Botrytis cinerea is considered a model organism for the study of plant–pathogen interaction showing great genetic diversity and a high degree of morphological variability depending on environmental conditions. The use of new compounds and plant-based elicitors may trigger the expression of different B. cinerea genes, providing new sources of virulence factors. This work is focused on elucidating the phenotypic effect in B. cinerea of different carbon sources such as glucose, cellulose and tomato cell walls (TCW). Production of botrydial and dihydrobotrydial toxins was evaluated using thin-layer chromatography (TLC), proton nuclear magnetic resonance spectroscopy (1H-NMR) and mass spectrometry (UPLC-HRESIMS). Expression of the toxin biosynthesis gene BcBOT2 was followed using RT-qPCR. Results show an inhibition of the toxin biosynthesis pathway when TCW are present as a sole carbon source, suggesting that the toxin is only produced when rich molecules, like glucose, are available for fungal metabolism. That suggests a connection between gene expression of virulence factors and environmental conditions, where the silent genes can be induced by different culture conditions.Electronic supplementary materialThe online version of this article (10.1007/s00284-017-1399-3) contains supplementary material, which is available to authorized users.
Intradermal tests were performed in the AP group as part of the inclusion criteria, using Pharmalgen (ALK Abelló SA, Madrid, Spain) at increasing concentrations from 0.00001 to 0.1 μg/mL. A positive result was established as the lowest concentration producing a wheal with mean diameter ≥5 mm [14].
Total serum IgE, sIgE and sIgG4 levelsTotal serum IgE levels were measured by sandwich immunoassay on an Advia Centaur analyser (Siemens Healthcare, USA); sIgE and sIgG4 levels to AmV, rApi m 1 (phospholipase A2), rApi m 2 (hyaluronidase), rApi m 3 (acidic phosphatase), rApi m 5 (dipeptidyl peptidase IV) and rApi m 10 (icarapin) were measured by fluoroimmunoassay with ImmunoCAP 250 (Thermofisher, Uppsala, Sweden), according to the manufacturer instructions. In order to quantify the sIgE and sIgG4 levels to Api m 4, melittin sequence H-GIGAVLKVLTTGLPALISWIKRKRQQ-OH (Schafer-N ApS, Denmark) was coupled to activated CAPs by Thermofisher Scientific [15].
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