Endophytic fungi from the roots of horseradish (Armoracia rusticana) and their interactions with the defensive metabolites of the glucosinolate myrosinase – isothiocyanate system

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Variability of Bioactive Glucosinolates, Isothiocyanates and Enzyme Patterns in Horseradish Hairy Root Cultures Initiated from Different Organs

Horseradish hairy root cultures are suitable plant tissue organs to study the glucosinolate–myrosinase–isothiocyanate system and also to produce the biologically active isothiocyanates and horseradish peroxidase, widely used in molecular biology. Fifty hairy root clones were isolated after Agrobacterium rhizogenes infection of surface sterilized Armoracia rusticana petioles and leaf blades, from which 21 were viable after antibiotic treatment. Biomass properties (e.g., dry weight %, daily growth index), glucosinolate content (analyzed by liquid chromatography-electronspray ionization-mass spectrometry (LC-ESI-MS/MS)), isothiocyanate and nitrile content (analyzed by gas chromatography-mass spectrometry (GC-MS)), myrosinase (on-gel detection) and horseradish peroxidase enzyme patterns (on-gel detection and spectrophotometry), and morphological features were examined with multi-variable statistical analysis. In addition to the several positive and negative correlations, the most outstanding phenomenon was many parameters of the hairy root clones showed dependence on the organ of origin. Among others, the daily growth index, sinigrin, glucobrassicin, 3-phenylpropionitrile, indole-3-acetonitrile and horseradish peroxidase values showed significantly higher levels in horseradish hairy root cultures initiated from leaf blades

A kerti kakukkfű szövettani és alaktani vizsgálata

Vizsgáltam a Lamiaceae családba tartozó Thymus vulgaris L. morfológiai, szövettani felépítését, elsősorban a szár differenciálódását valamint az epidermisz függelékeket. A kapott eredményeket összehasonlítottam a Thymus serpyllum L., valamint a szakirodalomból ismert szintén Lamiaceae családba tartozó Salvia sp. adataival. Továbbá elvégeztem a két vizsgált Thymus faj illóolaj-tartalom összehasonlítását, melyből a gyógyhatásra lehet következtetni.BscBiológiag

Glutathione protects Candida albicans against horseradish volatile oil

Horseradish essential oil (HREO; a natural mixture of different isothiocyanates) had strong fungicide effect against Candida albicans both in volatile and liquid phase. In liquid phase this antifungal effect was more significant than those of its main components allyl, and 2-phenylethyl isothiocyanate. HREO, at sublethal concentration, induced oxidative stress which was characterized with elevated superoxide content and up-regulated specific glutathione reductase, glutathione peroxidase, catalase and superoxide dismutase activities. Induction of specific glutathione S-transferase activities as marker of glutathione (GSH) dependent detoxification was also observed. At higher concentration, HREO depleted the GSH pool, increased heavily the superoxide production and killed the cells rapidly. HREO and the GSH pool depleting agent, 1-chlore-2,4-dinitrobenzene showed strong synergism when they were applied together to kill C. albicans cells. Based on all these, we assume that GSH metabolism protects fungi against isothiocyanates. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Additional file 3: of Endophytic fungi from the roots of horseradish (Armoracia rusticana) and their interactions with the defensive metabolites of the glucosinolate - myrosinase - isothiocyanate system

Table S2. Residual amount of sinigrin and minor glucosinolates in horseradish extract after 16 days of incubation with endophytic fungi from horseradish and soil fungi. (DOC 33 kb

Additional file 2: of Endophytic fungi from the roots of horseradish (Armoracia rusticana) and their interactions with the defensive metabolites of the glucosinolate - myrosinase - isothiocyanate system

Table S1. Changes in concentration of glucosinolates in horseradish extracts incubated with horseradish endophytes. Raw abundance data. (DOC 58 kb

Additional file 1: of Endophytic fungi from the roots of horseradish (Armoracia rusticana) and their interactions with the defensive metabolites of the glucosinolate - myrosinase - isothiocyanate system

Figures S1-S9. Supplementary Figures (including Compound structures; Decomposition of minor glucosinolates in horseradish extract by endophytic and soil fungi; Sinigrin decomposition by endophytes in sinigrin-supplemented Saboraud Glucose Broth; Growth curves; SPME-GC-MS chromatograms; LC-MS extracted ion chromatograms; Growth inhibition curves and IC50 calculation) and additional methodical details on SPME-GC-MS. (PDF 5696 kb