites reported to exhibit an incredible array of biological activities including antimicrobial, anticancer, antiviral, antioxidants, antiparasitics, immunosuppressants, immunomodulatory, antithrombotic, and biocontrol ability against plants pathogens and nematodes. This review comprehensively highlights over the period 1981-2019, the bioactive potential of metabolites produced by endophytes from Fusarium genus. Abbreviations: AIDS: Acquired immune deficiency syndrome; BAPT: C-13 phenylpropanoid side chain-CoA acyltransferase; CaBr2: Calcium bromide; DBAT: 10-deacetylbaccatin III-10-O-acetyl transferase; DNA: Deoxyribonucleic acid; EI-MS: Electron ionization mass spectrometer; EN: Enniatin; ERK: Extracellular regulated protein kinase; EtOAc: Ethyl acetate; FDA: Food and Drug Administration; GAE/g: Gallic acid equivalent per gram; GC-MS: Gas chromatography-mass spectrometry; HA: Hyperactivation; HCV: Hepatitis C Virus; HCVPR: Hepatitis C Virus protease; HeLa: Human cervical cancer cell line; HIV: Human immunodeficiency viruses; HPLC: High Performance Liquid Chromatography; IAA: Indole-3-acetic acid; IARC: International Agency for Research on Cancer; IC50: Half maximal inhibitory concentration; LC50: Concentration of the compound that is lethal for 50% of exposed population; LC-MS: Liquid chromatography-mass spectrometry; MCF-7: Human breast cancer cell line; MDR: Multidrug-resistant; MDRSA: Multidrug-resistant S. aureus; MFC: Minimum fungicidal concentration; MIC: Minimum inhibitory concentration; MRSA: Multidrug-resistant S. aureus; MTCC: Microbial type culture collection; PBMCs: Peripheral blood mononuclear cells; PCR: Polymerase chain reaction; TB: Tuberculosis; TLC: Thin layer chromatography; TNF: Tumor necrosis factor; WHO: World Health Organization http://www.zoobank.org/urn:lsid:zoobank.org:pub:D0A7B2D8-5952-436D-85C8-C79EAAD1013C.

Recent advances in inducing endophytic fungal specialized metabolites using small molecule elicitors including epigenetic modifiers.

Today when the quest of new lead molecules to supply the development pipeline is driving the course of drug discovery, endophytic fungi with their outstanding biosynthetic potential seem to be highly promising avenues for natural product scientists. However, challenges such as the production of inadequate quantities of compounds, the attenuation or loss of ability of endophytes to produce the compound of interest when grown in culture and the inability of fungal endophytes to express their full biosynthetic potential in laboratory conditions have been the major constraints. These have led to the application of small chemical elicitors that induce epigenetic changes in fungi to activate their silent gene clusters optimizing the amount of metabolites of interest or inducing the synthesis of hitherto undescribed compounds. In this respect small molecular weight compounds which are known to function as inhibitors of histone deacetylase (HDAC), DNA methyltransferase (DNMT) and proteasome have proven their efficacy in enhancing or inducing the production of specialized metabolites by fungi. Moreover, organic solvents, metals and plants extracts are also acknowledged for their ability to cause shifts in fungal metabolism. We highlight the successful studies from the past two decades reporting the ability of structurally diverse small molecular weight compounds to elicit the production of previously undescribed metabolites from endophytic fungi grown in culture. This mini review argues in favor of chemical elicitation as an effective strategy to optimize the production of fungal metabolites and invigorate the pipeline of drug discovery with new chemical entities.

Endophytic Penicillium species and their agricultural, biotechnological, and pharmaceutical applications.

Penicillium genus constituted by over 200 species is one of the largest and fascinating groups of fungi, particularly well established as a source of antibiotics. Endophytic Penicillium has been reported to colonize their ecological niches and protect their host plant against multiples stresses by exhibiting diverse biological functions that can be exploited for countless applications including agricultural, biotechnological, and pharmaceutical. Over the past 2 decades, endophytic Penicillium species have been investigated beyond their antibiotic potential and numerous applications have been reported. We comprehensively summarized in this review available data (2000-2019) regarding bioactive compounds isolated from endophytic Penicillium species as well as the application of these fungi in multiple agricultural and biotechnological processes. This review has shown that a very large number (131) of endophytes from this genus have been investigated so far and more than 280 compounds exhibiting antimicrobial, anticancer, antiviral, antioxidants, anti-inflammatory, antiparasitics, immunosuppressants, antidiabetic, anti-obesity, antifibrotic, neuroprotective effects, and insecticidal and biocontrol activities have been reported. Moreover, several endophytic Penicillium spp. have been characterized as biocatalysts, plant growth promoters, phytoremediators, and enzyme producers. We hope that this review summarizes the status of research on this genus and will stimulate further investigations.

Anti-leishmanial and Anti-inflammatory Agents from Endophytes: A Review.

Leishmaniases and chronic inflammatory diseases are the cause of millions of deaths in the world each year. The treatment of leishmaniasis is facing serious drawbacks particularly due to the limited number of effective medicines, the resistance, and the toxicity of available drugs. On the other hand, many drugs are used for the management of inflammatory disorders. However, the most commonly prescribed although efficient is highly toxic with multiples side effects. New leads compounds for the development of new anti-leishmanial and anti-inflammatory drugs are needed. Over the past decade, several studies on the potential of endophytes to produce bioactive metabolites have been reported. We are presenting in the present review the status of research from 2000 to 2019 on the anti-leishmanial and anti-inflammatory metabolites isolated from endophytes from diverse habitats. An emphasis was put on existing gaps in the literature to inspire and guide future investigations. We hope that this review will help accelerate the drug discovery against leishmaniases and inflammation-associated disorders.

Antiplasmodial potential and GC-MS fingerprint of endophytic fungal extracts derived from Cameroonian Annona muricata.

ETHNOPHARMACOLOGICAL RELEVANCE: Annona muricata (Annonaceae) is a commonly used medicinal plants in Cameroonian traditional medicines to treat various diseases including malaria. Previous studies have shown that extracts from this plant have antiplasmodial activity. AIM OF THE STUDY: This study aimed to explore the endophyic fungi associated with some parts of this plant for their ability to produce antiplasmodial metabolites. MATERIALS AND METHODS: One hundred and fifty-two endophytic fungi isolated from twelve different organs of A. muricata were cultured and the ethyl acetate extracts of conditioned media screened for antiplasmodial activity using the 96-well microtiter plate format SYBR green florescence assay against Chloroquine-sensitive Pf3D7 and Chloroquine-resistant PfINDO/PfDd2 strains of Plasmodium falciparum. RESULTS: Twenty-seven (17.76%) of fungi tested were found to completely inhibit the growth of Plasmodium parasites at 10 µg/mL. The 5.8S rDNA sequencing data revealed the strongly active (IC50 < 2 µg/mL against at least 2 P. falciparum strains) isolates to be Trichoderma afroharzianum AMrb7, Penicillium citrinum AMrb11, Neocosmospora rubicola AMb22, Penicillium tropicum AMb3, Penicillium citrinum AMrb23, Aspergillus versicolor AMb7, and Fusarium sp AMst1. Of these, the extracts from Penicillium citrinum AMrb11 (IC50 0.84-0.93 µg/mL) and Neocosmospora rubicola AMb22 (IC50 0.39-1.92 µg/mL) showed the highest promise against all three plasmodial strains with selectivity indices ranging from 34.71 to 180.97. Dynamic head space GC-MS analysis of ethyl acetate extracts of promising fungi revealed broad-spectrum antimicrobial compounds such as Penicidin, Aromadendrene, Ethyl p-methoxycinnamate, 2-Coumaranone and 2-Methyl resorcinol. CONCLUSION: These results have opened new avenues for discovery of novel antimalarial lead compounds from endophytic fungi associated with Annona muricata - a medicinally important plant.

Conditioned media and organic elicitors underpin the production of potent antiplasmodial metabolites by endophytic fungi from Cameroonian medicinal plants.

Plasmodial resistance to artemisinin-based combination therapies emphasizes the need for new drug development to control malaria. This paper describes the antiplasmodial activity of metabolites produced by endophytic fungi of three Cameroonian plants. Ethyl acetate extracts of fungi cultivated on three different media were tested against Plasmodium falciparum chloroquine-sensitive (Pf3D7) and chloroquine-resistant (PfINDO) strains using the SYBR green florescence assay. Selected endophytes were further grown in potato dextrose broth supplemented with small organic elicitors and their extracts tested for activity. The effect of elicitors on de novo metabolite synthesis was assessed by reverse-phase HPLC. Activity screening of 81 extracts indicated that Aspergillus niger 58 (IC50 2.25-6.69 µg/mL, Pf3D7), Fusarium sp. N240 (IC50 1.62-4.38 µg/mL, Pf3D7), Phomopsis sp. N114 (IC50 0.34-7.26 µg/mL, Pf3D7), and Xylaria sp. N120 (IC50 2.69-6.77 µg/mL, Pf3D7) produced potent extracts when grown in all three media. Further culture of these endophytes in potato dextrose broth supplemented with each of the eight small organic elicitors and subsequent extracts screening indicated the extract of Phomopsis sp. N114 grown with 1% 1-butanol to be highly selective and extremely potent (IC50 0.20-0.33 µg/mL; SI > 666). RPHPLC profiles of extracts of Phomopsis sp. N114 grown with or without 1-butanol showed some peaks of enhanced intensities in the former without any qualitative change in the chromatograms. This study showed the ability of selected endophytes to produce potent and selective antiplasmodial metabolites in varied culture conditions. It also showed how the production of desired metabolites can be enhanced by use of small molecular weight elicitors.