Metabolomics-Guided Discovery of New Dimeric Xanthones from Co-Cultures of Mangrove Endophytic Fungi Phomopsis asparagi DHS-48 and Phomopsis sp. DHS-11.

The co-culture strategy, which mimics natural ecology by constructing an artificial microbial community, is a useful tool for the activation of biosynthetic gene clusters (BGCs) to generate new metabolites, as well as to increase the yield of respective target metabolites. As part of our project aiming at the discovery of structurally novel and biologically active natural products from mangrove endophytic fungi, we selected the co-culture of a strain of Phomopsis asparagi DHS-48 with another Phomopsis genus fungus DHS-11, both endophyted in mangrove Rhizophora mangle considering the impart of the taxonomic criteria and ecological data. The competition interaction of the two strains was investigated through morphology observation and scanning electron microscopy (SEM), and it was found that the mycelia of the DHS-48 and DHS-11 compacted and tangled with each other with an interwoven pattern in the co-culture system. A new approach that integrates HPLC chromatogram, 1HNMR spectroscopy, UPLC-MS-PCA, and molecular networking enabled the targeted isolation of the induced metabolites, including three new dimeric xanthones phomoxanthones L-N (1-3), along with six known analogs (4-9). Their planar structures were elucidated by an analysis of their HRMS, MS/MS, and NMR spectroscopic data and the absolute configurations based on ECD calculations. These metabolites showed broad cytotoxic activity against the cancer cells assessed, of which compounds 7-9 displayed significant cytotoxicity towards human liver cells HepG-2 with IC50 values ranging from 4.83 µM to 12.06 µM. Compounds 1-6 exhibited weak immunosuppressive activity against the proliferation of ConA-induced (T-cell) and LPS-induced (B-cell) murine splenic lymphocytes. Therefore, combining co-cultivation with a metabolomics-guided strategy as a discovery tool will be implemented as a systematic strategy for the quick discovery of target bioactive compounds.

Identification of Mycoviruses in the Pathogens of Fragrant Pear Valsa Canker from Xinjiang in China.

As a common disease, canker seriously affects the yield and quality of fragrant pear due to the lack of effective control measures. Some fungi have been reported to harbor rich reservoirs of viral resources, and some mycoviruses can be used as biocontrol agents against plant diseases. In this study, 199 isolates were obtained from diseased branches of fragrant pear in the main production areas of Xinjiang. Among them, 134 belonged to Valsa spp., identified using morphological and molecular biological techniques, in which V. mali was the dominant species. The mycoviruses in Valsa spp. were further identified using metatranscriptomic sequencing and RT-PCR. The results revealed that a total of seven mycoviruses were identified, belonging to Botourmiaviridae, Endornaviridae, Fusariviridae, Hypoviridae, Mitoviridae, and Narnaviridae, among which Phomopsis longicolla hypovirus (PlHV) was dominant in all the sample collection regions. The Cryphonectria hypovirus 3-XJ1 (CHV3-XJ1), Botourmiaviridae sp.-XJ1 (BVsp-XJ1), and Fusariviridae sp.-XJ1 (Fvsp-XJ1) were new mycoviruses discovered within the Valsa spp. More importantly, compared with those in the virus-free Valsa spp. strain, the growth rate and virulence of the VN-5 strain co-infected with PlHV and CHV3-XJ1 were reduced by 59% and 75%, respectively, and the growth rate and virulence of the VN-34 strain infected with PlHV were reduced by 42% and 55%, respectively. On the other hand, the horizontal transmission efficiency of PlHV decreased when PlHV was co-infected with CHV3-XJ1, indicating that PlHV and CHV3-XJ1 were antagonistic. In summary, the mycoviruses in Valsa spp. were identified in Xinjiang for the first time, and three of them were newly discovered mycoviruses, with two strains yielding good results. These results will offer potential biocontrol resources for managing pear canker disease and provide a theoretical basis for the control of fruit tree Valsa canker disease.

Cytosporones with Anti-Inflammatory Activities from the Mangrove Endophytic Fungus Phomopsis sp. QYM-13.

Six previously undescribed cytosporone derivatives (phomotones A-E (1-5) and phomotone F (13)), two new spiro-alkanol phombistenes A-B (14-15), and seven known analogs (6-12) were isolated from the mangrove endophytic fungus Phomopsis sp. QYM-13. The structures of these compounds were elucidated using spectroscopic data analysis, electronic circular dichroism (ECD), and 13C NMR calculations. Compound 14 features an unprecedented 1,6-dioxaspiro[4.5]decane ring system. All isolates were evaluated for their inhibitory effect on nitric oxide (NO) in LPS-induced RAW264.7 cells. The results showed that compounds 1, 6, 8, and 11 exhibited potent bioactivities by comparing with positive control. Then, compound 1 displayed the anti-inflammatory effect by inhibiting the MAPK/NF-κB signaling pathways. Molecular docking further revealed the possible mechanism of compound 1 interaction with ERK protein.

Will the endophytic fungus Phomopsis liquidambari increase N-mineralization in maize soil?

Endophytes can be developed into biocontrol agents and can be fungi, bacteria, or archaea that live inside plant tissues without causing symptoms of disease. Phomopsis liquidambari is an endophytic fungus that plays an important ecosystem role as a biofertilizer by helping its host obtain soil nitrogen. How this fungus impacts N mineralization and microbial communities is little known. Our understanding of soil nutrient transformations and soil-plant-microbe interactions in Phomopsis liquidambari-crop versus conventional crop systems is incomplete. This study provided a better understanding of the effect of Phomopsis liquidambari on nitrogen mineralization and investigated the interaction between P. liquidambari and nitrogen, which in turn will be helpful to the farmer in reducing the required amount of soil N fertilizer. This change in N availability in maize soil will have significant implications for soil productivity and plant N utilization, especially in N-limited soils, and significantly reduce the required amount of soil N fertilizer. The effect of P. liquidambari on N mineralization in maize soil was investigated by treating it with four levels of N (urea) at rates of 0, 1.25, 2.5, and 3.75 g of nitrogen. N-mineralization was determined by the anaerobic incubation method. Were stored for 7 days in an incubator at a constant 37 C. A colorimetric microplate procedure was used for NH4+-N analysis. A significant increase in the available NH4+-N contents was reported in soil maize (Zea mays L.) inoculated with P. liquidambari, which increased by 80%. A significant increase in N-mineralization was observed under all N conditions. This work highlighted the importance of the fungal endophyte for soil N-mineralization with lower N input. Using this fungal agent will almost certainly help reduce fertilizer input.

Secondary Metabolite Analysis of Phomopsis sp. LGT-5 Based on the Dual Guidance of Whole-Genome Sequencing and HPLC-Q-ToF-MS/MS.

Microorganisms produce a wealth of structurally diverse specialized metabolites with a remarkable range of biological activities. The Phomopsis sp. LGT-5 was obtained through tissue block and repeatedly crossed methods from Tripterygium wilfordii Hook. F. The antibacterial experiments of LGT-5 showed that it has high inhibitory activity against Staphylococcus aureus and Pseudomonas aeruginosa, and moderate inhibitory activity against Candida albicans. To research the generation of the antibacterial phenomenon of LGT-5 and provide support for further research and application, the whole genome sequencing (WGS) of LGT-5 was obtained by single-molecule real-time DNA sequencing platform Pacific Biosciences (PacBio) sequencing and Illumina paired-end sequencing. The final assembled LGT-5 genome is 54.79 Mb with a contig N50 of 290.07 kb; in addition, its secondary metabolites were detected through HPLC-Q-ToF-MS/MS. By comparing its MS/MS data, the secondary metabolites were analyzed based on visual network maps obtained on the Global Natural Products Social Molecular Networking (GNPS). The analysis results showed that the secondary metabolites of LGT-5 were triterpenes and various cyclic dipeptides.

Multifaceted plant growth-promoting traits of indigenous rhizospheric microbes against Phomopsis theae, a causal agent of stem canker in tea plants.

Phomopsis canker is one of the major devastating stem diseases that occur in tea plants caused by the fungal pathogen Phomopsis theae. Rapid development of this disease leads to a capital loss in the tea industry which demands an ecofriendly disease management strategy to control this aggressive pathogen. A total of 245 isolates were recovered from the tea rhizosphere and screened for in vitro plant growth promoting (PGP) traits and antagonism against P. theae. Among them, twelve isolates exhibited multifarious PGP traits including phytohormones, siderophore, hydrogen cyanide, salicylic acid production, phosphate solubilization, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, and antifungal activity. In vitro studies on morphological, biochemical, and phylogenetic analyses classified the selected isolates as Pseudomonas fluorescens (VPF5), Bacillus subtilis (VBS3), Streptomyces griseus (VSG4) and Trichoderma viride (VTV7). Specifically, P. fluorescens VPF5 and B. subtilis VBS3 strains showed the highest level of PGP activities. On the other hand, VBS3 and VTV7 strains showed higher biocontrol efficacy in inhibiting mycelia growth and spore germination of P. theae. A detailed investigation on hydrolytic enzymes produced by antagonistic strains, which degrade the fungus cell wall, revealed that highest amount of chitinase and ß-1,3- glucanase in VTV7 and VBS3 strains. Further, the key antifungal secondary metabolites from these biocontrol agents associated with suppression of P. theae were identified using gas chromatography mass spectrometry. The above study clearly recognized the specific traits in the isolated microbes, which make them good candidates as plant growth-promoting rhizobacteria (PGPR) and biocontrol agents to improve plant growth and health. However, greenhouse trials and field application of these beneficial microbes is required to further confirm their efficacy for the management of stem canker in tea cultivation.

Evaluation of exotic soybean accessions and their use in developing improved soybean lines with resistance to Phomopsis seed decay.

Poor seed quality of soybean is often associated with Phomopsis seed decay (PSD), which is one of the most economically important seed diseases. Diaporthe longicolla (syn. Phomopsis longicolla) is the primary cause of PSD. Control of PSD is best accomplished by planting PSD-resistant cultivars. Sixteen exotic soybean accessions from the USDA soybean germplasm collection were screened for reaction to PSD at Stoneville, Mississippi. They consisted of maturity groups (MG) II, III and IV. Seeds from inoculated and non-inoculated plots harvested either promptly at maturity, or after a two-week delay in harvest, were assessed for infection by D. longicolla. Seed infection ranged from 0 to 36.7%. Overall, PI 417050 (MG II), PI 417017 (MG III), and PI 594692 (MG IV) had significantly (P ≤ 0.05) lower percentages of seed infected by D. longicolla and higher seed germinations than other genotypes in the same maturity groups. PI 587982A also performed well. As a result of these findings, these resistant accessions were used over multiple cycles of breeding to develop improved breeding lines with resistance to PSD and low seed damage. Breeding line 11043-225-72, with combined resistance from both PIs 417050 and 587982A, had low scores for PSD (6.7%) and seed damage (3.4%), while DS65-1, deriving resistance from PI 587982A, had the lowest seed damage score (1.1%) and the highest seed germination (85.6%) among all lines tested in 2017. DS65-1 and 11043-225-72, along with five other improved breeding lines, were provided to public soybean breeders for developing improved cultivars and germplasm lines. DS31-243 (PI 700941), derived from PI 587982A, was publicly released by the USDA in 2022. This research will lead to future releases of improved germplasm lines and cultivars with PSD resistance and high seed quality. It will also aid in disease management and be a benefit to soybean producers and the industry at large.

Specialized metabolites of the fungal genus Phomopsis: Structures, bioactivities and biosynthesis.

Fungi are important resources of novel bioactive compounds which have a high potential to be drug leads or candidates for further pharmacological applications. Phomopsis, a genus widely distributed in the environment, can produce various types of compounds including polyketides, alkaloids, terpenoids, cytochalasins, steroids and flavonoids. The metabolites of Phomopsis sp. showed diverse bioactivities such as antibacterial, anti-inflammatory, antimalarial, and so on, many of which may influence the physiological behaviour of the host plants. In this review, we focus on the chemical structures and biological activities of 183 specialized metabolites isolated from Phomopsis sp. in the decade (2013-2022). Moreover, the biosynthetic pathways of some typical components are summarized.

New Isocoumarin and Pyrone Derivatives from the Chinese Mangrove Plant Rhizophora mangle-Associated Fungus Phomopsis sp. DHS-11.

Mangrove-associated fungi are important sources for the discovery of new bioactive natural products. Three new isocoumarins (1-3) and one new pyrone derivative (4) were isolated from the ethyl acetate extract of the fermentation broth of the mangrove endophytic fungus Phomopsis sp. DHS-11. Nuclear magnetic resonance (NMR) spectroscopy (one-dimensional and two-dimensional) and mass spectrometry were used to determine the structures of these new compounds. The absolute configurations for the new isocoumarins 1-3 were determined by comparing their experimental and calculated electronic circular dichroism (ECD) spectra, while the configuration for the new pyrone-derivative 4 was tentatively solved by comparison of its 13C NMR data with reported data. In the biological activity test, compounds 1 and 3 showed cytotoxic activity against HeLa cells with IC50 values of 11.49 ± 1.64 µM and 8.70 ± 0.94 µM, respectively. The initial structure and activity relationship (SAR) analysis revealed that the length of the side chain at C-3 for isocoumarin-type compounds 1-3 could affect the cytotoxicity against HeLa cells. Compound 4 exhibited cytotoxic activities against human hepatoma cells HepG2 with an IC50 value of 34.10 ± 2.92 µM. All compounds have no immunosuppressive activity.

Biochemical, pathological and molecular characterisation of Phomopsis vexans: A causative of leaf blight and fruit rot in brinjal.

The pathogen Phomopsis vexans causes leaf blight, fruit rot, and damping off in brinjal plants, all of which are extremely detrimental. The pathogen affects host plant photosynthetic efficiency and fruit quantity and quality. An appreciation of the pathogenicity of P. vexans is essential for the effective control of infections in the field. Consequently, the goal of this study was to characterise P. vexans in terms of their biochemistry, molecular diversity, and pathogenicity. In terms of cellulase (97.7 U), catalase (12.2 U), and ascorbate peroxidase (147.3 U) activity, isolate PV1 performed best, followed by PV5 (CL-97.0 U, CAT-11.1 U and APX-144.4 U), and PV8 (CL-88.8 U, CAT-9.8 U and APX-141.9 U). In a greenhouse pathogenicity test, isolate PV1 had the highest incidence (97%) and severity (88.6%) of disease, whereas isolate PV6 showed the lowest incidence (57.2%) and severity (70%) of disease. The biochemical enzyme activity of P. vexans corresponds well with its greenhouse pathogenicity results, and its combination can be exploited to identify pathogenic P. vexans isolates. Using RAPD and ISSR primers, molecular characterisation indicated genetic diversity but could not distinguish isolates by geographical origin or pathogenicity. The pathogen P. vexans was verified by ITS1 and ITS4 molecular analysis, and the sequences were subsequently deposited in the NCBI database. In conclusion, the enzyme activity relevant to pathogenicity (CL, CAT and APX) in conjunction with the invivo pathogenicity assay might be utilised to differentiate between pathogenic (virulent) and non-pathogenic (avirulent) P. vexans isolates and develop suitable disease management strategies.

Secondary metabolites from a deep-sea derived fungal strain of Phomopsis lithocarpus FS508.

One new isoindolinone lactam lithocarlactam A (1) and one new triterpene lithocarin D (2), along with nine known natural products (3-11) were isolated from the broth extract of marine fungus Phomopsis lithocarpus FS508, which was isolated from a deep-sea sediment sample. Their structures were elucidated by interpretation of spectroscopic analysis and comparison with previously published literatures for the known compounds. Moreover, compounds 1 and 2 were evaluated for cytotoxic activity against SF-268, MCF-7, HepG-2, and A549 cell lines. However, both of them showed no activity against the tumor cell lines.

Foliar Fungicides Containing FRAC 11 Mitigate Phomopsis Stem Canker in Sunflower (Helianthus annuus).

Phomopsis stem canker reduces yield of sunflower (Helianthus annuus L.) up to or exceeding 40%; however, management recommendations have not been developed for U.S. farmers. Between 2009 and 2020, foliar fungicide trials were conducted in Minnesota, Nebraska, North Dakota, and South Dakota for a total of 49 location-years. Random effects meta-analyses were performed on the disease severity index (DSI) and yield data collected from the foliar fungicide trials to determine the overall and individual effectiveness of the tested fungicides. Effect sizes, Cohen's f or Hedges' g, were calculated as the difference in DSI or yield between the fungicide treatment and nontreated control (NTC) divided by the pooled SD. The pooled Cohen's f for DSI and yield was 0.40 (95% CI = [0.29, 0.42]), indicating a large effect size and that fungicide treatments had a significant effect on DSI and yield (P < 0.0001). Among the fungicide groups, quinone outside inhibitor (QoI) (DSI [k = 45; g = -0.47] and yield [k = 46; g = 0.41]) is moderately effective and premixes of demethylation inhibitors (DMI), succinate dehydrogenase inhibitors (SDHI), and QoI (DMI + SDHI + QoI) (DSI [k = 3; g = -0.79] and yield [k = 3; g = 0.94]) are largely effective in comparison with NTC. Upon performing prediction analyses, the probability of not recovering the fungicide application cost (Ploss) associated with QoI (pyraclostrobin) was <0.35 for a range of sunflower grain prices suggesting a greater probability of return on investment from a single application of fungicide. Overall, our study suggests that the use of QoI fungicides is likely to be profitable in the presence of Phomopsis stem canker (DSI > 5%).

Functional Characterization of Two Cell Wall Integrity Pathway Components of the MAPK Cascade in Phomopsis longicolla.

The pathogenic fungus Phomopsis longicolla causes numerous plant diseases, such as Phomopsis seed decay, pod and stem blight, and stem canker, which seriously affect the yield and quality of soybean production worldwide. Because of a lack of technology for efficient manipulation of genes for functional genomics, understanding of P. longicolla pathogenesis is limited. Here, we developed an efficient polyethylene glycol-mediated protoplast transformation system in P. longicolla that we used to characterize the functions of two genes involved in the cell wall integrity (CWI) pathway of the mitogen-activated protein kinase (MAPK) cascade, including PlMkk1, which encodes MAPK kinase, and its downstream gene PlSlt2, which encodes MAPK. Both gene knockout mutants ΔPlMkk1 and ΔPlSlt2 displayed a reduced growth rate, fragile aerial hyphae, abnormal polarized growth and pigmentation, defects in sporulation, inadequate CWI, enhanced sensitivity to abiotic stress agents, and significant deficiencies in virulence, although there were some differences in degree. The results suggest that PlMkk1 and PlSlt2 are crucial for a series of growth and development processes as well as pathogenicity. The developed transformation system will be a useful tool for additional gene function research and will aid in the elucidation of the pathogenic mechanisms of P. longicolla. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Genome-Wide Association Studies in Sunflower: Towards Sclerotinia sclerotiorum and Diaporthe/Phomopsis Resistance Breeding.

Diseases caused by necrotrophic fungi, such as the cosmopolitan Sclerotinia sclerotiorum and the Diaporthe/Phomopsis complex, are among the most destructive diseases of sunflower worldwide. The lack of complete resistance combined with the inefficiency of chemical control makes assisted breeding the best strategy for disease control. In this work, we present an integrated genome-wide association (GWA) study investigating the response of a diverse panel of sunflower inbred lines to both pathogens. Phenotypic data for Sclerotinia head rot (SHR) consisted of five disease descriptors (disease incidence, DI; disease severity, DS; area under the disease progress curve for DI, AUDPCI, and DS, AUDPCS; and incubation period, IP). Two disease descriptors (DI and DS) were evaluated for two manifestations of Diaporthe/Phomopsis: Phomopsis stem canker (PSC) and Phomopsis head rot (PHR). In addition, a principal component (PC) analysis was used to derive transformed phenotypes as inputs to a univariate GWA (PC-GWA). Genotypic data comprised a panel of 4269 single nucleotide polymorphisms (SNP), generated via genotyping-by-sequencing. The GWA analysis revealed 24 unique marker-trait associations for SHR, 19 unique marker-trait associations for Diaporthe/Phomopsis diseases, and 7 markers associated with PC1 and PC2. No common markers were found for the response to the two pathogens. Nevertheless, epistatic interactions were identified between markers significantly associated with the response to S. sclerotiorum and Diaporthe/Phomopsis. This suggests that, while the main determinants of resistance may differ for the two pathogens, there could be an underlying common genetic basis. The exploration of regions physically close to the associated markers yielded 364 genes, of which 19 were predicted as putative disease resistance genes. This work presents the first simultaneous evaluation of two manifestations of Diaporthe/Phomopsis in sunflower, and undertakes a comprehensive GWA study by integrating PSC, PHR, and SHR data. The multiple regions identified, and their exploration to identify candidate genes, contribute not only to the understanding of the genetic basis of resistance, but also to the development of tools for assisted breeding.

Endophytic fungus Phomopsis sp. as a source of 3-nitropropionic acid with larvicidal activity against Aedes aegypti (Linnaeus 1762, Diptera: Culicidae).

BACKGROUND: Aedes aegypti is the primary vector of viruses, such as Zika, chikungunya, yellow fever, and dengue. In this context, a biomonitored chemical study was conducted to evaluate the activity of the crude extract of the endophytic fungus Phomopsis sp. against the larvae of Aedes aegypti. METHODS: Crude extract, fractions, and isolated substances were evaluated in in-vitro assays against third-stage larvae of Aedes aegypti. RESULTS: We isolated 3-nitropropionic acid with an LC50 of 15.172 ppm and LC90 of 18.178 ppm after 24 hours of larval exposure. CONCLUSIONS: The results indicated that 3-nitropropionic acid exerted larvicidal activity.

DNP and ATP modulate the developments of pulp softening and breakdown in Phomopsis longanae Chi-infected fresh longan through regulating the cell wall polysaccharides metabolism.

Compared with P. longanae-infected longan, 2, 4-dinitrophenol (DNP) treatment for P. longanae-infected longan displayed the lower levels of pulp firmness, cell wall materials, ionic-soluble pectin, covalent-soluble pectin, hemicellulose, or cellulose, but the higher amount of water-soluble pectin, the higher activities of cell wall-degrading enzymes (CWDEs) (PG, ß-Gal, PME, Cx, and XET), and the higher transcript levels of CWDEs-related genes (DlPG1, DlPG2, Dlß-Gal1, DlPME1, DlPME2, DlPME3, DlCx1, and DlXET30). On the contrary, ATP treatment for P. longanae-infected longan exhibited opposite effects. The above results imply that DNP accelerated P. longanae-induced pulp softening and breakdown of fresh longan, which was because DNP up-regulated the transcript levels of CWDEs-related genes, enhanced the CWDEs activities, and accelerated the degradation of cell wall polysaccharides (CWP). However, ATP suppressed longan pulp softening and breakdown caused by P. longanae, because ATP down-regulated the transcript levels of CWDEs-related genes, lowered the CWDEs activities, and reduced the CWP degradation.

Immunosuppressive Cytochalasins from the Mangrove Endophytic Fungus Phomopsis asparagi DHS-48.

Three new cytochalasins, phomoparagins A-C (1-3), along with five known analogs (4-8), were isolated from Phomopsis asparagi DHS-48, a mangrove-derived endophytic fungus. Their structures, including their absolute configurations, were elucidated using a combination of detailed HRESIMS, NMR, and ECD techniques. Notably, 1 possessed an unprecedented 5/6/5/8/5-fused pentacyclic skeleton. These compounds were tested for their inhibitory activity against concanavalin A (ConA)/lipopolysaccharide (LPS)-induced spleen lymphocyte proliferation and calcineurin (CN) enzyme. Several metabolites (2 and 4-6) exhibited fascinating inhibitory activities with a relatively low toxicity. Furthermore, 2 was demonstrated to inhibit ConA-stimulated activation of NFAT1 dephosphorylation and block NFAT1 translocation in vitro, subsequently inhibiting the transcription of interleukin-2 (IL-2). Our results provide evidence that 2 may, at least partially, suppress the activation of spleen lymphocytes via the CN/NFAT signaling pathway, highlighting that it could serve as an effective immunosuppressant that is noncytotoxic and natural.

Pyrone Derivatives from a Mangrove Endophytic Fungus Phomopsis†asparagi LSLYZ-87.

Chemical investigation of the endophytic fungus Phomopsis asparagi LSLYZ-87 cultured on PDB medium led to the isolation of two new pyrone derivatives, phomasparapyrone A (1), and phomasparapyrone B (2), together with the known kojic acid (3). Their planar structures were connected through 1D and 2D NMR spectroscopic data. And the stereo structures of 1 and 2 were defined by comparison of the experimental ECD spectra to calculated one. All isolates were evaluated for their anti-neuroinflammatory activities. Among them, compound 2 showed moderate inhibition on NO accumulation induced by LPS on BV-2 cells in a dose dependent manner at 30, 40 and 50 µM, and without cytotoxicity in a concentration of 50.0 µM.

Cytotoxic Bromine- and Iodine-Containing Cytochalasins Produced by the Mangrove Endophytic Fungus Phomopsis sp. QYM-13 Using the OSMAC Approach.

Twelve new cytochalasins, phomopchalasins D-O (1-3, 5-12, and 14), including one brominated (2) and two iodinated cytochalasins (3 and 6), together with six known analogues (4, 13, and 15-18) were isolated from the mangrove-derived fungus Phomopsis sp. QYM-13 treated with 3% NaBr or 3% KI in potato liquid medium. Their structures and absolute configurations were established by extensive spectroscopic analysis (1D and 2D NMR, HRESIMS), electronic circular dichroism calculations, and a single-crystal X-ray diffraction experiment. Compounds 3 and 6 represent the first iodinated cytochalasins. Compounds 2, 15, 17, and 18 exhibited significant cytotoxicity against human cancer cell line MDA-MB-435 with IC50 values ranging from 0.2 to 8.2 µM.

Diaporthe/Phomopsis longicolla degrades an array of bisphenol analogues with secreted laccase.

With recent initiatives to ban bisphenol A (BPA) in certain commercial products, manufacturers shifted to the production and use of BPA analogues. However, some of these BPA alternatives still possess endocrine disruptive activities. Many fungal enzymes are known to biodegrade phenolic compounds, such as BPA. However, the activity of these enzymes on BPA analogues remains unexplored. This study reports a secreted laccase from the endophytic fungus Diaporthe longicolla capable of degrading an impressive range of bisphenol analogues. The secreted crude enzymes are optimally active at pH 5 from 39 °C to 60 °C, efficiently degrading BPA as well as BPA analogues BPB, BPC, BPE and BPF. A purified form of laccase was identified from the crude fungal extract using FPLC and peptide sequencing. Furthermore, BPA induced the expression of this D. longicolla laccase gene. Overall, this paper demonstrated that the crude laccase enzyme from D. longicolla metabolizes BPA and select analogues, implicating the potential role of this fungus to remove environmental bisphenols.