Rosellichalasins A-H, cytotoxic cytochalasans from the endophytic fungus Rosellinia sp. Glinf021.

Eight new cytochalasans rosellichalasins A-H (1-8), as well as two new shunt metabolites rosellinins A (9) and B (10) before intramolecular Diels-Alder cycloaddition reaction in cytochalasan biosynthesis, along with nine known cytochalsans (11-19) were isolated from the endophytic fungus Rosellinia sp. Glinf021, which was derived from the medicinal plant Glycyrrhiza inflata. Their structures were characterized by extensive analysis of 1D and 2D NMR as well as HRESIMS spectra and quantum chemical ECD calculations. The cytotoxic activities of these compounds were evaluated against four human cancer cell lines including HCT116, MDA-MB-231, BGC823, and PANC-1 with IC50 values ranging from 0.5 to 58.2 µM.

Diphenyl ethers from endophytic fungus Rhexocercosporidium sp. Dzf14 and their antibacterial activity by affecting homeostasis of cell membranes.

BACKGROUND: Phytopathogenic bacteria cause severe losses to crops every year. The management of crop bacterial diseases with chemical agents has been considered as the main strategy. In order to cope with the bactericide resistance made by the pathogens, new antibacterials need to be continuously developed. RESULTS: A chemical investigation from the endophytic fungus Rhexocercosporidium sp. Dzf14 has led to the isolation of 12 diphenyl ethers including two new ones named rhexocerin E (1) and rhexocercosporin G (2), along with two new depsides named rhexocerdepsides A (3) and B (4). The structures and absolute configurations of the new compounds were determined through comprehensive analysis of spectroscopic data and quantum chemical ECD calculations. Diphenyl ethers showed obviously antibacterial activity on Gram-positive bacteria. The structure-activity relationship of diphenyl ethers revealed that prenylation was critical to the antibacterial activity. Among them, rhexocercosporin D (12) possessed the strongest activity against Clavibacter michiganensis and Bacillus subtilis, and was selected for further mechanistic studies. It was found that rhexocercosporin D displayed bactericidal activity by affecting homeostasis of cell membranes. In addition to its rapid bactericidal effects on Gram-positive bacteria, rhexocercosporin D could restore the susceptibility against Gram-negative Agrobacterium tumefaciens by synergistic action with colistin. CONCLUSION: Twelve diphenyl ethers and two depsides were isolated from endophytic fungus Rhexocercosporidium sp. Dzf14. Isopentenyl was critical for diphenyl ethers against Gram-positive bacteria. Rhexocercosporin D could affect homeostasis of bacterial cell membrane to exert rapid bactericidal activity. These findings highlight the antibacterial potential of the diphenyl ethers in crop bacterial disease management. © 2024 Society of Chemical Industry.

A Genome-Wide Comparison of Rice False Smut Fungus Villosiclava virens Albino Strain LN02 Reveals the Genetic Diversity of Secondary Metabolites and the Cause of Albinism.

Rice false smut (RFS) caused by Villosiclava virens (anamorph: Ustilaginoidea virens) has become one of the most destructive fungal diseases to decrease the yield and quality of rice grains. An albino strain LN02 was isolated from the white RFS balls collected in the Liaoning Province of China in 2019. The strain LN02 was considered as a natural albino mutant of V. virens by analyzing its phenotypes, internal transcribed spacer (ITS) conserved sequence, and biosynthesis gene clusters (BGCs) for secondary metabolites. The total assembled genome of strain LN02 was 38.81 Mb, which was comprised of seven nuclear chromosomes and one mitochondrial genome with an N50 value of 6,326,845 bp and 9339 protein-encoding genes. In addition, the genome of strain LN02 encoded 19 gene clusters for biosynthesis of secondary metabolites mainly including polyketides, terpenoids and non-ribosomal peptides (NRPs). Four sorbicillinoid metabolites were isolated from the cultures of strain LN02. It was found that the polyketide synthase (PKS)-encoding gene uspks1 for ustilaginoidin biosynthesis in strain LN02 was inactivated due to the deletion of four bases in the promoter sequence of uvpks1. The normal uvpks1 complementary mutant of strain LN02 could restore the ability to synthesize ustilaginoidins. It demonstrated that deficiency of ustilaginoidin biosynthesis is the cause of albinism for RFS albino strain LN02, and V. virens should be a non-melanin-producing fungus. This study further confirmed strain LN02 as a white phenotype mutant of V. virens. The albino strain LN02 will have a great potential in the development and application of secondary metabolites. The physiological and ecological functions of ustilaginoidins in RFS fungus are needed for further investigation.

A radio pulsar phase from SGR J1935+2154 provides clues to the magnetar FRB mechanism.

The megajansky radio burst, FRB 20200428, and other bright radio bursts detected from the Galactic source SGR J1935+2154 suggest that magnetars can make fast radio bursts (FRBs), but the emission site and mechanism of FRB-like bursts are still unidentified. Here, we report the emergence of a radio pulsar phase of the magnetar 5 months after FRB 20200428. Pulses were detected in 16.5 hours over 13 days using the Five-hundred-meter Aperture Spherical radio Telescope, with luminosities of about eight decades fainter than FRB 20200428. The pulses were emitted in a narrow phase window anti-aligned with the x-ray pulsation profile observed using the x-ray telescopes. The bursts, conversely, appear in random phases. This dichotomy suggests that radio pulses originate from a fixed region within the magnetosphere, but bursts occur in random locations and are possibly associated with explosive events in a dynamically evolving magnetosphere. This picture reconciles the lack of periodicity in cosmological repeating FRBs within the magnetar engine model.

Magnetic field reversal in the turbulent environment around a repeating fast radio burst.

Fast radio bursts (FRBs) are brief, intense flashes of radio waves from unidentified extragalactic sources. Polarized FRBs originate in highly magnetized environments. We report observations of the repeating FRB 20190520B spanning 17 months, which show that the FRB's Faraday rotation is highly variable and twice changes sign. The FRB also depolarizes below radio frequencies of about 1 to 3 gigahertz. We interpret these properties as being due to changes in the parallel component of the magnetic field integrated along the line of sight, including reversing direction of the field. This could result from propagation through a turbulent magnetized screen of plasma, located 10-5 to [Formula: see text] parsecs from the FRB source. This is consistent with the bursts passing through the stellar wind of a binary companion of the FRB source.

Effects of Carbon, Nitrogen, Ambient pH and Light on Mycelial Growth, Sporulation, Sorbicillinoid Biosynthesis and Related Gene Expression in Ustilaginoidea virens.

Sorbicillinoids are a class of hexaketide metabolites produced by Ustilaginoidea virens (teleomorph: Villosiclava virens), an important fungal pathogen that causes a devastating rice disease. In this study, we investigated the effects of environmental factors, including carbon and nitrogen sources, ambient pH and light exposure, on mycelial growth, sporulation, as well as the accumulation of sorbicillinoids, and the expression of related genes involved in sorbicillinoid biosynthesis. It was found that the environmental factors had great influences on mycelial growth and sporulation of U. virens. Fructose and glucose, complex nitrogen sources, acidic conditions and light exposure were favorable for sorbicillinoid production. The relative transcript levels of sorbicillinoid biosynthesis genes were up-regulated when U. virens was separately treated with those environmental factors that favored sorbicillinoid production, indicating that sorbicillinoid biosynthesis was mainly regulated at the transcriptional level by different environmental factors. Two pathway-specific transcription factor genes, UvSorR1 and UvSorR2, were found to participate in the regulation of sorbicillinoid biosynthesis. These results will provide useful information to better understand the regulation mechanisms of sorbicillinoid biosynthesis, and be conducive to develop effective means for controlling sorbicillinoid production in U. virens.

Recent Advances in Search of Bioactive Secondary Metabolites from Fungi Triggered by Chemical Epigenetic Modifiers.

Genomic analysis has demonstrated that many fungi possess essential gene clusters for the production of previously unobserved secondary metabolites; however, these genes are normally reduced or silenced under most conditions. These cryptic biosynthetic gene clusters have become treasures of new bioactive secondary metabolites. The induction of these biosynthetic gene clusters under stress or special conditions can improve the titers of known compounds or the production of novel compounds. Among the inducing strategies, chemical-epigenetic regulation is considered a powerful approach, and it uses small-molecule epigenetic modifiers, which mainly act as the inhibitors of DNA methyltransferase, histone deacetylase, and histone acetyltransferase, to promote changes in the structure of DNA, histones, and proteasomes and to further activate cryptic biosynthetic gene clusters for the production of a wide variety of bioactive secondary metabolites. These epigenetic modifiers mainly include 5-azacytidine, suberoylanilide hydroxamic acid, suberoyl bishydroxamic acid, sodium butyrate, and nicotinamide. This review gives an overview on the method of chemical epigenetic modifiers to trigger silent or low-expressed biosynthetic pathways to yield bioactive natural products through external cues of fungi, mainly based on the research progress in the period from 2007 to 2022. The production of about 540 fungal secondary metabolites was found to be induced or enhanced by chemical epigenetic modifiers. Some of them exhibited significant biological activities such as cytotoxic, antimicrobial, anti-inflammatory, and antioxidant activity.

Activation of Ustilaginoidin Biosynthesis Gene uvpks1 in Villosiclava virens Albino Strain LN02 Influences Development, Stress Responses, and Inhibition of Rice Seed Germination.

Villosiclava virens (anamorph: Ustilaginoidea virens) is the pathogen of rice false smut (RFS), which is a destructive rice fungal disease. The albino strain LN02 is a natural white-phenotype mutant of V. virens due to its incapability to produce toxic ustilaginoidins. In this study, three strains including the normal strain P1, albino strain LN02, and complemented strain uvpks1C-1 of the LN02 strain were employed to investigate the activation of the ustilaginoidin biosynthesis gene uvpks1 in the albino strain LN02 to influence sporulation, conidia germination, pigment production, stress responses, and the inhibition of rice seed germination. The activation of the ustilaginoidin biosynthesis gene uvpks1 increased fungal tolerances to NaCl-induced osmotic stress, Congo-red-induced cell wall stress, SDS-induced cell membrane stress, and H2O2-induced oxidative stress. The activation of uvpks1 also increased sporulation, conidia germination, pigment production, and the inhibition of rice seed germination. In addition, the activation of uvpks1 was able to increase the mycelial growth of the V. virens albino strain LN02 at 23 °C and a pH from 5.5 to 7.5. The findings help in understanding the effects of the activation of uvpks1 in albino strain LN02 on development, pigment production, stress responses, and the inhibition of rice seed germination by controlling ustilaginoidin biosynthesis.

Frequency-dependent polarization of repeating fast radio bursts-implications for their origin.

The polarization of fast radio bursts (FRBs), which are bright astronomical transient phenomena, contains information about their environments. Using wide-band observations with two telescopes, we report polarization measurements of five repeating FRBs and find a trend of lower polarization at lower frequencies. This behavior is modeled as multipath scattering, characterized by a single parameter, σRM, the rotation measure (RM) scatter. Sources with higher σRM have higher RM magnitude and scattering time scales. The two sources with the highest σRM, FRB 20121102A and FRB 20190520B, are associated with compact persistent radio sources. These properties indicate a complex environment near the repeating FRBs, such as a supernova remnant or a pulsar wind nebula, consistent with their having arisen from young stellar populations.

Recent Advances in Sorbicillinoids from Fungi and Their Bioactivities (Covering 2016-2021).

Sorbicillinoids are a family of hexaketide metabolites with a characteristic sorbyl side chain residue. Sixty-nine sorbicillinoids from fungi, newly identified from 2016 to 2021, are summarized in this review, including their structures and bioactivities. They are classified into monomeric, dimeric, trimeric, and hybrid sorbicillinoids according to their basic structural features, with the main groups comprising both monomeric and dimeric sorbicillinoids. Some of the identified sorbicillinoids have special structures such as ustilobisorbicillinol A, and sorbicillasins A and B. The majority of sorbicillinoids have been reported from fungi genera such as Acremonium, Penicillium, Trichoderma, and Ustilaginoidea, with some sorbicillinoids exhibiting cytotoxic, antimicrobial, anti-inflammatory, phytotoxic, and α-glucosidase inhibitory activities. In recent years, marine-derived, extremophilic, plant endophytic, and phytopathogenic fungi have emerged as important resources for diverse sorbicillinoids with unique skeletons. The recently revealed biological activities of sorbicillinoids discovered before 2016 are also described in this review.

Phytotoxic Secondary Metabolites from Fungi.

Fungal phytotoxic secondary metabolites are poisonous substances to plants produced by fungi through naturally occurring biochemical reactions. These metabolites exhibit a high level of diversity in their properties, such as structures, phytotoxic activities, and modes of toxicity. They are mainly isolated from phytopathogenic fungal species in the genera of Alternaria, Botrytis, Colletotrichum, Fusarium, Helminthosporium, and Phoma. Phytotoxins are either host specific or non-host specific phytotoxins. Up to now, at least 545 fungal phytotoxic secondary metabolites, including 207 polyketides, 46 phenols and phenolic acids, 135 terpenoids, 146 nitrogen-containing metabolites, and 11 others, have been reported. Among them, aromatic polyketides and sesquiterpenoids are the main phytotoxic compounds. This review summarizes their chemical structures, sources, and phytotoxic activities. We also discuss their phytotoxic mechanisms and structure-activity relationships to lay the foundation for the future development and application of these promising metabolites as herbicides.

Preparation of a novel hydroxypropyl-γ-cyclodextrin functionalized monolith for separation of chiral drugs in capillary electrochromatography.

In this study, a novel hydroxypropyl-γ-cyclodextrin (HP-γ-CD) functionalized monolithic capillary column was prepared by one-pot sequential strategy and used for chiral separation in capillary electrochromatography for the first time. In one pot, GMA-HP-γ-CD as functional monomer was allowed to be formed via the ring opening reaction between HP-γ-CD and glycidyl methacrylate (GMA) catalyzed by 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and then copolymerized directly with ethylene dimethacrylate (EDMA) and 2-acrylamido-2-methyl propane sulfonic acid (AMPS) in the presence of porogenic solvents via thermally initiated free radical polymerization. The preparation conditions of monoliths were optimized. Enantiomer separations of six chiral drugs including pindolol, clorprenaline, tulobuterol, clenbuterol, propranolol, and tropicamide were achieved on the monolith. Among them, pindolol, clorprenaline, and tropicamide were baseline separated with resolution values of 1.62, 1.73, and 1.55, respectively. The mechanism of enantiomer separation was discussed by comparison of the HP-γ-CD and HP-ß-CD functionalized monoliths.

Kinetics of thermal oxidative degradation of poly (vinyl chloride) containing Ca and Sn at low temperature.

Calcium metal soap and polyol (dipentaerythritol) additives are replacing or partially replacing organotin in poly(vinyl chloride) (PVC) heat stabilizers due to their low cost, nontoxicity and safety. Therefore, investigating the low-temperature thermal oxidative degradation of stabilized plasticized PVC from the source is essential for recycling. This work uses isothermal thermogravimetry to investigate the thermal degradation process and isothermal discoloration of PVC/calcium metal soap/dipentaerythritol/organotin soft products with excellent heat resistance at 453-503 K and under air atmosphere. The chemical kinetics method is used to fit a single equation model of mass loss and time during the thermal oxidation degradation of PVC, and the kinetic equation obtained is: -ln(1 - α) = 3.83 × 103exp (-6834.4/T)t. When the temperature is 453-503 K, the calculation results are basically consistent with the experimental data and are independent of the heating rate and temperature changes. In addition, the isothermal discoloration of different PVC materials was tested under air atmosphere at 468 K. The results show that when the test material is PVC/calcium metal soap/dipentaerythritol/organotin, the heat aging time to become completely blackened is longer than that of the blank sample, which indicates a strong interaction occurs between Sn, Ca and dipentaerythritol complexes and PVC molecules, inhibiting the release of hydrogen chloride. At the same time, in order to recover PVC and prevent it from carbonization, if the temperature is set to 486 K, the thermal oxidation degradation time of PVC should be less than 130 min.

Enantioseparation and molecular modeling study of eight psychoactive drugs on a coated polysaccharide-based chiral stationary phase.

The enantioseparation of eight psychoactive drugs has been firstly performed on a coated cellulose-based chiral stationary phase (Chiralcel OJ-H). To obtain optimum separation conditions, the influences of alcohol modifiers and basic/acidic additives have been studied. As a result, except for the partial separation of oxybutynin enantiomers, the other seven drug enantiomers including mirtazapine, sulpiride, promethazine, citalopram, oxazepam, donepezil, and cyamemazine have been completely separated. Additionally, for gaining a better insight into the chiral recognition mechanisms, molecular docking was carried out using the Autodock software. Herein, binding energy and conformations of the chiral stationary phase complexes were provided, and it was found that the distinction in enantiomeric conformation determined the number and strength of intermolecular interactions between analytes and chiral stationary phase which resulted in the difference in binding energies of two enantiomers, and ultimately led to the different migration. These modeling results were in accordance with the observed enantioseparation results in high performance liquid chromatography experiments. At last, chiral separation mechanisms have been discussed in detail, and it has been confirmed that hydrogen bond, π-π, hydrophobic interactions, and some special interactions synergistically contributed to the enantioseparation of psychoactive drugs. This article is protected by copyright. All rights reserved.

New chlamydosporol derivatives from the endophytic fungus Pleosporales sp. Sigrf05 and their cytotoxic and antimicrobial activities.

Five new chlamydosporol derivatives, named pleospyrones A-E (1-5), together with one known congener (6), were isolated from the culture of the endophytic fungus Pleosporales sp. Sigrf05, obtained from the medicinal plant Siraitia grosvenorii. The structures of the new compounds were elucidated mainly by analysis of the HRESIMS, and (1D, 2D) NMR data, while ECD and optical rotation calculations were used to assign the absolute configurations. The plausible biosynthetic pathway of these compounds were proposed. The isolated compounds were evaluated for their cytotoxicity, antifungal and antibacterial activities. Compounds 1, and 4-6 were cytotoxic against the tested cancer cells with IC50 values of 1.26~47.5 µM. Compounds 1-3 showed moderate antifungal activities against Magnaporthe oryzae, while compound 5 displayed weak antibacterial activity.

Lophiostomin A-D: new 3,4-dihydroisocoumarin derivatives from the endophytic fungus Lophiostoma sp. Sigrf10.

Four new 3,4-dihydroisocoumarin congeners, named lophiostomin A-D (1-4), together with two known α-pyridones (5 and 6) were isolated from cultures of the endophytic fungus Lophiostoma sp. Sigrf10 obtained from Siraitia grosvenorii. The structures of the new compounds were determined via combined analysis involving 1D and 2D NMR, high-resolution electrospray ionization mass spectrometry (HRESIMS), and electronic circular dichroism (ECD) spectra, as well as quantum chemical ECD computations for assigning the absolute configurations. All the compounds were evaluated for their antibacterial and antifungal activities. Compounds 1 and 2 displayed moderate inhibitory activities against the spore germination of Magnaporthe oryzae, whereas 5 and 6 were active against the following tested pathogenic bacteria: Bacillus subtilis, Agrobacterium tumefaciens, Ralstonia solanacearum, and Xanthomonas vesicatoria.

Enantioseparation and determination of flumequine enantiomers in multiple food matrices with chiral liquid chromatography coupled with tandem mass spectrometry.

The present work firstly described the enantioseparation and determination of flumequine enantiomers in milk, yogurt, chicken, beef, egg, and honey samples by chiral liquid chromatography-tandem mass spectrometry. The enantioseparation was performed under reversed-phase conditions on a Chiralpak IC column at 20°C. The effects of chiral stationary phase, mobile phase components, and column temperature on the separation of flumequine enantiomers have been studied in detail. Target compounds were extracted from six different matrices with individual extraction procedure followed by cleanup using Cleanert C18 solid phase extraction cartridge. Good linearity (R2 >0.9913) was obtained over the concentration range of 0.125 to 12.5 ng g-1 for each enantiomer in matrix-matched standard calibration curves. The limits of detection and limits of quantification of two flumequine enantiomers were 0.015-0.024 and 0.045-0.063 ng g-1 , respectively. The average recoveries of the targeted compounds varied from 82.3 to 110.5%, with relative standard deviation less than 11.7%. The method was successfully applied to the determination of flumequine enantiomers in multiple food matrices, providing a reliable method for evaluating the potential risk in animal productions.

Enantioselective separation of eight antihistamines with α1-acid glycoprotein-based chiral stationary phase by HPLC: Development and validation for the enantiomeric quality control.

In the present paper, a sensitive and selective chiral LC method has been developed for enantiomeric separation and determination of eight antihistamines in pharmaceutical formulations. Enantiomeric separation was achieved on the α1-acid glycoprotein-based chiral stationary phase known as Chiralpak AGP. The effects of the mobile phase composition, type and proportion of organic modifier and the concentration and pH of the buffer on retention and resolution were fully investigated. All enantiomers of target analytes were successfully separated with the mobile phase consisting of methanol and 10 mM ammonium acetate buffer, at a flow rate of 0.9 mL min-1. Calibration curve of each enantiomer was linear over the concentration range of 5.0-100 µg mL-1, with the correlation coefficient above 0.9990. The limits of detection and quantification were below 0.80 and 3.18 µg mL-1, respectively. The recoveries were in the range of 98.0-102.4% with the relative standard deviation lower than 2.88% for all enantiomers. The validated method was demonstrated to be suitable for the quality control of the enantiomeric composition of antihistamines present in pharmaceuticals. Finally, the proposed chiral LC method was applied to the determination of antihistamines enantiomers in commercially available tablets containing racemic mixtures of compounds. Quantitative results of the commercial tablets demonstrated that the difference between the declared and determined values did not exceed 1.45%, and chiral antihistamines drugs were present in the form of racemates.

Enantioselective degradation of chiral fungicides triticonazole and prothioconazole in soils and their enantioselective accumulation in earthworms Eisenia fetida.

Triticonazole and prothioconazole are widely used systemic agricultural triazole fungicides both with a chiral center. In this work, the enantioselective degradation of triticonazole and prothioconazole in three types of soils were investigated under native conditions using reversed phase liquid chromatography-tandem mass spectrometry with a Chiralcel OD-RH column. The results indicated that the enantioselective degradation was observed with S-triticonazole and R-prothioconazole preferentially degraded and the degradation rate was fast with a half-life within 6 days. It was also found that the presence of earthworms can accelerate the degradation and further enhance degradation enantioselectivity of triticonazole and prothioconazole in soils. Moreover, the enantioselective of triticonazole and prothioconazole in earthworms were studied. The results showed that the bioaccumulation was enantioselective with R-triticonazole and S-prothioconazole preferentially accumulated, which was similar to the soil. Our findings suggest that the enantioselective toxicity and potential effects of the metabolites should be considered for more accurate assessment of ecological risks of triticonazole and prothioconazole to target and non-target species.

Lose-of-Function of a Rice Nucleolus-Localized Pentatricopeptide Repeat Protein Is Responsible for the floury endosperm14 Mutant Phenotypes.

BACKGROUND: The endosperm of rice (Oryza sativa) has been usually used for the study of starch synthesis. Although several related factors have been revealed, other unknown members remain to be identified, given that starch synthesis is a complicated and sophisticated process. RESULTS: Here, we identified and characterized a new rice seed mutant, floury endosperm14 (flo14), which showed chalked endosperm and seed-lethal phenotypes. Map-based cloning indicated FLO14 encodes a novel P-family PPR protein which contains ten PPR motifs. Afterwards the gene was named OsNPPR3. Subcellular localization showed OsNPPR3 was targeted to nucleolus. Quantitative RT-PCR analysis demonstrated that OsNPPR3 was universally expressed in various tissues, with pronounced levels during rice endosperm development. Molecular analysis further suggested that OsNPPR3 was involved in the regulation of expression levels and splicing of a few genes in mitochondria. CONCLUSION: The study demonstrates that the nucleolus-localized PPR protein is responsible for the flo14 mutant phenotypes through affecting nuclear and mitochondrial gene expression and splicing.