scholarly journals Blistering1 Modulates Penicillium expansum Virulence Via Vesicle-mediated Protein Secretion

2019 ◽  
Vol 19 (2) ◽  
pp. 344-361 ◽  
Author(s):  
Wayne M. Jurick ◽  
Hui Peng ◽  
Hunter S. Beard ◽  
Wesley M. Garrett ◽  
Franz J. Lichtner ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Secretion ◽  
Cross Sections ◽  
Fungal Growth ◽  
Single Copy ◽  
Protein Processing ◽  
Culture Broth ◽  
Penicillium Expansum ◽  
Mold Fungus ◽  
Host Infection

The blue mold fungus, Penicillium expansum, is a postharvest apple pathogen that contributes to food waste by rotting fruit and by producing harmful mycotoxins (e.g. patulin). To identify genes controlling pathogen virulence, a random T-DNA insertional library was created from wild-type P. expansum strain R19. One transformant, T625, had reduced virulence in apples, blistered mycelial hyphae, and a T-DNA insertion that abolished transcription of the single copy locus in which it was inserted. The gene, Blistering1, encodes a protein with a DnaJ domain, but otherwise has little homology outside the Aspergillaceae, a family of fungi known for producing antibiotics, mycotoxins, and cheese. Because protein secretion is critical for these processes and for host infection, mass spectrometry was used to monitor proteins secreted into liquid media during fungal growth. T625 failed to secrete a set of enzymes that degrade plant cell walls, along with ones that synthesize the three final biosynthetic steps of patulin. Consequently, the culture broth of T625 had significantly reduced capacity to degrade apple tissue and contained 30 times less patulin. Quantitative mass spectrometry of 3,282 mycelial proteins revealed that T625 had altered cellular networks controlling protein processing in the endoplasmic reticulum, protein export, vesicle-mediated transport, and endocytosis. T625 also had reduced proteins controlling mRNA surveillance and RNA processing. Transmission electron microscopy of hyphal cross sections confirmed that T625 formed abnormally enlarged endosomes or vacuoles. These data reveal that Blistering1 affects internal and external protein processing involving vesicle-mediated transport in a family of fungi with medical, commercial, and agricultural importance.

scholarly journals Two New Phomaligols from the Marine-Derived Fungus Aspergillus flocculosus and Their Anti-Neuroinflammatory Activity in BV-2 Microglial Cells

Marine Drugs ◽  
2021 ◽  
Vol 19 (2) ◽  
pp. 65
Author(s):  
Byeoung-Kyu Choi ◽  
Duk-Yeon Cho ◽  
Dong-Kug Choi ◽  
Phan Thi Hoai Trinh ◽  
Hee Jae Shin
Keyword(s):  
Nitric Oxide ◽  
Mass Spectrometry ◽  
Optical Rotation ◽  
Chemical Oxidation ◽  
Culture Broth ◽  
Membered Ring ◽  
Microglial Cells ◽  
No Production ◽  
Nmr Data ◽  
Ic50 Value

Two new phomaligols, deketo-phomaligol A (1) and phomaligol E (2), together with six known compounds (3–8) were isolated from the culture broth of the marine-derived fungus Aspergillus flocculosus. Compound 1 was first isolated as a phomaligol derivative possessing a five-membered ring. The structures and absolute configurations of the new phomaligols were determined by detailed analyses of mass spectrometry (MS), nuclear magnetic resonance (NMR) data, optical rotation values and electronic circular dichroism (ECD). In addition, the absolute configurations of the known compounds 3 and 4 were confirmed by chemical oxidation and comparison of optical rotation values. Isolated compounds at a concentration of 100 μM were screened for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced BV-2 microglial cells. Among the compounds, 4 showed moderate anti-neuroinflammatory effects with an IC50 value of 56.6 μM by suppressing the production of pro-inflammatory mediators in activated microglial cells without cytotoxicity.

scholarly journals New Isolated Metschnikowia pulcherrima Strains from Apples for Postharvest Biocontrol of Penicillium expansum and Patulin Accumulation

Toxins ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 397
Author(s):  
Laura Settier-Ramírez ◽  
Gracia López-Carballo ◽  
Pilar Hernández-Muñoz ◽  
Angélique Fontana ◽  
Caroline Strub ◽  
...  
Keyword(s):  
Liquid Medium ◽  
Fungal Growth ◽  
Its Region ◽  
Antagonistic Activity ◽  
Penicillium Expansum ◽  
Metschnikowia Pulcherrima ◽  
Potential Activity ◽  
Different Strains ◽  
Main Producer

Wild yeasts isolated from the surface of apples were screened for antagonistic activity against Penicillium expansum, the main producer of the mycotoxin patulin. Three antagonistic yeasts (Y33, Y29 and Y24) from a total of 90 were found to inhibit P. expansum growth. Identification by ITS region sequence and characterization showed that three selected isolates of yeast should be different strains of Metschnikowia pulcherrima. Several concentrations of the selected yeasts were used to study their in vitro antifungal effectivity against P. expansum on Petri dishes (plates with 63.6 cm2 surface) whereas their potential activity on patulin reduction was studied in liquid medium. Finally, the BCA that had the best in vitro antifungal capacity against P. and the best patulin degradation capacity was selected to be assessed directly on apples. All the selected strains demonstrated antifungal activity in vitro but the most efficient was the strain Y29. Isolated strains were able to reduce patulin content in liquid medium, Y29 being the only strain that completely reduced patulin levels within 120 h. The application of Y29 as biocontrol agent on the surface of apples inoculated with P. expansum, inhibited fungal growth and patulin production during storage. Therefore, the results shown that this yeast strain could be used for the reduction of P. expansum and its mycotoxin in apples or apple-based products by adapting the procedure application.

scholarly journals Time-of-Flight Secondary Ion Mass Spectrometry Imaging of Cross Sections from the Bacchanals Paintings of Nicolas Poussin

2021 ◽  
Vol 93 (10) ◽  
pp. 4463-4471
Author(s):  
Caroline Bouvier ◽  
Helen Glanville ◽  
Laurence de Viguerie ◽  
Chiara Merucci ◽  
Philippe Walter ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cross Sections ◽  
Mass Spectrometry Imaging ◽  
Secondary Ion Mass Spectrometry ◽  
Time Of Flight ◽  
Ion Mass Spectrometry ◽  
Nicolas Poussin ◽  
Secondary Ion

Control of Aspergillus flavus and aflatoxin production with spices in culture medium and rice

2013 ◽  
Vol 6 (1) ◽  
pp. 43-50 ◽  
Author(s):  
V. Aiko ◽  
A. Mehta
Keyword(s):  
Aspergillus Flavus ◽  
High Performance ◽  
Fungal Growth ◽  
Aflatoxin Production ◽  
Culture Broth ◽  
Minimum Inhibitory Concentrations ◽  
Food Grains ◽  
Star Anise ◽  
Aflatoxin B ◽  
Layer Chromatography

Cinnamon, cardamom, star anise and clove were studied for their effect on growth of Aspergillus flavus and aflatoxin B1 (AFB1) synthesis. The experiments were carried out in yeast extract sucrose culture broth as well as in rice supplemented with spices. AFB1 produced was analysed qualitatively and quantitatively using thin layer chromatography and high performance liquid chromatography, respectively. At a concentration of 10 mg/ml, cardamom and star anise did not exhibit any antifungal or anti-aflatoxigenic activity in culture broth, whereas cinnamon and clove inhibited A. flavus growth completely. The minimum inhibitory concentrations of cinnamon and clove were 4 and 2 mg/ml, respectively. Concentrations of cinnamon and clove below their minimum inhibitory concentrations showed enhanced fungal growth, while AFB1 synthesis was reduced. Clove inhibited the synthesis of AFB1 significantly up to 99% at concentrations ≥1.0 mg/ml. The spices also inhibited AFB1 synthesis in rice at 5 mg/g, although fungal growth was not inhibited. Clove and cinnamon inhibited AFB1 synthesis significantly up to 99 and 92%, respectively, and star anise and cardamom by 41 and 23%, respectively. The results of this study suggest the use of whole spices rather than their essential oils for controlling fungal and mycotoxin contamination in food grains.

scholarly journals Natalenamides A–C, Cyclic Tripeptides from the Termite-Associated Actinomadura sp. RB99

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 3003 ◽  
Author(s):  
Seoung Rak Lee ◽  
Dahae Lee ◽  
Jae Sik Yu ◽  
René Benndorf ◽  
Sullim Lee ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cyclic Peptides ◽  
Culture Broth ◽  
Resonance Spectroscopy ◽  
Ionization Mass ◽  
Inhibitory Effects ◽  
Associated Bacteria ◽  
Chemical Structures ◽  
New Compounds ◽  
Powerful Strategy

In recent years, investigations into the biochemistry of insect-associated bacteria have increased. When combined with analytical dereplication processes, these studies provide a powerful strategy to identify structurally and/or biologically novel compounds. Non-ribosomally synthesized cyclic peptides have a broad bioactivity spectrum with high medicinal potential. Here, we report the discovery of three new cyclic tripeptides: natalenamides A–C (compounds 1–3). These compounds were identified from the culture broth of the fungus-growing termite-associated Actinomadura sp. RB99 using a liquid chromatography (LC)/ultraviolet (UV)/mass spectrometry (MS)-based dereplication method. Chemical structures of the new compounds (1–3) were established by analysis of comprehensive spectroscopic methods, including one-dimensional (1H and 13C) and two-dimensional (1H-1H-COSY, HSQC, HMBC) nuclear magnetic resonance spectroscopy (NMR), together with high-resolution electrospray ionization mass spectrometry (HR-ESIMS) data. The absolute configurations of the new compounds were elucidated using Marfey’s analysis. Through several bioactivity tests for the tripeptides, we found that compound 3 exhibited significant inhibitory effects on 3-isobutyl-1-methylxanthine (IBMX)-induced melanin production. The effect of compound 3 was similar to that of kojic acid, a compound extensively used as a cosmetic material with a skin-whitening effect.

scholarly journals A Novel Approach to Characterize the Lipidome of Marine Archaeon Nitrosopumilus maritimus by Ion Mobility Mass Spectrometry

2021 ◽  
Vol 12 ◽  
Author(s):  
Kai P. Law ◽  
Wei He ◽  
Jianchang Tao ◽  
Chuanlun Zhang
Keyword(s):  
Mass Spectrometry ◽  
Cross Sections ◽  
Ion Mobility ◽  
Resolving Power ◽  
Ion Mobility Mass Spectrometry ◽  
Ion Chemistry ◽  
Lipid Species ◽  
Mobility System ◽  
Archaeal Lipids

Archaea are differentiated from the other two domains of life by their biomolecular characteristics. One such characteristic is the unique structure and composition of their lipids. Characterization of the whole set of lipids in a biological system (the lipidome) remains technologically challenging. This is because the lipidome is innately complex, and not all lipid species are extractable, separable, or ionizable by a single analytical method. Furthermore, lipids are structurally and chemically diverse. Many lipids are isobaric or isomeric and often indistinguishable by the measurement of mass or even their fragmentation spectra. Here we developed a novel analytical protocol based on liquid chromatography ion mobility mass spectrometry to enhance the coverage of the lipidome and characterize the conformations of archaeal lipids by their collision cross-sections (CCSs). The measurements of ion mobility revealed the gas-phase ion chemistry of representative archaeal lipids and provided further insights into their attributions to the adaptability of archaea to environmental stresses. A comprehensive characterization of the lipidome of mesophilic marine thaumarchaeon, Nitrosopumilus maritimus (strain SCM1) revealed potentially an unreported phosphate- and sulfate-containing lipid candidate by negative ionization analysis. It was the first time that experimentally derived CCS values of archaeal lipids were reported. Discrimination of crenarchaeol and its proposed stereoisomer was, however, not achieved with the resolving power of the SYNAPT G2 ion mobility system, and a high-resolution ion mobility system may be required for future work. Structural and spectral libraries of archaeal lipids were constructed in non-vendor-specific formats and are being made available to the community to promote research of Archaea by lipidomics.

scholarly journals Mass Spectrometry Imaging of Specialized Metabolites for Predicting Lichen Fitness and Snail Foraging

Plants ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 70 ◽  
Author(s):  
Alice Gadea ◽  
Mathieu Fanuel ◽  
Anne-Cécile Le Lamer ◽  
Joël Boustie ◽  
Hélène Rogniaux ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cross Sections ◽  
Mass Spectrometry Imaging ◽  
Ecological Studies ◽  
Optimal Defense Theory ◽  
Specialized Metabolites ◽  
Reproductive Tissues ◽  
Feeding Experiments ◽  
Slow Growing ◽  
Defense Theory

Lichens are slow-growing organisms supposed to synthetize specialized metabolites to protect themselves against diverse grazers. As predicted by the optimal defense theory (ODT), lichens are expected to invest specialized metabolites in higher levels in reproductive tissues compared to thallus. We investigated whether Laser Desorption Ionization coupled to Mass Spectrometry Imaging (LDI-MSI) could be a relevant tool for chemical ecology issues such as ODT. In the present study, this method was applied to cross-sections of thalli and reproductive tissues of the lichen Pseudocyphellaria crocata. Spatial mapping revealed phenolic families of metabolites. A quantification of these metabolites was carried out in addition to spatial imaging. By this method, accumulation of specialized metabolites was observed in both reproductive parts (apothecia and soralia) of P. crocata, but their nature depended on the lichen organs: apothecia concentrated norstictic acid, tenuiorin, and pulvinic acid derivatives, whereas soralia mainly contained tenuiorin and pulvinic acid. Stictic acid, tenuiorin and calycin, tested in no-choices feeding experiments, were deterrent for N. hookeri while entire thalli were consumed by the snail. To improve better knowledge in relationships between grazed and grazing organisms, LDI-MSI appears to be a complementary tool in ecological studies

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