Overexpression of a Freesia hybrida flavonoid 3-O-glycosyltransferase gene, Fh3GT1, enhances transcription of key anthocyanin genes and accumulation of anthocyanin and flavonol in transgenic petunia (Petunia hybrida)

Sun, Wei; Meng, Xiangyu; Liang, Lingjie; Li, Yueqing; Zhou, Tongtong; Cai, Xinquan; Wang, Li; Gao, Xiang

doi:10.1007/s11627-017-9836-3

Cited by 21 publications

(8 citation statements)

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“…To isolate the potential glucosyltransferases participating in flavonoid especially flavonol biosynthesis, the amino acid sequence of Fh3GT1 hitherto described in our earlier studies was used as a bait probe to thoroughly search the would-be flavonoid 3- O -glucosyltransferases in F. hybrida (Sui et al, 2011; Sun et al, 2016; Sun et al, 2017). Afterwards, another 38 putative unigenes were mined from the Freesia floral transcriptomic database and predicted as glycosyltransferases.…”

Section: Resultsmentioning

confidence: 99%

“…‘Red River ® ’ and ‘Ambiance’, cultivars of F. hybrida with red and white flowers, were grown in a greenhouse with a 14-h:10-h (light/dark) photoperiod at about 15°C. For detecting spatiotemporal expression profiles, five floral developmental stages and eight kinds of tissues or organs of Red River ® were sampled as mentioned in earlier studies (Sui et al, 2011; Sun et al, 2015; Sun et al, 2016; Sun et al, 2017; Li, Y. et al, 2016; Li et al, 2017; Gao et al, 2018; Ju et al, 2018; Li et al, 2019). For gene expression analysis between Red River ® and Ambiance, flowers fully bloomed on the first day were collected.…”

Section: Methodsmentioning

confidence: 99%

“…Till now, a versatile transient protoplast transfection assay was established based on the protoplasts isolated from Freesia callus (Shan et al, 2019a). Moreover, four regulatory genes and seven anthocyanin biosynthetic genes, including one glycosyltransferase gene Fh3GT1 shown to prefer UDP-glucose as sugar donor to glycosylate anthocyanidins and flavonols, were cloned and characterized (Sui et al, 2011; Sun et al, 2015; Sun et al, 2016, Sun et al, 2017; Li, Y. et al, 2016; Li et al, 2017; Li et al, 2019; Ju et al, 2018; Shan et al, 2019b). However, the expression patterns and enzymatic characteristics of Fh3GT1 could not coincide well with flavonol accumulations during flower development, which suggested the existence of other potentially uncharacterized UF3GT genes preferentially linked to flavonol glycosylation.…”

Section: Introductionmentioning

confidence: 99%

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Functional Differentiation of Duplicated Flavonoid 3-O-Glycosyltransferases in the Flavonol and Anthocyanin Biosynthesis of Freesia hybrida

Meng

Zhou

et al. 2019

Front. Plant Sci.

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Flavonols and anthocyanins are two widely distributed groups of flavonoids that occurred apart during plant evolution and biosynthesized by shared specific enzymes involved in flavonoid metabolism. UDP-glucose, flavonoid 3-O-glycosyltransferase (UF3GT), is one of the common enzymes which could catalyze the glycosylation of both flavonol and anthocyanidin aglycons simultaneously in vitro. However, whether and how UF3GT paralogous genes function diversely at the biochemical and transcriptional levels are largely unknown. Recently, Fh3GT1 was identified to be a member of UF3GTs in Freesia hybrida. However, its expression patterns and enzymatic characteristics could not coincide well with flavonol accumulation. In an attempt to characterize other flavonoids, especially flavonol glycosyltransferase genes in Freesia, three closest candidate UFGT genes—Fh3GT2, Fh3GT3, and Fh3GT4—were mined from the Freesia transcriptomic database and isolated from the flowers of the widely distributed Freesia cultivar, Red River®. Based on bioinformatic analysis and enzymatic assays, Fh3GT2 turned out to be another bona fide glycosyltransferase gene. Biochemical analysis further proved that Fh3GT2 preferentially glucosylated kaempferol while Fh3GT1 controlled the glucosylation of quercetin and anthocyanidins. In addition, transfection assays demonstrated that Fh3GT2 could be mainly activated by the flavonol regulator FhMYBF1 or the anthocyanin regulator FhPAP1, whereas Fh3GT1 could only be activated by FhPAP1. These findings suggested that Fh3GTs might have functionally diverged in flavonoid biosynthesis at both the biochemical and transcriptional levels.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functional Differentiation of Duplicated Flavonoid 3-O-Glycosyltransferases in the Flavonol and Anthocyanin Biosynthesis of Freesia hybrida

Meng

Zhou

et al. 2019

Front. Plant Sci.

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“…Recent studies revealed that there were five kinds of anthocyanin aglycons, namely delphinidin, petunidin, malvinidin, peonidin, and cyanidin, and two kinds of flavonols, namely kaempferol and quercetin, as well as PAs in F. hybrida “Red River ® ” (Sun et al, 2015, 2016; Li et al, 2016). To date, six structural genes and two IIIf bHLH regulatory genes have been functionally characterized (Sui et al, 2011; Sun et al, 2015, 2016, 2017; Li et al, 2016, 2017; Ju et al, 2018). In this study, the first MYB regulatory gene FhMYB5 was isolated and performed as candidate gene involved in flavonoid biosynthesis in the Freesia flowers.…”

Section: Introductionmentioning

confidence: 99%

The R2R3-MYB Factor FhMYB5 From Freesia hybrida Contributes to the Regulation of Anthocyanin and Proanthocyanidin Biosynthesis

Shan

Zhou

et al. 2019

Front. Plant Sci.

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The flavonoids are important and nourishing compounds for plants and human. The transcription regulation of anthocyanin and proanthocyanidin (PA) biosynthesis was extensively studied in dicot compared with monocot plants. In this study, we characterized the functionality of an R2R3-MYB gene FhMYB5 from the monocotyledonous flowering plant of Iridaceae, Freesia hybrida. Multiple sequence alignment and phylogenetic analysis implied that FhMYB5 was clustered into grapevine VvMYB5b subclade. Correlation analysis indicated that the spatio-temporal expression patterns of FhMYB5 coincided well with anthocyanin and PA accumulations in Freesia per se. Furthermore, transient transfection assays in Freesia protoplasts revealed that the late flavonoid biosynthetic genes (e.g., DFR and LDOX) were slightly up-regulated by FhMYB5 alone, whereas both early and late biosynthetic genes were significantly activated when FhMYB5 were co-infected with either of the two IIIf clade bHLH genes, FhTT8L and FhGL3L. Moreover, these results were further confirmed by co-transfection of FhMYB5 with either of the bHLH genes aforementioned into protoplasts expressing GUS reporter gene driven by Freesia promoters. In addition, the overexpression of FhMYB5 in tobacco and Arabidopsis could also significantly up-regulate the expression of genes participating in the general flavonoid pathway. In conclusion, FhMYB5 was proved to function in the general flavonoid pathway in Freesia. The results implied a function conservation of flavonoid biosynthesis related MYB regulators in angiosperm plants.

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“…Flavonoids comprise five major subgroups in higher plants beside anthocyanins: chalcones, flavones, flavonols, flavandiols or proanthocyanidins, and aurones (Winkel-Shirley, 2001a). Most of the structural genes encoding the enzymes responsible for anthocyanin biosynthesis have been isolated from various plants, for example, Arabidopsis (Arabidopsis thaliana), apple (Malus domestica), petunia (Petunia hybrida), grape (Vitis vinifera), and other species [5][6][7][8]. Conversion of the initial precursor phenylalanine is catalyzed in a stepwise manner by phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3hydroxylase (F3′H) or flavonoid 3′,5′-hydroxylase (F3′5′H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), and O-methyltransferase (OMT) [ 9].…”

Section: Introductionmentioning

confidence: 99%

Comprehensive influences of overexpression of a MYB transcriptor regulating anthocyanin biosynthesis on transcriptome and metabolome of tobacco leaves

Zong¹,

Li²,

Xi³

et al. 2019

Preprint

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Background Overexpression of MYB transcription factors can induce the expression of structural genes for anthocyanin biosynthesis and increase the anthocyanin content of plant tissues. However, it remains unclear whether MYB transcription factor overexpression effects the activation of other genes and the concomitant accumulation of chemical compounds. Results Overexpression of LrAN2 promoted anthocyanin accumulation in a variety of tissues in tobacco cultivar Samsun. Only 185 unigenes, from total of 160,965, were expressed differently in leaves and 241 chemical compounds exhibited differences in accumulation. Four anthocyanins, including apigeninidin chloride, cyanidin 3-O-malonylhexoside, pelargonidin 3-O-beta-D-glucoside, and cyanidin 3,5-O-diglucoside were detected only in transgenic lines, which could explain the purple leaf phenotype. Beside anthocyanins, the phenylpropanoids, polyphenols (catechins), flavonoids, flavones, and flavonols were also upregulated. Overexpression of LrAN2 activated the basic helix-loop-helix transcription factor AN1b, and the MYB transcription factor MYB3. Additionally, structural genes associated with the phenylpropanoid biosynthetic pathway were activated, which lead to the upregulated accumulation of phenylpropanoid, polyphenol (catechin), flavonoid, flavone, flavonol, and anthocyanin. The MYB transcription factor CPC, a negative regulator of anthocyanin biosynthesis, was also expressed at increased levels in transgenic lines, which implie that a negative regulation mechanism existed in the anthocyanin biosynthesis pathway. The relative contents of all 19 differently accumulated amino groups and derivatives were decreased in transgenic lines, which meant that the phenylalanine biosynthesis pathway used other amino acids as substrates. Interestingly, the expression of acetylalkylglycerol acetylhydrolase was suppressed in transgenic lines, which caused the accumulation of 19 lyso-phosphatidylcholine derivatives and a decrease in production of eight octodecane derivatives. Conclusions Overexpression of LrAN2 activates the pathway of anthocyanin synthesis and metabolism in tobacco. Four anthocyanins lead to the purple leaf phenotype The main pathways of flavonoid biosynthesis were up-regulated. This research provides more information about the function of MYB transcription factors in anthocyanin biosynthesis and the production of other chemical compounds. This work will help breeders to obtain new plant cultivars with high anthocyanin contents using biotechnology.

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Overexpression of a Freesia hybrida flavonoid 3-O-glycosyltransferase gene, Fh3GT1, enhances transcription of key anthocyanin genes and accumulation of anthocyanin and flavonol in transgenic petunia (Petunia hybrida)

Cited by 21 publications

References 58 publications

Functional Differentiation of Duplicated Flavonoid 3-O-Glycosyltransferases in the Flavonol and Anthocyanin Biosynthesis of Freesia hybrida

Functional Differentiation of Duplicated Flavonoid 3-O-Glycosyltransferases in the Flavonol and Anthocyanin Biosynthesis of Freesia hybrida

The R2R3-MYB Factor FhMYB5 From Freesia hybrida Contributes to the Regulation of Anthocyanin and Proanthocyanidin Biosynthesis

Comprehensive influences of overexpression of a MYB transcriptor regulating anthocyanin biosynthesis on transcriptome and metabolome of tobacco leaves

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