Molecular and Biochemical Analysis of Chalcone Synthase from Freesia hybrid in Flavonoid Biosynthetic Pathway

Sun, Wei; Meng, Xiangyu; Liang, Lingjie; Jiang, Wangshu; Huang, Yafei; He, Jie; Hu, Haiyan; Almqvist, Jonas; Gao, Xiang; Wang, Li

doi:10.1371/journal.pone.0119054

Cited by 112 publications

(98 citation statements)

References 67 publications

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“…Zhang et al (2015), Yin et al (2015), and Sun et al (2015) suggested that the CHS gene plays an essential role in anthocyanin biosynthesis. Guo et al (2015) suggest that IbCHI is a key enzyme in the in the anthocyanin biosynthesis of sweet potato.…”

Section: Discussionmentioning

confidence: 99%

“…The interference decreased CHS expression and anthocyanin content by 88.8% and 87% in Prunus persica, respectively . Overexpression of FhCHS1 in Arabidopsis thaliana changed the pigmentation phenotype of the seed coats, cotyledons, and hypocotyls (Sun et al, 2015). Chalcone isomerase (CHI) catalyzes the stereospecific isomerization of chalcones into their corresponding (2S)-flavanones (Guo et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

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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“…Therefore, the complicated flavonoid compounds in freesia flowers indicated a sophisticated biosynthetic pathway and transcription regulation network for the flavonoid accumulation. So far, two anthocyanin biosynthetic genes, Fh3GT1 and FhCHS1 , as well as two bHLH regulatory genes, FhGL3L and FhTT8L , were isolated and functionally verified (Sui et al, 2011; Sun et al, 2015, 2016; Li et al, 2016). However, no DFR -like genes have been isolated and functionally characterized, which is worthy of further concerns because their particular positions in flavonoid biosynthetic pathway.…”

Section: Introductionmentioning

confidence: 99%

Dihydroflavonol 4-Reductase Genes from Freesia hybrida Play Important and Partially Overlapping Roles in the Biosynthesis of Flavonoids

Liu

Cai

et al. 2017

Front. Plant Sci.

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Dihydroflavonol-4-reductase (DFR) is a key enzyme in the reduction of dihydroflavonols to leucoanthocyanidins in both anthocyanin biosynthesis and proanthocyanidin accumulation. In many plant species, it is encoded by a gene family, however, how the different copies evolve either to function in different tissues or at different times or to specialize in the use of different but related substrates needs to be further investigated, especially in monocot plants. In this study, a total of eight putative DFR-like genes were firstly cloned from Freesia hybrida. Phylogenetic analysis showed that they were classified into different branches, and FhDFR1, FhDFR2, and FhDFR3 were clustered into DFR subgroup, whereas others fell into the group with cinnamoyl-CoA reductase (CCR) proteins. Then, the functions of the three FhDFR genes were further characterized. Different spatio-temporal transcription patterns and levels were observed, indicating that the duplicated FhDFR genes might function divergently. After introducing them into Arabidopsis dfr (tt3-1) mutant plants, partial complementation of the loss of cyanidin derivative synthesis was observed, implying that FhDFRs could convert dihydroquercetin to leucocyanidin in planta. Biochemical assays also showed that FhDFR1, FhDFR2, and FhDFR3 could utilize dihydromyricetin to generate leucodelphinidin, while FhDFR2 could also catalyze the formation of leucocyanidin from dihydrocyanidin. On the contrary, neither transgenic nor biochemical analysis demonstrated that FhDFR proteins could reduce dihydrokaempferol to leucopelargonidin. These results were consistent with the freesia flower anthocyanin profiles, among which delphinidin derivatives were predominant, with minor quantities of cyanidin derivatives and undetectable pelargonidin derivatives. Thus, it can be deduced that substrate specificities of DFRs were the determinant for the categories of anthocyanins aglycons accumulated in F. hybrida. Furthermore, we also found that the divergence of the expression patterns for FhDFR genes might be controlled at transcriptional level, as the expression of FhDFR1/FhDFR2 and FhDFR3 was controlled by a potential MBW regulatory complex with different activation efficiencies. Therefore, it can be concluded that the DFR-like genes from F. hybrida have diverged during evolution to play partially overlapping roles in the flavonoid biosynthesis, and the results will contribute to the study of evolution of DFR gene families in angiosperms, especially for monocot plants.

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“…CHS is a key enzyme in the anthocyanin biosynthesis pathway, catalysing the first step of the pathway that leads to the synthesis of flavonoids (Sun et al 2015). Therefore, it is not surprising that we found higher levels of its transcript in red tissues both in the RNAseq and in the qPCR analysis.…”

Section: Reverse Transcription and Quantitative Pcr Of Selected Genesmentioning

confidence: 66%

Developing a new variety of kentia palms (Howea forsteriana): up-regulation of cytochrome b561 and chalcone synthase is associated with red colouration of the stems

et al. 2018

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Savolainen (2018): Developing a new variety of kentia palms (Howea forsteriana): up-regulation of cytochrome b561 and chalcone synthase is associated with red colouration of the stems, Botany Letters, DOI: 10.1080DOI: 10. /23818107.2017 The kentia palm (Howea forsteriana) is endemic to a 12 km 2 volcanic island in the Tasman Sea, Lord Howe Island. It is one of the most traded houseplants in the world. The typical kentia palm presents a dark green-coloured stem. Note that the stem is made of leaf rachis and petioles, which are intermingled towards the base. However, we discovered on Lord Howe Island a new biological variety that has a red stem. Red-stemmed palm species are known and highly demanded as decorative plants. However, these red palm horticultural varieties require tropical or subtropical conditions to grow, hence commercialization is limited. Thus, a red-stemmed variety of H. forsteriana may have tremendous market potential. Nonetheless, palm trees grow generally slowly and often reach maturity at 15-20 years old or later, which may make conventional strategies unsuitable for the propagation of a new variety. This difficulty needs to be addressed before commercialization can be achieved. Here, we found that anthocyanin is responsible for the red colouration of the stem in the new variety. Using RNA sequencing and quantitative PCR, we identified two gene isoforms displaying altered expression associated with this red colouration, encoding a cytochrome b561 and a chalcone synthase. The latter protein is known to be part of the anthocyanin biosynthesis pathway, which plays a central role in pigmentation in plants. The levels of cytochrome b561 transcripts accumulated were found to be well correlated with an increased anthocyanin concentration in the red stems.

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Molecular and Biochemical Analysis of Chalcone Synthase from Freesia hybrid in Flavonoid Biosynthetic Pathway

Cited by 112 publications

References 67 publications

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

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

Dihydroflavonol 4-Reductase Genes from Freesia hybrida Play Important and Partially Overlapping Roles in the Biosynthesis of Flavonoids

Developing a new variety of kentia palms (Howea forsteriana): up-regulation of cytochrome b561 and chalcone synthase is associated with red colouration of the stems

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