scholarly journals Induction of Anthocyanin Accumulation in Crabapple (Malus cv.) Leaves by Low Temperatures

HortScience ◽  
2015 ◽  
Vol 50 (5) ◽  
pp. 640-649 ◽  
Author(s):  
Ji Tian ◽  
Zhen-yun Han ◽  
Li-ru Zhang ◽  
Ting-Ting Song ◽  
Jie Zhang ◽  
...  
Keyword(s):  
Low Temperatures ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Human Diet ◽  
Helix Loop Helix ◽  
Leaf Coloration ◽  
Regulatory Complex ◽  
R2r3 Myb ◽  
Reverse Transcript ◽  
Anthocyanin Biosynthetic Genes

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. There is thus considerable interest in the factors that regulate synthesis. Malus crabapple leaves are rich sources of these compounds, and in this study we analyzed leaf coloration, anthocyanin levels, and the expression levels of anthocyanin biosynthetic and regulatory genes in three crabapple cultivars (Royalty, Prairifire, and Flame) following various temperature treatments. We found that low temperatures (LTs) promoted anthocyanin accumulation in ‘Royalty’ and ‘Prairifire’, leading to red leaves, but not in ‘Flame’, which accumulated abundant colorless flavonols and retained green colored leaves. Quantitative reverse transcript PCR (RT-PCR) analyses indicated that the expression of several anthocyanin biosynthetic genes was induced by LTs, as were members of the R2R3-MYB, basic helix–loop–helix (bHLH) and WD40 transcription factor families that are thought to act in a complex. We propose that anthocyanin biosynthesis is differentially regulated in the three cultivars by LTs via the expression of members of this anthocyanin regulatory complex.

scholarly journals CsMYB3 and CsRuby1 form an ‘Activator-and-Repressor’ Loop for the Regulation of Anthocyanin Biosynthesis in Citrus

2019 ◽  
Vol 61 (2) ◽  
pp. 318-330 ◽  
Author(s):  
Ding Huang ◽  
Zhouzhou Tang ◽  
Jialing Fu ◽  
Yue Yuan ◽  
Xiuxin Deng ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Myb Transcription Factor ◽  
Nitrogen Stress ◽  
Loop Model ◽  
Anthocyanin Accumulation ◽  
Promoter Activation ◽  
R2r3 Myb ◽  
Activation Capacity ◽  
Anthocyanin Biosynthetic Genes

Abstract Anthocyanins are preferentially accumulated in certain tissues of particular species of citrus. A R2R3-MYB transcription factor (named Ruby1) has been well documented as an activator of citrus anthocyanin biosynthesis. In this study, we characterized CsMYB3, a transcriptional repressor that regulates anthocyanin biosynthesis in citrus. CsMYB3 was expressed in anthocyanin-pigmented tissues, and the expression was closely associated with that of Ruby1, which is a key anthocyanin activator. Overexpression of CsMYB3 in Arabidopsis resulted in a decrease in anthocyanins under nitrogen stress. Overexpression of CsMYB3 in the background of CsRuby1-overexpressing strawberry and Arabidopsis reduced the anthocyanin accumulation level. Transient promoter activation assays revealed that CsMYB3 could repress the activation capacity of the complex formed by CsRuby1/CsbHLH1 for the anthocyanin biosynthetic genes. Moreover, CsMYB3 could be transcriptionally activated by CsRuby1 via promoter binding, thus forming an ‘activator-and-repressor’ loop to regulate anthocyanin biosynthesis in citrus. This study shows that CsMYB3 plays a repressor role in the regulation of anthocyanin biosynthesis and proposes an ‘activator-and-repressor’ loop model constituted by CsRuby1 and CsMYB3 in the regulation of anthocyanin biosynthesis in citrus.

scholarly journals The Structure and Methylation Level of the McMYB10 Promoter Determine the Leaf Color of Malus Crabapple

HortScience ◽  
2017 ◽  
Vol 52 (4) ◽  
pp. 520-526 ◽  
Author(s):  
Ji Tian ◽  
Ke-ting Li ◽  
Shi-ya Zhang ◽  
Jie Zhang ◽  
Ting-ting Song ◽  
...  
Keyword(s):  
Methylation Level ◽  
Anthocyanin Biosynthesis ◽  
Anthocyanin Accumulation ◽  
Substantial Variation ◽  
Leaf Color ◽  
Human Diet ◽  
Development Stages ◽  
Leaf Coloration ◽  
The Relationship ◽  
Myb10 Transcription Factor

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. The MYB10 transcription factor (TF) plays an important role in regulating anthocyanin biosynthesis in Malus crabapple leaves. However, little is known about how the promoter regulates McMYB10 expression and influences the substantial variation in leaf anthocyanin accumulation and coloration that is observed in different crabapple cultivars. In this study, we analyzed leaf coloration, anthocyanin levels, and the expression levels of McMYB10 in the leaves of 15 crabapple cultivars with three leaf colors at various development stages, and showed that the expression of McMYB10 correlates positively with anthocyanin accumulation. We also examined the relationship between the number of R6 and R1 elements in the McMYB10 promoters of the different cultivars and the pigmentation of the new buds of spring-red cultivars, as well as the methylation level of the McMYB10 promoters at different development stages in three representative crabapple cultivars. The ratio of R6/R1 minisatellites in the promoters correlated with the color and anthocyanin accumulation in new crabapple buds, and we concluded that the differences in promoter structure and methylation level of the McMYB10 promoters coordinately affect the leaf color of crabapple cultivars.

scholarly journals Nighttime Temperatures and Sunlight Intensities Interact to Influence Anthocyanin Biosynthesis and Photooxidative Sunburn in “Fuji” Apple

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaomin Xue ◽  
Ying Duan ◽  
Jinzheng Wang ◽  
Fengwang Ma ◽  
Pengmin Li
Keyword(s):  
Light Intensity ◽  
Low Temperatures ◽  
Anthocyanin Biosynthesis ◽  
High Light ◽  
Anthocyanin Accumulation ◽  
Low Light ◽  
Low Light Intensity ◽  
Apple Peel ◽  
Fuji Apple ◽  
Nighttime Temperatures

Light and low temperatures induce anthocyanin accumulation, but intense sunlight causes photooxidative sunburn. Nonetheless, there have been few studies of anthocyanin synthesis under different sunlight intensities and low nighttime temperatures. Here, low nighttime temperatures followed by low light intensity were associated with greater anthocyanin accumulation and the expression of anthocyanin biosynthesis genes in “Fuji” apple peel. UDP-glucose flavonoid-3-O-glucosyltransferase (UFGT) activity was positively associated with anthocyanin enrichment. Ascorbic acid can be used as an electron donor of APX to scavenge H2O2 in plants, which makes it play an important role in oxidative defense. Exogenous ascorbate altered the anthocyanin accumulation and reduced the occurrence of high light–induced photooxidative sunburn by removing hydrogen peroxide from the peel. Overall, low light intensity was beneficial for the accumulation of anthocyanin and did not cause photooxidative sunburn, whereas natural light had the opposite effect on the apple peel at low nighttime temperatures. This study provides an insight into the mechanisms by which low temperatures induce apple coloration and high light intensity causes photooxidative sunburn.

MYB44 competitively inhibits the formation of the MYB340-bHLH2-NAC56 complex to regulate anthocyanin biosynthesis in purple-fleshed sweet potato

2020 ◽  
Author(s):  
Zeng-Zheng Wei ◽  
Kang-Di Hu ◽  
Dong-Lan Zhao ◽  
Jun Tang ◽  
Zhong-Qin Huang ◽  
...  
Keyword(s):  
Sweet Potato ◽  
Regulatory Network ◽  
Anthocyanin Biosynthesis ◽  
Expression System ◽  
Firefly Luciferase ◽  
Anthocyanin Accumulation ◽  
Tuberous Roots ◽  
Regulatory Complex ◽  
Transcriptional Complex ◽  
Insight Into

Abstract Background: Anthocyanins, which have important biological functions and have a beneficial effect on human health, notably account for pigmentation in purple-fleshed sweet potato tuberous roots. Individual regulatory factors of anthocyanin biosynthesis have been identified; however, the regulatory network of anthocyanin biosynthesis in purple-fleshed sweet potato is unclear. Results: We functionally determined that IbMYB340 cotransformed with IbbHLH2 in tobacco and strawberry receptacles induced anthocyanin accumulation, and the addition of IbNAC56a or IbNAC56b caused increased pigmentation. Furthermore, we confirmed the interaction of IbMYB340 with IbbHLH2 and IbNAC56a or IbNAC56b via yeast two-hybrid and firefly luciferase complementation assays; these proteins could form a MYB340-bHLH2-NAC56a or MYB340-bHLH2-NAC56b transcriptional complex to regulate anthocyanin biosynthesis by binding to the IbANS promoter rather than the IbUFGT promoter. Furthermore, it was found by a transient expression system in tobacco leaves that IbMYB44 could decrease anthocyanin accumulation. Moreover, the interaction of IbMYB44 with IbMYB340 and IbNAC56a or IbNAC56b was verified. This result suggested that IbMYB44 acts as a repressor of anthocyanin in sweet potato.Conclusions: The repressor IbMYB44 affected anthocyanin biosynthesis by competitively inhibiting the IbMYB340-IbbHLH2-IbNAC56a or IbMYB340-IbbHLH2-IbNAC56b regulatory complex formation. Overall, the present study proposed a novel regulatory network whereby several vital TFs play key roles in regulating anthocyanin biosynthesis, and it provides strong insight into the potential mechanism underlying anthocyanin biosynthesis in sweet potato tuberous roots with purple color.

scholarly journals Loss of the R2R3 MYB Transcription Factor RsMYB1 Shapes Anthocyanin Biosynthesis and Accumulation in Raphanus sativus

2021 ◽  
Vol 22 (20) ◽  
pp. 10927
Author(s):  
Da-Hye Kim ◽  
Jundae Lee ◽  
JuHee Rhee ◽  
Jong-Yeol Lee ◽  
Sun-Hyung Lim
Keyword(s):  
Transcription Factor ◽  
Raphanus Sativus ◽  
Anthocyanin Biosynthesis ◽  
Antioxidant Properties ◽  
Myb Transcription Factor ◽  
Anthocyanin Accumulation ◽  
Truncated Protein ◽  
Frame Shift ◽  
Frame Shift Mutation ◽  
R2r3 Myb

The red or purple color of radish (Raphanus sativus L.) taproots is due to anthocyanins, which have nutritional and aesthetic value, as well as antioxidant properties. Moreover, the varied patterns and levels of anthocyanin accumulation in radish roots make them an interesting system for studying the transcriptional regulation of anthocyanin biosynthesis. The R2R3 MYB transcription factor RsMYB1 is a key positive regulator of anthocyanin biosynthesis in radish. Here, we isolated an allele of RsMYB1, named RsMYB1Short, in radish cultivars with white taproots. The RsMYB1Short allele carried a 4 bp insertion in the first exon causing a frame-shift mutation of RsMYB1, generating a truncated protein with only a partial R2 domain at the N-terminus. Unlike RsMYB1Full, RsMYB1Short was localized to the nucleus and the cytoplasm and failed to interact with their cognate partner RsTT8. Transient expression of genomic or cDNA sequences for RsMYB1Short in radish cotyledons failed to induce anthocyanin accumulation, but that for RsMYB1Full activated it. Additionally, RsMYB1Short showed the lost ability to induce pigment accumulation and to enhance the transcript level of anthocyanin biosynthetic genes, while RsMYB1Full promoted both processes when co-expressed with RsTT8 in tobacco leaves. As the result of the transient assay, co-expressing RsTT8 and RsMYB1Full, but not RsMYB1Short, also enhanced the promoter activity of RsCHS and RsDFR. We designed a molecular marker for RsMYB1 genotyping, and revealed that the RsMYB1Short allele is common in white radish cultivars, underscoring the importance of variation at the RsMYB1 locus in anthocyanin biosynthesis in the radish taproot. Together, these results indicate that the nonsense mutation of RsMYB1 generated the truncated protein, RsMYB1Short, that had the loss of ability to regulate anthocyanin biosynthesis. Our findings highlight that the frame shift mutation of RsMYB1 plays a key role in anthocyanin biosynthesis in the radish taproot.

scholarly journals The Basic Helix-Loop-Helix Transcription Factor SmbHLH1 Represses Anthocyanin Biosynthesis in Eggplant

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhaofei Duan ◽  
Shiyu Tian ◽  
Guobin Yang ◽  
Min Wei ◽  
Jing Li ◽  
...  
Keyword(s):  
Plant Species ◽  
Anthocyanin Biosynthesis ◽  
Solanum Melongena ◽  
Amino Acid Sequences ◽  
Negative Regulator ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Assay ◽  
Anthocyanin Accumulation ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

Many basic helix-loop-helix transcription factors (TFs) have been reported to promote anthocyanin biosynthesis in numerous plant species, but little is known about bHLH TFs that inhibit anthocyanin accumulation. In this study, SmbHLH1 from Solanum melongena was identified as a negative regulator of anthocyanin biosynthesis. However, SmbHLH1 showed high identity with SmTT8, which acts as a SmMYB113-dependent positive regulator of anthocyanin-biosynthesis in plants. Overexpression of SmbHLH1 in eggplant caused a dramatic decrease in anthocyanin accumulation. Only the amino acid sequences at the N and C termini of SmbHLH1 differed from the SmTT8 sequence. Expression analysis revealed that the expression pattern of SmbHLH1 was opposite to that of anthocyanin accumulation. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays showed that SmbHLH1 could not interact with SmMYB113. Dual-luciferase assay demonstrated that SmbHLH1 directly repressed the expression of SmDFR and SmANS. Our results demonstrate that the biological function of bHLHs in anthocyanin biosynthesis may have evolved and provide new insight into the molecular functions of orthologous genes from different plant species.

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 Ectopic Expression of Multiple Chrysanthemum (Chrysanthemum × morifolium) R2R3-MYB Transcription Factor Genes Regulates Anthocyanin Accumulation in Tobacco

Genes ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 777 ◽  
Author(s):  
Yan Hong ◽  
Mengling Li ◽  
Silan Dai
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Functional Divergence ◽  
Ectopic Expression ◽  
Chrysanthemum Morifolium ◽  
Amino Acid Sequences ◽  
Regulatory Function ◽  
Myb Transcription Factor ◽  
Anthocyanin Accumulation ◽  
R2r3 Myb

The generation of chrysanthemum (Chrysanthemum × morifolium) flower color is mainly attributed to the accumulation of anthocyanins. In the anthocyanin biosynthetic pathway in chrysanthemum, although all of the structural genes have been cloned, the regulatory function of R2R3-MYB transcription factor (TF) genes, which play a crucial role in determining anthocyanin accumulation in many ornamental crops, still remains unclear. In our previous study, four light-induced R2R3-MYB TF genes in chrysanthemum were identified using transcriptomic sequencing. In the present study, we further investigated the regulatory functions of these genes via phylogenetic and alignment analyses of amino acid sequences, which were subsequently verified by phenotypic, pigmental, and structural gene expression analyses in transgenic tobacco lines. As revealed by phylogenetic and alignment analyses, CmMYB4 and CmMYB5 were phenylpropanoid and flavonoid repressor R2R3-MYB genes, respectively, while CmMYB6 was an activator of anthocyanin biosynthesis, and CmMYB7 was involved in regulating flavonol biosynthesis. Compared with wild-type plants, the relative anthocyanin contents in the 35S:CmMYB4 and 35S:CmMYB5 tobacco lines significantly decreased (p < 0.05), while for 35S:CmMYB6 and 35S:CmMYB7, the opposite result was obtained. Both in the 35S:CmMYB4 and 35S:CmMYB5 lines, the relative expression of several anthocyanin biosynthetic genes in tobacco was significantly downregulated (p < 0.05); on the contrary, several genes were upregulated in the 35S:CmMYB6 and 35S:CmMYB7 lines. These results indicate that CmMYB4 and CmMYB5 negatively regulate anthocyanin biosynthesis in chrysanthemum, while CmMYB6 and CmMYB7 play a positive role, which will aid in understanding the complex mechanism regulating floral pigmentation in chrysanthemum and the functional divergence of the R2R3-MYB gene family in higher plants.

scholarly journals Light- and Temperature-Induced Expression of an R2R3-MYB Gene Regulates Anthocyanin Biosynthesis in Red-Fleshed Kiwifruit

2019 ◽  
Vol 20 (20) ◽  
pp. 5228 ◽  
Author(s):  
Min Yu ◽  
Yuping Man ◽  
Yanchang Wang
Keyword(s):  
Transcriptional Activation ◽  
Anthocyanin Biosynthesis ◽  
Physiological Role ◽  
Regulatory Function ◽  
Anthocyanin Content ◽  
Anthocyanin Accumulation ◽  
Virus Induced Gene Silencing ◽  
Highly Correlated ◽  
Myb Genes ◽  
R2r3 Myb

The R2R3 MYB genes associated with the flavonoid/anthocyanidin pathway feature two repeats, and represent the most abundant classes of MYB genes in plants; however, the physiological role and regulatory function of most R2R3 MYBs remain poorly understood in kiwifruit (Actinidia). Here, genome-wide analysis identified 155 R2R3-MYBs in the ‘Red 5′ version of the Actinidia chinensis genome. Out of 36 anthocyanin-related AccR2R3-MYBs, AcMYB10 was the most highly expressed in inner pericarp of red-fleshed kiwifruit. The expression of AcMYB10 was highly correlated with anthocyanin accumulation in natural pigmentation during fruit ripening and light-/temperature-induced pigmentation in the callus. AcMYB10 is localized in the nuclei and has transcriptional activation activity. Overexpression of AcMYB10 elevates anthocyanin accumulation in transgenic A. chinensis. In comparison, A. chinensis fruit infiltrated with virus-induced gene silencing showed delayed red coloration, lower anthocyanin content, and lower expression of AcMYB10. The transient expression experiment in Nicotiana tabacum leaves and Actinidia arguta fruit indicated the interaction of AcMYB10 with AcbHLH42 might strongly activate anthocyanin biosynthesis by activating the transcription of AcLDOX and AcF3GT. In conclusion, this study provides novel molecular information about R2R3-MYBs in kiwifruit, advances our understanding of light- and temperature-induced anthocyanin accumulation, and demonstrates the important function of AcMYB10 in the biosynthesis of anthocyanin in kiwifruit.

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