Anthocyanin Accumulation and Expression of Anthocyanin Biosynthetic Genes in Radish (Raphanus sativus)

Park, Nam Il; Xu, Hui; Li, Xiaohua; Jang, In Hyuk; Park, Suhyoung; Ahn, Gil Hwan; Lim, Yong Pyo; Kim, Sun-Ju; Park, Sang Un

doi:10.1021/jf200824c

Cited by 102 publications

(91 citation statements)

References 37 publications

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“…Radishes are cultivated to produce seed oil and sprouts, as well as edible taproots, which contain valuable phytochemicals such as glucosinolates and flavonoids (Blažević and Mastelić 2009;Park et al 2011). Some radish varieties have red skin and/or flesh due to anthocyanin accumulation.…”

Section: Introductionmentioning

confidence: 99%

Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1

et al. 2015

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RsMYB1, a MYB TF of red radish origin, was characterized as a positive regulator to transcriptionally activate the anthocyanin biosynthetic machinery by itself in Arabidopsis and tobacco plants. Anthocyanins, providing the bright red-orange to blue-violet colors, are flavonoid-derived pigments with strong antioxidant activity that have benefits for human health. We isolated RsMYB1, which encodes an R2R3-MYB transcription factor (TF), from red radish plants (Raphanus sativus L.) that accumulate high levels of anthocyanins. RsMYB1 shows higher expression in red radish than in common white radish, in both leaves and roots, at different growth stages. Consistent with RsMYB1 function as an anthocyanin-promoting TF, red radishes showed higher expression of all six anthocyanin biosynthetic and two anthocyanin regulatory genes. Transient expression of RsMYB1 in tobacco showed that RsMYB1 is a positive regulator of anthocyanin production with better efficiency than the basic helix-loop-helix (bHLH) TF gene B-Peru. Also, the synergistic effect of RsMYB1 with B-Peru was larger than the effect of the MYB TF gene mPAP1D with B-peru. Arabidopsis plants stably expressing RsMYB1 produced red pigmentation throughout the plant, accompanied by up-regulation of the six structural and two regulatory genes for anthocyanin production. This broad transcriptional activation of anthocyanin biosynthetic machinery in Arabidopsis included up-regulation of TRANSPARENT TESTA8, which encodes a bHLH TF. These results suggest that overexpression of RsMYB1 promotes anthocyanin production by triggering the expression of endogenous bHLH genes as potential binding partners for RsMYB1. In addition, RsMYB1-overexpressing Arabidopsis plants had a higher antioxidant capacity than did non-transgenic control plants. Taken together, RsMYB1 is an actively positive regulator for anthocyanins biosynthesis in radish plants and it might be one of the best targets for anthocyanin production by single gene manipulation being applicable in diverse plant species.

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Section: Introductionmentioning

confidence: 99%

Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1

et al. 2015

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“…LcMYB1 controls anthocyanin biosynthesis in litchi, and LcUFGT might be the structural gene that is targeted and regulated by LcMYB1 (Lai et al, 2014). Concerning the anthocyanin biosynthesis in radish, Park et al (2011) reported that RsDFR and RsANS were found to accumulate in the flesh or skin, and radish skin contained higher CHS, CHI, and F3H transcript levels than radish flesh. Lim et al (2015) isolated the RsMYB1 gene from red radish (red skin) plants and suggested that RsMYB1 is an actively positive regulator for anthocyanin biosynthesis in radish plants.…”

Section: Introductionmentioning

confidence: 99%

Red Pigment Content and Expression of Genes Related to Anthocyanin Biosynthesis in Radishes (Raphanus sativus L.) with Different Colored Flesh

Chen¹,

Chen²,

Huo³

et al. 2016

JAS

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Radish with red skin and red flesh (Raphanus sativus) is a unique vegetable containing large amounts of a red pigment, which is widely used in foods, wine, and cosmetics. To investigate the gene or genes that play a key role in anthocyanin biosynthesis in radish with red skin and red flesh, the red pigment content and expression of genes involved in anthocyanin biosynthesis of 16 varieties with different colored flesh were studied. The expression level of the genes RsPAL, RsCHS, RsCHI, RsDFR, RsF3H, RsF3'H, and RsANS in radish with red skin and red flesh are all significantly higher than that of radish with white skin and white flesh, radish with red skin and white flesh, radish with green skin and pinky flesh, and radish with red skin and pinky flesh. Correlation analysis indicated that the gene expression level of RsDFR, RsF3H, RsCHS, RsANS, RsF3'H, RsCHI, and RsPAL showed remarkable positive and significant correlation to red pigment content of radish. Stepwise regression analysis showed that the gene expression level of RsDFR had the highest and significant direct effect (0.8932) on red pigment content of radish. The results indicated that (1) the red pigment content of radish is closely related to the increased expression of a number of structural genes in anthocyanin biosynthesis, (2) the RsDFR gene plays a key role in anthocyanin biosynthesis in red radish with red flesh, and (3) RsDFR might be one of the best targets in genetic engineering for anthocyanin production from radish and other plants.

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“…Expression of F3'H in root skin and flesh was assayed by RT-qPCR assay using a 20 μL reaction mix containing 1 μL of 10X diluted cDNA templates, 10 μL of SYBR Premix Ex-Taq II (TaKaRa Bio), and 0.25 μM of 26S ribosomal RNA (26S) primer (Park et al, 2011; forward 5'-AACACCCTTTGTGGGTTCTAGGT-3' and reverse 5'-GCCCTCGACCTATTCTCAAACTT-3') and the above F3'H primers. Each RT-qPCR assay was conducted using a Thermal Cycler Dice Real-Time System (TaKaRa Bio) with an initial denaturation of 30 s at 95°C, followed by 50 cycles of denaturation for 5 s at 95°C, annealing for 10 s at 58°C (F3'H)/56°C (26S), and elongation for 20 s at 72°C.…”

Section: Expression Analysis Of Radish Anthocyanin Biosynthesis Genesmentioning

confidence: 99%

“…Radish anthocyanins are brightly colored over a wide pH range and their multiple acylation with hydroxycinnamic acids contributes to their remarkable heat stability in acidic environments (Jing et al, 2012). Radish anthocyanins have been widely used as natural colorants and ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), CHS, CHI, F3H, DFR, and ANS, was investigated in white and red radishes (Park et al, 2011). However, little attention has been paid to the relationship between F3'H expression and root color in radishes.…”

Section: Introductionmentioning

confidence: 99%

“…sativus) called 'Hatsuka Daikon' in Japan, white radishes were recessive to colored radishes (such as purple and red colors) and the white color was controlled by a single gene because the ratio between colored and white radishes was 3:1 in the F 2 population (Tatebe, 1938). Park et al (2011) showed that the genes RsDFR and RsANS of the anthocyanin biosynthesis pathway were expressed more highly in the red tissues of red Chinese radishes, and were not expressed in white radishes or white tissues of red Chinese radishes. In the present study, we confirmed the expression patterns of these two genes (Fig.…”

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Insertion of a Retrotransposon into a Flavonoid 3'-hydroxylase Homolog Confers the Red Root Character in the Radish (Raphanus sativus L. var. longipinnatus L. H. Bailey)

et al. 2018

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Flavonoid 3'-hydroxylase (F3'H) has an important role in determining anthocyanin patterns in plants. Here, we analyzed root coloration characteristics in the radish by investigating anthocyanidin and expression of F3'H genes in cultivars with purple, red, or white roots. Cyanidin was detected in the roots of the purple radish, while pelargonidin was found in the red radish; however, neither of these anthocyanidins was detected in white radish. We isolated the RsF3'H gene of the purple root radish and found that it showed a relatively higher level of expression compared to the RsF3'H of the red radish. Moreover, we identified a retrotransposon, gypsy-Ty3, in the first exon of the F3'H homolog in the red radish. These results suggested that the F3'H enzyme may determine cyanidin-based anthocyanin in the purple radish, and that the lack of F3'H function due to the retrotransposon insertion, contributed to pelargonidin-based anthocyanin accumulation in the red radish.

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Anthocyanin Accumulation and Expression of Anthocyanin Biosynthetic Genes in Radish (Raphanus sativus)

Cited by 102 publications

References 37 publications

Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1

Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1

Red Pigment Content and Expression of Genes Related to Anthocyanin Biosynthesis in Radishes (Raphanus sativus L.) with Different Colored Flesh

Insertion of a Retrotransposon into a <i>Flavonoid 3'-hydroxylase</i> Homolog Confers the Red Root Character in the Radish (<i>Raphanus sativus</i> L. var. <i>longipinnatus</i> L. H. Bailey)

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