2 6 dimethoxyphenol

Glucose, fructose, gluconic acid, and glycerol in must were determined using commercial enzymatic kits (Megazyme, Bray, Ireland). Laccase activity was determined using as substrate five mM 2,6-dimethoxyphenol (Sigma-Aldrich, St. Louis, MO, USA) according to Tosi et al. [4 (link)] and expressed as unit per mL (U mL⁻¹), where one unit corresponds to the amount of enzyme converting one μmol of substrate over one min.
The detection of B. cinerea in the grapes was confirmed by performing a species-specific polymerase chain reaction (PCR) analysis using as template DNA extracted from an amount of 50 g of berries. DNA extraction was carried out according to Rezaian and Krake [12 (link)], while PCR assay was performed using Bot-F and Bot-R primers as previously described by Lorenzini and Zapparoli [13 (link)].

Octan-2-ol (97%), 1-hexanol (99%), cis-3-hexenol (98%), trans-3-hexenol (97%), vanillin (99%), 2,6-dimethoxyphenol (99%), linalool (97%), terpinen-4-ol (≥95%), α-terpineol (90%), nerol (≥97%), geraniol (98%), linalool oxide (≥97%), β-citronellol (95%), p-cymene (99%), terpinolene (≥85%), γ-terpinene (≥97%), limonene (97%), 1,8-cineole (99%), 1,4-cineole(≥98.5%), β-damascenone (≥98%), isoamyl alcohol (98%), benzyl alcohol (≥99%), 2-phenylethanol (≥99%), ethyl acetate (99%), ethyl butanoate (99%), ethyl 3-methyl butanoate (≥98%), isoamyl acetate (≥95%), ethyl hexanoate (≥95%), phenylethyl acetate (99%), n-hexyl acetate (≥98%), ethyl lactate (≥98%), ethyl octanoate (≥98%), ethyl decanoate (≥98%), hexanoic acid (≥99%), octanoic acid (≥98%), α-phellandrene (95%), p-menthane-1,8-diol (97%), 3-methylbutanoic acid (99%), α-ionone (90%), 1-pentanol (99%), 1-butanol (≥99%), 2-butanol (≥99%), ethyl guaiacol (≥99%), vinyl guaiacol (≥98%), methyl-vanillate (99%), ethyl vanillate (99%), were supplied by Sigma Aldrich (Milan, Italy). Dichloromethane (≥99.8%) and methanol (≥99.8%), were provided by Honeywell (Seelze, Germany). Sodium chloride (≥99.5%) was supplied by Sigma Aldrich (Milan, Italy).

Aflatoxin B₁ (AFB₁), zearalenone (ZEN), deoxynivalenol (DON), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS), 2,6-dimethoxy phenol (DMP), syringaldazine (SGZ), and methyl syringate were purchased from Sigma-Aldrich (St. Louis, MO, USA). FB₁ (fumonisin B₁) and OTA (ocharatoxin A) were purchased from Pribolab (Beijing, China). DNA polymerase, T4 ligase, acetonitrile, and trifluoroacetic acid were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Vanillin, p-coumaric, syringic acid, syringaldehyde, caffeic acid, 1-hydroxybenzotriazole (HBT), gallic acid, isopropyl-β-D-thiogalactoside (IPTG), and kanamycin were purchased from Solarbio (Beijing, China). Ni-NTA agarose was purchased from QIAGEN (Hilden, Germany). The fungal laccase from Ganoderma sp. was purchased from Sunson (Yinchuan, Ningxia, China). Plant extracts from E. brevicornu, C. sativus L., L. angustifolia, A. officinalis, and S. tenuifolia were purchased from Ciyuan Biotech (Xi’an, Shanxi, China). All other chemicals were of analytical grade or chromatographically pure, and were commercially available.

Daunomycin hydrochlorine (≥90%), doxorubicin hydrochlorine (≥98%), streptomycin (≥98%), phosphate-buffered saline (PBS) without Ca²⁺ and Mg²⁺, fetal bovine serum (FBS), resazurin, MEM medium without phenol red (≥98%), o-dianisidine (≥95%), veratryl alcohol (96%), 2,6-dimethoxyphenol (99%), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (≥98%), DPPH^• (2,2-diphenyl-1-picrylhydrazyl), trolox (97%), peroxidase, type II from horseradish (200 KU) and catechol (≥99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals and reagents were of analytical grade.

Octan-2-ol (97%), 1-hexanol (99%), cis-3-hexenol (98%), trans-3-hexenol (97%), vanillin (99%), 2,6-dimethoxyphenol (99%), linalool (97%), terpinen-4-ol (≥95%), α-terpineol (90%), nerol (≥97%), geraniol (98%), linalool oxide (≥97%), β-citronellol (95%), p-cymene (99%), terpinolene (≥85%), γ-terpinene (≥97%), limonene (97%), 1,8-cineole (99%), 1,4-cineole(≥98.5%), β-damascenone (≥98%), isoamyl alcohol (98%), benzyl alcohol (≥99%), 2-phenylethanol (≥99%), ethyl acetate (99%), ethyl butanoate (99%), ethyl 3-methyl butanoate ((≥98%), isoamyl acetate (≥95%), ethyl hexanoate (≥95%), phenylethyl acetate (99%), n-hexyl acetate (≥98%), ethyl lactate (≥98%), ethyl octanoate (≥98%), ethyl decanoate (≥98%), hexanoic acid (≥99%), octanoic acid (≥98%), α-phellandrene (95%), p-menthane-1,8-diol (97%), 3-methylbutanoic acid (99%), α-ionone (90%), 1-pentanol (99%), 1-butanol (≥99%), 2-butanol (≥99%), ethyl guaiacol (≥99%), vinyl guaiacol (≥98%), methyl-vanillate (99%) and ethyl vanillate (99%), were supplied by Sigma Aldrich (Milan, Italy). Dichloromethane (≥99.8%) and methanol (≥99.8%), were provided by Honeywell (Seelze, Germany). Sodium chloride (≥99.5%) was supplied by Sigma Aldrich (Milan, Italy).

All chemicals were reagent-grade purity. ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), 2.6-dimethoxyphenol (DMP), sinapic acid, guaiacol, potassium octacyanomolybdate (IV) (K₄[Mo(CN)₈]), violuric acid and the Yeast Transformation Kit were purchased from Sigma (St. Louis, MO, United States). Zymoprep Yeast Plasmid Miniprep and Zymoclean Gel DNA Recovery Kit were from Zymo Research (Orange, CA, United States). NucleoSpin Plasmid kit was purchased from Macherey-Nagel (Düren, Germany). The uracil independent and ampicillin resistance shuttle vector pJRoC30 was obtained from the California Institute of Technology (Caltech, CA, United States). The protease deficient S. cerevisiae strain BJ5465 (α ura3-52 trp1 leu2Δ1 his3Δ200 pep4:HIS3 prb1Δ1.6R can1 GAL) was from LGC Promochem (Barcelona, Spain). The Escherichia coli strain XL2-Blue competent cells and Pfu DNA Polymerase were obtained from Agilent Technologies (Santa Clara, CA, United States). Restriction endonucleases BamHI and XhoI were purchased from New England Biolabs (Ipswich, MA, United States). Oligonucleotide primers were acquired from Isogen Life Science (Barcelona, Spain).