Anthocyanin Production and Enzymatic Degradation during the Development of Dark Purple and Lilac Paprika Fruit

in Journal of the American Society for Horticultural Science

During development, the fruit of some paprika (Capsicum annuum L.) cultivars shows a change in color from green to dark purple (e.g., ‘Mavras’) or lilac (e.g., ‘Tequila’). However, this purple coloration is rare among paprika cultivars and disappears in ripened fruit, which are red. Therefore, we investigated the mechanism causing this color change in the cultivars Mavras and Tequila to better understand how purple ripened fruit could be generated. High-performance liquid chromatography (HPLC) analyses of the anthocyanin contents of the fruit indicated that anthocyanin was undetectable in green fruit, accumulated in dark purple or lilac ones, and then decreased again in red ones in both cultivars. Furthermore, expressions of most of the analyzed anthocyanin biosynthesis–related genes and genes for their transcription factors increased in dark purple or lilac fruit and decreased in red ones, i.e., it was synchronized with the changes in anthocyanin contents. Furthermore, anthocyanin degradation activity as a result of peroxidases was detected at all stages but increased when the lilac or dark purple color started to fade. Thus, the development of purple coloration is caused by increased anthocyanin biosynthesis, whereas the fading of this coloration is a result of both a decrease in anthocyanin biosynthesis and an increase in anthocyanin degradation. At the ripening stage, the green pigment (chlorophyll) contents decreased, whereas the red pigment (carotenoid, particularly capsanthin) contents increased. However, these timings did not completely coincide with the timing of anthocyanin degradation, suggesting that the content of each pigment is individually regulated, and so purple, green, and red coloration could be freely expressed in mature paprika fruit.

Contributor Notes

We thank Kumi Yamanaka and Manami Akai for technical assistance.

Corresponding author. E-mail: agrisys@meiji.ac.jp.

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Article Figures

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    Fruit of the paprika cultivars Mavras and Tequila at the indicated number of days after pollination.

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    Content of (A, B) anthocyanins, (C, D) chlorophylls, and (E, F) carotenoids in fruit of the paprika cultivars Mavras and Tequila at the indicated number of days after pollination (DAP) in Expts. 1 (A, C, E) and 2 (B, D, F). Data were recorded on a fresh weight basis and are mean ± se (A-B: n = 3 at 10, 50, and 60 DAP and n = 6 at 15, 20, and 40 DAP; C–F: n = 3). Data for the anthocyanin contents were obtained from Table 1, whereas those for the chlorophyll contents are the sum of chlorophylls a and b from Table 2 and those for the carotenoid contents are the sum of β-carotene, lutein, zeaxanthin, violaxanthin, capsanthin, and capsorubin from Table 3. Asterisks indicate significant differences between ‘Mavras’ and ‘Tequila’ fruit at the same DAP (Student’s t test at P < 0.05).

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    Relative expression levels of anthocyanin biosynthesis–related genes and transcription factor genes in fruit of the paprika cultivars Mavras and Tequila at the indicated numbers of days after pollination (DAP) in Expt. 1. Data are means ± se (n = 3 at 10, 50, and 60 DAP; n = 6 at 15, 20, and 40 DAP). Different letters indicate significant differences among DAPs for each cultivar (Tukey–Kramer test, P < 0.05). PAL = phenylalanine ammonia-lyase, CHS = chalcone synthase, CHI = chalcone isomerase, F3H = flavanone 3-hydroxylase, F3ʹ5ʹH = flavonoid 3′5′-hydroxylase, DFR = dihydroflavonol 4-reductase, ANS = anthocyanidin synthase, UFGT = UDP glucose: flavonoid 3-O-glucosyltransferase, bHLH = basic helix-loop-helix, and WDR = WD40-repeat.

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    Relative expression levels of anthocyanin biosynthesis–related genes and transcription factor genes in the fruit of the paprika cultivars Mavras and Tequila at the indicated number of days after pollination (DAP) in Expt. 2. See Fig. 3 for additional details.

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    Anthocyanin degradation by enzyme extracts from the fruit of the paprika cultivars Mavras and Tequila. Reaction mixture contained anthocyanin and enzyme extract from the pericarps of the paprika fruit at 40 d after pollination (AE); anthocyanin and H2O2 (AH); anthocyanin, enzyme extract, and H2O2 (AEH); or anthocyanin, boiled enzyme extract, and H2O2 (AboEH). Δ anthocyanin content is the difference in anthocyanin content before and after the reaction. Data are means ± se (n = 3 for AE, AH, and AboEH; n = 6 for AEH). Different letters indicate significant differences among the reaction mixtures for the same cultivar (Tukey–Kramer test at P < 0.05).

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    Anthocyanin degradation by enzyme extracts from the fruit of the paprika cultivars Mavras and Tequila in the presence of H2O2 at different number of days after pollination (DAP) in Expts. 1 and 2. Data are means ± se (n = 3 at 10, 15, and 20 DAP; n = 6 at 40, 50, and 60 DAP). Δ anthocyanin content is the difference in anthocyanin content before and after the reaction. Different letters indicate significant differences among DAP for each cultivar (Tukey–Kramer test at P < 0.05).

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