GC-MS-Guided Antimicrobial Defense Responsive Secondary Metabolites from the Endophytic Fusarium solani Isolated from Tinospora cordifolia and Their Multifaceted Biological Properties.

Medicinal plants are hosts to an infinite number of microorganisms, commonly referred to as endophytes which are rich in bioactive metabolites yielding favorable biological activities. The endophytes are known to have a profound impact on their host plant by promoting the accumulation of secondary metabolites which are beneficial to humankind. In the present study, the fungal endophyte, Fusarium solani (ABR4) from the medicinal plant Tinospora cordifolia, was assessed for its bioactive secondary metabolites employing fermentation on a solid rice medium. The crude ABR4 fungal extract was sequentially purified using the solvent extraction method and characterized using different spectroscopic and analytical techniques namely TLC, UV spectroscopic analysis, HRESI-MS, FTIR, and GC-MS analysis. The GC-MS analysis revealed the presence of pyridine, benzoic acid, 4-[(trimethylsilyl)oxy]-trimethylsilyl ester, hexadecanoic acid trimethylsilyl ester, and oleic acid trimethylsilyl ester. The cytotoxic ability of ABR4 was evaluated by MTT assay against lung cancer (A549) and breast cancer (MCF-7) cell lines. The compounds did not exhibit significant cytotoxicity against the tested cell lines. The endophytic ABR4 extract was evaluated for its antimicrobial potential against human pathogens (S. aureus, B. cereus, E. coli, S. typhimurium, P. aeruginosa, and C. albicans) by recording 47 to 54% inhibition. Taken together, the endophytic fungal strain ABR4 demonstrated a remarkable antimicrobial activity against the tested pathogens. Furthermore, the functional metabolites isolated from the endophytic strain ABR4 reveal its broader usage as antimicrobial agents for newer drug development in the pharmaceutical industry.

Trichoderma-Mediated ZnO Nanoparticles and Their Antibiofilm and Antibacterial Activities.

Antimicrobial resistance is a major global health concern and one of the gravest challenges to humanity today. Antibiotic resistance has been acquired by certain bacterial strains. As a result, new antibacterial drugs are urgently required to combat resistant microorganisms. Species of Trichoderma are known to produce a wide range of enzymes and secondary metabolites that can be exploited for the synthesis of nanoparticles. In the present study, Trichoderma asperellum was isolated from rhizosphere soil and used for the biosynthesis of ZnO NPs. To examine the antibacterial activity of ZnO NPs against human pathogens, Escherichia coli and Staphylococcus aureus were used. The obtained antibacterial results show that the biosynthesized ZnO NPs were efficient antibacterial agents against the pathogens E. coli and S. aureus, with an inhibition zone of 3-9 mm. The ZnO NPs were also effective in the prevention of S. aureus biofilm formation and adherence. The current work shows that the MIC dosages of ZnO NPs (25, 50, and 75 µg/mL) have effective antibacterial activity and antibiofilm action against S. aureus. As a result, ZnO NPs can be used as a part of combination therapy for drug-resistant S. aureus infections, where biofilm development is critical for disease progression.

Recent advances in nanoremediation: Carving sustainable solution to clean-up polluted agriculture soils.

Agriculture is one of the foremost significant human activities, which symbolizes the key source for food, fuel and fibers. This activity results in a lot of ecological harms particularly with the excessive usage of chemical fertilizers and pesticides. Different agricultural practices have remained industrialized to advance food production, due to the growth in the world population and to meet the food demand through the routine use of more effective fertilizers and pesticides. Soil is intensely embellished by environmental contamination and it can be stated as "universal incline." Soil pollution usually occurs from sewage wastes, accidental discharges or as byproducts of chemical residues of unrestrained production of numerous materials. Soil pollution with hazardous materials alters the physical, chemical, and biological properties, causing undesirable changes in soil fertility and ecosystem. Engineered nanomaterials offer various solutions for remediation of contaminated soils. Engineered nanomaterial-enable technologies are able to prevent the uncontrolled release of harmful materials into the environment along with capabilities to combat soil and groundwater borne pollutants. Currently, nanobiotechnology signifies a hopeful attitude to advance agronomic production and remediate polluted soils. Studies have outlined the way of nanomaterial applications to restore the eminence of the environment and assist the detection of polluted sites, along with potential remedies. This review focuses on the latest developments in agricultural nanobiotechnology and the tools developed to combat soil or land and or terrestrial pollution, as well as the benefits of using these tools to increase soil fertility and reduce potential toxicity.

Ameliorated Antibacterial and Antioxidant Properties by Trichoderma harzianum Mediated Green Synthesis of Silver Nanoparticles.

Biosynthesis of silver nanoparticles using beneficial Trichoderma harzianum is a simple, eco-friendly and cost-effective route. Secondary metabolites secreted by T. harzianum act as capping and reducing agents that can offer constancy and can contribute to biological activity. The present study aimed to synthesize silver nanoparticles using T. harzianum cell filtrate and investigate different bioactive metabolites based on LC-MS/MS analysis. The synthesized silver nanoparticles (AgNPs) from T. harzianum were characterized by ultraviolet-visible spectrophotometry, Fourier transform infrared spectrometry (FT-IR), energy-dispersive spectroscopy (EDS), dynamic light scattering (DLS), X-ray powder diffraction (XRD) and scanning electron microscopy (SEM). The surface plasmon resonance of synthesized particles formed a peak centered near 438 nm. The DLS study determined the average size of AgNPs to be 21.49 nm. The average size of AgNPs was measured to be 72 nm by SEM. The cubic crystal structure from XRD analysis confirmed the synthesized particles as silver nanoparticles. The AgNPs exhibited remarkable antioxidant properties, as determined by DPPH and ferric reducing antioxidant power (FRAP) assay. The AgNPs also exhibited broad-spectrum antibacterial activity against two Gram-positive bacteria (S. aureus and B. subtilis) and two Gram-negative bacteria (E. coli and R. solanacearum). The minimum inhibitory concentration (MIC) of AgNPs towards bacterial growth was evaluated. The antibacterial activity of AgNPs was further confirmed by fluorescence microscopy and SEM analysis.

In vitro therapeutic evaluation of nanoliposome loaded with Xyloglucans polysaccharides from Tamarindus flower extract.

Nanoparticles are interesting area of research developed for several diagnostic and therapeutic applications. Tamarind flower extract is rich in Xyloglucan, a starch like polysaccharide which promotes proliferation and various application areas like drug-delivery technology. In recent years researchers are evaluating nanoliposome using in vitro and in vivo studies to discover their biomedical applications. Considering the importance and feasibility of nanoliposome, the present study is focused on synthesis of liposomes via biological method. The biological molecules of Tamarindus indica flower were used for the synthesis of nanoliposome. The synthesized Tamarindus indica flower extract lipid nanoparticles (TifeLiNPs) loaded with xyloglucans were characterized and evaluated for therapeutic applications (antibacterial, antioxidant, antidiabetic, anticancer and anti-inflammatory activities) under in vitro condition. UV-Vis spectral analysis revealed the emission of peak at 232 nm. Further, the chemical characterization using FTIR revealed the presence of components in the functional group. EDX analysis exhibited the presence of O, Na, P and Cl, while DLS confirmed bilayer formation of xyloglucan and liposomes with uniform size (70-80 nm) and spherical shape. The Physicochemical characterization of tamarind flower extract for its chemical composition revealed the presence of carbohydrates, alkaloids, terpenoids, glycosides, saponins, tannins and flavonoids in confirmatory test. Presence of carbohydrate polymers such as rhamnose, arabinose, galactose, glucose and xylose revealed using high performance anion exchange (HPAE) chromatography under basic conditions on an ion chromatographic system were measured using Pulsed Amperometric Detection (PAD). The synthesized nanoliposome evaluated against Gram negative and Gram positive bacteria showed potential antibacterial activity. TifeLiNPs demonstrated significant in vitro antioxidant potential, antidiabetic, anti-cancer and anti-inflammatory activity. Overall, the present study exhibited the potential application of TifeLiNPs for biomedical purposes.

Bioactive compounds guided diversity of endophytic fungi from Baliospermum montanum and their potential extracellular enzymes.

Baliospermum montanum (Willd.) Muell. Arg, a medicinal plant distributed throughout India from Kashmir to peninsular-Indian region is extensively used to treat jaundice, asthma, and constipation. In the current study, 203 endophytic fungi representing twenty-nine species were isolated from tissues of B. montanum. The colonization and isolation rate of endophytes were higher in stem followed by seed, root, leaf and flower. The phytochemical analysis revealed 70% endophytic isolates showed alkaloids and flavonoids, 13% were positive for phenols, saponins and terpenoids. Further, these endophytes produced remarkable extracellular enzymes such as amylase, cellulase, phosphates, protease and lipase. The most promisive three endophytic fungi were identified by ITS region and secreted metabolites were identified by gas chromatography-mass spectrometry (GC-MS/MS). The GC-MS profile detected twenty-five bioactive compounds from ethyl acetate extracts. Among endophytic fungi, Trichoderma reesei isolated from flower exhibited nine bioactive compounds namely, 2-Cyclopentenone, 2-(4-chloroanilino)-4-piperidino, Oxime-methoxy-Phenyl, Methanamine N-hydroxy-N-methyl, Strychane, Cyclotetrasiloxane, Octamethyl and 1-Acetyl-20a-hydroxy-16-methylene. The endophyte, Aspergillus brasiliensis isolated from root and Fusarium oxysporum isolated from seed produced nine and seven bioactive compounds, respectively. Overall, a significant contribution of bioactive compounds was noticed from the diverse endophytic fungi associated with B. montanum and could be explored for development of novel drug with commercial values.

GC-MS analysis of phytoconstituents from Amomum nilgiricum and molecular docking interactions of bioactive serverogenin acetate with target proteins.

Amomum nilgiricum is one of the plant species reported from Western Ghats of India, belonging to the family Zingiberaceae, with ethno-botanical values, and is well-known for their ethno medicinal applications. In the present investigation, ethyl acetate and methanol extracts of A. nilgiricum were analyzed by Fourier transform infrared spectrometer (FTIR) and gas chromatography-mass spectrometry (GC-MS) to identify the important functional groups and phytochemical constituents. The FTIR spectra revealed the occurrence of functional characteristic peaks of aromatic amines, carboxylic acids, ketones, phenols and alkyl halides group from leaf and rhizome extracts. The GC-MS analysis of ethyl acetate and methanol extracts from leaves, and methanol extract from rhizomes of A. nilgiricum detected the presence of 25 phytochemical compounds. Further, the leaf and rhizome extracts of A. nilgiricum showed remarkable antibacterial and antifungal activities at 100 mg/mL. The results of DPPH and ferric reducing antioxidant power assay recorded maximum antioxidant activity in A. nilgiricum methanolic leaf extract. While, ethyl acetate leaf extract exhibited maximum α-amylase inhibition activity, followed by methanolic leaf extract exhibiting aldose reductase inhibition. Subsequently, these 25 identified compounds were analyzed for their bioactivity through in silico molecular docking studies. Results revealed that among the phytochemical compounds identified, serverogenin acetate might have maximum antibacterial, antifungal, antiviral, antioxidant and antidiabetic properties followed by 2,4-dimethyl-1,3-dioxane and (1,3-13C2)propanedioic acid. To our best knowledge, this is the first description on the phytochemical constituents of the leaves and rhizomes of A. nilgiricum, which show pharmacological significance, as there has been no literature available yet on GC-MS and phytochemical studies of this plant species. The in silico molecular docking of serverogenin acetate was also performed to confirm its broad spectrum activities based on the binding interactions with the antibacterial, antifungal, antiviral, antioxidant and antidiabetic target proteins. The results of the present study will create a way for the invention of herbal medicines for several ailments by using A. nilgiricum plants, which may lead to the development of novel drugs.

Detection and Characterization of Antibacterial Siderophores Secreted by Endophytic Fungi from Cymbidium aloifolium.

Endophytic fungi from orchid plants are reported to secrete secondary metabolites which include bioactive antimicrobial siderophores. In this study endophytic fungi capable of secreting siderophores were isolated from Cymbidium aloifolium, a medicinal orchid plant. The isolated extracellular siderophores from orchidaceous fungi act as chelating agents forming soluble complexes with Fe3+. The 60% endophytic fungi of Cymbidium aloifolium produced hydroxamate siderophore on CAS agar. The highest siderophore percentage was 57% in Penicillium chrysogenum (CAL1), 49% in Aspergillus sydowii (CAR12), 46% in Aspergillus terreus (CAR14) by CAS liquid assay. The optimum culture parameters for siderophore production were 30 °C, pH 6.5, maltose and ammonium nitrate and the highest resulting siderophore content was 73% in P. chrysogenum. The total protein content of solvent-purified siderophore increased four-fold compared with crude filtrate. The percent Fe3+ scavenged was detected by atomic absorption spectra analysis and the highest scavenging value was 83% by P. chrysogenum. Thin layer chromatography of purified P. chrysogenum siderophore showed a wine-colored spot with Rf value of 0.54. HPLC peaks with Rts of 10.5 and 12.5 min were obtained for iron-free and iron-bound P. chrysogenum siderophore, respectively. The iron-free P. chrysogenum siderophore revealed an exact mass-to-charge ratio (m/z) of 400.46 and iron-bound P. chrysogenum siderophore revealed a m/z of 453.35. The solvent-extracted siderophores inhibited the virulent plant pathogens Ralstonia solanacearum, that causes bacterial wilt in groundnut and Xanthomonas oryzae pv. oryzae which causes bacterial blight disease in rice. Thus, bioactive siderophore-producing endophytic P. chrysogenum can be exploited in the form of formulations for development of resistance against other phytopathogens in crop plants.

Mycosynthesis of ZnO Nanoparticles Using Trichoderma spp. Isolated from Rhizosphere Soils and Its Synergistic Antibacterial Effect against Xanthomonas oryzae pv. oryzae.

The Plant Growth Promoting Fungi (PGPF) is used as a source of biofertilizers due to their production of secondary metabolites and beneficial effects on plants. The present work is focused on the co-cultivation of Trichoderma spp. (T. harzianum (PGT4), T. reesei (PGT5) and T. reesei (PGT13)) and the production of secondary metabolites from mono and co-culture and mycosynthesis of zinc oxide nanoparticles (ZnO NPs), which were characterized by a UV visible spectrophotometer, Powder X-ray Diffraction (PXRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM) with Energy Dispersive Spectroscopy (EDAX) and Transmission Electron Microscope (TEM) and Selected Area (Electron) Diffraction (SAED) patterns. The fungal secondary metabolite crude was extracted from the mono and co-culture of Trichoderma spp. And were analyzed by GC-MS, which was further subjected for antibacterial activity against Xanthomonas oryzae pv. Oryzae, the causative organism for Bacterial Leaf Blight (BLB) in rice. Our results showed that the maximum zone of inhibition was recorded from the co-culture of Trichoderma spp. rather than mono cultures, which indicates that co-cultivation of beneficial fungi can stimulate the synthesis of novel secondary metabolites better than in monocultures. ZnO NPs were synthesized from fungal secondary metabolites of mono cultures of Trichoderma harzianum (PGT4), Trichoderma reesei (PGT5), Trichoderma reesei (PGT13) and co-culture (PGT4 + PGT5 + PGT13). These ZnO NPs were checked for antibacterial activity against Xoo, which was found to be of a dose-dependent manner. In summary, the biosynthesized ZnO NPs and secondary metabolites from co-culture of Trichoderma spp. are ecofriendly and can be used as an alternative for chemical fertilizers in agriculture.

Endophytic Fungi-Alternative Sources of Cytotoxic Compounds: A Review.

Cancer is a major cause of death worldwide, with an increasing number of cases being reported annually. The elevated rate of mortality necessitates a global challenge to explore newer sources of anticancer drugs. Recent advancements in cancer treatment involve the discovery and development of new and improved chemotherapeutics derived from natural or synthetic sources. Natural sources offer the potential of finding new structural classes with unique bioactivities for cancer therapy. Endophytic fungi represent a rich source of bioactive metabolites that can be manipulated to produce desirable novel analogs for chemotherapy. This review offers a current and integrative account of clinically used anticancer drugs such as taxol, podophyllotoxin, camptothecin, and vinca alkaloids in terms of their mechanism of action, isolation from endophytic fungi and their characterization, yield obtained, and fungal strain improvement strategies. It also covers recent literature on endophytic fungal metabolites from terrestrial, mangrove, and marine sources as potential anticancer agents and emphasizes the findings for cytotoxic bioactive compounds tested against specific cancer cell lines.

Characterisation and bioactivity of oosporein produced by endophytic fungus Cochliobolus kusanoi isolated from Nerium oleander L.

Bioactive compounds comprising secondary metabolites produced by endophytic fungi have wide applications in pharmacology and agriculture. Isolation, characterisation and evaluation of biological activities of secondary metabolites were carried out from Cochliobolus kusanoi an endophytic fungus of Nerium oleander L. The fungus was identified based on 18S rDNA sequence analysis. There are no reports available on the compounds of C.kusanoi hence, antimicrobial metabolite produced by this fungus was extracted and purified by fractionation using hexane, diethyl ether, dichloromethane, ethyl acetate and methanol. Out of all the solvent fractions, the methanol fraction exhibited better antimicrobial activity which was further purified and characterised as oosporein. Oosporein from C.kusanoi exhibited broad spectrum in vitro antimicrobial, antioxidant and cytotoxic activities. The characterisation and antioxidant activity of oosporein from C. kusanoi are reported for the first time.