Molecular cloning and functional characterization of dihydroflavonol-4-reductase gene from Calibrachoa hybrida

Chu, Yunxia; Pan, Junsong; Wu, Aiming; Cai, Run; Chen, Hairong

doi:10.1016/j.scienta.2013.08.017

Cited by 17 publications

(18 citation statements)

References 20 publications

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“…Because only one transgenic line (Line 9) showed very higher expression level of IbDHKR , the definitive conclusion of overexpressing IbDHKR on anthocyanin accumulation needed more independent transgenic lines. In fact, overexpression of DFR definitively promoted anthocyanin biosynthesis in many plants ( Xie et al, 2004 ; Lee et al, 2008 ; Huang et al, 2012 ; Chu et al, 2014 ).…”

Section: Resultsmentioning

confidence: 99%

A Root-Preferential DFR-Like Gene Encoding Dihydrokaempferol Reductase Involved in Anthocyanin Biosynthesis of Purple-Fleshed Sweet Potato

et al. 2017

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Purple-fleshed sweet potato is good for health due to rich anthocyanins in tubers. Although the anthocyanin biosynthetic pathway is well understood in up-ground organs of plants, the knowledge on anthocyanin biosynthesis in underground tubers is limited. In the present study, we isolated and functionally characterized a root-preferential gene encoding dihydrokaempferol reductase (IbDHKR) from purple-fleshed sweet potato. IbDHKR showed highly similarity with the reported dihydroflavonol reductases in other plant species at the sequence levels and the NADPH-binding motif and the substrate-binding domain were also found in IbDHKR. The tissue profile showed that IbDHKR was expressed in all the tested organs, but with much higher level in tuber roots. The expression level of IbDHKR was consistent with the anthocyanin content in sweet potato organs, suggesting that tuber roots were the main organs to synthesize anthocyanins. The recombinant 44 kD IbDHKR was purified and fed by three different dihydroflavonol substrates including dihydrokaempferol (DHK), dihydroquerctin, and dihydromyrecetin. The substrate feeding assay indicated that only DHK could be accepted as substrate by IbDHKR, which was reduced to leucopelargonidin confirmed by LC-MS. Finally, IbDHKR was overexpressed in transgenic tobacco. The IbDHKR-overexpression tobacco corolla was more highly pigmented and contained higher level of anthocyanins than the wild-type tobacco corolla. In summary, IbDHKR was a root-preferential gene involved in anthocyanin biosynthesis and its encoding protein, specifically catalyzing DHK reduction to yield leucopelargonidin, was a candidate gene for engineering anthocyanin biosynthetic pathway.

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Section: Resultsmentioning

confidence: 99%

A Root-Preferential DFR-Like Gene Encoding Dihydrokaempferol Reductase Involved in Anthocyanin Biosynthesis of Purple-Fleshed Sweet Potato

et al. 2017

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“…Dihydroflavonol-4-reductase (DFR), which is a downstream enzyme in anthocyanin biosynthesis, could regulate the critical rate-limiting step of anthocyanin biosynthesis processes by catalyzing dihydroflavonols to leucoanthocyanidins [4,36]. Recently, a series of DFR gene homologs have been identified in various plant species such as in Rosa rugosa [4], Camellia sinensis [37], Populus trichocarpa [38], and Calibrachoa hybrida [39]. The previous works have confirmed that overexpression of DFR genes within tobacco and petunia could accelerate the anthocyanin accumulation and promote the red coloration of flowers [4,39].…”

Section: Discussionmentioning

confidence: 99%

“…Recently, a series of DFR gene homologs have been identified in various plant species such as in Rosa rugosa [4], Camellia sinensis [37], Populus trichocarpa [38], and Calibrachoa hybrida [39]. The previous works have confirmed that overexpression of DFR genes within tobacco and petunia could accelerate the anthocyanin accumulation and promote the red coloration of flowers [4,39]. Here, two LlDFRs ( DFR2-1 and DFR2-2 ) were isolated from DEGs.…”

Section: Discussionmentioning

confidence: 99%

Integrating Transcriptomic and GC-MS Metabolomic Analysis to Characterize Color and Aroma Formation during Tepal Development in Lycoris longituba

Yue

Liu

Shi

et al. 2019

Plants

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Lycoris longituba, belonging to the Amaryllidaceae family, is a perennial bulb bearing flowers with diverse colors and fragrance. Selection of cultivars with excellent colored and scented flowers has always been the breeding aim for ornamental plants. However, the molecular mechanisms underlying color fading and aroma production during flower expansion in L. longituba remain unclear. Therefore, to systematically investigate these important biological phenomena, the tepals of L. longituba from different developmental stages were used to screen and analyze the metabolic components and relevant genes. Utilizing the Illumina platform, a total of 144,922 unigenes were obtained from the RNA-Seq libraries. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that the phenylpropanoid biosynthesis and flavonoid biosynthesis pathways might play important roles during color and aroma changes. Metabolomic analysis identified 29 volatile organic components (VOCs) from different developmental stages of L. longituba tepals, and orthogonal partial least-squares discriminate analysis (OPLS-DA) revealed that trans-β-ocimene—a terpene—was the most important aroma compound. Meanwhile, we found the content of anthocyanin was significantly reduced during the tepal color fading process. Then, we identified two dihydroflavonol-4-reductase (DFR) and three terpene synthase (TPS) genes, for which expression changes coincided with the production patterns of anthocyanins and trans-β-ocimene, respectively. Furthermore, a number of MYB and bHLH transcription factors (TFs) which might be involved in color- and aroma-formation were also identified in L. longituba tepal transcriptomes. Taken together, this is the first comprehensive report of the color and fragrance in tepals of L. longituba and these results could be helpful in understanding these characteristics and their regulation networks.

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“…D85102) and C. hybrida (GenBank No. KC140106) that was under the control of the 35S promoter was transformed into petunia variety 9702, as described by Chu et al (2014). Three 9H transgenic plants (9H30, 9H31, and 9H33) and three 9R plants (9R8, 9R9, and 9R12) were grown in a greenhouse with the original petunia cultivar 9702.…”

Section: Plant Materialsmentioning

confidence: 99%

Expression analysis of dihydroflavonol 4-reductase genes in Petunia hybrida

Chu

Chen

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Dihydroflavonol 4-reductase (DFR) genes from Rosa chinensis (Asn type) and Calibrachoa hybrida (Asp type), driven by a CaMV 35S promoter, were integrated into the petunia (Petunia hybrida) cultivar 9702. Exogenous DFR gene expression characteristics were similar to flower-color changes, and effects on anthocyanin concentration were observed in both types of DFR gene transformants. Expression analysis showed that exogenous DFR genes were expressed in all of the tissues, but the expression levels were significantly different. However, both of them exhibited a high expression level in petals that were starting to open. The introgression of DFR genes may significantly change DFR enzyme activity. Anthocyanin ultraperformance liquid chromatography results showed that anthocyanin concentrations changed according to DFR enzyme activity. Therefore, the change in flower color was probably the result of a DFR enzyme change. Pelargonidin 3-O-glucoside was found in two different transgenic petunias, indicating that both CaDFR and RoDFR could catalyze dihydrokaempferol. Our results also suggest that transgenic 5011 Expression analysis of DFRs in Petunia hybrida©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (2): 5010-5021 (2015) petunias with DFR gene of Asp type could biosynthesize pelargonidin 3-O-glucoside.

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Molecular cloning and functional characterization of dihydroflavonol-4-reductase gene from Calibrachoa hybrida

Cited by 17 publications

References 20 publications

A Root-Preferential DFR-Like Gene Encoding Dihydrokaempferol Reductase Involved in Anthocyanin Biosynthesis of Purple-Fleshed Sweet Potato

A Root-Preferential DFR-Like Gene Encoding Dihydrokaempferol Reductase Involved in Anthocyanin Biosynthesis of Purple-Fleshed Sweet Potato

Integrating Transcriptomic and GC-MS Metabolomic Analysis to Characterize Color and Aroma Formation during Tepal Development in Lycoris longituba

Expression analysis of dihydroflavonol 4-reductase genes in Petunia hybrida

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