Proliferacorins A-M: Thirteen undescribed acorane sesquiterpenoids isolated from the fungus Fusarium proliferatum.

In this study, 13 previously undescribed acorane sesquiterpenoids, namely, proliferacorins A-M, were isolated from the solid fermentation of Fusarium proliferatum. Their structures and absolute configurations were confirmed via spectroscopic analyses, quantum-chemical NMR calculations with DP4+ probability analyses, ECD calculations and comparisons, and single-crystal X-ray diffraction techniques. Proliferacorins A-E (1-5) have a 7-oxa-tricyclo[6.3.1.01,5]tridecane decorated with a rare ether bridge between C-7 and C-11, while proliferacorin F (6) possesses a 7-oxa-tricyclic[6.4.0.01,5]dodecane skeleton with an unusual ether bond between C-6 and C-11. Proliferacorins C and D (3 and 4) are a pair of isomers at the carbon bridge between C-5 and C-7, whereas proliferacorins H and I (8 and 9) are a pair of spiro carbon isomers. All isolates were tested for their cytotoxic, anti-inflammatory, and immunosuppressive activities.

Chemical Constituents of a Marine-derived Fungus Fusarium oxysporum F0888 and their Antibacterial Activity.

Five new compounds, including four hydroxyphenylacetic acid derivatives, stachylines H-K (1-4), a derivative of hydroxyphenylethanol (5), as well as seven known compounds were obtained from a marine-derived fungus Fusarium oxysporum F0888 isolated from sediments in the South China Sea. The structures and absolute configurations of new compounds were determined by spectroscopic (IR, NMR, and HR-ESI-MS) analyses, comparison of optical rotations, and the modified Mosher's MTPA ester method. Antimicrobial and anti-inflammatory activities of compounds 1-12 were tested. Unfortunately, all of isolated compounds were inactivity.

A novel core-shell up-conversion nanoparticles immunochromatographic assay for the detection of deoxynivalenol in cereals.

Deoxynivalenol (DON) toxin is a type B group of trichothecene mycotoxins mainly originating from specific Fusarium fungi, seriously harming human and livestock health. Herein, a novel core-shell up-conversion nanoparticles immunochromatographic assay (CS-UCNPs-ICA) was developed for deoxynivalenol based on the competitive reaction principle. By exploiting the fluorescence intensity of the T and C lines of CS-UCNPs-ICA, the concentrations of DON were obtained sensitively and precisely under optimized conditions in 5 min with a detection limit of 0.1 ng/mL. The CS-UCNPs-ICA strips only specifically detect DON and its derivatives (3-Ac-DON and 15-Ac-DON), with no cross-reaction with other mycotoxins. The low CV values illustrated a modest intra- and inter-assay variation, confirming the superior precision of this method. In the spiked experiment, the mean recoveries of corn and wheat ranged from 94.74% to 100.90% and 96.21%-104.81%, respectively. Furthermore, the approach generated results that were in good agreement with data from HPLC and ELISA analyses of naturally contaminated feed and cereals, confirming that the significant advantages of proposed strips were their high practicality, rapidness, and simplicity. Therefore, the CS-UCNPs-ICA strips platform serves as a promising candidate for developing new approaches for rapid testing or high throughput screening from DON in food products.

Improving the productivity of gibberellic acid by combining small-molecule compounds-based targeting technology and transcriptomics analysis in Fusarium fujikuroi.

Gibberellic acid (GA3), produced industrially by Fusarium fujikuroi, stands as a crucial plant growth regulator extensively employed in the agriculture filed while limited understanding of the global metabolic network hinders researchers from conducting rapid targeted modifications. In this study, a small-molecule compounds-based targeting technology was developed to increase GA3 production. Firstly, various small molecules were used to target key nodes of different pathways and the result displayed that supplement of terbinafine improved significantly GA3 accumulation, which reached to 1.08 g/L. Subsequently, lipid and squalene biosynthesis pathway were identified as the key pathways influencing GA3 biosynthesis by transcriptomic analysis. Thus, the strategies including in vivo metabolic engineering modification and in vitro supplementation of lipid substrates were adopted, both contributed to an enhanced GA3 yield. Finally, the engineered strain demonstrated the ability to achieve a GA3 yield of 3.24 g/L in 5 L bioreactor when utilizing WCO as carbon source and feed.

Toxicity of the emerging mycotoxins beauvericin and enniatins: A mini-review.

Beauvericin and enniatins, emerging mycotoxins produced mainly by Fusarium species, are natural contaminants of cereals and cereal products. These mycotoxins are cyclic hexadepsipeptides with ionophore properties and their toxicity mechanism is related to their ability to transport cations across the cell membrane. Beauvericin and enniatins are cytotoxic, as they decrease cell viability, promote cell cycle arrest, and increase apoptosis and the generation of reactive oxygen species in several cell lines. They also cause changes at the transcriptomic level and have immunomodulatory effects in vitro and in vivo. Toxicokinetic results are scarce, and, despite its proven toxic effects in vitro, no regulation or risk assessment has yet been performed due to a lack of in vivo data. This mini-review aims to report the information available in the literature on studies of in vitro and in vivo toxic effects with beauvericin and enniatins, which are mycotoxins of increasing interest to animal and human health.

The antifungal metabolites isolated from maize endophytic fungus Fusarium sp. induced by OSMAC strategy.

Six new sesquiterpenes, fusarchlamols A-F (1, 2, 4-7); one new natural product of sesquiterpenoid, methyltricinonoate (3); and ten known compounds were found from Fusarium sp. cultured in two different media by the one strain many compounds strategy. The compounds (1, 2, and 4-11) were isolated from Fusarium sp. in PDB medium, and compounds (3-5, 8, and 10-17) were discovered from Fusarium sp. in coffee medium. Additionally, the configuration of 8 was first reported in the research by Mosher's method. The structures were established by 1D, 2D NMR, mass spectrometry, calculated ECD spectra, and Mosher's method. Compounds 1, 2, 6/7, 12, and 16 indicated significant antifungal activities against the phytopathogen Alternaria alternata isolated from Coffea arabica with MICs of 1 µg/mL. The investigation on the anti-phytopathogen activity of metabolites can provide lead compounds for agrochemicals.

Mid-Level Data Fusion Combined with the Fingerprint Region for Classification DON Levels Defect of Fusarium Head Blight Wheat.

In this study, a method of mid-level data fusion with the fingerprint region was proposed, which was combined with the characteristic wavelengths that contain fingerprint information in NIR and FT-MIR spectra to detect the DON level in FHB wheat during wheat processing. NIR and FT-MIR raw spectroscopy data on normal wheat and FHB wheat were obtained in the experiment. MSC was used for pretreatment, and characteristic wavelengths were extracted by CARS, MGS and XLW. The variables that can effectively reflect fingerprint information were retained to build the mid-level data fusion matrix. LS-SVM and PLS-DA were applied to investigate the performance of the single spectroscopic model, mid-level data fusion model and mid-level data fusion with fingerprint information model, respectively. The experimental results show that mid-level data fusion with a fingerprint information strategy based on fused NIR and FT-MIR spectra represents an effective method for the classification of DON levels in FHB wheat samples.

Occurrence of resorcyclic acid lactones in porcine urine: discrimination between illegal use and contamination.

Zeranol (α-zearalanol, α-ZAL), is a resorcyclic acid lactone (RAL). Its administration to farm animals to improve meat production has been prohibited in the European Union due to the potential risk to human health. However, it has been demonstrated that α-ZAL may be present in livestock animals due to Fusarium fungi that produce fusarium acid lactones contamination in feed. The fungi produce a small amount of zearalenone (ZEN), which is metabolized to zeranol. The potential endogenous origin of α-ZAL makes it difficult to correlate positive samples to a potential illicit treatment with α-ZAL. We present two experimental studies that investigated the origin of natural and synthetic RALs in porcine urine. Urine samples from pigs that were either fed with ZEN-contaminated feed or administered α-ZAL by injection were analyzed by liquid chromatography coupled to tandem mass spectrometry, with the method validated according to Commission Implementing Regulation (EU) 2021/808. The data show that although the concentration of α-ZAL in the ZEN feed-contaminated samples is significantly lower than in the illicit administration samples, α-ZAL can occur in porcine urine via natural metabolism. Additionally, the feasibility of using the ratio of forbidden/fusarium RALs in porcine urine as a reliable biomarker for illicit treatment with α-ZAL administration was evaluated for the first time. This study demonstrated that the obtained ratio in the contaminated ZEN feed study was close to 1, while in the illegally administered α-ZAL samples the ratio is always higher than 1 (up to 135). Therefore, this study proves that the ratio criteria (already used when a forbidden RAL is detected in bovine urine) may also be used for porcine urine.

Occurrence of Fusarium mycotoxins in freshly harvested highland barley (qingke) grains from Tibet, China.

Highland barley, also called "qingke" in Tibetan, is mainly cultivated in the Tibetan Plateau of China and has been used as a major staple food for Tibetans. Recently, Fusarium head blight (FHB) of qingke was frequently observed around the Brahmaputra River in Tibet. Considering the importance of qingke for Tibetans, the assessment of Fusarium mycotoxin contamination is essential for food safety. In this study, a total of 150 freshly harvested qingke grain samples were obtained from three regions around the Brahmaputra River in Tibet (China) in 2020. The samples were investigated for the occurrence of 20 Fusarium mycotoxins using high-performance liquid chromatography-tandem mass spectrometry (HPLC‒MS/MS). The most frequently occurring mycotoxin was enniatin B (ENB) (46%), followed by enniatin B1 (ENB1) (14.7%), zearalenone (ZEN) (6.0%), enniatin A1 (ENA1) (3.3%), enniatin A (ENA) (1.3%), beauvericin (BEA) (0.7%), and nivalenol (NIV) (0.7%). Due to the increase in altitude, the cumulative precipitation level and average temperature decreased from the downstream to the upstream of the Brahmaputra River; this directly correlated to the contamination level of ENB in qingke, which gradually decreased from downstream to upstream. In addition, the level of ENB in qingke obtained from qingke-rape rotation was significantly lower than that from qingke-wheat and qingke-qingke rotations (p < 0.05). These results disseminated the occurrence of Fusarium mycotoxins and provided further understanding of the effect of environmental factors and crop rotation on Fusarium mycotoxins.

Exploring the chemical diversity of phytopathogenic fungi infecting edible fruits.

Two fungi, Fusarium guttiforme and Colletotrichum horii, were cultured under different conditions to obtain fourteen compounds. The axenic cultures of F. guttiforme and C. horii in potato dextrose broth (PDB) medium yielded fusaric acid (1), 9,10-dehydrofusaric acid (2), and tyrosol, whereas their co-cultivation produced fusarinol (5), a fusaric acid complex with magnesium (3), 9,10-dehydrofusaric acid complex with magnesium (4), and 5-butyl-5-(hydroxymethyl) dihydrofuranone (9). Upon changing the medium from PDB to Czapek, different compounds (uracil, p-hydroxy acetophenone, and cyclo(L-Leu-L-Pro) were obtained. Fusaric acid (1) was biotransformed into fusarinol (5) by C. horii, suggesting a detoxification process, and three other compounds were obtained: 7-hydroxyfusarinol (7), 9,10-dehydrofusarinol (6), and fusarinyl acetate (8). Epigenetic modulation of suberohydroxamic acid against F. guttiforme afforded gibepyrone B (10). These compounds were subjected to a papain inhibition enzymatic assay; the highest inhibitory activity was displayed by the two magnesium complexes, at 56 and 54% inhibition, respectively.

Utilization of industrial waste - Liquid cheese whey for the batch fermentation of lovastatin using Fusarium nectrioides (MH173849) an endophytic fungus: Screening, production and characterization.

In this current research, a novel way of utilizing the plant weed and dairy industrial waste for the cost-effective production of Lovastatin by the novel fungus Fusarium nectrioides (MH173849) under controlled conditions was reported for the first time with scientific evidence. A total of 25 endophytic fungi were isolated from the 90 tissue fragments of Euphorbia hirta (L) and identified based on morphological and microscopical characteristics. All the fungal isolates were screened for Lovastatin production using Neurospora crassa bioassay. Among the 25 fungal isolates, Fusarium sp2, Nigrospora sphaerica, and Fusarium sp 4 showed maximum zone of inhibition and they were further verified by Thin Layer Chromatography. Since the Rf values of Fusarium sp 4 and standard Lovastatin were the same, further characterization was preceded only with Fusarium sp 4. An evolutionary relationship of two positive isolates, Fusarium sp 2 and Fusarium sp 4 was studied with other Lovastatin-producing fungi. Gene sequencing and BLAST revealed that a novel fungus, Fusarium sp 4 was found to be Fusarium nectrioides (MH173849) and it was further used for batch fermentation of Lovastatin in the modified media using liquid cheese whey under controlled conditions, which enhanced the productivity up to 43.40 µg/mL with the minimum purification steps. LC-MS-MS and NMR studies confirmed the production of Lovastatin by F. nectrioides (MH173849) due to the presence of Pyran molecule hydrogen, Hydrogen fusing two molecules as intermediate with triplet signal groups, methylbutanoic acid, and hexahydro naphthalene. Therefore, this fungus may be utilized by industries for the cost-effective production of Lovastatin.

Acuminatums E and F, two new cyclic lipopeptides from Fusarium lateritium HU0053 and their antifungal activity.

Two new cyclic lipopeptides, acuminatums E (1) and F (2), together with four known cyclic lipopeptides, acuminatums A-D (3-6) were isolated from the corn culture of endophytic Fusarium lateritium HU0053. Their structures were elucidated by spectroscopic and advanced Marfey's amino acid analysis. All compounds were found to exhibit antifungal activities against Penicillium digitatum. Acuminatum F (2), a new cyclic lipopeptide containing an unusual 3, 4-dihydroxy-phenylalanine unit exhibited the strongest antifungal activities with inhibition zone of 6.5 mm at the dose of 6.25 µg. Therefore, acuminatum F might be a potential environmental-friendly preservative for citrus fruits.

Antitumor potential of tricyclic pyridone analogues from an entomopathogenic fungus, Fusarium avenaceum SYKC02-P-1.

Two new tricyclic pyridone analogues, fusapyridons C (1) and D (2), were isolated along with structurally related known compounds 3 - 5 from the entomopathogenic fungus, F. avenaceum SYKC02-P-1. The structures of compounds 1 and 2 were elucidated by analyzing the spectral data of UV, 1 D and 2 D NMR as well as HRESIMS. The absolute configurations of fusapyridons C and D were established by means of single crystal X-ray diffraction and electronic circular dichroism calculation. And antitumor testing of all the isolates showed that compounds 4 and 5 exhibited significant inhibitory activity against the human prostate cancer cells (PC-3 cell lines) with IC50 values of 2.76 and 1.86 µM, respectively.

Cyclic hexadepsipeptides from the fermentation of Fusarium sp. DCJ-A and their cytotoxic activities.

Beauvercin H (1), a new cyclic hexadepsipeptide, and two known ones (2 and 3) were isolated from the EtOH extract of the solid culture of Fusarium sp. Their structures were elucidated by spectroscopic analysis, including extensive 1D and 2D NMR techniques, as well as comparison with literature values. Additionally, compounds 1-3 were tested for their cytotoxic activities. The results showed that all isolated compounds exhibited cytotoxic activities against five human cancer cell lines with IC50 values ranging from 1.379 to 13.12 µM.

The control of Fusarium growth and decontamination of produced mycotoxins by lactic acid bacteria.

Global crop and food contamination with mycotoxins are one of the primary worldwide concerns, while there are several restrictions regarding approaching conventional physical and chemical mycotoxins decontamination methods due to nutrition loss, sensory attribute reduction in foods, chemical residual, inconvenient operation, high cost of equipment, and high energy consumption of some methods. In this regard, the overarching challenges of mycotoxin contamination in food and food crops require the development of biological decontamination strategies. Using certain lactic acid bacteria (LAB) as generally recognized safe (GRAS) compounds is one of the most effective alternatives due to their potential to release antifungal metabolites against various fungal factors species. This review highlights the potential applications of LAB as biodetoxificant agents and summarizes their decontamination activities against Fusarium growth and Fusarium mycotoxins released into food/feed. Firstly, the occurrence of Fusarium and the instrumental and bioanalytical methods for the analysis of mycotoxins were in-depth discussed. Upgraded knowledge on the biosynthesis pathway of mycotoxins produced by Fusarium offers new insightful ideas clarifying the function of these secondary metabolites. Moreover, the characterization of LAB metabolites and their impact on the decontamination of the mycotoxin from Fusarium, besides the main mechanisms of mycotoxin decontamination, are covered. While the thematic growth inhibition of Fusarium and decontamination of their mycotoxin by LAB is very complex, approaching certain lactic acid bacteria (LAB) is worth deeper investigations.

Extraction, characterization, and biological activities of exopolysaccharides from plant root soil fungus Fusarium merismoides A6.

The exploration of polysaccharides from microorganisms is of great importance. In this study, a new type of exopolysaccharide excreted by Fusarium merismoides A6 (FM-EPS) was isolated, and the extraction conditions were optimized using a response surface methodology (RSM). The extraction temperature at 0 °C, a precipitation time of 7.83 h, and an ethanol precipitation concentration of 77.64% were predicted and proved to be the best extraction conditions with the maximum extraction yield of 0.74 g/mL. Then, two fractions of F. merismoides A6 exopolysaccharides (FM-EPS1 and FM-EPS2) were obtained through DEAE Sepharose fast flow column chromatography. As indicated by monosaccharide composition analysis, both fractions mainly consisted of mannose, glucose, galactose, and ribose, with an average molecular weight of 5.14 × 104 and 6.50 × 104 g/mol, respectively. FT-IR and NMR spectroscopy indicated the FM-EPSs had both α- and ß-glycosidic bonds. Moreover, the determination of antioxidant and antiproliferative activities in vitro proved that FM-EPSs had good antioxidant activities and antiproliferation activities. FM-EPS1 showed stronger antioxidant activities than FM-EPS2. FM-EPS2 showed antiproliferation activities on HeLa and HepG2 cells, while FM-EPS1 had no obvious antiproliferative activity. Therefore, FM-EPSs could be explored as potential antioxidant and anticancer agent applied in food, feed, nutraceutical, pharmaceutical, cosmetics, and chemical industries.

Cytotoxic Cyclodepsipeptides and Cyclopentane Derivatives from a Plant-Associated Fungus Fusarium sp.

In this work, four new cyclodepsipeptides, fusarihexins C-E (1-3) and enniatin Q (4), four new cyclopentane derivatives, fusarilins A-D (5-8), together with eight known compounds (9-16), were isolated from cultures of the endophytic fungus Fusarium sp. The structures of the isolated compounds were elucidated by analysis of HRMS and NMR spectroscopic data. The absolute configurations were determined using Marfey's method, a modified Mosher's method, single-crystal X-ray diffraction analysis, and ECD analysis. The antitumor activities of the isolated compounds in vitro were evaluated. Cyclodepsipeptides displayed cytotoxicities against the Huh-7, MRMT-1, and HepG-2 cell lines. Compounds 4, 9, 10, and 12 with IC50 values of 1.0-9.1 µM exhibited the most potent cytotoxicities against the three cell lines as compared to the positive control-5-fluorouracil. Compounds 1-3 and 11 exhibited moderate cytotoxic activities (IC50 values of 10.7-20.1 µM).

Preparation of 3-O-, 5-O- and N-palmitoyl derivatives of fumonisin B1 toxin and their characterisation with NMR and LC-HRMS methods.

We have previously published six esterified O-acyl (EFB1) and three N-acyl fumonisin B1 derivatives extracted from rice cultures inoculated with Fusarium verticillioides, amongst these the identification of N-palmitoyl-FB1 has been clearly established in a spiking experiment. At that time, it was assumed that as in the case of O-acyl-FB1 derivatives, linoleic-, oleic- or palmitic acid esterify through the OH group on the 3C or 5C atom of the carbon chain of the fumonisins. In our most recent experiments, we have synthetically acylated the FB1 toxin and subsequently purified 3-O-palmitoyl- and 5-O-palmitoyl-FB1 toxins in addition to the N-palmitoyl-FB1 toxin. They were identified and characterised using 1H and 13C NMR as well as LC-HRMS. Our aim was the identification of the previously detected O-acyl-FB1 derivatives over the course of a spiking experiment, which were obtained through the solid-phase fermentation of Fusarium verticillioides. By spiking the three synthesized and identified components one-by-one into the fungal culture extract and analysing these cultures using LC-MS, it was clearly demonstrated that the F. verticillioides strain produced both the 5-O-palmitoyl-FB1 and N-palmitoyl-FB1 toxins, but did not produce 3-O-palmitoyl-FB1. Thus, it is highly probable that the components thought to be 3-O-acyl-(linoleoyl-, oleoyl-, palmitoyl-) FB1 derivatives in our previous communication are presumably 10-O-acyl-FB1 derivatives. Since these acylated FB1 derivatives can occur naturally in e.g. maize, the use of these synthesized components as reference materials is of great importance in order to obtain accurate qualitative and quantitative data on the occurrence of acylated fumonisins in different matrices including maize based feed samples. The production of these substances has also made it possible to test their toxicity in cell culture and small animal experiments.

Global regulatory factor VeA upregulates the production of antitumor substances in endophytic Fusarium solani.

A number of studies have demonstrated that endophytic fungi have the potential to produce antitumor active substances with novel structures and significant activities. In our previous studies, we isolated a Fusarium strain from the stem of the medicinal plant Nothapodytes pittosporoides (Oliv.). In this study, we identified this strain as Fusarium solani and found that its crude extract has significant antitumor activity against human alveolar adenocarcinoma cells (A549). We overexpressed the global regulatory factor VeA in F. solani (VeAOE), resulting in a significant increase in antitumor activity. The MTT assay results showed that the inhibition rate of the VeAOE mutant extract on A549 cancer cells was significantly higher than that of the WT extract, as the IC50 decreased from 369.22 to 285.89 µg/mL, and the apoptosis ratio was significantly increased by approximately 4.86-fold. In VeAOE, accumulation of alkaloids, terpenoids, carboxylic acid derivatives, phenols and flavonoid metabolites with potential antitumor activity was significantly increased compared with WT based on metabolomic analysis. Additionally, transcriptome analysis found that the expression patterns of 48 genes related to antitumor activity were significantly changed in VeAOE, mainly involving glycosyl hydrolases, the Zn(2)-Cys(6) class, cytochrome P450 monooxygenase, 3-isopropylmalate dehydratase, and polyketide synthases. These results suggested that VeA mediated the antitumor activity of the metabolites in F. solani HB1-J1 by regulating multiple metabolic pathways.

The Effect of Foliar Fungicide and Insecticide Application on the Contamination of Fumonisins, Moniliformin and Deoxynivalenol in Maize Used for Food Purposes.

The fungal ear rot of maize cultivated in temperate areas is mainly due to the Fusarium species. The use of insecticides against European Corn Borer (ECB) reduces the severity of fungal ear rot as well as the fumonisin (FB) and moniliformin (MON) levels in maize kernels at harvest, which in turn results in a lowering of their effect on deoxynivalenol (DON) control. However, the direct fungicidal control of ear rot has rarely been implemented for maize, and the first studies reported conflicting results on the reduction of mycotoxins. In the present experiment, field trials were carried out in North Italy over three growing seasons to study the effect of fungicide application timings on maize to control mycotoxins, considering the interaction of the application with the insecticide treatment, according to a full factorial split plot design. The mycotoxin content was determined through LC-MS/MS analysis. The field trials showed a significant reduction in ECB severity (75%), fungal ear rot severity (68%), Fusarium Liseola section infection (46%), FBs (75%) and MON (79%) as a result of the insecticide application for all the years, while the DON content increased by 60%. On the other hand, a fungicide application alone or applied in plots protected by an insecticide was never effective for the fungal symptoms, infection or mycotoxin content. The results confirm that a correct insecticide application to control ECB damage is the most effective agrochemical solution for the control of fungal ear rot, FBs and MON.