scholarly journals MaMYB4, an R2R3-MYB Repressor Transcription Factor, Negatively Regulates the Biosynthesis of Anthocyanin in Banana

2021 ◽  
Vol 11 ◽  
Author(s):  
Gui-Ming Deng ◽  
Sen Zhang ◽  
Qiao-Song Yang ◽  
Hui-Jun Gao ◽  
Ou Sheng ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Anthocyanin Synthesis ◽  
Structural Genes ◽  
Banana Plant ◽  
Binding Factor ◽  
Responsive Element ◽  
Banana Leaves ◽  
R2r3 Myb

Anthocyanins spatiotemporally accumulate in certain tissues of particular species in the banana plant, and MYB transcription factors (TFs) serve as their primary regulators. However, the precise regulatory mechanism in banana remains to be determined. Here, we report the identification and characterization of MaMYB4, an R2R3-MYB repressor TF, characterized by the presence of EAR (ethylene-responsive element binding factor–associated amphiphilic repression) and TLLLFR motifs. MaMYB4 expression was induced by the accumulation of anthocyanins. Transgenic banana plants overexpressing MaMYB4 displayed a significant reduction in anthocyanin compared to wild type. Consistent with the above results, metabolome results showed that there was a decrease in all three identified cyanidins and one delphinidin, the main anthocyanins that determine the color of banana leaves, whereas both transcriptome and reverse transcription–quantitative polymerase chain reaction analysis showed that many key anthocyanin synthesis structural genes and TF regulators were downregulated in MaMYB4 overexpressors. Furthermore, dual-luciferase assays showed that MaMYB4 was able to bind to the CHS, ANS, DFR, and bHLH promoters, leading to inhibition of their expression. Yeast two-hybrid analysis verified that MaMYB4 did not interact with bHLH, which ruled out the possibility that MaMYB4 could be incorporated into the MYB-bHLH-WD40 complex. Our results indicated that MaMYB4 acts as a repressor of anthocyanin biosynthesis in banana, likely due to a two-level repression mechanism that consists of reduced expression of anthocyanin synthesis structural genes and the parallel downregulation of bHLH to interfere with the proper assembly of the MYB-bHLH-WD40 activation complex. To the best of our knowledge, this is the first MYB TF that regulates anthocyanin synthesis that was identified by genetic methods in bananas, which will be helpful for manipulating anthocyanin coloration in banana programs in the future.

CBFs Function in Anthocyanin Biosynthesis by Interacting with MYB113 in Eggplant (Solanum melongena L.)

2019 ◽  
Vol 61 (2) ◽  
pp. 416-426 ◽  
Author(s):  
Lu Zhou ◽  
Yongjun He ◽  
Jing Li ◽  
Yang Liu ◽  
Huoying Chen
Keyword(s):  
Low Temperature ◽  
Anthocyanin Biosynthesis ◽  
Solanum Melongena ◽  
Structural Genes ◽  
Cold Response ◽  
Anthocyanin Pathway ◽  
R2r3 Myb ◽  
Dependent Pathway ◽  
Shed Light ◽  
R2r3 Myb Transcription Factors

Abstract Eggplant is rich in anthocyanins. R2R3-MYB transcription factors play a key role in the anthocyanin pathway. Low temperature is vital abiotic stress that affects the anthocyanin biosynthesis in plants. CBFs (C-repeat binding factors) act as central regulators in cold response. In this study, we found that SmCBF1, SmCBF2 and SmCBF3, via their C-terminal, physically interacted with SmMYB113, a key regulator of anthocyanin biosynthesis in eggplant. SmCBF2 and SmCBF3 upregulated the expression of SmCHS and SmDFR via a SmMYB113-dependent pathway. In addition, the transient expression assays demonstrated that co-infiltrating SmCBFs and SmMYB113 significantly improved the contents of anthocyanin and the expression levels of anthocyanin structural genes in tobacco. When SmTT8, a bHLH partner of SmMYB113, coexpressed with SmCBFs and SmMYB113, the anthocyanin contents were significantly enhanced compared with SmCBFs and SmMYB113. Furthermore, overexpression of SmCBF2 and SmCBF3 could facilitate the anthocyanin accumulation under cold conditions in Arabidopsis. Taken together, these results shed light on the functions of SmCBFs and potential mechanisms of low-temperature-induced anthocyanin biosynthesis in eggplant.

scholarly journals Transcription Factor VviMYB86 Oppositely Regulates Proanthocyanidin and Anthocyanin Biosynthesis in Grape Berries

2021 ◽  
Vol 11 ◽  
Author(s):  
Jing Cheng ◽  
Keji Yu ◽  
Ying Shi ◽  
Jun Wang ◽  
Changqing Duan
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Gene Expression Pattern ◽  
Anthocyanin Synthesis ◽  
Grape Berries ◽  
Flavonoid Synthesis ◽  
Leucoanthocyanidin Reductase ◽  
Arabidopsis Protoplast ◽  
R2r3 Myb

Proanthocyanidins (PAs) and anthocyanins are two vital groups of flavonoid compounds for grape berries and red wines. Several transcription factors (TFs) have been identified to be involved in regulating PA and anthocyanin biosynthesis in grape berries. However, research on TFs with different regulatory mechanisms for these two biosynthesis branches in grapes remains limited. In this study, we identified an R2R3-MYB TF, VviMYB86, whose spatiotemporal gene expression pattern in grape berries coincided well with PA accumulation but contrasted with anthocyanin synthesis. Both in vivo and in vitro experiments verified that VviMYB86 positively regulated PA biosynthesis, primarily by upregulating the expression of the two leucoanthocyanidin reductase (LAR) genes in the Arabidopsis protoplast system, as well as in VviMYB86-overexpressing grape callus cultured under 24 h of darkness. Moreover, VviMYB86 was observed to repress the anthocyanin biosynthesis branch in grapes by downregulating the transcript levels of VviANS and VviUFGT. Overall, VviMYB86 is indicated to have a broad effect on flavonoid synthesis in grape berries. The results of this study will help elucidate the regulatory mechanism governing the expression of the two LAR genes in grape berries and provide new insights into the regulation of PA and anthocyanin biosynthesis in grape berries.

scholarly journals Regulation of MYB Transcription Factors of Anthocyanin Synthesis in Lily Flowers

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaojuan Yin ◽  
Yibing Zhang ◽  
Li Zhang ◽  
Baohua Wang ◽  
Yidi Zhao ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Transient Expression ◽  
Anthocyanin Biosynthesis ◽  
Gene Promoter ◽  
Flower Color ◽  
Preliminary Evidence ◽  
Anthocyanin Synthesis ◽  
Myb Transcription Factors ◽  
Color Formation ◽  
R2r3 Myb

Flower color is the decisive factor that affects the commercial value of ornamental flowers. Therefore, it is important to study the regulation of flower color formation in lily to discover the positive and negative factors that regulate this important trait. In this study, MYB transcription factors (TFs) were characterized to understand the regulatory mechanism of anthocyanin biosynthesis in lily. Two R2R3-MYB TFs, LvMYB5, and LvMYB1, were found to regulate anthocyanin biosynthesis in lily flowers. LvMYB5, which has an activation motif, belongs to the SG6 MYB protein subgroup of Arabidopsis thaliana. Transient expression of LvMYB5 indicated that LvMYB5 can promote coloration in Nicotiana benthamiana leaves, and that expression of LvMYB5 increases the expression levels of NbCHS, NbDFR, and NbANS. VIGS experiments in lily petals showed that the accumulation of anthocyanins was reduced when LvMYB5 was silenced. Luciferase assays showed that LvMYB5 can promote anthocyanin synthesis by activating the ANS gene promoter. Therefore, LvMYB5 plays an important role in flower coloration in lily. In addition, the transient expression experiment provided preliminary evidence that LvMYB1 (an R2R3-MYB TF) inhibits anthocyanin synthesis in lily flowers. The discovery of activating and inhibitory factors related to anthocyanin biosynthesis in lily provides a theoretical basis for improving flower color through genetic engineering. The results of our study provide a new direction for the further study of the mechanisms of flower color formation in lilies.

scholarly journals The combination of R2R3-MYB gene AmRosea1 and hairy root culture is a useful tool for rapidly induction and production of anthocyanins in Antirrhinum majus L

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chunlan Piao ◽  
Jinguo Wu ◽  
Min-Long Cui
Keyword(s):  
Hairy Roots ◽  
Anthocyanin Biosynthesis ◽  
Ectopic Expression ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Anthocyanin Accumulation ◽  
Structural Genes ◽  
Myb Gene ◽  
Transformed Hairy Roots ◽  
R2r3 Myb

AbstractAnthocyanins are the largest group of water-soluble pigments and beneficial for human health. Although most plants roots have the potential to express natural biosynthesis pathways required to produce specialized metabolites such as anthocyanins, the anthocyanin synthesis is specifically silenced in roots. To explore the molecular mechanism of absence and production ability of anthocyanin in the roots, investigated the effect of a bHLH gene AmDelila, and an R2R3-MYB gene AmRosea1, which are the master regulators of anthocyanin biosynthesis in Antirrhinum majus flowers, by expressing these genes in transformed hairy roots of A. majus. Co-ectopic expression of both AmDelila and AmRosea1 significantly upregulated the expression of the key target structural genes in the anthocyanin biosynthesis pathway. Furthermore, this resulted in strongly enhanced anthocyanin accumulation in transformed hairy roots. Ectopic expression of AmDelila alone did not gives rise to any significant anthocyanin accumulation, however, ectopic expression of AmRosea1 alone clearly upregulated expression of the main structural genes as well as greatly promoted anthocyanin accumulation in transformed hairy roots, where the contents reached 0.773–2.064 mg/g fresh weight. These results suggest that AmRosea1 plays a key role in the regulatory network in controlling the initiation of anthocyanin biosynthesis in roots, and the combination of AmRosea1 and hairy root culture is a powerful tool to study and production of anthocyanins in the roots of A. majus.

scholarly journals Identification of the Eutrema Salsugineum EsMYB90 gene important for anthocyanin biosynthesis

2020 ◽  
Author(s):  
Yuting Qi ◽  
Caihong Gu ◽  
Xingjun Wang ◽  
Shiqing Gao ◽  
Changsheng Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transgenic Plants ◽  
Anthocyanin Biosynthesis ◽  
Ectopic Expression ◽  
Myb Transcription Factor ◽  
Anthocyanin Synthesis ◽  
Eutrema Salsugineum ◽  
Tobacco Transgenic Plants ◽  
Myb Genes ◽  
R2r3 Myb

Abstract Abstract Background: Anthocyanins contribute to coloration and antioxidation effects in different plant tissues. MYB transcription factors have been demonstrated to be a key regulator for anthocyanin synthesis in many plants. However, little information was available about the MYB genes in the halophyte species Eutrema salsugineum . Result: Here we report the identification of an important anthocyanin biosynthesis regulator Es MYB90 from Eutrema salsugineum , which is a halophyte tolerant to multiple abiotic stresses. Our phylogenetic and localization analyses supported that Es MYB90 is an R2R3 type of MYB transcription factor. Ectopic expression of EsMYB90 in tobacco and Arabidopsis enhanced pigmentation and anthocyanin accumulation in various organs. The transcriptome analysis revealed that 42 genes upregulated by Es MYB90 in 35S : EsMYB90 tobacco transgenic plants are required for anthocyanin biosynthesis. Moreover, our qRT-PCR results showed that Es MYB90 promoted expression of early ( PAL , CHS , and CHI ) and late ( DFR , ANS , and UFGT ) anthocyanin biosynthesis genes in stems, leaves, and flowers of 35S : EsMYB90 tobacco transgenic plants. Conclusions: Our results indicated that Es MYB90 is a MYB transcription factor, which regulates anthocyanin biosynthesis genes to control anthocyanin biosynthesis. Our work provides a new tool to enhance anthocyanin production in various plants. Keywords : Anthocyanin, flavonoid, Eutrema salsugineum , R2R3 MYB transcription factor, Es MYB90, transcriptional regulation, anthocyanin biosynthesis genes.

Functional R2R3-MYB Transcription Factor NsMYB1, Regulating Anthocyanin Biosynthesis, Was Relative To The Fruit Color Differentiation in Nitraria Sibirica Pall

2021 ◽  
Author(s):  
Xuemei Bao ◽  
Yuan Zong ◽  
Na Hu ◽  
Shiming Li ◽  
Baolong Liu ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Chemical Analysis ◽  
Anthocyanin Biosynthesis ◽  
Fruit Color ◽  
Tibet Plateau ◽  
Myb Transcription Factor ◽  
Structural Genes ◽  
Transcript Factor ◽  
Nitraria Sibirica ◽  
R2r3 Myb

Abstract Background Nitraria sibirica Pall. is an economic plant with two kinds of fruit color, widely spreads in the Qinghai Tibet Plateau. The chemical analysis and pharmacological evaluation had been carried out for several tens of years, the mechanism behind the fruit color differentiation is still unclear. Results In this manuscript, the chemical analysis of the extractions showed that the chemical composition of fruit color was anthocyanin, and two kind of Nitraria sibirica Pall. were caused by the content differentiation with the same anthocyanin kinds. Cya-nidin-3-[2ʹ’-(6ʹ’’-coumaroyl)-glucosyl]-glucoside (C3G) was the major anthocyanin. Transcriptome analysis and the qRT-PCR revealed that the structural genes relative to anthocyanin biosynthesis except CHS, F3’5’H and ANS were up-regulated in BF compared with RF, which indicated that transcript factor should be the reason for the expression difference of the structure genes. In the unigenes of the transcript factor MYB and bHLH, relative to anthocyanin, only NsMYB1 (Clus-ter-8422.10600), was high-expression and up-expression in the BF. NsMYB1 encoded the same length protein with four amino acid differences in the RF and BF, and both contained the intact DNA, HTH-MYB and SANT domains. NsMYB1 was close to the AtMYB114, AtMYB113 and AtPAP1, regulating anthocyanin biosynthesis, in phylogenetic relationship. Both NsMYB1r and NsMYB1b could promote the transcript of the structural genes, and induced the anthocyanin accumulation in all tissues of transgenic tobacco. The insertion of ‘TATA’ in the promoter of NsMYB1r gave one more promoter region, and was the reason for higher transcripts in black fruit possibly. Conclusions NsMYB1 was a functional R2R3-MYB transcription factor, regulated the anthocyanin biosynthesis, and leaded to the fruit color differentiation in Nitraria sibirica Pall.

scholarly journals Metabolome and Transcriptome Sequencing Analysis Reveals Anthocyanin Metabolism in Pink Flowers of Anthocyanin-Rich Tea (Camellia sinensis)

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1064 ◽  
Author(s):  
Dylan Rothenberg ◽  
Haijun Yang ◽  
Meiban Chen ◽  
Wenting Zhang ◽  
Lingyun Zhang
Keyword(s):  
Camellia Sinensis ◽  
Flower Development ◽  
Molecular Mechanisms ◽  
Anthocyanin Biosynthesis ◽  
Functional Enrichment ◽  
Anthocyanin Synthesis ◽  
Sequencing Analysis ◽  
Anthocyanin Accumulation ◽  
Structural Genes ◽  
Anthocyanin Pathway

Almost all flowers of the tea plant (Camellia sinensis) are white, which has caused few researchers to pay attention to anthocyanin accumulation and color changing in tea flowers. A new purple-leaf cultivar, Baitang purple tea (BTP) was discovered in the Baitang Mountains of Guangdong, whose flowers are naturally pink, and can provide an opportunity to understand anthocyanin metabolic networks and flower color development in tea flowers. In the present study, twelve anthocyanin components were identified in the pink tea flowers, namely cyanidin O-syringic acid, petunidin 3-O-glucoside, pelargonidin 3-O-beta-d-glucoside, which marks the first time these compounds have been found in the tea flowers. The presence of these anthocyanins seem most likely to be the reason for the pink coloration of the flowers. Twenty-one differentially expressed genes (DEGs) involved in anthocyanin pathway were identified using KEGG pathway functional enrichment, and ten of these DEG’s screened using venn and KEGG functional enrichment analysis during five subsequent stages of flower development. By comparing DEGs and their expression levels across multiple flower development stages, we found that anthocyanin biosynthesis and accumulation in BTP flowers mainly occurred between the third and fourth stages (BTP3 to BTP4). Particularly, during the period of peak anthocyanin synthesis 17 structural genes were upregulated, and four structural genes were downregulated only. Ultimately, eight critical genes were identified using weighted gene co-expression network analysis (WGCNA), which were found to have direct impact on biosynthesis and accumulation of three flavonoid compounds, namely cyanidin 3-O-glucoside, petunidin 3-O-glucoside and epicatechin gallate. These results provide useful information about the molecular mechanisms of coloration in rare pink tea flower of anthocyanin-rich tea, enriching the gene resource and guiding further research on anthocyanin accumulation in purple tea.

scholarly journals Transcriptomic Profiling of Apple Calli With a Focus on the Key Genes for ALA-Induced Anthocyanin Accumulation

2021 ◽  
Vol 12 ◽  
Author(s):  
Jie Zheng ◽  
Longbo Liu ◽  
Huihui Tao ◽  
Yuyan An ◽  
Liangju Wang
Keyword(s):  
Molecular Mechanisms ◽  
Aminolevulinic Acid ◽  
Anthocyanin Biosynthesis ◽  
Transcript Level ◽  
Flavonoid Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Structural Genes ◽  
Metabolism Regulation ◽  
Positive Regulators ◽  
R2r3 Myb

The red color is an attractive trait of fruit and determines its market acceptance. 5-Aminolevulinic acid (ALA), an eco-friendly plant growth regulator, has played a universal role in plant secondary metabolism regulation, particularly in flavonoid biosynthesis. It has been widely reported that ALA can up-regulate expression levels of several structural genes related to flavonoid metabolism and anthocyanin accumulation. However, the molecular mechanisms behind ALA-induced expression of these genes are complicated and still far from being completely understood. In this study, transcriptome analysis identified the differentially expressed genes (DEGs) associated with ALA-induced anthocyanin accumulation. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the flavonoid biosynthesis (ko00941) pathway was significantly enhanced in the ALA-treated apple calli at 24, 48, and 72 h after the treatment. Expression pattern revealed that ALA up-regulated the expression of the structural genes related to not only anthocyanin biosynthesis (MdCHS, MdCHI, MdF3’H, MdDFR, MdANS, and MdUFGT) but also anthocyanin transport (MdGST and MdMATE). Two R2R3-MYB transcription factors (MdMYB10 and MdMYB9), which are the known positive regulators of anthocyanin biosynthesis, were significantly induced by ALA. Gene overexpression and RNA interference assays demonstrated that MdMYB10 and MdMYB9 were involved in ALA-induced anthocyanin biosynthesis. Moreover, MdMYB10 and MdMYB9 might positively regulate the transcription of MdMATE8 by binding to the promoter region. These results indicate that MdMYB10 and MdMYB9 modulated structural gene expression of anthocyanin biosynthesis and transport in response to ALA-mediated apple calli coloration at the transcript level. We herein provide new details regarding transcriptional regulation of ALA-induced color development.

scholarly journals Identification of the Eutrema Salsugineum EsMYB90 gene important for anthocyanin biosynthesis

2020 ◽  
Author(s):  
Yuting Qi ◽  
Caihong Gu ◽  
Xingjun Wang ◽  
Shiqing Gao ◽  
Changsheng Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transgenic Plants ◽  
Anthocyanin Biosynthesis ◽  
Ectopic Expression ◽  
Myb Transcription Factor ◽  
Anthocyanin Synthesis ◽  
Eutrema Salsugineum ◽  
Tobacco Transgenic Plants ◽  
Myb Genes ◽  
R2r3 Myb

Abstract Background: Anthocyanins contribute to coloration and antioxidation effects in different plant tissues. MYB transcription factors have been demonstrated to be a key regulator for anthocyanin synthesis in many plants. However, little information was available about the MYB genes in the halophyte species Eutrema salsugineum . Result: Here we report the identification of an important anthocyanin biosynthesis regulator Es MYB90 from Eutrema salsugineum , which is a halophyte tolerant to multiple abiotic stresses. Our phylogenetic and localization analyses supported that Es MYB90 is an R2R3 type of MYB transcription factor. Ectopic expression of EsMYB90 in tobacco and Arabidopsis enhanced pigmentation and anthocyanin accumulation in various organs. The transcriptome analysis revealed that 42 genes upregulated by Es MYB90 in 35S : EsMYB90 tobacco transgenic plants are required for anthocyanin biosynthesis. Moreover, our qRT-PCR results showed that Es MYB90 promoted expression of early ( PAL , CHS , and CHI ) and late ( DFR , ANS , and UFGT ) anthocyanin biosynthesis genes in stems, leaves, and flowers of 35S : EsMYB90 tobacco transgenic plants. Conclusions: Our results indicated that Es MYB90 is a MYB transcription factor, which regulates anthocyanin biosynthesis genes to control anthocyanin biosynthesis. Our work provides a new tool to enhance anthocyanin production in various plants. Keywords : Anthocyanin, flavonoid, Eutrema salsugineum , R2R3 MYB transcription factor, Es MYB90, transcriptional regulation, anthocyanin biosynthesis genes.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

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