ABSTRACT
Fungi exhibit a wide range of ecological guilds, but those that live within the inner tissues of plants (also known as endophytes) are particularly relevant due to the benefits they sometimes provide to their hosts, such as herbivory deterrence, disease protection, and growth promotion. Recently, endophytes have gained interest as potential biocontrol agents against crop pathogens, for example, coffee plants (Coffea arabica). Published results from research performed in our laboratory showed that endophytic fungi isolated from wild Rubiaceae plants were effective in reducing the effects of the American leaf spot of coffee (Mycena citricolor). One of these isolates (GU11N) from the plant Randia grandifolia was identified as Daldinia eschscholtzii (Xylariales). Its antagonism mechanisms, effects, and chemistry against M. citricolor were investigated by analyzing its volatile profile alone and in the presence of the pathogen in contactless and dual culture assays. The experimental design involved direct sampling of agar plugs in vials for headspace (HS) and headspace solid-phase microextraction (HS-SPME) gas chromatography-mass spectrometry (GC-MS) analysis. Additionally, we used ultrahigh-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS/MS) to identify nonvolatile compounds from organic extracts of the mycelia involved in the interaction. Results showed that more volatile compounds were identified using HS-SPME (39 components) than those by the HS technique (13 components), sharing only 12 compounds. Statistical tests suggest that D. eschscholtzii inhibited the growth of M. citricolor through the release of VOCs containing a combination of 1,8-dimethoxynapththalene and terpene compounds affecting M. citricolor pseudopilei. The damaging effects of 1,8-dimethoxynaphthalene were corroborated in an in vitro test against M. citricolor pseudopilei; scanning electron microscopy (SEM) photographs confirmed structural damage. After analyzing the UHPLC-HRMS/MS data, a predominance of fatty acid derivatives was found among the putatively identified compounds. However, a considerable proportion of features (37.3%) remained unannotated. In conclusion, our study suggests that D. eschscholtzii has potential as a biocontrol agent against M. citricolor and that 1,8-dimethoxynaphthalene contributes to the observed damage to the pathogen's reproductive structures.
ABSTRACT
In Costa Rica, two species of Psidium fruits, P. guajava and P. friedrichsthalianum, are widely consumed as food and used in folk medicine. Although studies have revealed the health effects of these fruits, there has been little research showing the antiplatelet activity of these species. This work evaluated the antiplatelet potential of aqueous extracts made from leaves and fruits of pink guava and Costa Rican guava. Platelet aggregation was induced by the platelet agonists ADP, TRAP-6, collagen and PMA. Platelet activation and secretion were studied using flow cytometry. The chemical profiles of the four extracts were characterized using UPLC-DAD-ESI-QTOF-MS. The studies revealed that the aqueous extracts of leaves and fruits of P. guajava and P. friedrichsthalianum inhibited platelet aggregation induced by ADP (4 µM), TRAP-6 (5 µM), collagen (1 µg mL-1) and PMA (100 nM), and the effect was dependent on the extract concentration. Extracts of leaves and fruits of pink guava and Costa Rican guava reduced secretion of P-selectin and activation of GP IIb/IIIa. The extracts of leaves and fruits of pink guava and Costa Rican guava proved to be a rich source of phenolic compounds, mainly quercetin aglycones and proanthocyanidins derived from (epi) catechin units. Other compounds such as ellagitannins, and benzophenones were also putatively identified. This research showed that P. guajava and P. friedrichsthalianum could potentially be used for the prevention of thrombotic events.
Subject(s)
Psidium , Costa Rica , Fruit , Plant Extracts/pharmacology , Plant Leaves , Polyphenols/pharmacologyABSTRACT
Global Natural Product Social Molecular Networking (GNPS) is an interactive online small molecule-focused tandem mass spectrometry (MS2) data curation and analysis infrastructure. It is intended to provide as much chemical insight as possible into an untargeted MS2 dataset and to connect this chemical insight to the user's underlying biological questions. This can be performed within one liquid chromatography (LC)-MS2 experiment or at the repository scale. GNPS-MassIVE is a public data repository for untargeted MS2 data with sample information (metadata) and annotated MS2 spectra. These publicly accessible data can be annotated and updated with the GNPS infrastructure keeping a continuous record of all changes. This knowledge is disseminated across all public data; it is a living dataset. Molecular networking-one of the main analysis tools used within the GNPS platform-creates a structured data table that reflects the molecular diversity captured in tandem mass spectrometry experiments by computing the relationships of the MS2 spectra as spectral similarity. This protocol provides step-by-step instructions for creating reproducible, high-quality molecular networks. For training purposes, the reader is led through a 90- to 120-min procedure that starts by recalling an example public dataset and its sample information and proceeds to creating and interpreting a molecular network. Each data analysis job can be shared or cloned to disseminate the knowledge gained, thus propagating information that can lead to the discovery of molecules, metabolic pathways, and ecosystem/community interactions.
