Isolation of Daucus carota ethylene insensitive3-like (DcEIL) involved in stress-inducible DcMYB1 expression in suspension-cultured carrot cells

Miyahara, Taira; Satoh, Shunsuke; Maeda, Kazuhiro; Kimura, Soichi; Sasaki, Nobuhiro; Ozeki, Yoshihiro

doi:10.5511/plantbiotechnology.27.91

Cited by 4 publications

(3 citation statements)

References 22 publications

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“…An EIL1 was isolated and its involvement in stress‐inducible DcMYB1 expression in suspension‐cultured carrot cells identified. DcEIL bound to a region of the DcMYB1 promoter containing a putative cis ‐element, which suppressed DcMYB1 promoter transcription (Miyahara et al, 2010 ). A recent study reported that MdEIL1 could promote anthocyanin accumulation by directly inducing MdMYB1 transcription in apple fruit (An et al, 2018 ).…”

Section: Discussionmentioning

confidence: 99%

Ethylene‐induced NbMYB4L is involved in resistance against tobacco mosaic virus in Nicotiana benthamiana

Zhu

Zhou

Zhang

et al. 2021

Molecular Plant Pathology

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Several MYB transcription factors are known to play important roles in plant resistance to environmental stressors. However, the mechanism governing the involvement of MYBs in regulating tobacco mosaic virus (TMV) resistance in plants is still unclear. In this study, we found that not only is Nicotiana benthamiana MYB4-like involved in defence against TMV, but also that the ethylene pathway participates in MYB4L-mediated resistance. Transcription of NbMYB4L was up-regulated in N. benthamiana infected with TMV. Silencing of NbMYB4L led to intensified TMV replication, whereas overexpression of NbMYB4L induced significant resistance to TMV. Transcription of NbMYB4L was greater in 1-aminocyclopropanecarboxylic acid (ACC, ethylene precursor)pretreated plants but lower when the ethylene signalling pathway was blocked during TMV infection. Gene expression analysis showed that the transcription of NbMYB4L was largely suppressed in ETHYLENE INSENSITIVE 3-like 1(EIL1)-silenced plants. The results of electrophoretic mobility shift assay and chromatin immunoprecipitationquantitative PCR (ChIP-qPCR) experiments indicated that NbEIL1 could directly bind to two specific regions of the NbMYB4L promoter. Furthermore, a luciferase assay revealed that NbEIL1 significantly induced the reporter activity of the MYB4L promoter in N. benthamiana. These results point to NbEIL1 functioning as a positive regulator of NbMYB4L transcription in N. benthamiana against TMV. Collectively, our work reveals that EIL1 and MYB4L constitute a coherent feed-forward loop involved in the robust regulation of resistance to TMV in N. benthamiana.

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Section: Discussionmentioning

confidence: 99%

Ethylene‐induced NbMYB4L is involved in resistance against tobacco mosaic virus in Nicotiana benthamiana

Zhu

Zhou

Zhang

et al. 2021

Molecular Plant Pathology

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“…DcPAL1 gene expression has been reported to be involved mainly in stress- and elicitor-induced biosynthesis of phenolic compounds, whereas DcPAL3 is involved in anthocyanin synthesis [46, 47]. In cultured carrot cells, Dc EIL negatively regulates Dc MYB1 by binding to the DcMYB1 promoter and suppressing its expression under non-stress conditions [48]. However, no significant variation in DcEIL expression was detected in the present study, indicating that degradation of the pool of Dc EIL protein occurs prior to Dc MYB1 expression.…”

Section: Discussionmentioning

confidence: 99%

Foliar-applied ethephon enhances the content of anthocyanin of black carrot roots (Daucus carota ssp. sativus var. atrorubens Alef.)

et al. 2017

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BackgroundBlack carrots (Daucus carota ssp. sativus var. atrorubens Alef.) constitute a valuable source of anthocyanins, which are used as natural red, blue and purple food colourants. Anthocyanins and phenolic compounds are specialised metabolites, accumulation of which often requires elicitors, which act as molecular signals in plant stress responses. In the present study, ethephon, an ethylene-generating compound was explored as enhancer of anthocyanin and phenolic contents during growth of ‘Deep Purple’ black carrots. The effects of ethephon on several parameters were investigated, and the expression of biosynthetic anthocyanin genes was studied during growth and anthocyanin accumulation.ResultsRoots of ethephon-treated carrot plants exhibited an increase in anthocyanin content of approximately 25%, with values ranging from 2.25 to 3.10 mg g−1 fresh weight, compared with values ranging from 1.50 to 1.90 mg g−1 fresh weight in untreated roots. The most rapid accumulation rate for anthocyanins, phenolic compounds, soluble solids and dry matter was observed between 10 and 13 weeks after sowing in both untreated and ethephon-treated carrots. The differences in anthocyanin contents between untreated and treated carrots increased for several weeks after the ethephon treatment was terminated. Five cyanidin-based anthocyanin forms were identified, with variable relative abundance values detected during root growth. Overall, the expression of the anthocyanin biosynthetic genes analysed (PAL1, PAL3, F3H1, DFR1, LDOX2) increased in response to ethephon treatment, as did the expression of the MYB1 transcription factor, which is associated with activation of the phenylpropanoid pathway under stress conditions. In addition, a correlation was proposed between ethylene and sugar contents and the induction of anthocyanin synthesis.ConclusionsThis study presents a novel method for enhancing anthocyanin content in black carrots. This finding is of economic importance as increased pigment concentration per unit of biomass implies improved profitability parameters in food colour production. We provide new insight into the accumulation patterns of the different cyanidin-based anthocyanins and phenolic compounds during root growth. Moreover, we show that enhanced anthocyanin content in ethephon-treated carrots is accompanied by increased expression of anthocyanin biosynthetic genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-017-1021-7) contains supplementary material, which is available to authorized users.

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“…Total RNA was extracted using an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). The cDNAs were synthesized and real-time RT-PCR was performed as previously reported (Miyahara et al 2010). RT-PCR was performed using Go Taq Hot Start DNA polymerase (Promega, Tokyo, Japan) with an Applied Biosystems 2720 Thermal Cycler (Life Technologies Japan, Tokyo, Japan).…”

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confidence: 99%

Identification of genes in Arabidopsis thaliana with homology to a novel acyl-glucose dependent glucosyltransferase of carnations

Miyahara¹,

Matsuba²,

Ozeki³

et al. 2011

Plant Biotechnology

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Glycoside hydrolases (GHs) are categorized as enzymes that catalyze the hydrolysis of the glycosidic bond between two or more carbohydrates or a carbohydrate and a non-carbohydrate moiety. They are classified into various families based on their sequence similarities. Protein folding similarities and conserved amino acid residues may be responsible for the homologous functions of enzymes within each family. At present, the GH classification in the Carbohydrate-Active enZYmes Database (http://www.cazy.org/) lists 118 families, and b -glucosidases are found in families 1, 3, 5, 9, and 30. Those in the GH family 1 (GH1) are enzymes that catalyze the hydrolysis of the b -glycosidic bond at the non-reducing end of their substrates. The GH1 enzymes belong to the GH-A clan and have an (a /b ) 8 barrel super secondary structure. They also have both the catalytic acid/base and the nucleophile consensus sequences. Both consensus sequences contain glutamate residues (Opassiri et al. 2006). In a recent b -glycosidase mutagenesis study, new enzymes were developed by replacing the nucleophile glutamate residues with different non-electric charge residues of other bglycosidases (Hancock et al. 2005). These new enzymes were found to show a partial decrease to complete loss of hydrolysis activity and an increase in reverse hydrolysis activity, thereby generating catalysts for the synthesis of new glycosidic linkages. These mutant glycosidases have been termed 'glycosynthases' (Mayer et al. 2000). Glycosynthases are expected to have considerable potential as tools in the production of pharmaceuticals by enzymatic synthesis. Synthesis of oligosaccharides by glycosynthases is more advantageous than chemical synthesis because of the stereoselectivity of the sugar acceptors. However, the sugar donors for glycosynthases need to be carefully selected to maintain high glycosyltransferase activity (Shaikh and Withers 2008).In a previous study, we were able to detect the activity of acyl-glucose:anthocyanin 5-O-glucosyltransferase (DcAA5GT) in Dianthus caryophyllus and of a homologue in Delphinium grandiflorum (DgAA7GT), and we also isolated the relevant cDNA in each species (Matsuba et al. 2010). Although DcAA5GT and DgAA7GT have glucosyltransferase activity, they are nevertheless classified as GH1 b -glucosidases. In addition, DcAA5GT and DgAA7GT do not use UDPglucose as a donor. GH1 family covers a wide range of b -glycoside hydrolases with different sugar specificity and thus includes b -glycosidases of different EC numbers, such as b -glucosides (EC 3. 2. Abstract Recently, we characterized an acyl-glucose : anthocyanin 5-O-glucosyltransferase (DcAA5GT) in Dianthus caryophyllus that has novel characteristics. We also showed that a homologous enzyme, acyl-glucose:anthocyanin 7-Oglucosyltransferase (DgAA7GT), is present in Delphinium grandiflorum. These enzymes are both classified as members of glycoside hydrolase family 1 (GH1). Here, we searched the Arabidopsis thaliana database for genes with homology to DcAA5GT and DgAA7GT and iden...

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Isolation of Daucus carota ethylene insensitive3-like (DcEIL) involved in stress-inducible DcMYB1 expression in suspension-cultured carrot cells

Cited by 4 publications

References 22 publications

Ethylene‐induced NbMYB4L is involved in resistance against tobacco mosaic virus in Nicotiana benthamiana

Ethylene‐induced NbMYB4L is involved in resistance against tobacco mosaic virus in Nicotiana benthamiana

Foliar-applied ethephon enhances the content of anthocyanin of black carrot roots (Daucus carota ssp. sativus var. atrorubens Alef.)

Identification of genes in Arabidopsis thaliana with homology to a novel acyl-glucose dependent glucosyltransferase of carnations

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