MYB_SH[AL]QKY[RF] transcription factors MdLUX and MdPCL-like promote anthocyanin accumulation through DNA hypomethylation and MdF3H activation in apple

2020 ◽  
Author(s):  
Wen-Fang Li ◽  
Gai-Xing Ning ◽  
Cun-Wu Zuo ◽  
Ming-Yu Chu ◽  
Shi-Jin Yang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Transcription Factors ◽  
Promoter Region ◽  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Anthocyanin Synthesis ◽  
Mrna Levels ◽  
Anthocyanin Accumulation ◽  
Dna Hypomethylation ◽  
Fruit Skin

Abstract Heritable DNA methylation is a highly conserved epigenetic mark that is important for many biological processes. In a previous transcriptomic study on the fruit skin pigmentation of apple (Malus domestica Borkh.) cv. ‘Red Delicious’ (G0) and its four continuous-generation bud sport mutants including ‘Starking Red’ (G1), ‘Starkrimson’ (G2), ‘Campbell Redchief’ (G3) and ‘Vallee spur’ (G4), we identified MYB transcription factors (TFs) MdLUX and MdPCL-like involved in regulating anthocyanin synthesis. However, how these TFs ultimately determine the fruit skin colour traits remain elusive. Here, bioinformatics analysis revealed that MdLUX and MdPCL-like contained a well-conserved motif SH[AL]QKY[RF] in their C-terminal region and were located in the nucleus of onion epidermal cells. Overexpression of MdLUX and MdPCL-like in ‘Golden Delicious’ fruits, ‘Gala’ calli and Arabidopsis thaliana promoted the accumulation of anthocyanin, whereas MdLUX and MdPCL-like suppression inhibited anthocyanin accumulation in ‘Red Fuji’ apple fruit skin. Yeast one-hybrid assays revealed that MdLUX and MdPCL-like may bind to the promoter region of the anthocyanin biosynthesis gene MdF3H. Dual-luciferase assays indicated that MdLUX and MdPCL-like activated MdF3H. The whole-genome DNA methylation study revealed that the methylation levels of the mCG context at the upstream (i.e., promoter region) of MdLUX and MdPCL-like were inversely correlated with their mRNA levels and anthocyanin accumulation. Hence, the data suggest that MYB_SH[AL]QKY[RF] TFs MdLUX and MdPCL-like promote anthocyanin biosynthesis in apple fruit skins through the DNA hypomethylation of their promoter regions and the activation of the structural flavonoid gene MdF3H.

scholarly journals Improving the Anthocyanin Accumulation of Hypocotyls in Radish Sprouts by Hemin-induced NO

2020 ◽  
Author(s):  
Nana Su ◽  
Ze Liu ◽  
Hui Chen ◽  
Mengyang Niu ◽  
Jin Cui
Keyword(s):  
Nitric Oxide ◽  
Transcription Factors ◽  
Anthocyanin Biosynthesis ◽  
Reductase Activity ◽  
Specific Inhibitor ◽  
Anthocyanin Synthesis ◽  
Zinc Protoporphyrin ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Radish Sprouts

Abstract Background: The biosynthesis of anthocyanin in the hypocotyls of radish (Raphanus sativus L.) sprouts was enhanced by hemin in our preliminary experiments, but the underlying mechanism is unclear. Here, we found that NO (nitric oxide) exerted an essential role in Hemin-regulated anthocyanin biosynthesis, which was supported by the following results.Results: Hemin boosted anthocyanin as well as NO content. NO-scavenger cPTIO (carboxy-PTIO) significantly attenuated hemin-induced increase of anthocyanin content, transcripts of anthocyanin synthesis related genes and positive transcription factors, implying that NO played a prominent role during hemin-induced anthocyanin biosynthesis. Hemin specific inhibitor ZnPP (Zinc Protoporphyrin) strongly reduced anthocyanin content, while, NO donor SNP (Sodium Nitroprusside) addition considerably reversed this inhibition and by contrast, resulted in a significant increase in anthocyanin accumulation, closely paralleling the transcripts of structural genes and transcription factors. Moreover, NO content, NR (nitrate reductase) activity and expression level of NOA (nitric oxide associated factor) were up-regulated by Hemin. Conclusions:Those consequences indicated that NO might work downstream in Hemin-heightened anthocyanin accumulation in radish sprouts.

scholarly journals The RNA Directed DNA Methylation (RdDM) Pathway Regulates Anthocyanin Biosynthesis in Crabapple (Malus cv. spp.) Leaves by Methylating the McCOP1 Promoter

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2466
Author(s):  
Yifan Xing ◽  
Ziyi Xie ◽  
Weilei Sun ◽  
Yuying Sun ◽  
Zhenyun Han ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Methylation Level ◽  
Bisulfite Sequencing ◽  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Bimolecular Fluorescence Complementation ◽  
Rna Expression ◽  
Anthocyanin Accumulation ◽  
Transcript Levels ◽  
Direct Target

The synthesis of anthocyanin pigments in plants is known to be regulated by multiple mechanisms, including epigenetic regulation; however, the contribution of the RNA-directed DNA methylation (RdDM) pathway is not well understood. Here, we used bisulfite sequencing and Real Time (RT)-quantitative (q) PCR to analyze the methylation level of the promoter of constitutively photomorphogenic 1 (McCOP1) from Malus cv. spp, a gene involved in regulating anthocyanin biosynthesis. The CHH methylation level of the McCOP1 promoter was negatively correlated with McCOP1 RNA expression, and inhibiting DNA methylation caused decreased methylation of the McCOP1 promoter and asymmetric cytosine CHH methylation. We observed that the McCOP1 promoter was a direct target of the RdDM pathway argonaute RISC component 4 (McAGO4) protein, which bound to a McCOP1 promoter GGTTCGG site. Bimolecular fluorescence complementation (BIFC) analysis showed that RNA-directed DNA methylation (McRDM1) interacted with McAGO4 and another RdDM protein, domains rearranged methyltransferase 2 (McDRM2), to regulate the CHH methylation of the McCOP1 promoter. Detection of CHH methylation and COP1 gene expression in the Arabidopsis thalianaatago4, atdrm2 and atrdm1 mutants showed that RDM1 is the effector of the RdDM pathway. This was confirmed by silencing McRDM1 in crabapple leaves or apple fruit, which resulted in a decrease in McCOP1 CHH methylation and an increase in McCOP1 transcript levels, as well as in anthocyanin accumulation. In conclusion, these results show that the RdDM pathway is involved in regulating anthocyanin accumulation through CHH methylation of the McCOP1 promoter.

FvbHLH9 Functions as a Positive Regulator of Anthocyanin Biosynthesis by Forming a HY5–bHLH9 Transcription Complex in Strawberry Fruits

2020 ◽  
Vol 61 (4) ◽  
pp. 826-837 ◽  
Author(s):  
Yang Li ◽  
Pengbo Xu ◽  
Guanqun Chen ◽  
Jun Wu ◽  
Zhongchi Liu ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Promoter Region ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Synthesis ◽  
Bhlh Transcription Factor ◽  
Anthocyanin Accumulation ◽  
Positive Regulator ◽  
Key Enzyme ◽  
Synergistic Regulation ◽  
Transient Overexpression

Abstract Anthocyanin accumulation is transcriptionally regulated by the MYB–bHLH–WD40 complex. Light is indispensable for anthocyanin accumulation, and light-inducible MYB and HY5 were considered to promote anthocyanin accumulation in many fruits. Whether and how light-inducible bHLH transcription factor and HY5 regulate anthocyanin synthesis in strawberry is unknown. In this study, we identified a bHLH transcription factor, FvbHLH9, which was induced by light as well as FvHY5, and found that, similar to FvHY5, the transient overexpression and interference FvbHLH9 in strawberry fruits can promote and decrease anthocyanin accumulation, respectively, indicating FvbHLH9 functions as a positive regulator of anthocyanin biosynthesis. Furthermore, we confirmed that both FvHY5 and FvbHLH9 specifically bind to the promoter region of some key enzyme genes, including FvDFR, and the expression of FvDFR was activated through the heterodimer formation between FvHY5 and FvbHLH9. Finally, we confirmed that FvbHLH9-promoted anthocyanin accumulation is dependent on HY5–bHLH heterodimerisation in Arabidopsis. Our findings provide insights into a mechanism involving the synergistic regulation of light-dependent coloration and anthocyanin biosynthesis via a HY5–bHLH heterodimer formed by the interaction of FvHY5 and FvbHLH9 in strawberry fruits.

Expression of Anthocyanin Biosynthetic Genes during Fruit Development in ‘Fengxiang’ Strawberry

2012 ◽  
Vol 455-456 ◽  
pp. 443-448
Author(s):  
Bo Zhou ◽  
Shu Hua Yan ◽  
Yu Hua Li
Keyword(s):  
Fruit Development ◽  
Anthocyanin Biosynthesis ◽  
Northern Analysis ◽  
Anthocyanin Synthesis ◽  
Mrna Levels ◽  
Anthocyanin Accumulation ◽  
Flavonoid Pathway ◽  
Biosynthetic Genes ◽  
Pigment Accumulation ◽  
Anthocyanin Biosynthetic Genes

Anthocyanins are the main pigments in flowers and fruits. In most cases, anthocyanin accumulation in fruit is highly controlled by the developmental level. In this study, the cDNA fragments of three genes, chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS), which are involved in the flavonoid pathway, were isolated from total RNA of strawberry ripe fruit by using polymerase chain reaction technique and labeled as probes to determine the expression of anthocyanin biosynthetic genes. Northern analysis showed that a correlation between anthocyanin accumulation and expression of the flavonoid pathway genes during the ripening of strawberry fruits. At the early stages of fruit development, the mRNA levels encoding CHS, DFR, ANS were high probably responsible for the accumulation of condensed tannins, but the levels decreased dramatically when fruits turned white from green. During the stage of pigment accumulation, their mRNA levels increased strongly to be involved anthocyanin biosynthesis. Difference of CHS in mRNA abundance was correlated with differential accumulation of anthocyanins throughout the process of fruit development. Therefore, CHS could be a key structure gene involved in anthocyanin synthesis. Furthermore, the co-ordination of expression of anthocyanin biosynthetic genes implied a common regulatory mechanism controlling the expression of structural genes in the flavonoid pathway.

Whole-genome DNA methylation patterns and complex associations with gene expression associated with anthocyanin biosynthesis in apple fruit skin

Planta ◽  
2019 ◽  
Vol 250 (6) ◽  
pp. 1833-1847 ◽  
Author(s):  
Wen-Fang Li ◽  
Gai-Xing Ning ◽  
Juan Mao ◽  
Zhi-Gang Guo ◽  
Qi Zhou ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Whole Genome ◽  
Fruit Skin ◽  
Methylation Patterns

scholarly journals REVEILLE Transcription Factors Contribute to the Nighttime Accumulation of Anthocyanins in ‘Red Zaosu’ (Pyrus Bretschneideri Rehd.) Pear Fruit Skin

2020 ◽  
Vol 21 (5) ◽  
pp. 1634
Author(s):  
Xieyu Li ◽  
Ting Wu ◽  
Hanting Liu ◽  
Rui Zhai ◽  
Yao Wen ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Anthocyanin Biosynthesis ◽  
Expression Patterns ◽  
Luciferase Reporter ◽  
Anthocyanin Accumulation ◽  
Pear Fruit ◽  
Biosynthetic Genes ◽  
Expression Levels ◽  
Fruit Skin ◽  
Genes Encoding

Anthocyanin biosynthesis exhibits a rhythmic oscillation pattern in some plants. To investigate the correlation between the oscillatory regulatory network and anthocyanin biosynthesis in pear, the anthocyanin accumulation and the expression patterns of anthocyanin late biosynthetic genes (ALBGs) were investigated in fruit skin of ‘Red Zaosu’ (Pyrus bretschneideri Rehd.). The anthocyanin accumulated mainly during the night over three continuous days in the fruit skin, and the ALBGs’ expression patterns in ‘Red Zaosu’ fruit skin were oscillatory. However, the expression levels of typical anthocyanin-related transcription factors did not follow this pattern. Here, we found that the expression patterns of four PbREVEILLEs (PbRVEs), members of a class of atypical anthocyanin-regulated MYBs, were consistent with those of ALBGs in ‘Red Zaosu’ fruit skin over three continuous days. Additionally, transient expression assays indicated that the four PbRVEs promoted anthocyanin biosynthesis by regulating the expression of the anthocyanin biosynthetic genes encoding dihydroflavonol-4-reductase (DFR) and anthocyanidin synthase (ANS) in red pear fruit skin, which was verified using a dual-luciferase reporter assay. Moreover, a yeast one-hybrid assay indicated that PbRVE1a, 1b and 7 directly bound to PbDFR and PbANS promoters. Thus, PbRVEs promote anthocyanin accumulation at night by up-regulating the expression levels of PbDFR and PbANS in ‘Red Zaosu’ fruit skin.

scholarly journals Transposon-induced methylation of the RsMYB1 promoter disturbs anthocyanin accumulation in red-fleshed radish

2020 ◽  
Vol 71 (9) ◽  
pp. 2537-2550 ◽  
Author(s):  
Qingbiao Wang ◽  
Yanping Wang ◽  
Honghe Sun ◽  
Liang Sun ◽  
Li Zhang
Keyword(s):  
Dna Methylation ◽  
Raphanus Sativus ◽  
Promoter Region ◽  
Sequence Variation ◽  
Anthocyanin Biosynthesis ◽  
Underlying Mechanism ◽  
Anthocyanin Accumulation ◽  
Virus Induced Gene Silencing ◽  
Genome Resequencing ◽  
Raphanus Sativus L

Abstract Red-fleshed radish (Raphanus sativus L.) is a unique cultivar whose taproot is rich in anthocyanins beneficial to human health. However, the frequent occurrence of white-fleshed mutants affects the purity of commercially produced radish and the underlying mechanism has puzzled breeders for many years. In this study, we combined quantitative trait location by genome resequencing and transcriptome analyses to identify a candidate gene (RsMYB1) responsible for anthocyanin accumulation in red-fleshed radish. However, no sequence variation was found in the coding and regulatory regions of the RsMYB1 genes of red-fleshed (MTH01) and white-fleshed (JC01) lines, and a 7372 bp CACTA transposon in the RsMYB1 promoter region occurred in both lines. A subsequent analysis suggested that the white-fleshed mutant was the result of altered DNA methylation in the RsMYB1 promoter. This heritable epigenetic change was due to the hypermethylated CACTA transposon, which induced the spreading of DNA methylation to the promoter region of RsMYB1. Thus, RsMYB1 expression was considerably down-regulated, which inhibited anthocyanin biosynthesis in the white-fleshed mutant. An examination of transgenic radish calli and the results of a virus-induced gene silencing experiment confirmed that RsMYB1 is responsible for anthocyanin accumulation. Moreover, the mutant phenotype was partially eliminated by treatment with a demethylating agent. This study explains the molecular mechanism regulating the appearance of white-fleshed mutants of red-fleshed radish.

scholarly journals Effect of Temperature on Anthocyanin Synthesis and Ethylene Production in the Fruit of Early- and Medium-maturing Apple Cultivars during Ripening Stages

HortScience ◽  
2014 ◽  
Vol 49 (12) ◽  
pp. 1510-1517 ◽  
Author(s):  
Chikako Honda ◽  
Hideo Bessho ◽  
Mari Murai ◽  
Hiroshi Iwanami ◽  
Shigeki Moriya ◽  
...  
Keyword(s):  
Ethylene Production ◽  
Climatic Condition ◽  
Maximum Level ◽  
Apple Fruit ◽  
Climatic Conditions ◽  
Effect Of Temperature ◽  
Anthocyanin Synthesis ◽  
Anthocyanin Accumulation ◽  
Fruit Skin ◽  
Anthocyanin Concentration

The objective of this study was to investigate the effects of temperature treatments on anthocyanin accumulation and ethylene production in the fruit of early- and medium-maturing cultivars that were harvested early during fruit ripening. We first investigated the effects of various temperature treatments on anthocyanin accumulation in detached apples of ‘Tsugaru’, ‘Tsugaru Hime’, ‘Akane’ and ‘Akibae’ using an incubator. Three years of experiments demonstrated that at harvest, the lower-temperature treatments induced anthocyanin accumulation in ‘Tsugaru’, ‘Tsugaru Hime’, and ‘Akibae’ fruits, whereas the increases in anthocyanin accumulation under the 25 °C treatment were similar to those under the 15 and 20 °C treatments in ‘Akane’ fruit. The rate of ethylene production did not increase substantially during the temperature treatments in any of the four cultivars, except after the treatments of ‘Tsugaru’ fruit at harvest. The inhibition of ethylene action by the application of 1-methylcyclopropene (1-MCP) to detached fruits at harvest suppressed anthocyanin development under 15 and 20 °C temperature treatments in ‘Tsugaru’, ‘Tsugaru Hime’, and ‘Akibae’, but not in ‘Akane’. In the second experiment, we investigated changes in the anthocyanin concentration in attached fruit of ‘Misuzu Tsugaru’ under different temperature conditions in a greenhouse. At harvest, the anthocyanin concentration in fruit under the hotter climatic condition (29 °C 12 hours/19 °C 12 hours) was lower than that under the control condition (25 °C 12 hours/15 °C 12 hours). During the last week before harvest, anthocyanin development proceeded rapidly in fruit skin not only under the control condition, but also under the hotter climatic condition. The rapid accumulation of anthocyanin in the fruit skin of ‘Misuzu Tsugaru’ at harvest under a relatively high temperature (25 °C) condition was confirmed by the experiment using an incubator. At harvest, the maximum level of ethylene production in fruits sampled from trees grown under the hotter climatic condition was 9-fold higher than that in fruits from trees grown under the control condition. These results indicate that the comparison of pigmentation potential after the 15 or 25 °C treatments using detached fruit was effective for estimating anthocyanin accumulation in fruit skins under hotter climatic conditions in early- and medium-maturing cultivars that were harvested early and that a hotter climatic condition during ripening increased ethylene production in apple fruit after harvest.

scholarly journals PbGA2ox8 induces vascular-related anthocyanin accumulation and contributes to red stripe formation on pear fruit

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Rui Zhai ◽  
Zhigang Wang ◽  
Chengquan Yang ◽  
Kui Lin-Wang ◽  
Richard Espley ◽  
...  
Keyword(s):  
Promoter Region ◽  
Vascular Bundles ◽  
Anthocyanin Accumulation ◽  
Comparative Transcriptome ◽  
Wild Type ◽  
Pear Fruit ◽  
Gene Encoding ◽  
Fruit Skin ◽  
Bud Sport ◽  
Transient Overexpression

AbstractFruit with stripes, which are generally longitudinal, can occur naturally, but the bioprocesses underlying this phenomenon are unclear. Previously, we observed an atypical anthocyanin distribution that caused red-striped fruit on the spontaneous pear bud sport “Red Zaosu” (Pyrus bretschneideri Rehd.). In this study, comparative transcriptome analysis of the sport and wild-type “Zaosu” revealed that this atypical anthocyanin accumulation was tightly correlated with abnormal overexpression of the gene-encoding gibberellin (GA) 2-beta-dioxygenase 8, PbGA2ox8. Consistently, decreased methylation was also observed in the promoter region of PbGA2ox8 from “Red Zaosu” compared with “Zaosu”. Moreover, the GA levels in “Red Zaosu” seedlings were lower than those in “Zaosu” seedlings, and the application of exogenous GA4 reduced abnormal anthocyanin accumulation in “Red Zaosu”. Transient overexpression of PbGA2ox8 reduced the GA4 level and caused anthocyanin accumulation in pear fruit skin. Moreover, the presence of red stripes indicated anthocyanin accumulation in the hypanthial epidermal layer near vascular branches (VBs) in “Red Zaosu”. Transient overexpression of PbGA2ox8 resulting from vacuum infiltration induced anthocyanin accumulation preferentially in calcium-enriched areas near the vascular bundles in pear leaves. We propose a fruit-striping mechanism, in which the abnormal overexpression of PbGA2ox8 in “Red Zaosu” induces the formation of a longitudinal array of anthocyanin stripes near vascular bundles in fruit.

scholarly journals Calcium Positively Mediates Blue Light-Induced Anthocyanin Accumulation in Hypocotyl of Soybean Sprouts

2021 ◽  
Vol 12 ◽  
Author(s):  
Gang Hu ◽  
Xiaomeng Yue ◽  
Jinxue Song ◽  
Guipei Xing ◽  
Jun Chen ◽  
...  
Keyword(s):  
Blue Light ◽  
Anthocyanin Biosynthesis ◽  
Calcium Supplementation ◽  
Distribution Patterns ◽  
Anthocyanin Synthesis ◽  
Regulation Mechanism ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Soybean Sprouts ◽  
Expression Of Genes

Soybean sprouts are a flavorful microgreen that can be eaten all year round and are widely favored in Southeast Asia. In this study, the regulatory mechanism of calcium on anthocyanin biosynthesis in soybean sprouts under blue light was investigated. The results showed that blue light, with a short wavelength, effectively induced anthocyanin accumulation in the hypocotyl of soybean sprout cultivar “Dongnong 690.” Calcium supplementation further enhanced anthocyanin content, which was obviously inhibited by LaCl3 and neomycin treatment. Moreover, exogenous calcium changed the metabolism of anthocyanins, and seven anthocyanin compounds were detected. The trend of calcium fluorescence intensity in hypocotyl cells, as well as that of the inositol 1,4,5-trisphosphate and calmodulin content, was consistent with that of anthocyanins content. Specific spatial distribution patterns of calcium antimonate precipitation were observed in the ultrastructure of hypocotyl cells under different conditions. Furthermore, calcium application upregulated the expression of genes related to anthocyanin biosynthesis, and calcium inhibitors suppressed these genes. Finally, transcriptomics was performed to gain global insights into the molecular regulation mechanism of calcium-associated anthocyanin production. Genes from the flavonoid biosynthesis pathway were distinctly enriched among the differentially expressed genes, and weighted gene co-expression network analysis showed that two MYBs were related to the accumulation of anthocyanins. These results indicated that calcium released from apoplast and intracellular stores in specific spatial-temporal features promote blue light-induced anthocyanin accumulation by upregulation of the expression of genes related to anthocyanin synthesis of “Dongnong 690” hypocotyl. The findings deepen the understanding of the calcium regulation mechanism of blue light-induced anthocyanin accumulation in soybean sprouts, which will help growers produce high-quality foods beneficial for human health.

Sign in / Sign up

Export Citation Format

Share Document