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2 protocols using dihydromyricetin

1

Petunia Flower Developmental Stages

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Flowers (stage 1: buds of 0.6–3 cm length, stage 2: buds of 3–5 cm length, stage 3: open flowers) of cv. Salmon Ray (Danziger, Moshav Mishmar Hashiva, Israel), cv. Viva Orange (Florensis, Ambacht, The Netherlands), and cv. Electric Orange (Selecta One, Stuttgart, Germany) were harvested in the summers 2015–2017. Non-transgenic control plants of Petunia × hybrida cv. BabyDoll were obtained from Selecta One, cvs. Corso Rot, Corso Blau and Blackberry were purchased from Austrosaat (Vienna, Austria). The plant material was harvested from balcony pots or garden beddings, shock-frozen and kept at −80°C until analysis. Images of the petunia varieties are found in Figures 1, 2 and Supplementary Figure S3.
Reference compounds (cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, dihydromyricetin, dihydroquercetin, kaempferol, myricetin, and quercetin) were purchased from Extrasynthese (Genay, France), dihydrokaempferol from Sigma Aldrich (Vienna, Austria). Radiolabeled substrates were synthesized as previously described (Halbwirth et al., 2006 (link)).
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2

Polyphenol Characterization and Preparation

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Figure 1 shows the structure of the 19 polyphenols included in this study. Quercetin dihydrate (≥98%), morin hydrate, (±)-naringenin (~95% ), galangin, apigenin (≥95%), rutin hydrate (≥94%), chrysin (97%), (-)-epigallocatechin gallate [(-)-EGCG] (≥95%), (-)-epigallocatechin [(-)-EGC] (≥95%), (-)-epicatechin gallate [(-)-ECG] (≥95%), (-)-epicatechin [(-)-EC] (≥95%) and gallic acid monohydrate [GA] (≥99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Myricetin, dihydroMyricetin (both ≥95%) and kaempferide were obtained from Extrasynthese (Genay, France). KNO 3 and absolute ethanol (both pro analysis) were purchased from Kemika (Zagreb, Croatia). Buffer solution pH 3 was obtained from Reagecon (Shannon, Co. Clare, Ireland). Water was deionised by the Millipore Milli-Q system to the resistivity ≥18 MΩ cm.
Stock standard solutions of polyphenols (1.0x10 -2 mol L -1 ) were prepared from the dry pure substances. The stock solutions of catechins (EGCG, EGC, ECG, and EC) and gallic acid were prepared in deionised water obtained from a Millipore Milli-Q purification system. All other flavonoid stock standard solutions were prepared in absolute ethanol. The stock solutions were protected from light with aluminium foil and kept in a refrigerator.
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