Rice plants were treated with 10 or 20 mM of DHA (L-dehydroascorbic acid, Sigma-Aldrich, Cat. No. 261,556) (Chavan et al. 2022 (link)) and/or 300 µM PA (piperonylic acid, Sigma-Aldrich, Cat. No. P49805) (Desmedt et al. 2021 (link)). The chosen 20 mM concentration of DHA was previously optimized for best efficacy against M. graminicola and lack of phytotoxicity (Chavan et al. 2022 (link)). In order to potentially reduce the dose of DHA, the half dose, i.e., 10 mM was used while combining it with PA. Since PA was dissolved in DMSO, an additional mock treatment with only DMSO was included. Each plant was treated with 6.25 ml solution or distilled water containing 0.02% (v/v) of Tween20 (Sigma-Aldrich, Cat. No. P1379) for efficient spread and uptake of chemicals. Plants were treated by spraying the above-ground parts using a hand automizer sprayer.
L dehydroascorbic acid
Manufactured by Merck Group
Sourced in Spain
L-dehydroascorbic acid is a chemical compound that is used in various laboratory applications. It is a form of ascorbic acid, also known as vitamin C. The core function of L-dehydroascorbic acid is to serve as a reducing agent and an antioxidant in scientific research and experiments.
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Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (3 mentions)
Piperonylic acid, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Diamide, by Merck Group (1 mentions)
Catalase, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Anti gfp, by Thermo Fisher Scientific (1 mentions)
5 bromo 2 deoxyuridine brdu, by Thermo Fisher Scientific (1 mentions)
Anti prdx1, by Cell Signaling Technology (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Thermo Fisher Scientific (1 mentions)
Anti prdx3, by Abcam (1 mentions)
N ethylmaleimide, by Merck Group (1 mentions)
L ascorbic acid, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
N acetyl l cysteine, by Merck Group (1 mentions)
Hydrogen peroxide h2o2, by Merck Group (1 mentions)
3 protocols using l dehydroascorbic acid
1
Rice plants treated with DHA and PA
2
Oxidative Stress Assay Reagents
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l -ascorbic acid (AA), l -dehydroascorbic acid (DHA), N-ethylmaleimide (NEM), crystal violet, N-acetyl-l -cysteine (NAC), glutathione, DMSO, dithiothreitol (DTT), hydrogen peroxide (H2O2), catalase, diamide, trichloroacetic acid (TCA) and cycloheximide were purchased from Sigma-Aldrich. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA), 5-bromo-2′-deoxy-uridine (BrdU) and propidium iodide (PI) were from Thermo Fisher Scientific. The following antibodies were used: anti-PRDX3 (Abcam), anti-GFP (Life Technologies), anti-β-actin (Sigma-Aldrich), anti-PRDX1 (Cell Signaling Technology), FITC Mouse anti-BrdU (BD Biosciences).
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3
Grapevine Cell Elicitation with B. cinerea
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Azure A (AzA) (80%), chloroplatinic acid hexahydrate (≥99.9%), citric acid (trisodium salt), L-dehydroascorbic acid (DHA), ethanol (≥ 99.5% v/v), D(+)-glucose, H2O2 (35%), D-mannitol, L-methionine, trans-resveratrol, salicylic acid, sodium Lascorbate, sodium lauryl sulfate (SDS 95%), ascorbate oxidase (AO) from Cucurbita sp.
(156.60 units/mg solid), catalase (CAT) from bovine liver (2,200 units/mg protein) and peroxidase from horseradish (HRP, 313 units/mg solid) were obtained from Merck-Sigma-Aldrich (Spain). MeJa was purchased from Duchefa (Spain). KCl, KNO3, K2HPO4, KH2PO4 and K4Fe(CN)6 were acquired from Merck. H2SO4 and HCl were supplied by Panreac. Clopyralid 72% came from Dow AgroSciences (US).
The fungus B. cinerea was grown on potato dextrose agar plates for 20 days at 28 ºC until extensive sporulation. Spores were recovered by adding 10 ml of distilled sterile water to each plate. The suspension was filtered through nylon mesh in order to remove contaminating hyphae. A haemocytometer was used to determine the spore concentration.
For the elicitation experiments, grapevine cell cultures were cultivated with B. cinerea spores at a final concentration of 6 × 10 4 spores/mL. All the measurements were taken with freshly prepared solutions. The concentration of the H2O2 stock solutions was spectrophotometrically checked at 240 nm (ε = 43.6 M -1 cm -1 ) [28] .
(156.60 units/mg solid), catalase (CAT) from bovine liver (2,200 units/mg protein) and peroxidase from horseradish (HRP, 313 units/mg solid) were obtained from Merck-Sigma-Aldrich (Spain). MeJa was purchased from Duchefa (Spain). KCl, KNO3, K2HPO4, KH2PO4 and K4Fe(CN)6 were acquired from Merck. H2SO4 and HCl were supplied by Panreac. Clopyralid 72% came from Dow AgroSciences (US).
The fungus B. cinerea was grown on potato dextrose agar plates for 20 days at 28 ºC until extensive sporulation. Spores were recovered by adding 10 ml of distilled sterile water to each plate. The suspension was filtered through nylon mesh in order to remove contaminating hyphae. A haemocytometer was used to determine the spore concentration.
For the elicitation experiments, grapevine cell cultures were cultivated with B. cinerea spores at a final concentration of 6 × 10 4 spores/mL. All the measurements were taken with freshly prepared solutions. The concentration of the H2O2 stock solutions was spectrophotometrically checked at 240 nm (ε = 43.6 M -1 cm -1 ) [28] .
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