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Fig. 7 | BMC Plant Biology

Fig. 7

From: In planta high levels of hydrolysable tannins inhibit peroxidase mediated anthocyanin degradation and maintain abaxially red leaves of Excoecaria Cochinchinensis

Fig. 7

Localization of hydrolysable tannins in leaves and their copigmentation with anthocyanins and inhibition on POD activity. A-B Cross-sections of the stage 2 leaf blades of Excoecaria (A) and Osmanthus (B) for anthocyanin localization. Scale bars indicate 50 μm. C-D The combination of ruthenium red and toluidine blue staining of the sections (5-μm-thick) of the stage 2 leaf blades of Excoecaria (C) and Osmanthus (D). Scale bars indicate 100 μm. E-F The vanillin staining of the sections (5-μm-thick) of the stage 2 leaf blades of Excoecaria (E) and Osmanthus (F). Scale bars indicate 100 μm. G-H Histochemical localization of POD activity in the leaves. Cross-sections of Excoecaria (G) and Osmanthus (H) leaf blades at stage 2 were incubated with guaiacol and H2O2 at pH 7.0. Scale bars indicate 100 μm. I-J Control cross-sections for the POD histochemical localization. Cross-sections of Excoecaria (I) and Osmanthus (J) leaf blades as in (I-J) was boiled for 15 min for enzyme denature and served as the control. Scale bars indicate 100 μm. K Total tannin content as TAE (tannic acid equivalent) in the leaves of Excoecaria and Osmanthus. L Gallic acid content in the leaves determined by HPLC using gallic acid as standard (Additional file 3: Figure S3). The statistical details of the values presented in (K-L) were as described in Fig. 1. M-O Copigmentation of galloylglucoses/ellagitannins on the main anthocyanins in Excoecaria and Osmanthus. The relative absorbance at 510 nm of Cy-3-(2″-galloylglucoside) (M) and Cy-3-O-glucoside (N) from Excoecaria leaves, of Cy-3-O-rutinoside from Osmanthus leaves (O) after addition of 0, 100, 300 μg/mL GAE of the tannins. An asterisk (* p < 0.05) represents the significance of difference between the addition of 0 and 100 or 300 μg/mL GAE of the tannins. Two asterisks represent p < 0.01, three asterisks represent p < 0.001, four asterisks represent p < 0.0001. All the values were means of the measurements of three individual repeats

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