scholarly journals CRISPR/Cas9-mediated targeted mutation reveals a role for AN4 rather than DPL in regulating venation formation in the corolla tube of Petunia hybrida

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Bin Zhang ◽  
Xiaojing Xu ◽  
Renwei Huang ◽  
Sha Yang ◽  
Mingyang Li ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Anthocyanin Biosynthesis ◽  
Corolla Tube ◽  
Myb Transcription Factor ◽  
Pcr Analysis ◽  
Flower Bud ◽  
Plant Trait ◽  
Targeted Mutation ◽  
R2r3 Myb ◽  
Ornamental Traits

AbstractVenation is a common anthocyanin pattern displayed in flowers that confers important ornamental traits to plants. An anthocyanin-related R2R3-MYB transcription factor, DPL, has been proposed to regulate corolla tube venation in petunia plants. Here, however, we provide evidence redefining the role of DPL in petunia. A CRISPR/Cas9-mediated mutation of DPL resulted in the absence of the vein-associated anthocyanin pattern above the abaxial surface of the flower bud, but not corolla tube venation, thus indicating that DPL did not regulate the formation of corolla tube venation. Alternately, quantitative real-time PCR analysis demonstrated that the spatiotemporal expression pattern of another R2R3-MYB gene, AN4, coincided with the formation of corolla tube venation in petunia. Furthermore, overexpression of AN4 promoted anthocyanin accumulation by increasing the expression of anthocyanin biosynthesis genes. CRISPR/Cas9-mediated mutation of AN4 led to an absence of corolla tube venation, suggesting that this gene in fact determines this key plant trait. Taken together, the results presented here redefine the prime regulator of corolla tube venation, paving the way for further studies on the molecular mechanisms underlying the various venation patterns in petunia.

Identification of R2R3-MYB gene family reveal candidate genes for anthocyanin biosynthesis in Lonicera caerulea fruit based on RNA-seq data

2021 ◽  
pp. 1-19
Author(s):  
Huixin Gang ◽  
Qian Zhang ◽  
Jing Chen ◽  
Dong Qin ◽  
Junwei Huo
Keyword(s):  
Candidate Genes ◽  
Molecular Mechanisms ◽  
Anthocyanin Biosynthesis ◽  
Expression Profiles ◽  
Myb Transcription Factor ◽  
Anthocyanin Content ◽  
Rna Seq ◽  
Lonicera Caerulea ◽  
Conserved Sequence ◽  
R2r3 Myb

BACKGROUND: R2R3-MYB transcription factor (TF) family plays important roles in various biological processes in many plants, especially in the regulation of plant flavonoid accumulation. The fruit of Lonicera caerulea contains abundant anthocyanin. OBJECTIVE: The R2R3-MYB TF family was systematically analyzed according to the RNA-seq data, and the R2R3-MYB candidate genes that were involved in anthocyanin biosynthesis in the fruit of Lonicera caerulea were screened. METHODS: The R2R3-MYB TFs in Lonicera caerulea were identified, and the physical and chemical properties, protein conserved sequence alignment and motifs of each R2R3-MYB TFs were analyzed using bioinformatics methods. The expression levels of these genes and anthocyanin levels in different tissues and different developmental stages of fruit were determined by RT-qPCR and pH shift method. RESULTS: A total of 59 genes encoding R2R3-MYB TFs in Lonicera caerulea were identified and clustered into 20 subgroups (C1 to C20) based on the relationship to AtR2R3-MYBs. Expression profiles showed that the expression of CL6086 and CL552 in fruit were higher than other tissues, and upregulated in the veraison fruit compared to the green ripe fruit. As the expression of the two genes was concurrent with the anthocyanin content, and showed high correlation with anthocyanin biosynthetic structural genes, they were considered as closely related to anthocyanin biosynthesis in the fruit. CONCLUSION: The results provide a systematic analysis of LcR2R3-MYBs, and the foundation for further molecular mechanisms research of anthocyanin biosynthesis regulated by R2R3-MYB in the fruit of Lonicera caerulea.

An R2R3-MYB transcription factor CmMYB21 represses anthocyanin biosynthesis in color fading petals of chrysanthemum

2022 ◽  
Vol 293 ◽  
pp. 110674
Author(s):  
Yiguang Wang ◽  
Li-Jie Zhou ◽  
Yuxi Wang ◽  
Zhiqiang Geng ◽  
Baoqing Ding ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Anthocyanin Biosynthesis ◽  
Myb Transcription Factor ◽  
Color Fading ◽  
R2r3 Myb

scholarly journals PbCOP1.1 Contributes to the Negative Regulation of Anthocyanin Biosynthesis in Pear

Plants ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 39 ◽  
Author(s):  
Meng Wu ◽  
Min Si ◽  
Xieyu Li ◽  
Linyan Song ◽  
Jianlong Liu ◽  
...  
Keyword(s):  
Protein Sequence ◽  
Molecular Mechanisms ◽  
Anthocyanin Biosynthesis ◽  
Pcr Analysis ◽  
Anthocyanin Synthesis ◽  
Light Signaling ◽  
Pear Fruit ◽  
Phylogenetic Tree Analysis ◽  
Expression Levels ◽  
Protein Sequence Alignment

The synthesis of anthocyanin in pear (Pyrus bretschneideri) fruit is regulated by light. However, little is known about the molecular mechanisms of pear fruit coloring mediated by upstream light-signaling regulators. Here, the photoresponse factors CONSTITUTIVE PHOTOMORPHOGENIC (COP) 1.1 and 1.2 were cloned from ‘Red Zaosu’ peel to study their functions in pear fruit coloring. The overexpression vectors pBI121-PbCOP1.1 and pBI121-PbCOP1.2 were constructed to analyze their effects on anthocyanin synthesis in pear fruit. A protein sequence alignment and phylogenetic tree analysis revealed that PbCOP1 proteins are highly homologous with those of other species. An analysis of tissue differential expression showed that the greatest expression levels of PbCOP1s occurred in the leaves. Their expression levels increased in the leaves during development, when the leaves changed from red to green. The overexpression of PbCOP1s in the peel resulted in reduced anthocyanin synthesis at the injection sites. A quantitative PCR analysis of the injection sites showed that PbCOP1.1 significantly inhibited the expression of the anthocyanin synthesis-related genes CHI, DFR, UFGT2, bHLH3, HY5 and GST. Based on the above results, we hypothesize that PbCOP1.1 is an anthocyanin synthetic inhibitory factor of pear coloration.

scholarly journals A R2R3-MYB Transcription Factor, VvMYBC2L2, Functions as a Transcriptional Repressor of Anthocyanin Biosynthesis in Grapevine (Vitis vinifera L.)

Molecules ◽  
2018 ◽  
Vol 24 (1) ◽  
pp. 92 ◽  
Author(s):  
Ziguo Zhu ◽  
Guirong Li ◽  
Li Liu ◽  
Qingtian Zhang ◽  
Zhen Han ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Vitis Vinifera ◽  
Anthocyanin Biosynthesis ◽  
Transcriptional Repressor ◽  
Negative Regulator ◽  
Myb Transcription Factor ◽  
Vitis Vinifera L ◽  
Flavonoid Pathway ◽  
Leucoanthocyanidin Reductase ◽  
R2r3 Myb

In grapevine, the MYB transcription factors play an important role in the flavonoid pathway. Here, a R2R3-MYB transcription factor, VvMYBC2L2, isolated from Vitis vinifera cultivar Yatomi Rose, may be involved in anthocyanin biosynthesis as a transcriptional repressor. VvMYBC2L2 was shown to be a nuclear protein. The gene was shown to be strongly expressed in root, flower and seed tissue, but weakly expressed during the fruit development in grapevine. Overexpressing the VvMYBC2L2 gene in tobacco resulted in a very marked decrease in petal anthocyanin concentration. Expression analysis of flavonoid biosynthesis structural genes revealed that chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductase (LAR) and UDP glucose flavonoid 3-O-glucosyl transferase (UFGT) were strongly down-regulated in the VvMYBC2L2-overexpressed tobacco. In addition, transcription of the regulatory genes AN1a and AN1b was completely suppressed in transgenic plants. These results suggested that VvMYBC2L2 plays a role as a negative regulator of anthocyanin biosynthesis.

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 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.

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