Isolation and functional characterization of two 5-O-glucosyltransferases related to anthocyanin biosynthesis from Freesia hybrida

Ju, Zhigang; Sun, Wei; Meng, Xiangyu; Liang, Lingjie; Li, Yueqing; Zhou, Tongtong; Shen, Huan; Gao, Xiang; Wang, Li

doi:10.1007/s11240-018-1447-0

Cited by 11 publications

(7 citation statements)

References 55 publications

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“…‘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: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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“…Additionally, flower anthocyanin accumulation patterns were tightly regulated in a spatio-temporal manner, making F. hybrida , a representative and powerful system to explore chemical ecology, genetics and regulation of anthocyanin biosynthesis. In Freesia flowers, five anthocyanin aglycons (delphinidin, cyanidin, petunidin, peonidin and malvinidin) and the ABGs have been investigated in the widely cultivated red flowered cultivar Red River® 40 – 46 . Moreover, the versatile transient expression system based on the flower petals or protoplasts isolated from Freesia callus partially overcomes the obstacle of the recalcitrant characters of monocots against Agrobacterium and immensely accelerates the molecular investigations in Freesia per se 47 , 48 .…”

Section: Introductionmentioning

confidence: 99%

MYB repressors and MBW activation complex collaborate to fine-tune flower coloration in Freesia hybrida

Shan

Gao

et al. 2020

Commun Biol

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Floral anthocyanin has multiple ecological and economic values, its biosynthesis largely depends on the conserved MYB-bHLH-WD40 (MBW) activation complex and MYB repressors hierarchically with the MBW complex. In contrast to eudicots, the MBW regulatory network model has not been addressed in monocots because of the lack of a suitable system, as grass plants exhibit monotonous floral pigmentation patterns. Presently, the MBW regulatory network was investigated in a non-grass monocot plant, Freesia hybrida . FhMYB27 and FhMYBx with different functional manners were confirmed to be anthocyanin related R2R3 and R3 MYB repressors, respectively. Particularly, FhMYBx could obstruct the formation of positive MBW complex by titrating bHLH proteins, whereas FhMYB27 mainly defected the activator complex into suppressor via its repression domains in C-terminus. Furthermore, the hierarchical and feedback regulatory loop was verified, indicating the synergistic and sophisticated regulatory network underlying Freesia anthocyanin biosynthesis was quite similar to that reported in eudicot plants.

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Isolation and functional characterization of two 5-O-glucosyltransferases related to anthocyanin biosynthesis from Freesia hybrida

Cited by 11 publications

References 55 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

MYB repressors and MBW activation complex collaborate to fine-tune flower coloration in Freesia hybrida

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