Weibo Sun

Determination and Analysis of Ustiloxins A and B by LC-ESI-MS and HPLC in False Smut Balls of Rice

Ustiloxins are cyclopeptide mycotoxins produced by the pathogenic fungus Villosiclava virens of rice false smut. Ustiloxins A and B as two main mycotoxins were determined conveniently by LC-ESI-MS in the water extract from rice false smut balls which were mostly composed of the chlamydospores and mycelia of the pathogen. Both ustiloxins A and B in the water extract were also quantitatively analyzed by HPLC. This is the first report on the determination and analysis of ustiloxins A and B simultaneously by LC-ESI-MS and HPLC in false smut balls of rice.

Bis-naphtho-γ-pyrones from Fungi and Their Bioactivities

Bis-naphtho-γ-pyrones are an important group of aromatic polyketides derived from fungi. They have a variety of biological activities including cytotoxic, antitumor, antimicrobial, tyrosine kinase and HIV-1 integrase inhibition properties, demonstrating their potential applications in medicine and agriculture. At least 59 bis-naphtho-γ-pyrones from fungi have been reported in the past few decades. This mini-review aims to briefly summarize their occurrence, biosynthesis, and structure, as well as their biological activities. Some considerations regarding to synthesis, production, and medicinal and agricultural applications of bis-naphtho-γ-pyrones are also discussed.

Natural Dibenzo-α-Pyrones and Their Bioactivities

Natural dibenzo-α-pyrones are an important group of metabolites derived from fungi, mycobionts, plants and animal feces. They exhibit a variety of biological activities such as toxicity on human and animals, phytotoxicity as well as cytotoxic, antioxidant, antiallergic, antimicrobial, antinematodal, and acetylcholinesterase inhibitory properties. Dibenzo-α-pyrones are biosynthesized via the polyketide pathway in microorganisms or metabolized from plant-derived ellagitannins and ellagic acid by intestinal bacteria. At least 53 dibenzo-α-pyrones have been reported in the past few decades. This mini-review aims to briefly summarize the occurrence, biosynthesis, biotransformation, as well as their biological activities and functions. Some considerations related to synthesis, production and applications of dibenzo-α-pyrones are also discussed.

Extraction Optimization of Water-Extracted Mycelial Polysaccharide from Endophytic Fungus Fusarium oxysporum Dzf17 by Response Surface Methodology

Water-extracted mycelial polysaccharide (WPS) from the endophytic fungus Fusarium oxysporum Dzf17 isolated from Dioscorea zingiberensis was found to be an efficient elicitor to enhance diosgenin accumulation in D. zingigerensis cultures, and also demonstrated antioxidant activity. In this study, response surface methodology (RSM) was employed to optimize the extraction process of WPS from F. oxysporum Dzf17 using Box-Behnken design (BBD). The ranges of the factors investigated were 1–3 h for extraction time (X1), 80–100 °C for extraction temperature (X2), and 20–40 (v/w) for ratio of water volume (mL) to raw material weight (g) (X3). The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis. Statistical analysis showed that the polynomial regression model was in good agreement with the experimental results with the determination coefficient (R2) of 0.9978. By solving the regression equation and analyzing the response surface contour plots, the extraction parameters were optimized as 1.7 h for extraction time, 95 °C for extraction temperature, 39 (v/w) for ratio of water volume (mL) to raw material weight (g), and with 2 extractions. The maximum value (10.862%) of WPS yield was obtained when the WPS extraction process was conducted under the optimal conditions.

Bioactive Bis-naphtho-γ-pyrones from Rice False Smut Pathogen Ustilaginoidea virens

Ustilaginoidins were bis-naphtho-γ-pyrones mycotoxins possessing an aR configuration of the chiral axis previously reported from the false smut balls of rice infected by the fungal pathogen Ustilaginoidea virens. To investigate the chemical diversity of these metabolites and their bioactivities, we fermented this fungus on solid rice media, which afforded the isolation of 13 ustilaginoidins, including seven new compounds, namely ustilaginoidins K-P, 1-6, and E1, 7, together with the known ustilaginoidins A, 8, D, 9, E, 10, F, 11, and G, 12, and isochaetochromin B2, 13. The structures of the new compounds were elucidated by using (1D, 2D) NMR, high-resolution mass spectrometry, UV, and circular dichroism, as well as by comparison with the literature data. A plausible biosynthesis pathway was proposed for these dimeric polyketides. The isolated compounds were evaluated for their antibacterial, cytotoxic, and radicle elongation inhibitory activities. Ustilaginoidins K, 1 and L, 2 showed cytotoxic activities on the A2780 human ovarian cancer cell line with IC50 values of 4.18 and 7.26 μM, respectively. Ustilaginoidins N, 4, D, 9, E, 10, and G, 12 were active against the tested pathogenic bacteria with MIC values in the range of 16-64 μg/mL. Ustilaginoidins O, 5, E, 10, and F, 11, and isochaetochromin B2, 13 displayed moderate inhibitory activity on the radicle elongation of rice seeds.

Purification of Ustiloxins A and B from Rice False Smut Balls by Macroporous Resins

Ustiloxins are cyclopeptide mycotoxins produced by Villosiclava virens, the pathogenic fungus of rice false smut disease. Both resins SP207 and SP700 were screened to show the best adsorption and desorption properties for ustiloxins A and B among 20 commercial macroporous resins. Dynamic adsorption and desorption tests were carried out to optimize the process parameters. The optimal conditions for adsorption of resin SP207 were a processing volume as 32 bed volumes (BV), pH value of 4, and flow rate of 2 BV/h; and those for desorption of resin SP207 were a 40:60 (v/v) ratio of ethanol to water, an eluent volume of 4 BV, pH value of 4 and a flow rate of 3 BV/h. The optimal conditions for adsorption of resin SP700 were a processing volume of 26 BV, pH value as 4, flow rate of 2 BV/h; and those for desorption of resin SP700 were a 30:70 (v/v) ratio of ethanol to water solution as eluent, volume of 4 BV, pH value as 4 and flow rate of 2 BV/h. Under the optimal conditions; the purities of ustiloxins A and B obtained with resin SP207 increased 23.06-fold and 19.78-fold, respectively; and their recoveries were 96.67% and 81.25%; respectively. Similarly; the purities of ustiloxins A and B obtained with resin SP700 increased 14.75-fold and 15.33-fold and their recoveries were 93.65% and 88.64%; respectively. The results show that adsorption and desorption on SP207 and SP700 resins are effective strategies for purifying ustiloxins A and B. The developed methods are beneficial for large-scale preparation and purification of ustiloxins A and B from rice false smut balls.

Enhancement of Palmarumycins C12 and C13 Production in Liquid Culture of Endophytic Fungus Berkleasmium sp. Dzf12 after Treatments with Metal Ions

The influences of eight metal ions (i.e., Na+, Ca2+, Ag+, Co2+, Cu2+, Al3+, Zn2+, and Mn4+) on mycelia growth and palmarumycins C12 and C13 production in liquid culture of the endophytic fungus Berkleasmium sp. Dzf12 were investigated. Three metal ions, Ca2+, Cu2+ and Al3+ were exhibited as the most effective to enhance mycelia growth and palmarumycin production. When calcium ion (Ca2+) was applied to the medium at 10.0 mmol/L on day 3, copper ion (Cu2+) to the medium at 1.0 mmol/L on day 3, aluminum ion (Al3+) to the medium at 2.0 mmol/L on day 6, the maximal yields of palmarumycins C12 plus C13 were obtained as 137.57 mg/L, 146.28 mg/L and 156.77 mg/L, which were 3.94-fold, 4.19-fold and 4.49-fold in comparison with that (34.91 mg/L) of the control, respectively. Al3+ favored palmarumycin C12 production when its concentration was higher than 4 mmol/L. Ca2+ had an improving effect on mycelia growth of Berkleasmium sp. Dzf12. The combination effects of Ca2+, Cu2+ and Al3+ on palmarumycin C13 production were further studied by employing a statistical method based on the central composite design (CCD) and response surface methodology (RSM). By solving the quadratic regression equation between palmarumycin C13 and three metal ions, the optimal concentrations of Ca2+, Cu2+ and Al3+ in medium for palmarumycin C13 production were determined as 7.58, 1.36 and 2.05 mmol/L, respectively. Under the optimum conditions, the predicted maximum palmarumycin C13 yield reached 208.49 mg/L. By optimizing the combination of Ca2+, Cu2+ and Al3+ in medium, palmarumycin C13 yield was increased to 203.85 mg/L, which was 6.00-fold in comparison with that (33.98 mg/L) in the original basal medium. The results indicate that appropriate metal ions (i.e., Ca2+, Cu2+ and Al3+) could enhance palmarumycin production. Application of the metal ions should be an effective strategy for palmarumycin production in liquid culture of the endophytic fungus Berkleasmium sp. Dzf12.

In vitro antioxidant activities of polysaccharides from endophytic fungus Fusarium oxysporum Dzf17

Three polysaccharides, namely exopolysaccharide (EPS), water-extracted mycelial polysaccharide (WPS) and sodium hydroxide-extracted mycelial polysaccharide (SPS), from the endophytic fungus Fusarium oxysporum Dzf17 were investigated for their in vitro antioxidant activities. Among them, SPS was the most active antioxidant component, and WPS exhibited moderate antioxidant activity. The median effective concentration (EC 50 ) values of the polysaccharides were 162.38 µg/ml (for WPS), 63.37 µg/ml (for SPS) by DPPH radical scavenging activity assay, and 54.54 µg/ml (for WPS) and 44.91 µg/ml (for SPS) by using ferrous ions chelating activity assay. The polysaccharides from F. oxysporum Dzf17 could be an alternative source as the antioxidant components.

Main Ustilaginoidins and Their Distribution in Rice False Smut Balls

Rice false smut has become an increasingly serious fungal disease in rice (Oryza sativa L.) production worldwide. Ustilaginoidins are bis-naphtho-γ-pyrone mycotoxins previously isolated from the rice false smut balls (FSBs) infected by the pathogen Villosiclava virens in rice spikelets on panicles. To investigate the main ustilaginoidins and their distribution in rice FSBs, five main bis-naphtho-γ-pyrones, namely ustilaginoidins A (1), G (2), B (3), I (4) and C (5), were isolated and identified by NMR and high-resolution mass spectrometry as well as by comparison with the data in the literature. The rice FSBs at early, middle and late maturity stages were divided into their different parts and the contents of five main ustilaginoidins for each part were determined by HPLC analysis. The results revealed that the highest levels of ustilaginoidins were in late stage rice FSBs, followed by those at middle stage. Most ustilaginoidins, 96.4% of the total quantity, were distributed in the middle layer at early stage. However, ustilaginoidins were mainly distributed in the outer and middle layers at middle and late stages. Small amounts of ustilaginoidins A (1) and G (2) were found in the inner part of rice FSBs at each maturity stage. The contents of ustilaginoidins A (1) and G (2) without hydroxymethyl groups at C-2 and C-2’ of the γ-pyrone rings in rice FSBs were relatively high at early stage, while the contents of ustilaginoidins B (3), I (4), and C (5) with hydroxymethyl groups at C-2 or C-2’ were relatively high at late stage.

Antimicrobial metabolites from the endophytic fungus Gliomastix murorum Ppf8 associated with the medicinal plant Paris polyphylla var. yunnanensis

Two antimicrobial metabolites were obtained by bioassay-guided fractionation from the endophytic fungus Gliomastix murorum Ppf8 associated with the medicinal plant Paris polyphylla var. yunnanensis. By means of physicochemical and spectrometric analysis, they were identified as (1) ergosta-5,7,22-trien-3-ol and (2) 2,3-dihydro-5-hydroxy-α,α-dimethyl-2-benzofuranmethanol. Both microdilution-colorimetric and spore germination assays were employed to evaluate their antimicrobial activity. Two compounds were screened to be active against the test microorganisms with the minimum inhibitory concentration (MIC) values ranging from 100 to 200 μg/ml, and the median inhibitory concentration (IC50) values ranged from 55.65 to 145.36 μg/ml. The results provide promising baseline information for the potential application of the metabolites of this endophytic fungus as antimicrobial agents.