Discovery of Novel Antioomycete Metabolites from the Marine-Derived Fungus Paraconiothyrium sporulosum.

In our screening program for natural products that are effective in controlling plant diseases, we found that the culture filtrate of Paraconiothyrium sporulosum SFC20160907-M11 effectively suppressed the development of tomato late blight disease caused by Phytophthora infestans. Using a bioassay-guided fractionation of antioomycete activity, 12 active compounds (1-12) were obtained from an ethyl acetate extract of the culture filtrate. Chemical structures of five new compounds 1-5 were determined by the extensive analyses of nuclear magnetic resonance (NMR), high resolution mass spectrometry (HRMS), and circular dichroism (CD) data. Interestingly, mycosporulonol (1) and botrallin (8) completely inhibited the growth of P. infestans at concentrations of 8 and 16 µg/mL, respectively. Furthermore, the spray treatment of 1 and 8 (500 µg/mL) successfully protected tomato seedlings against P. infestans with disease control values of 92%. Taken together, these results suggest that the culture filtrates of P. sporulosum SFC20160907-M11 and their bioactive metabolites can be used as new antioomycete agents for Phytophthora late blight control.

Edeine B1 produced by Brevibacillus brevis reduces the virulence of a plant pathogenic fungus by inhibiting mitochondrial respiration.

Plant pathogenic fungi cause serious diseases, which result in the loss of crop yields and reduce the quality of crops worldwide. To counteract the escalating risks of chemical fungicides, interest in biological control agents to manage plant diseases has significantly increased. In this study, we comprehensively screened microbial culture filtrates using a yeast screening system to find microbes exhibiting respiratory inhibition activity. Consequently, we found a soil-borne microbe Brevibacillus brevis HK544 strain exhibiting a respiration inhibitory activity and identified edeine B1 (EB1) from the culture filtrate of HK544 as the active compound of the respiration inhibition activity. Furthermore, against a plant pathogenic fungus Fusarium graminearum, our results showed that EB1 has effects on multiple aspects of respiration with the downregulation of most of the mitochondrial-related genes based on transcriptome analysis, differential EB1-sensitivity from targeted mutagenesis, and the synergistic effects of EB1 with electron transport chain complex inhibitors. With the promising plant disease control efficacy of B. brevis HK544 producing EB1, our results suggest that B. brevis HK544 has potential as a biocontrol agent for Fusarium head blight.IMPORTANCEAs a necrotrophic fungus, Fusarium graminearum is a highly destructive pathogen causing severe diseases in cereal crops and mycotoxin contamination in grains. Although chemical control is considered the primary approach to control plant disease caused by F. graminearum, fungicide-resistant strains have been detected in the field after long-term continuous application of fungicides. Moreover, applying chemical fungicides that trigger mycotoxin biosynthesis is a great concern for many researchers. Biocontrol of Fusarium head blight (FHB) by biological control agents (BCAs) represents an alternative approach and could be used as part of the integrated management of FHB and mycotoxin production. The most extensive studies on bacterial BCAs-fungal communications in agroecosystems have focused on antibiosis. Although many BCAs in agricultural ecology have already been used for fungal disease control, the molecular mechanisms of antibiotics produced by BCAs remain to be elucidated. Here, we found a potential BCA (Brevibacillus brevis HK544) with a strong antifungal activity based on the respiration inhibition activity with its active compound edeine B1 (EB1). Furthermore, our results showed that EB1 secreted by HK544 suppresses the expression of the mitochondria-related genes of F. graminearum, subsequently suppressing fungal development and the virulence of F. graminearum. In addition, EB1 exhibited a synergism with complex I inhibitors such as rotenone and fenazaquin. Our work extends our understanding of how B. brevis HK544 exhibits antifungal activity and suggests that the B. brevis HK544 strain could be a valuable source for developing new crop protectants to control F. graminearum.

Phenyl Ethers from the Marine-Derived Fungus Aspergillus tabacinus and Their Antimicrobial Activity Against Plant Pathogenic Fungi and Bacteria.

Marine fungi produce various secondary metabolites with unique chemical structures and diverse biological activities. In the continuing search for new antifungal agents from fungi isolated from marine environments, the culture filtrate of a fungus Aspergillus tabacinus SFC20160407-M11 exhibited the potential to control plant diseases caused by fungi. From the culture filtrate of A. tabacinus SFC20160407-M11, a total of seven compounds were isolated and identified by activity-guided column chromatography and spectroscopic analysis: violaceol I (1), violaceol II (2), diorcinol (3), versinol (4), orcinol (5), orsellinic acid (6), and sydowiol C (7). Based on in vitro bioassays against 17 plant pathogenic fungi and bacteria, violaceols and diorcinol (1-3) showed a broad spectrum of antimicrobial activity with minimum inhibitory concentration values in the range of 6.3-200 µg mL-1. These compounds also effectively reduced the development of rice blast, tomato late blight, and pepper anthracnose caused by plant pathogenic fungi in a dose-dependent manner. Our results suggest that A. tabacinus SFC20160407-M11 and its phenyl ether compounds could be used for developing new antimicrobial agents to protect crops from plant pathogens.

Curvicollide D, a new modified γ-lactone from the culture broth of Albifimbria verrucaria and its antifungal activity against plant pathogenic fungi.

In our screening program for new antifungal active compounds, a new modified γ-lactone curvicollide D (1) together with five known trichothecenes (2-6) were isolated from the culture filtrate of fungus Albifimbria verrucaria based on the in vitro antifungal assay. The chemical structure of new compound 1 was elucidated by NMR and HR-MS spectroscopic analyses, and the relative configurations of 1 were deduced from NOE experiments and coupling constant analysis. Compound 1 exhibited moderate antifungal activities against plant pathogenic fungi Botrytis cinerea, Colletotrichum coccodes, and Magnaporthe oryzae with MIC value in a range of 100-200 µg ml-1. Moreover, trichothecene compounds (2-6) displayed a broad spectrum of antifungal activities with MIC values in a range of 6.3-100 µg ml-1.

Machine-learning reinforcement for optimizing multilayered thin films: applications in designing broadband antireflection coatings.

The design and fabrication of nanoscale multilayered thin films play an essential role in regulating the operation efficiency of sensitive optical sensors and filters. In this paper, we introduce a packaged tool that employs flexible electromagnetic calculation software with machine learning in order to find the optimized double-band antireflection coatings in intervals of wavelength from 3 to 5 µm and 8 to 12 µm. Instead of computing or modeling an extremely enormous set of thin film structures, this tool enhanced with machine learning can swiftly predict the optical properties of a given structure with >99.7% accuracy and a substantial reduction in computation costs. Furthermore, the tool includes two learning methods that can infer a global optimal structure or suitable local optimal ones. Specifically, these well-trained models provide the highest accurate double-band average transmission coefficient combined with the lowest number of layers or the thinnest total thickness starting from a reference multilayered structure. Finally, the more sophisticated enhancement method, called the double deep Q-learning network, exhibited the best performance in finding optimal antireflective multilayered structures with the highest double-band average transmission coefficient of about 98.95%.

Clerodane Diterpenoids Identified from Polyalthia longifolia Showing Antifungal Activity against Plant Pathogens.

In the search for new natural resources showing plant disease control effects, we found that the methanol extract of Polyalthia longifolia suppressed fungal disease development in plants. To identify the bioactive substances, the methanol extract of P. longifolia was extracted by organic solvents, and consequently, four new 2-oxo-clerodane diterpenes (1-4), a new 4(3 → 2)-abeo-clerodane diterpene (5), together with ten known compounds (6-16) were isolated and identified from the extracts. Of the new compounds, compound 2 showed a broad spectrum of antifungal activity with moderated minimum inhibitory concentration (MIC) values in a range of 50-100 µg/mL against tested fungal pathogens. Considering with the known compounds, compound 6 showed the most potent antifungal activity with an MIC value in the range of 6.3-12.5 µg/mL. When compound 6 was evaluated for an in vivo antifungal activity against rice blast, tomato late blight, and pepper anthracnose, compound 6 reduced the plant disease by at least 60% compared to the untreated control at concentrations of 250 and 500 µg/mL. Together, our results suggested that the methanol extract of twigs and leaves of P. longifolia and its major compound 6 could be used as a source for the development of eco-friendly plant protection agents.

In Vitro and In Vivo Antifungal Activity of Sorbicillinoids Produced by Trichoderma longibrachiatum.

In the search for antifungal agents from marine resources, we recently found that the culture filtrate of Trichoderma longibrachiatum SFC100166 effectively suppressed the development of tomato gray mold, rice blast, and tomato late blight. The culture filtrate was then successively extracted with ethyl acetate and n-butanol to identify the fungicidal metabolites. Consequently, a new compound, spirosorbicillinol D (1), and a new natural compound, 2',3'-dihydro-epoxysorbicillinol (2), together with 11 known compounds (3-13), were obtained from the solvent extracts. The chemical structures were determined by spectroscopic analyses and comparison with literature values. The results of the in vitro antifungal assay showed that of the tested fungal pathogens, Phytophthora infestans was the fungus most sensitive to the isolated compounds, with MIC values ranging from 6.3 to 400 µg/mL, except for trichotetronine (9) and trichodimerol (10). When tomato plants were treated with the representative compounds (4, 6, 7, and 11), bisvertinolone (6) strongly reduced the development of tomato late blight disease compared to the untreated control. Taken together, our results revealed that the culture filtrate of T. longibrachiatum SFC100166 and its metabolites could be useful sources for the development of new natural agents to control late blight caused by P. infestans.

The role of dietary methionine concentrations on growth, metabolism and N-retention in cobia (Rachycentron canadum) at elevated water temperatures.

This study determined impacts of dietary methionine concentrations at two temperatures on growth, feeding efficiency and N-metabolites in juvenile cobia. Methionine concentrations of the experimental diets were deficient (M9; 9 g/kg), sufficient (M12; 12 g/kg) and surplus (M16, 16 g/kg). Water temperature was normal (30°C) or elevated (34°C). Twenty cobia in triplicate tanks were fed the experimental diets for 6 weeks. Both methionine and temperature affected cobia's growth and feeding efficiency. Cobia fed M9 performed lower than the fish fed M12 and M16 diets. Additionally, cobia reared at 34°C performed poorer than at 30°C, probably due to lower voluntary feed intake in the fish reared at 34°C. Protein efficiency ratio and protein productive value in cobia fed M9 diet were less than M12 or M16 diets. This was confirmed with the improved retentions of indispensable amino acids (AAs). No interactions between methionine and temperature were observed in growth and protein accretion. At 30°C, CF improved, while HSI and VSI declined upon methionine supplementation levels. Of which an interaction between temperature and methionine was present. Plasma, muscle and liver free AA and N-metabolites were affected by methionine and temperature. Furthermore, temperature affected cobia's lipid class composition, resulting in increased phospholipids and cholesterol at 34°C.

Antidiabetic and antioxidant activities of red seaweed Laurencia dendroidea

Objective: To investigate antidiabetic and antioxidant activities of the extract and fractions from Vietnamese red seaweed Laurencia dendroidea. Methods: The seaweed Laurencia dendroidea was extracted by using microwave-assisted extraction method in 80％ methanol. The seaweed extract was then fractionated using different solvents (n-hexane, chloroform, ethyl acetate, butanol and water). These obtained fractions were evaluated for α-glucosidase inhibitory and antioxidant activities. Antioxidant activities were tested using DPPH, nitric oxide radical scavenging and metal chelating assays. The enzyme inhibition mode was determined using Lineweaver-Burk plot. For acidic and thermal stabilities, the ethyl acetate fraction was treated at pH 2.0 and 100 ℃, respectively. The residual inhibitory activity of the fraction was calculated based on the initial inhibitory activity. For in vivo antidiabetic activity, mice were divided into four groups, including normal control, diabetic control, diabetic mice treated with ethyl acetate fraction and diabetic mice treated with gliclazide. Blood glucose level of treated mice during acute and prolonged treatments was measured. To evaluate the toxicity of the ethyl acetate fraction, the body weight changes and activities of liver function enzymes (aspartate transaminase, alanine transaminase and gammaglutamyl transferase) were carried out. Results: The extract of Laurencia dendroidea showed strong α-glucosidase inhibitory and DPPH radical scavenging activities. Methanolic concentrations affected both α-glucosidase inhibitory and antioxidant activities. A 80％ aqueous methanol was the suitable solvent for extraction of enzyme inhibitors and antioxidants. Among solvent fractions, ethyl acetate fraction had the highest inhibitory activities against α-glucosidase with a mixed type of inhibition and the strongest antioxidant activities, and was stable under acidic and thermal conditions. The ethyl acetate fraction treated diabetic mice significantly reduced blood glucose level compared with the diabetic control group (13.16 mmol/L vs. 22.75 mmol/L after 3 hours of treatment). Oral administration of ethyl acetate fraction did not exhibit toxicity at a dose of 100 mg/kg body weight as determined by body weight changes and liver biochemical parameters. Conclusions: Laurencia dendroidea could be a potential source for production of antidiabetic and antioxidative agents.

25 nm Single-Crystal Silicon Nanowires Fabricated by Anisotropic Wet Etching.

We report a top-down method for fabricating ultra-high aspect ratio single-crystal silicon nanowires. The fabrication method is based on the standard photolithography technique and anisotropic wet etching of the single-crystal silicon in KOH solution. SiO2 mask nanolines used for patterning single-crystal silicon nanowires are formed by the undercut etching of thin SiO2 layer in buffered hydrofluoric solution. The minimum width of the SiO2 mask nanolines are 50 nm. The length of SiO2 mask nanolines is 2 cm. The single-crystal silicon nanowires have been successfully transferred from the SiO2 mask nanolines by KOH anisotropic wet-chemical etching. The minimum width of the silicon nanowire has obtained to be 25 nm. The fabricated single-crystal silicon nanowires have trapezoidal and triangular cross sections, which are useful for applications in nanoelectronic and nanophotonic elements.

Feed intake and brain neuropeptide Y (NPY) and cholecystokinin (CCK) gene expression in juvenile cobia fed plant-based protein diets with different lysine to arginine ratios.

Cobia (Rachycentron canadum, Actinopterygii, Perciformes;10.5±0.1g) were fed to satiation with three plant-based protein test diets with different lysine (L) to arginine (A) ratios (LL/A, 0.8; BL/A, 1.1; and HL/A, 1.8), using a commercial diet as control for six weeks. The test diets contained 730 g kg(-1) plant ingredients with 505-529 g protein, 90.2-93.9 g lipid kg(-1) dry matter; control diet contained 550 g protein and 95 g lipid kg(-1) dry matter. Periprandial expression of brain NPY and CCK (npy and cck) was measured twice (weeks 1 and 6). At week one, npy levels were higher in pre-feeding than postfeeding cobia for all diets, except LL/A. At week six, npy levels in pre-feeding were higher than in postfeeding cobia for all diets. cck in pre-feeding cobia did not differ from that in postfeeding for all diets, at either time point. Cobia fed LL/A had lower feed intake (FI) than cobia fed BL/A and control diet, but no clear correlations between dietary L/A ratio and FI, growth and expression of npy and cck were detected. The data suggest that NPY serves as an orexigenic factor, but further studies are necessary to describe links between dietary L/A and regulation of appetite and FI in cobia.